CN110215516A - It is a kind of to inhibit the treatment of CDK5 synergetic immunity in the application inhibited in breast cancer - Google Patents
It is a kind of to inhibit the treatment of CDK5 synergetic immunity in the application inhibited in breast cancer Download PDFInfo
- Publication number
- CN110215516A CN110215516A CN201810180887.5A CN201810180887A CN110215516A CN 110215516 A CN110215516 A CN 110215516A CN 201810180887 A CN201810180887 A CN 201810180887A CN 110215516 A CN110215516 A CN 110215516A
- Authority
- CN
- China
- Prior art keywords
- breast cancer
- cdk5
- antibody
- cancer immunotherapy
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000013717 Cyclin-Dependent Kinase 5 Human genes 0.000 title claims abstract description 89
- 108010025454 Cyclin-Dependent Kinase 5 Proteins 0.000 title claims abstract description 89
- 206010006187 Breast cancer Diseases 0.000 title claims abstract description 40
- 208000026310 Breast neoplasm Diseases 0.000 title claims abstract description 40
- 230000036039 immunity Effects 0.000 title abstract description 6
- 238000011282 treatment Methods 0.000 title description 9
- 230000002195 synergetic effect Effects 0.000 title description 5
- 239000003112 inhibitor Substances 0.000 claims abstract description 56
- 230000000903 blocking effect Effects 0.000 claims abstract description 35
- 238000000034 method Methods 0.000 claims abstract description 31
- 239000003814 drug Substances 0.000 claims abstract description 24
- 238000012360 testing method Methods 0.000 claims abstract description 17
- 230000001900 immune effect Effects 0.000 claims abstract description 16
- 239000000890 drug combination Substances 0.000 claims abstract description 11
- 238000002619 cancer immunotherapy Methods 0.000 claims abstract 13
- 230000014509 gene expression Effects 0.000 claims description 27
- 230000000694 effects Effects 0.000 claims description 25
- BTIHMVBBUGXLCJ-OAHLLOKOSA-N seliciclib Chemical compound C=12N=CN(C(C)C)C2=NC(N[C@@H](CO)CC)=NC=1NCC1=CC=CC=C1 BTIHMVBBUGXLCJ-OAHLLOKOSA-N 0.000 claims description 23
- 210000004027 cell Anatomy 0.000 claims description 21
- 210000002540 macrophage Anatomy 0.000 claims description 9
- 101150029707 ERBB2 gene Proteins 0.000 claims description 8
- 238000011284 combination treatment Methods 0.000 claims description 8
- PIMQWRZWLQKKBJ-SFHVURJKSA-N 2-[(2S)-1-[3-ethyl-7-[(1-oxido-3-pyridin-1-iumyl)methylamino]-5-pyrazolo[1,5-a]pyrimidinyl]-2-piperidinyl]ethanol Chemical compound C=1C(N2[C@@H](CCCC2)CCO)=NC2=C(CC)C=NN2C=1NCC1=CC=C[N+]([O-])=C1 PIMQWRZWLQKKBJ-SFHVURJKSA-N 0.000 claims description 7
- 238000002649 immunization Methods 0.000 claims description 7
- 230000003053 immunization Effects 0.000 claims description 7
- 238000013518 transcription Methods 0.000 claims description 7
- 230000035897 transcription Effects 0.000 claims description 7
- 230000002068 genetic effect Effects 0.000 claims description 6
- 238000002560 therapeutic procedure Methods 0.000 claims description 6
- 108091027967 Small hairpin RNA Proteins 0.000 claims description 5
- 108020004459 Small interfering RNA Proteins 0.000 claims description 5
- 229950009859 dinaciclib Drugs 0.000 claims description 5
- -1 small molecule compound Chemical class 0.000 claims description 5
- 229940045513 CTLA4 antagonist Drugs 0.000 claims description 4
- 241000124008 Mammalia Species 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- GTVPOLSIJWJJNY-UHFFFAOYSA-N olomoucine Chemical compound N1=C(NCCO)N=C2N(C)C=NC2=C1NCC1=CC=CC=C1 GTVPOLSIJWJJNY-UHFFFAOYSA-N 0.000 claims description 4
- 239000004055 small Interfering RNA Substances 0.000 claims description 4
- 101100108853 Mus musculus Anp32e gene Proteins 0.000 claims description 3
- 208000003721 Triple Negative Breast Neoplasms Diseases 0.000 claims description 3
- 208000026535 luminal A breast carcinoma Diseases 0.000 claims description 3
- 208000026534 luminal B breast carcinoma Diseases 0.000 claims description 3
- 208000022679 triple-negative breast carcinoma Diseases 0.000 claims description 3
- 230000006651 lactation Effects 0.000 claims description 2
- 108010033276 Peptide Fragments Proteins 0.000 claims 2
- 102000007079 Peptide Fragments Human genes 0.000 claims 2
- 102000037982 Immune checkpoint proteins Human genes 0.000 claims 1
- 108091008036 Immune checkpoint proteins Proteins 0.000 claims 1
- 230000001360 synchronised effect Effects 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 abstract description 68
- 238000009169 immunotherapy Methods 0.000 abstract description 16
- 210000001744 T-lymphocyte Anatomy 0.000 abstract description 11
- 230000002401 inhibitory effect Effects 0.000 abstract description 9
- 201000011510 cancer Diseases 0.000 abstract description 8
- 230000005764 inhibitory process Effects 0.000 abstract description 8
- 230000008595 infiltration Effects 0.000 abstract description 7
- 238000001764 infiltration Methods 0.000 abstract description 7
- 210000004981 tumor-associated macrophage Anatomy 0.000 abstract description 6
- 230000036210 malignancy Effects 0.000 abstract description 3
- 229940125644 antibody drug Drugs 0.000 abstract description 2
- 230000008859 change Effects 0.000 abstract description 2
- 230000002441 reversible effect Effects 0.000 abstract description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 17
- 210000001519 tissue Anatomy 0.000 description 16
- 210000004881 tumor cell Anatomy 0.000 description 15
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 14
- 102100040678 Programmed cell death protein 1 Human genes 0.000 description 14
- 229940079593 drug Drugs 0.000 description 14
- 238000002648 combination therapy Methods 0.000 description 12
- 238000001514 detection method Methods 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 11
- 238000002360 preparation method Methods 0.000 description 9
- 238000012545 processing Methods 0.000 description 8
- 210000000130 stem cell Anatomy 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 7
- 239000002552 dosage form Substances 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 230000007705 epithelial mesenchymal transition Effects 0.000 description 6
- 210000004072 lung Anatomy 0.000 description 6
- 239000008194 pharmaceutical composition Substances 0.000 description 6
- 238000009097 single-agent therapy Methods 0.000 description 6
- 238000012546 transfer Methods 0.000 description 6
- 230000000118 anti-neoplastic effect Effects 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 102000000905 Cadherin Human genes 0.000 description 4
- 108050007957 Cadherin Proteins 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 4
- 230000006044 T cell activation Effects 0.000 description 4
- 102000013127 Vimentin Human genes 0.000 description 4
- 108010065472 Vimentin Proteins 0.000 description 4
- 230000034994 death Effects 0.000 description 4
- 238000011065 in-situ storage Methods 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- 210000005048 vimentin Anatomy 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 101150053721 Cdk5 gene Proteins 0.000 description 3
- 108010077544 Chromatin Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 101150106931 IFNG gene Proteins 0.000 description 3
- 210000003483 chromatin Anatomy 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 238000003209 gene knockout Methods 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 230000009545 invasion Effects 0.000 description 3
- 239000004005 microsphere Substances 0.000 description 3
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 101100378744 Rattus norvegicus Aldh1a7 gene Proteins 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 210000000577 adipose tissue Anatomy 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000005809 anti-tumor immunity Effects 0.000 description 2
- 238000009175 antibody therapy Methods 0.000 description 2
- 201000008275 breast carcinoma Diseases 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 201000001441 melanoma Diseases 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000006201 parenteral dosage form Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- 238000010200 validation analysis Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 101150113296 ACAT1 gene Proteins 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 108010074708 B7-H1 Antigen Proteins 0.000 description 1
- 102100032912 CD44 antigen Human genes 0.000 description 1
- 210000001239 CD8-positive, alpha-beta cytotoxic T lymphocyte Anatomy 0.000 description 1
- 108010021064 CTLA-4 Antigen Proteins 0.000 description 1
- 102000008203 CTLA-4 Antigen Human genes 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 102100035189 GPI ethanolamine phosphate transferase 1 Human genes 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 101000598552 Homo sapiens Acetyl-CoA acetyltransferase, mitochondrial Proteins 0.000 description 1
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 1
- 101500025419 Homo sapiens Epidermal growth factor Proteins 0.000 description 1
- 101001093751 Homo sapiens GPI ethanolamine phosphate transferase 1 Proteins 0.000 description 1
- 101000884271 Homo sapiens Signal transducer CD24 Proteins 0.000 description 1
- 101000617830 Homo sapiens Sterol O-acyltransferase 1 Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 206010062016 Immunosuppression Diseases 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 241000283953 Lagomorpha Species 0.000 description 1
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 1
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 101100519207 Mus musculus Pdcd1 gene Proteins 0.000 description 1
- 101100310657 Mus musculus Sox1 gene Proteins 0.000 description 1
- 208000002454 Nasopharyngeal Carcinoma Diseases 0.000 description 1
- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 108010029755 Notch1 Receptor Proteins 0.000 description 1
- 239000012270 PD-1 inhibitor Substances 0.000 description 1
- 239000012668 PD-1-inhibitor Substances 0.000 description 1
- 101710126211 POU domain, class 5, transcription factor 1 Proteins 0.000 description 1
- 241000283089 Perissodactyla Species 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 102100024216 Programmed cell death 1 ligand 1 Human genes 0.000 description 1
- 108091007744 Programmed cell death receptors Proteins 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 102100038081 Signal transducer CD24 Human genes 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 102100021993 Sterol O-acyltransferase 1 Human genes 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229940123237 Taxane Drugs 0.000 description 1
- 208000024770 Thyroid neoplasm Diseases 0.000 description 1
- 102100023935 Transmembrane glycoprotein NMB Human genes 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001720 carbohydrates Chemical group 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 229940099112 cornstarch Drugs 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000009261 endocrine therapy Methods 0.000 description 1
- 229940034984 endocrine therapy antineoplastic and immunomodulating agent Drugs 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 239000001761 ethyl methyl cellulose Substances 0.000 description 1
- 235000010944 ethyl methyl cellulose Nutrition 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 101150016624 fgfr1 gene Proteins 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 210000004013 groin Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 108091008039 hormone receptors Proteins 0.000 description 1
- 229940116978 human epidermal growth factor Drugs 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 238000012151 immunohistochemical method Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000012308 immunohistochemistry method Methods 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- 108010082117 matrigel Proteins 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- GVUGOAYIVIDWIO-UFWWTJHBSA-N nepidermin Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CS)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CS)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C(C)C)C(C)C)C1=CC=C(O)C=C1 GVUGOAYIVIDWIO-UFWWTJHBSA-N 0.000 description 1
- 229960003301 nivolumab Drugs 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 229940100688 oral solution Drugs 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229940121655 pd-1 inhibitor Drugs 0.000 description 1
- 229960002621 pembrolizumab Drugs 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000000505 pernicious effect Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 229920001592 potato starch Polymers 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 102000003998 progesterone receptors Human genes 0.000 description 1
- 108090000468 progesterone receptors Proteins 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 230000022983 regulation of cell cycle Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 238000009938 salting Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000007910 systemic administration Methods 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 1
- 201000002510 thyroid cancer Diseases 0.000 description 1
- 108091007466 transmembrane glycoproteins Proteins 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 238000012762 unpaired Student’s t-test Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Endocrinology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention provides a kind of combination medicine combinations for Breast cancer immunotherapy, including effective quantity CDK5 inhibitor and a effective amount of immunologic test point blocking antibody.The present invention still further provides a kind of immunotherapy of tumors drug combination method.Present invention discover that CDK5 is inhibited malignancy of tumor can be inhibited to be in progress, reverse inhibitive ability of immunity tumor microenvironment, change tumor-associated macrophage phenotype and function, to improve T cell infiltration, with in immunologic test point blocking antibody drug combination, the two can play synergy, further enhance the inhibition for tumour.
Description
Technical field
The invention belongs to technical field of pharmaceutical biotechnology, and in particular to a kind for the treatment of of inhibition CDK5 synergetic immunity is inhibiting mammary gland
Application in cancer.
Background technique
Breast cancer is common one of the malignant tumour of women, and disease incidence cumulative year after year has become Chinese women cancer
The first cause of associated death.On the basis of gene expression difference, the immunohistochemistry marker of selected characteristic (estrogen by
Body ER, progesterone receptor PR, human epidermal growth factor acceptor Her2), breast cancer is clinically divided into 4 seed types i.e.:
Luminal A (ER+/PR+Her2-)、Luminal B(ER+/PR+Her2+Or ER+/PR+Ki67 > 14%), Her2+(ER-PR-
Her2+) and three yin breast cancer (Triple-negative breast cancer, TNBC) (ER-PR-Her2-).Wherein, three
Negative patient with breast cancer accounts for about the 15%-20% of breast cancer, and is mainly in young women.Differentiation degree is low, growth is fast, tumour body
Product is the feature of TNBC greatly.Because of TNBC inside tumor, the heterogeneity between different tumours is big, still without effective targeted therapy
Method.Simultaneously as not expressing hormone receptor, endocrine therapy is not reacted.Current clinically drug therapy first choice chemotherapy,
Chemotherapeutics includes anthracycline, taxanes and/or platinum-like compounds.With patient with breast cancer, especially three negative breast cancer are suffered from
Person increases year by year, needs a kind of effective therapeutic strategy.
Different from traditional tumour therapeutic strategy, immunization therapy targets patients immune system, itself antitumor exempts to activate
Epidemic disease reaction.Programmed death receptor -1 (programmed death-1, PD-1) is expressed in T cell, belongs to immunoglobulin
Superfamily I type transmembrane glycoprotein.Through FDA approval listing two kinds targeting PD-1 blocking antibody (Pembrolizumab,
Nivolumab) the significant effect in melanoma treatment.However, targeting PD-1 Immunotherapy Strategy is in other solid tumors, such as cream
Offer limited effectiveness in gland cancer, liver cancer etc., and different degrees of immunization therapy occur and resist and recur.How immune control is improved
Curative effect is treated, one of current immunization therapy research hotspot is become.
Cell cycle dependent kinases 5 (cyclin-dependent kinase 5, CDK5) belong to CDK family, with it
Unlike his CDK: CDK5 not cell cycle regulation.Clinical data is also shown: kinds of tumor cells (breast cancer, it is non-small thin
Born of the same parents' lung cancer, thyroid cancer, nasopharyngeal carcinoma, gastric cancer etc.) in, CDK5 expression activation is positively correlated with malignancy of tumor progress, and and patient
It is negatively correlated that OS, PFS etc. treat terminal.Especially in May, 2017, Science report CDK5 inhibitor can significantly inhibit it is swollen
The expression of oncocyte PD-L1 has further been pushed to target CDK5 as the research of the antineoplastic new strategy of starting point.Currently,
There are several Cdk5 inhibitor to enter clinical experimental stage, such as the first and second generation ATP competitive inhibitor R-roscovitine,
Dinaciclib and AT7519 etc..And non ATP competitiveness CDK5 inhibitor such as Cpd1, p5 peptide etc. also has become antineoplastic new medicine source
Molecule.But there is no disclose being associated between CDK5 and the immunization therapy of tumour for the prior art.
Summary of the invention
In view of the foregoing deficiencies of prior art, the purpose of the present invention is to provide the connection that can be used for immunotherapy of tumors
Combination with medication combination.
First aspect present invention provides a kind of combination medicine combination for immunotherapy of tumors, including a effective amount of
CDK5 inhibitor and a effective amount of immunologic test point blocking antibody.
The present invention provides CDK5 inhibitor in the pharmaceutical composition of preparation and immunologic test point blocking antibody drug combination
Purposes.
The CDK5 inhibitor refers to the compound for having inhibitory effect for CDK5 kinase activity.
Include but is not limited to inhibitory effect for CDK5: inhibiting CDK5 kinase activity, or CDK5 gene is inhibited to turn
Record or expression.
The CDK5 inhibitor can be siRNA, shRNA, antibody, small molecule compound, small peptide.
What the embodiment of the present invention was enumerated, the CDK5 inhibitor is R-Roscovitine, and the CDK5 inhibitor is also optional
From: olomoucine (olomoucine) and its derivative, Dinaciclib (MK-7965, SCH727965), AT7519, CIP,
P5, Cpd1 etc..
The immunologic test point blocking antibody can be selected from one of anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody etc.
Or combinations thereof.In the preferred embodiment of the invention, the immunologic test point blocking antibody therapy medicine is anti-PD-1 antibody.
The combination therapy pharmaceutical composition can be any one in following form:
1) independent preparation is respectively prepared in CDK5 inhibitor and immunologic test point blocking antibody, the dosage form of preparation can be identical
Or it is different, administration route may be the same or different.
2) CDK5 inhibitor and immunologic test point blocking antibody are configured to compound preparation.In CDK5 inhibitor and immune inspection
Blocking antibody is made an inventory of using the administration of identical administration route and when applying simultaneously, the shape that the two is configured to compound preparation can be used
Formula.
Second aspect of the present invention provides a kind of immunotherapy of tumors drug combination treatment method, to apply to Object associates
A effective amount of CDK5 inhibitor and a effective amount of immunologic test point blocking antibody.
The object is mammal or the T cell of the mammal, macrophage, tumour cell.The lactation
Animal is preferably rodent, artiodactylous animals, Perissodactyla animal, Lagomorph, primate etc..The Primates
Animal is preferably monkey, ape or homo sapiens.
The object can be the patient for suffering from tumour or expect to improve the individual of anti-tumor immunity, or swollen to suffer from
The patient of tumor or the individual in vitro CTL cell for expecting to improve anti-tumor immunity.
It is can synchronizing or sequentially give a effective amount of CDK5 inhibitor and a effective amount of immunologic test point blocking antibody.
Based on CDK5 be newfound immunotherapy of tumors target spot, the present invention it has been investigated that, with immunologic test point hinder
Break in antibody combined medication, the two can play synergy, further enhance the inhibition for tumour.
The tumour that the immunotherapy of tumors is directed to includes the various hypotypes of breast cancer, including but not limited to: Luminal A
(ER+/PR+ Her2-)、Luminal B(ER+/PR+Her2+Or ER+/PR+Ki67 > 14%), Her2+(ER-PR-Her2+), with
And three yin breast cancer (Triple-negative breast cancer, TNBC) (ER-PR-Her2-)。
The CDK5 inhibitor and immunologic test point blocking antibody therapy medicine can before receiving immunotherapy of tumors,
In, backward object application.
The present invention has the advantages that
(1) by a series of experiment in vitro, discovery CDK5 inhibitor plays in modulate tumor cell stemness conversion process
Vital effect inhibits CDK5 activity that can significantly inhibit the conversion of tumour cell stemness, EMT, invasion, each mistake such as transfer
Journey.
(2) in animal level, CDK5 activity is inhibited to significantly improve tumour immunity microenvironment: on the one hand, dramatically increased swollen
T cell infiltrates in tumor tissue, this process may be with tumor-associated macrophage phenotype and function conversion correlation (from immunosupress
M2 type macrophage to promote antineoplastic immune M1 type macrophage)；On the other hand, it is pernicious to significantly inhibit tumour cell
Conversion.In the combination therapy mouse in-situ inoculating breast cancer experiment with anti-PD-1 antibody, CDK5 inhibitor Roscovitine can
To cooperate with the anti-tumor activity for promoting immunocyte (cd8 t cell, tumor-associated macrophage etc.) to mediate with PD-1 antibody.
(3) although the prior art demonstrates the drug safety of CDK5 inhibitor, but CDK5 inhibitor original indication
Clinical effectiveness is unsatisfactory.The present invention provides new clinical value to CDK5 inhibitor in immunotherapy of tumors field,
Not only its further market application provides wide prospect, also provides significantly more efficient therapeutic scheme for tumor patient.
The above description is only an overview of the technical scheme of the present invention, in order to better understand the technical means of the present invention,
And can be implemented in accordance with the contents of the specification, below by with preferable case study on implementation of the invention and cooperate attached drawing carry out specifically
It is bright.A specific embodiment of the invention is shown in detail by following embodiment and its attached drawing.
Detailed description of the invention
The drawings described herein are used to provide a further understanding of the present invention, constitutes a part of the invention, this hair
Bright illustrative embodiments and their description are used to explain the present invention, and are not constituted improper limitations of the present invention, in the accompanying drawings:
Fig. 1 .CDK5 inhibits (micromolecular inhibitor, CDK5 gene knockout) to inhibit tumour cell EMT, invasion, stemness conversion
A. it is inoculated with breast carcinoma cell strain (CDK5-WT and CDK5-KO) in Balb/C female mice fat pad, building cream
Gland cancer in-situ inoculating breast cancer mouse model, bioluminescence imaging technology detect breast cancer lung tissue transfer case.
B. the cell Transwell Matrigel is used, Validation in vitro CDK5 inhibitor and gene knockout are thin to breast cancer
The influence of born of the same parents' invasive ability.
C. pass through scratch experiment, influence of the Validation in vitro CDK5 inhibitor to breast cancer cell transfer ability.
D. it is expressed under the effect of CDK5 inhibitor by WB detection EMT related gene (E-cadherin and Vimentin)
Situation.
E. external microsphere forms experiment, verifies influence of the CDK5 inhibitor to breast cancer cell stem cell properties.
F. differential expression of the breast carcinoma stem cell related gene under the effect of CDK5 inhibitor is detected by RT-qPCR.
Fig. 2 .CDK5 inhibitor Roscovitine and PD-1 blocking antibody Combined Treatment inhibit mouse breast cancer development process
A. to 4T1, plantation breast cancer mouse carries out Roscovitine and the combination therapy of PD-1 blocking antibody in situ, and detection is swollen
Tumor growth curve and mouse survival rate compare the difference of combination therapy Yu monotherapy effect, administration routes and administration number of days
See on figure A with dosage.
B. after being administered 20 days, put to death mouse and separate tumor tissues, weigh tumor weight and weight, compare combination therapy with
The difference on effect of monotherapy.
C. each processing group mouse lung tissue is separated, dyeing is fixed with picric acid saturation dyeing liquor, counts transfer stove number, and
Compare combination therapy and monotherapy difference on effect.
D. mouse survival curve is analyzed (n=10) with log-rank (Mantel-Cox) statistical method
E. by cd8 t cell PD-1, IFNg expression in each processing group mouse tumor tissue of flow cytometer showed, to compare
The influence of combination therapy and monotherapy to tumor-infiltrated killer T cell activation degree.
F. pass through the cd8 t cell infiltrated in each processing group mouse tumor tissue of flow cytometer showed and Treg cellular infiltration situation,
And compare the influence of combination therapy and monotherapy to T cell infiltration degree.Cd8 t cell and Treg ratio are analyzed simultaneously.
G. by the tumor-infiltrated macrophage ratio of flow cytometer showed, phenotypic alternation, and compare combination therapy and monotherapy
Influence to tumor-associated macrophage phenotypic function.
Specific embodiment
The present invention it has been investigated that, the adjustable tumour immunity microenvironment of CDK5 inhibitor.Therefore, it is considered that CDK5 inhibitor
It can be used as immunotherapy of tumors regulator and other immunotherapy of tumors agent be used in combination to improve immunotherapy of tumors curative effect.Into
One step studies have shown that with the antibody combined medication of anti-PD-L1, the two can play synergy, can further enhance
Inhibition for tumour.
CDK5 inhibitor
Refer to the compound that there is inhibitory effect for CDK5.Include but is not limited to inhibitory effect for CDK5: suppression
CDK5 activity processed, or inhibit the genetic transcription or expression of CDK5.
The CDK5 inhibitor include but is not limited to be siRNA, shRNA, antibody, small molecule compound.
CDK5 activity is inhibited to refer to that CDK5 enzyme activity declines.Preferably, before compared to inhibiting, CDK5 enzyme activity is reduced at least
10%, at least 30%, then good reduction at least 50% are preferably reduced, more preferably reduces at least 70%, optimal reduction is at least
90%.
The genetic transcription or expression for inhibiting CDK5 refer to: transcribing the gene of CDK5 not, or reduce turn of the gene of CDK5
Record activity, or express that the gene of CDK5 not, or reduce the activity of gene expression of CDK5.
Those skilled in the art can be used conventional method and CDK5 genetic transcription or expression be adjusted, such as clpp gene
It removes, homologous recombination, RNA interfering etc..
The inhibition of genetic transcription or the expression of CDK5 can detect expression quantity verifying by PCR and Western Blot.
Preferably, compared with wild type, ACAT1 genetic transcription or expression reduce at least 10%, preferably reduce at least
30%, then good reduction at least 50%, more preferably reduce at least 70%, and good reduction at least 90%, optimal ACAT1 gene
Absolutely not express.
Combination therapy pharmaceutical composition and method of administration
The combination therapy pharmaceutical composition can make any one in following form:
One) independent pharmaceutical preparation, the dosage form of preparation is respectively prepared in CDK5 inhibitor and immune detection point blocking antibody
It may be the same or different, administration route also may be the same or different.In use, can two kinds of medicines use simultaneously, can also two kinds of medicines successively make
With.Formerly still it should apply other drugs to body in body effective period with drug when consecutive administration.
Two) CDK5 inhibitor and immune detection point blocking antibody are configured to compound preparation.CDK5 inhibitor and it will exempt from
Epidemic disease test point blocking antibody can be used the two being configured to compound preparation using the administration of identical administration route and when applying simultaneously
Form.
The immune detection common administrated method of point blocking antibody is intravenous injection, intravenous drip or arterial perfusion.Its usage
Dosage can refer to the prior art.
The common administrated method of small molecule compound can make gastrointestinal administration either parenteral.siRNA,
ShRNA, antibody then generally use parenteral.Can be local administration can also be Formulations for systemic administration.
It is can synchronizing or sequentially give a effective amount of CDK5 inhibitor and a effective amount of immune detection point blocking antibody.
In some embodiments, CDK5 inhibitor uses gastrointestinal administration dosage form, and immune detection point blocking antibody uses parenteral
Dosage form.Alternatively, CDK5 inhibitor and immune detection point blocking antibody are all made of parenteral dosage forms.In use, can two kinds of medicines
Use simultaneously, can also two kinds of medicines successively use.It, should be formerly still introversive to organism effective period with drug when consecutive administration
Organism applies other drugs.
When preparing drug or pharmaceutical composition as one of main active or main active using CDK5 inhibitor.It is logical
Often, in drug other than effective component, according to the needs of different dosage forms, one or more pharmaceutically acceptable loads be will also include
Body or auxiliary material.
" pharmaceutically acceptable " refer to when biomolecule ontology and composition is appropriate give animal or people when, they will not be produced
Raw unfavorable, allergy or other adverse reactions.
" pharmaceutically acceptable carrier or auxiliary material " should be compatible with CDK5 inhibitor, can be blended without logical
The effect of pharmaceutical composition is greatly lowered in normal situation.It can be used as some substances of pharmaceutically acceptable carrier or auxiliary material
Specific example is carbohydrate, such as lactose, dextrose and saccharose；Starch, such as cornstarch and potato starch；Cellulose and its derivative
Object, such as sodium carboxymethylcellulose pyce, ethyl cellulose and methylcellulose；Gelatin；Talcum；Solid lubricant；Polyalcohol；Emulsifier；
Colorant；Flavoring agent；Tablet agent, stabilizer；Antioxidant；Preservative；Apirogen water；Isotonic salting liquid.These substances according to
It needs to be used to help the stability of formula or helps to improve activity or its biological effectiveness or generate in oral situation can
The mouthfeel or smell of receiving.
In the present invention, unless stated otherwise, pharmaceutical dosage form is not particularly limited, and can be made into injection, oral solution, piece
The dosage forms such as agent, capsule, dripping pill, spray can be prepared by conventional method.The selection of pharmaceutical dosage form should be with administration mode phase
Match.
Before further describing the specific embodiments of the present invention, it should be appreciated that protection scope of the present invention is not limited to down
State specific specific embodiment；It is also understood that term used in the embodiment of the present invention is specific specific in order to describe
Embodiment, rather than limiting the scope of protection of the present invention.
Unless otherwise defined, all technical and scientific terms and those skilled in the art of the present technique used in the present invention are usual
The meaning of understanding is identical.In addition to specific method, equipment, material used in the embodiment, according to those skilled in the art
Grasp and record of the invention to the prior art, can also use and method described in the embodiment of the present invention, equipment, material
Any method, equipment and the material of the similar or equivalent prior art realizes the present invention.
Unless otherwise stated, disclosed in this invention experimental method, detection method, preparation method be all made of this technology neck
Molecular biology, biochemistry, chromatin Structure and the analysis of domain routine, analytical chemistry, cell culture, recombinant DNA technology with
And the routine techniques of related fields.These technologies also have perfect explanation in the prior art, and for details, reference can be made to Sambrook etc.
MOLECULAR CLONING:A LABORATORY MANUAL, Second edition, Cold Spring Harbor
Laboratory Press, 1989 andThird edition, 2001；Ausubel etc., CURRENT PROTOCOLS IN
MOLECULAR BIOLOGY, John Wiley&Sons, New York, 1987 and periodic updates；the
Series METHODS IN ENZYMOLOGY, Academic Press, San Diego；Wolffe, CHROMATIN
STRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998；METHODS IN
ENZYMOLOGH, Vol.304, Chromation (P.M.Wassarman and A.P.Wolffe, eds.), Academic
Press, San Diego, 1999；With METHODS IN MOLECULAR BIOLOGY, Vol.119, Chromatin
Protocols (P.B.Becker, ed.) Humana Press, Totowa, 1999 etc..
1. material and reagent
Cell culture medium (DMEM), fetal calf serum (FBS) are purchased from Life Technologies；Flow cytometer showed antibody α-
MCD4, α-mCD8, α-mCD3 ε, α-IFN γ, α-mCD45, α-mF4/80, α-mPD-1, α-mCD25 are purchased from eBioscience；
Western-blotting antibody α-E-cadherin, α-Vimentin are purchased from Bo Aosen.R-Roscovitine is purchased from MCE；
2. cell of the present invention and cultural method:
The initial source of 4T1 cell line and ATCC, are provided by Cell Bank of Chinese Academy of Sciences.With containing 10% fetal calf serum and
The RPMI1640 culture medium inoculated of 1% Pen .- Strep is placed in 37 DEG C, 5%CO in culture dish2Saturated humidity incubator
Middle adhere-wall culture.With the cell dissociation buffer vitellophag containing 0.25% pancreatin and 0.02%EDTA, cell is resuspended, is then inoculated with
Into new culture dish, passage cell.
3. animal model building of the present invention:
Balb/C female mice is purchased from dimension tonneau China.By above-mentioned cell culture to logarithmic growth phase.And disappeared with above-mentioned pancreatin
Change liquid and cell is resuspended into serum free medium, density 107Cell/ml is placed in spare on ice.It is drawn using micro syringe
20 μ l (about 105A cell), in-situ inoculating is in 20g or so female balb/c mouse (possessing complete immune system) groin
Fat pad in.Construct breast cancer orthotopic Mice Inoculated model.
4. inhibitor configuration of the present invention and administration mode:
CDK5 inhibitor, Roscovitine (HY-30237) are purchased from MCE company, are dissolved in the physiological saline containing 1%DMSO
In, final concentration of 0.2mg/ml.Intraperitoneal injection, dosage 2mg/kg, dosage period are two days 1 time；PD-1 inhibitor,
5. pharmacodynamic assessment method of the present invention:
1) survivorship curve: tumor growth curve uses vernier caliper, measures tumor tissues volume in each processing group, formula
Are as follows: V=0.5x long x wide x wide.It is primary every 2 days records；It is administered continuously in each processing group to natural death, and records dead day
Number draws survivorship curve.
2) lung tissue transfer stove is analyzed: each processing group mouse peritoneal injected fluorescein (10 μ g/mouse), with toy
Living imaging instrument (IVIS Lumina XR) carries out living imaging analysis；After administration 28 days, the neck that breaks puts to death mouse, separates each processing
Group mouse lung tissue, using saturation picric acid solution (25% formaldehyde, 5% acetic acid, 70% saturation picric acid) fixed, dyeing, solution
Cut open branch on count stove number under mirror；Isolated lung tissue is subjected to paraffin embedding, prepares histotomy, HE staining analysis is carried out and turns
Shift one's love condition.
3) serological analysis: separation peripheral blood serum, by ELISA detection method to wherein IL-10, IL-12, TGF β,
IFN γ expression is tested and analyzed.
4) tumor microenvironment is analyzed: being prepared tumor tissues single cell suspension by clostridiopetidase A enzymatic isolation method, is passed through fluidic cell
Instrument analyzes each immunocyte infiltration and activation situation in tumor tissues: CD8+T cell (CD45-FITC, CD3-PE, CD8-
CY5.5), Treg (CD3-PE, CD4-FITC, CD25-CY5.5), t cell activation (CD3-PE, CD8-CY5.5, IFN γ-
APC), macrophages infiltration and phenotype (F4/80-APC, CD206-PE, MHC-II-FITC), PD-1 expression (CD45-FITC,
CD3-PE, PD-1-APC) and Isotype control (IgG-FITC, IgG-PE, IgG-CY5.5, IgG-APC)；Pass through immunohistochemistry
Method, analyze tumor tissues in cell factor (IL-10, IL-12, IFNg, TGFb) expression, tumor cell proliferation
(Ki67), apoptosis (TUNEL) situation, extracellular matrix remodeling (collagen, MMPs)；By magnetic bead sorting, separate in tumor tissues
Tumor-associated macrophage, and related gene expression situation is analyzed by the method for RT-qPCR.
5) tumor stem cell related gene expression feelings tumour cell stemness transformation assay: are analyzed using immunohistochemical method
Condition (Oct4, Aldh1)；Flow cytometer showed tumor stem cell feature shows mark molecule (CD24, CD44) expression；It is external micro-
Sphere forms experiment, detects tumour cell stem cell properties in each processing group.
6. data statistic analysis
Be in figure release in especially indicate outer, all data statistics herein are all with GraphPad Prism (GraphPad
Software, Inc.) it is for statistical analysis, analysis method uses two-tailed unpaired Student ' s t-
test；* P < 0.05, * * P < 0.01, * * * P < 0.001, ns: without significant difference (no significant
difference)。
Following embodiment will the present invention is further illustrated in conjunction with attached drawing.
Embodiment 1: CDK5 (micromolecular inhibitor, CDK5 gene knockout) is inhibited to check tumour cell EMT, invasion, stemness
Conversion
The purpose of the present embodiment is the effect for verifying CDK5 inhibitor in tumour cell vicious transformation.CDK5 inhibitor
Roscovitine treatment non-small cell lung cancer has been subjected to clinical I phase experiment, has verified that its safety in clinical trial, but
It is to be not improved to tumor patient survival rate due to it so terminating experiment.Therefore, Roscovitine is selected to verify its conduct
A possibility that immunotherapy of tumors target spot.Experiment in vivo shows that Roscovitine can significantly inhibit breast cancer cell lung
The generation (Fig. 1 .A) of transfer.It is swollen that experiment in vitro shows that either Roscovitine or CDK5 knockout can significantly inhibit
Oncocyte invasive ability (Fig. 1 .B).E-cadherin expression decline, Vimentin expression increases can be with pointer tumour cell EMT
Process detects E-cadherin and Vimentin expression by WB, and CDK5 inhibitor can significantly inhibit as the result is shown
The expression of Vimentin raises simultaneously the expression of E-cadherin, illustrates that CDK5 inhibitor Roscovitine can be significantly inhibited
Tumour cell EMT process, and along with dose dependent.External microsphere forms experiment and further illustrates, CDK5 inhibitor
Roscovitine can significantly inhibit tumor stem cell characteristic.Simultaneously by being detected to tumor stem cell related gene expression
It was found that Roscovitine can significantly inhibit tumor stem cell related gene transcription (Aldh1, Fgfr1, Notch1,
Sox1, Oct4), and dose dependent is presented.
Embodiment 2:CDK5 inhibitor Roscovitine and PD-1 blocking antibody Combined Treatment inhibit mouse breast cancer development
Process.
At present in immunotherapy of tumors method, PD-1 blocking antibody and CTLA-4 blocking antibody are in melanoma and non-small
Preferable therapeutic effect is achieved in cell lung cancer, but complexity and the limitation of hyperimmunization inhibition due to tumor microenvironment
The effect of immunotherapy of tumors.The purpose of the present embodiment is verifying CDK5 as tumour cell itself grade malignancy is improved, and is adjusted
Save immune synergistic effect of microenvironment inhibition feature during combined immunization checkpoint blocking antibody is in immunotherapy of tumors.
The result shows that Roscovitine with obtain better curative effect in PD-1 blocking antibody combination therapy, the growth of mouse tumor is aobvious
Work slows down, while the life span of mouse significantly extends (Fig. 2 .A, B, D).The number of Pulmonary metastasis focuses significantly reduces, and implies tumour
Process has received significant inhibition (Fig. 2 .C).It is found by carrying out analysis to tumor-infiltrated T cell, Roscovitine treatment
Afterwards, the infiltration of T cell in tumor tissues is dramatically increased, and T cell infiltration is one of limitation most important factor of immunization therapy less.
To tumor-infiltrated t cell activation marker flow cytometer detection the results show that Roscovitine exclusive use cannot activate T cell, no
PD-1 and IFNg expression can be raised.It is only used in combination with PD-1 blocking antibody, T cell activation degree (figure could be enhanced
2.E).In addition, tumor-associated macrophage is one of immunocyte important in tumor tissues, immunosupress type M2 type macrophage is thin
Born of the same parents have extremely strong immunosuppression capability, and can significantly reverse M2 type macrophage to promote to M1 type after Roscovitine administration
(Fig. 2 .F) is converted into antineoplastic immune macrophage phenotype.In conclusion Roscovitine and PD-1 blocking antibody are combined
Treatment has synergistic effect, can inhibit to influence tumor microenvironment immune state, combined PD-while tumour cell vicious transformation
The antineoplastic immune that 1 blocking antibody further promotes T cell to mediate, to greatly extend tumor-bearing mice survival rate.
The foregoing is merely preferred embodiments of the invention, are not intended to limit the present invention, for those skilled in the art,
The invention may be variously modified and varied, any modification equivalent replacement done within the spirit and principles of the present invention,
Improve etc., it should all be included in the protection scope of the present invention.
Claims (12)
1. a kind of combination medicine for Breast cancer immunotherapy combines, including a effective amount of CDK5 inhibitor and a effective amount of
Immunologic test point blocking antibody.
2. being combined as described in claim 1 for the combination medicine of Breast cancer immunotherapy, which is characterized in that the CDK5
Inhibitor inhibits CDK5 activity, or inhibits CDK5 genetic transcription or expression.
3. being combined as described in claim 1 for the combination medicine of Breast cancer immunotherapy, which is characterized in that described
CDK5 inhibitor is selected from siRNA, shRNA, antibody, small molecule compound, peptide fragment.
4. being combined as described in claim 1 for the combination medicine of Breast cancer immunotherapy, which is characterized in that the CDK5
Inhibitor is selected from: R-Roscovitine, olomoucine (olomoucine) and its derivative, Dinaciclib (MK-7965,
SCH727965), AT7519, CIP, p5, Cpd1 etc..
5. being combined as described in claim 1 for the combination medicine of Breast cancer immunotherapy, which is characterized in that described immune
Checkpoint blocking antibody is selected from anti-PD-1 antibody, anti-PD-L1 antibody or anti-CTLA-4 antibody.
6. a kind of Breast cancer immunotherapy drug combination treatment method, to apply a effective amount of CDK5 inhibitor to Object associates,
And a effective amount of immunologic test point blocking antibody.
7. Breast cancer immunotherapy drug combination treatment method as claimed in claim 6, which is characterized in that synchronous or sequentially
Give a effective amount of CDK5 inhibitor and a effective amount of immunologic test point blocking antibody.
8. Breast cancer immunotherapy drug combination treatment method as claimed in claim 6, which is characterized in that the immunization therapy needle
Pair each hypotype of breast cancer: Luminal A (ER+/PR+Her2-)、Luminal B(ER+/PR+Her2+Or ER+/PR+Ki67 >
14%), Her2+(ER-PR-Her2+) and three yin breast cancer (Triple-negative breast cancer, TNBC) (ER-
PR-Her2-)。
9. Breast cancer immunotherapy drug combination treatment method as claimed in claim 6, which is characterized in that the CDK5 inhibits
Agent is selected from siRNA, shRNA, antibody, small molecule compound, peptide fragment, preferably R-Roscovitine.
10. Breast cancer immunotherapy drug combination treatment method as claimed in claim 6, which is characterized in that the immunologic test
Point blocking antibody is selected from anti-PD-1 antibody, anti-PD-L1 antibody or anti-CTLA-4 antibody.
11. Breast cancer immunotherapy drug combination treatment method as claimed in claim 6, which is characterized in that the object is
The immunocyte of mammal or the mammal, preferably cd8 t cell, macrophage.
12. Breast cancer immunotherapy drug combination treatment method as claimed in claim 6, which is characterized in that the lactation is dynamic
Object is behaved.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810180887.5A CN110215516A (en) | 2018-03-02 | 2018-03-02 | It is a kind of to inhibit the treatment of CDK5 synergetic immunity in the application inhibited in breast cancer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810180887.5A CN110215516A (en) | 2018-03-02 | 2018-03-02 | It is a kind of to inhibit the treatment of CDK5 synergetic immunity in the application inhibited in breast cancer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110215516A true CN110215516A (en) | 2019-09-10 |
Family
ID=67822295
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810180887.5A Pending CN110215516A (en) | 2018-03-02 | 2018-03-02 | It is a kind of to inhibit the treatment of CDK5 synergetic immunity in the application inhibited in breast cancer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110215516A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111410693A (en) * | 2020-04-15 | 2020-07-14 | 山西农业大学 | CDK5 resistant nano antibody and application thereof |
| WO2022179507A1 (en) * | 2021-02-23 | 2022-09-01 | Jiangsu Alphamab Biopharmaceuticals Co., Ltd. | Methods of preventing, alleviating or treating tumors |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105451770A (en) * | 2013-08-20 | 2016-03-30 | 默沙东公司 | Treating cancer with a combination of a pd-1 antagonist and dinaciclib |
| CN106955354A (en) * | 2016-01-11 | 2017-07-18 | 中国科学院上海生命科学研究院 | Combination medicine for immunotherapy of tumors is combined |
| WO2017205514A1 (en) * | 2016-05-25 | 2017-11-30 | Case Western Reserve University | Methods of sensitizing cancer to immunotherapy |
-
2018
- 2018-03-02 CN CN201810180887.5A patent/CN110215516A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105451770A (en) * | 2013-08-20 | 2016-03-30 | 默沙东公司 | Treating cancer with a combination of a pd-1 antagonist and dinaciclib |
| CN106955354A (en) * | 2016-01-11 | 2017-07-18 | 中国科学院上海生命科学研究院 | Combination medicine for immunotherapy of tumors is combined |
| WO2017205514A1 (en) * | 2016-05-25 | 2017-11-30 | Case Western Reserve University | Methods of sensitizing cancer to immunotherapy |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111410693A (en) * | 2020-04-15 | 2020-07-14 | 山西农业大学 | CDK5 resistant nano antibody and application thereof |
| CN111410693B (en) * | 2020-04-15 | 2020-12-22 | 山西农业大学 | CDK5 resistant nano antibody and application thereof |
| WO2022179507A1 (en) * | 2021-02-23 | 2022-09-01 | Jiangsu Alphamab Biopharmaceuticals Co., Ltd. | Methods of preventing, alleviating or treating tumors |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Agliano et al. | The challenge of targeting cancer stem cells to halt metastasis | |
| Kesari | Understanding glioblastoma tumor biology: the potential to improve current diagnosis and treatments | |
| Florio et al. | The status of the art of human malignant glioma management: the promising role of targeting tumor-initiating cells | |
| CN105899223A (en) | Treatment of metastatic prostate cancer | |
| Tian et al. | HYD-PEP06 suppresses hepatocellular carcinoma metastasis, epithelial–mesenchymal transition and cancer stem cell-like properties by inhibiting PI3K/AKT and WNT/β-catenin signaling activation | |
| Fu et al. | Combination foretinib and anti-PD-1 antibody immunotherapy for colorectal carcinoma | |
| Chang et al. | PI3Kαδ inhibitor combined with radiation enhances the antitumor immune effect of anti-PD1 in a syngeneic murine triple-negative breast cancer model | |
| CN102227635A (en) | Na/k-atpase expression as indicator for treatment of cancer | |
| CN110215516A (en) | It is a kind of to inhibit the treatment of CDK5 synergetic immunity in the application inhibited in breast cancer | |
| Carter et al. | A three-dimensional organoid model of primary breast cancer to investigate the effects of oncolytic virotherapy | |
| Zhao et al. | Distinct EphB4-mediated mechanisms of apoptotic and resistance to dasatinib in human chronic myeloid leukemia and K562 cell lines | |
| Juric et al. | Monocytes promote liver carcinogenesis in an oncogene-specific manner | |
| CN110464845A (en) | A kind of combination medicine combination and its application for Breast cancer immunotherapy | |
| Qin et al. | Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma therapy: Challenges and opportunities | |
| CN111297943A (en) | Supercritical CO2Application of fluid extraction of salvia miltiorrhiza extract | |
| CN114917359A (en) | PROTAC composition aiming at cell cycle multi-space-time distribution anti-cancer targets | |
| US11179425B2 (en) | Method of activating tumor-infiltrating lymphocytes (TILs) | |
| Wang et al. | Tumor-associated macrophages facilitate oral squamous cell carcinomas migration and invasion by MIF/NLRP3/IL-1β circuit: A crosstalk interrupted by melatonin | |
| KR20220123033A (en) | Cancer Treatment Methods and Medicines | |
| CN106333951B (en) | A kind of application of mTOR kinase inhibitors and the composition of mapk kinase inhibitor | |
| CN111419832A (en) | Pharmaceutical composition and application thereof in preparing medicines for treating tumors | |
| CN116983310B (en) | Pharmaceutical composition for treating liver cancer and application thereof | |
| CN115068487B (en) | Antitumor combination preparation containing hydroxyprogesterone caproate and application thereof | |
| CN113769096B (en) | Medical application of glucose 6-phosphate dehydrogenase inhibitor | |
| CN108795945A (en) | DNA nanometers of trains of self assembly aptamer and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190910 |