CN112543651A - 使car t细胞恢复活力 - Google Patents
使car t细胞恢复活力 Download PDFInfo
- Publication number
- CN112543651A CN112543651A CN201980052586.7A CN201980052586A CN112543651A CN 112543651 A CN112543651 A CN 112543651A CN 201980052586 A CN201980052586 A CN 201980052586A CN 112543651 A CN112543651 A CN 112543651A
- Authority
- CN
- China
- Prior art keywords
- car
- component
- cells
- drug
- targeting ligand
- 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.)
- Granted
Links
- 108091007741 Chimeric antigen receptor T cells Proteins 0.000 title claims abstract description 57
- 230000003716 rejuvenation Effects 0.000 title claims description 14
- 239000003814 drug Substances 0.000 claims abstract description 63
- 239000003446 ligand Substances 0.000 claims abstract description 63
- 229940079593 drug Drugs 0.000 claims abstract description 61
- 230000008685 targeting Effects 0.000 claims abstract description 58
- 108020003175 receptors Proteins 0.000 claims abstract description 36
- 230000004927 fusion Effects 0.000 claims abstract description 25
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 55
- 238000000034 method Methods 0.000 claims description 43
- 229940044616 toll-like receptor 7 agonist Drugs 0.000 claims description 38
- 102000005962 receptors Human genes 0.000 claims description 35
- 230000027455 binding Effects 0.000 claims description 29
- 239000003112 inhibitor Substances 0.000 claims description 29
- 239000000427 antigen Substances 0.000 claims description 19
- 108091007433 antigens Proteins 0.000 claims description 19
- 102000036639 antigens Human genes 0.000 claims description 19
- 239000000556 agonist Substances 0.000 claims description 18
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 claims description 18
- QJJXYPPXXYFBGM-LFZNUXCKSA-N Tacrolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1\C=C(/C)[C@@H]1[C@H](C)[C@@H](O)CC(=O)[C@H](CC=C)/C=C(C)/C[C@H](C)C[C@H](OC)[C@H]([C@H](C[C@H]2C)OC)O[C@@]2(O)C(=O)C(=O)N2CCCC[C@H]2C(=O)O1 QJJXYPPXXYFBGM-LFZNUXCKSA-N 0.000 claims description 17
- 108010015832 Non-Receptor Type 2 Protein Tyrosine Phosphatase Proteins 0.000 claims description 16
- 102100033141 Tyrosine-protein phosphatase non-receptor type 2 Human genes 0.000 claims description 16
- 102100039390 Toll-like receptor 7 Human genes 0.000 claims description 15
- 230000002401 inhibitory effect Effects 0.000 claims description 14
- 230000011664 signaling Effects 0.000 claims description 14
- 101000669402 Homo sapiens Toll-like receptor 7 Proteins 0.000 claims description 11
- 238000011068 loading method Methods 0.000 claims description 11
- 125000006850 spacer group Chemical group 0.000 claims description 11
- 108010062677 Diacylglycerol Kinase Proteins 0.000 claims description 10
- 239000012528 membrane Substances 0.000 claims description 10
- 230000037361 pathway Effects 0.000 claims description 10
- 102000010451 Folate receptor alpha Human genes 0.000 claims description 9
- 108050001931 Folate receptor alpha Proteins 0.000 claims description 9
- 101000617285 Homo sapiens Tyrosine-protein phosphatase non-receptor type 6 Proteins 0.000 claims description 9
- 229940044665 STING agonist Drugs 0.000 claims description 9
- 102000018679 Tacrolimus Binding Proteins Human genes 0.000 claims description 9
- 108010027179 Tacrolimus Binding Proteins Proteins 0.000 claims description 9
- 102100021657 Tyrosine-protein phosphatase non-receptor type 6 Human genes 0.000 claims description 9
- 210000001163 endosome Anatomy 0.000 claims description 9
- 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 claims description 9
- 238000004873 anchoring Methods 0.000 claims description 8
- 102000011107 Diacylglycerol Kinase Human genes 0.000 claims description 7
- 102000006815 folate receptor Human genes 0.000 claims description 7
- 108020005243 folate receptor Proteins 0.000 claims description 7
- 229940014144 folate Drugs 0.000 claims description 6
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 6
- 235000019152 folic acid Nutrition 0.000 claims description 6
- 239000011724 folic acid Substances 0.000 claims description 6
- 108010050904 Interferons Proteins 0.000 claims description 5
- 229940124615 TLR 7 agonist Drugs 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 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 claims description 5
- 229940122907 Phosphatase inhibitor Drugs 0.000 claims description 4
- 108010060825 Toll-Like Receptor 7 Proteins 0.000 claims description 4
- 210000000172 cytosol Anatomy 0.000 claims description 4
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims description 2
- 230000035899 viability Effects 0.000 claims description 2
- 230000009711 regulatory function Effects 0.000 abstract 1
- 108010019670 Chimeric Antigen Receptors Proteins 0.000 description 62
- 210000004027 cell Anatomy 0.000 description 51
- 206010028980 Neoplasm Diseases 0.000 description 39
- 230000000694 effects Effects 0.000 description 26
- 230000006870 function Effects 0.000 description 13
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 12
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 12
- 102000002689 Toll-like receptor Human genes 0.000 description 11
- 108020000411 Toll-like receptor Proteins 0.000 description 11
- 230000002147 killing effect Effects 0.000 description 11
- 201000011510 cancer Diseases 0.000 description 9
- 230000009089 cytolysis Effects 0.000 description 9
- 238000013461 design Methods 0.000 description 9
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 8
- 101710196623 Stimulator of interferon genes protein Proteins 0.000 description 8
- 230000014509 gene expression Effects 0.000 description 8
- 230000001965 increasing effect Effects 0.000 description 8
- QJJXYPPXXYFBGM-SHYZHZOCSA-N tacrolimus Natural products CO[C@H]1C[C@H](CC[C@@H]1O)C=C(C)[C@H]2OC(=O)[C@H]3CCCCN3C(=O)C(=O)[C@@]4(O)O[C@@H]([C@H](C[C@H]4C)OC)[C@@H](C[C@H](C)CC(=C[C@@H](CC=C)C(=O)C[C@H](O)[C@H]2C)C)OC QJJXYPPXXYFBGM-SHYZHZOCSA-N 0.000 description 8
- 238000002560 therapeutic procedure Methods 0.000 description 8
- 238000000338 in vitro Methods 0.000 description 7
- 230000002829 reductive effect Effects 0.000 description 7
- 210000003171 tumor-infiltrating lymphocyte Anatomy 0.000 description 7
- 102100034458 Hepatitis A virus cellular receptor 2 Human genes 0.000 description 6
- 102100038970 Histone-lysine N-methyltransferase EZH2 Human genes 0.000 description 6
- 238000011156 evaluation Methods 0.000 description 6
- 230000001506 immunosuppresive effect Effects 0.000 description 6
- 230000000638 stimulation Effects 0.000 description 6
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 5
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 5
- 101000882127 Homo sapiens Histone-lysine N-methyltransferase EZH2 Proteins 0.000 description 5
- 230000001472 cytotoxic effect Effects 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 230000028327 secretion Effects 0.000 description 5
- 150000003384 small molecules Chemical class 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- 238000011357 CAR T-cell therapy Methods 0.000 description 4
- 101150046249 Havcr2 gene Proteins 0.000 description 4
- 102000017578 LAG3 Human genes 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 238000007792 addition Methods 0.000 description 4
- 238000013459 approach Methods 0.000 description 4
- 238000002659 cell therapy Methods 0.000 description 4
- 231100000433 cytotoxic Toxicity 0.000 description 4
- 150000001982 diacylglycerols Chemical class 0.000 description 4
- 231100000673 dose–response relationship Toxicity 0.000 description 4
- 210000004698 lymphocyte Anatomy 0.000 description 4
- 230000003389 potentiating effect Effects 0.000 description 4
- 230000000770 proinflammatory effect Effects 0.000 description 4
- 239000003801 protein tyrosine phosphatase 1B inhibitor Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 230000002441 reversible effect Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 description 3
- 102100039498 Cytotoxic T-lymphocyte protein 4 Human genes 0.000 description 3
- 102100022735 Diacylglycerol kinase alpha Human genes 0.000 description 3
- 101710181033 Diacylglycerol kinase alpha Proteins 0.000 description 3
- 229940076838 Immune checkpoint inhibitor Drugs 0.000 description 3
- 101150030213 Lag3 gene Proteins 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000002648 combination therapy Methods 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 229940044680 immune agonist Drugs 0.000 description 3
- 239000012651 immune agonist Substances 0.000 description 3
- 239000012274 immune-checkpoint protein inhibitor Substances 0.000 description 3
- 230000002101 lytic effect Effects 0.000 description 3
- 210000002540 macrophage Anatomy 0.000 description 3
- 210000000066 myeloid cell Anatomy 0.000 description 3
- 230000035515 penetration Effects 0.000 description 3
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 102100023990 60S ribosomal protein L17 Human genes 0.000 description 2
- 108010021064 CTLA-4 Antigen Proteins 0.000 description 2
- 229940045513 CTLA4 antagonist Drugs 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 101001068133 Homo sapiens Hepatitis A virus cellular receptor 2 Proteins 0.000 description 2
- 206010025323 Lymphomas Diseases 0.000 description 2
- 102100024193 Mitogen-activated protein kinase 1 Human genes 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical group C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 2
- 208000036142 Viral infection Diseases 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 238000009175 antibody therapy Methods 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 238000003501 co-culture Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000001086 cytosolic effect Effects 0.000 description 2
- 238000002716 delivery method Methods 0.000 description 2
- 238000001212 derivatisation Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000010874 in vitro model Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 108091070501 miRNA Proteins 0.000 description 2
- 230000002018 overexpression Effects 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000003909 pattern recognition Methods 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- BXNMTOQRYBFHNZ-UHFFFAOYSA-N resiquimod Chemical compound C1=CC=CC2=C(N(C(COCC)=N3)CC(C)(C)O)C3=C(N)N=C21 BXNMTOQRYBFHNZ-UHFFFAOYSA-N 0.000 description 2
- 230000000284 resting effect Effects 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 230000002459 sustained effect Effects 0.000 description 2
- 238000007910 systemic administration Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- FKSFKBQGSFSOSM-QFIPXVFZSA-N 1-[(2S)-butan-2-yl]-N-[(4,6-dimethyl-2-oxo-1H-pyridin-3-yl)methyl]-3-methyl-6-[6-(1-piperazinyl)-3-pyridinyl]-4-indolecarboxamide Chemical compound C1=C2N([C@@H](C)CC)C=C(C)C2=C(C(=O)NCC=2C(NC(C)=CC=2C)=O)C=C1C(C=N1)=CC=C1N1CCNCC1 FKSFKBQGSFSOSM-QFIPXVFZSA-N 0.000 description 1
- KXSKAZFMTGADIV-UHFFFAOYSA-N 2-[3-(2-hydroxyethoxy)propoxy]ethanol Chemical compound OCCOCCCOCCO KXSKAZFMTGADIV-UHFFFAOYSA-N 0.000 description 1
- UZOSBNQFZUJWFP-UHFFFAOYSA-N 4-[2-(6-methylpyridin-2-yl)-5,6-dihydro-4h-pyrrolo[1,2-b]pyrazol-3-yl]quinoline-6-carboxamide;hydrate Chemical compound O.CC1=CC=CC(C=2C(=C3CCCN3N=2)C=2C3=CC(=CC=C3N=CC=2)C(N)=O)=N1 UZOSBNQFZUJWFP-UHFFFAOYSA-N 0.000 description 1
- 229940126253 ADU-S100 Drugs 0.000 description 1
- 208000004736 B-Cell Leukemia Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 108091033409 CRISPR Proteins 0.000 description 1
- 238000010354 CRISPR gene editing Methods 0.000 description 1
- 101100043687 Caenorhabditis elegans stim-1 gene Proteins 0.000 description 1
- 102100025570 Cancer/testis antigen 1 Human genes 0.000 description 1
- 206010050685 Cytokine storm Diseases 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 108010058222 Deoxyguanosine kinase Proteins 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 229920001917 Ficoll Polymers 0.000 description 1
- 101710196274 Histone-lysine N-methyltransferase EZH2 Proteins 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000856237 Homo sapiens Cancer/testis antigen 1 Proteins 0.000 description 1
- 101000889276 Homo sapiens Cytotoxic T-lymphocyte protein 4 Proteins 0.000 description 1
- 101001137987 Homo sapiens Lymphocyte activation gene 3 protein Proteins 0.000 description 1
- 101000693243 Homo sapiens Paternally-expressed gene 3 protein Proteins 0.000 description 1
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 1
- 101000831567 Homo sapiens Toll-like receptor 2 Proteins 0.000 description 1
- 101150103227 IFN gene Proteins 0.000 description 1
- 206010062016 Immunosuppression Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- IVRXNBXKWIJUQB-UHFFFAOYSA-N LY-2157299 Chemical compound CC1=CC=CC(C=2C(=C3CCCN3N=2)C=2C3=CC(=CC=C3N=CC=2)C(N)=O)=N1 IVRXNBXKWIJUQB-UHFFFAOYSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 210000004322 M2 macrophage Anatomy 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 102000016397 Methyltransferase Human genes 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 102000038030 PI3Ks Human genes 0.000 description 1
- 108091007960 PI3Ks Proteins 0.000 description 1
- 102100025757 Paternally-expressed gene 3 protein Human genes 0.000 description 1
- 230000006044 T cell activation Effects 0.000 description 1
- 108010092262 T-Cell Antigen Receptors Proteins 0.000 description 1
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 1
- 102100033456 TGF-beta receptor type-1 Human genes 0.000 description 1
- 229940124613 TLR 7/8 agonist Drugs 0.000 description 1
- 102100024333 Toll-like receptor 2 Human genes 0.000 description 1
- 108010011702 Transforming Growth Factor-beta Type I Receptor Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 238000011467 adoptive cell therapy Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- RFCBNSCSPXMEBK-INFSMZHSSA-N c-GMP-AMP Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]3[C@@H](O)[C@H](N4C5=NC=NC(N)=C5N=C4)O[C@@H]3COP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=C(NC2=O)N)=C2N=C1 RFCBNSCSPXMEBK-INFSMZHSSA-N 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000002619 cancer immunotherapy Methods 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 230000009137 competitive binding Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 206010052015 cytokine release syndrome Diseases 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 238000000432 density-gradient centrifugation Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000008995 epigenetic change Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 229950000456 galunisertib Drugs 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 201000005787 hematologic cancer Diseases 0.000 description 1
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 1
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 1
- 239000012216 imaging agent Substances 0.000 description 1
- 229960002751 imiquimod Drugs 0.000 description 1
- DOUYETYNHWVLEO-UHFFFAOYSA-N imiquimod Chemical compound C1=CC=CC2=C3N(CC(C)C)C=NC3=C(N)N=C21 DOUYETYNHWVLEO-UHFFFAOYSA-N 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000005746 immune checkpoint blockade Effects 0.000 description 1
- 239000012642 immune effector Substances 0.000 description 1
- 229940121354 immunomodulator Drugs 0.000 description 1
- 229960001438 immunostimulant agent Drugs 0.000 description 1
- 239000003022 immunostimulating agent Substances 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 108091005434 innate immune receptors Proteins 0.000 description 1
- 230000011542 interferon-beta production Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000009191 jumping Effects 0.000 description 1
- 229940043355 kinase inhibitor Drugs 0.000 description 1
- 238000000670 ligand binding assay Methods 0.000 description 1
- 108020001756 ligand binding domains Proteins 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- NSQSAUGJQHDYNO-UHFFFAOYSA-N n-[(4,6-dimethyl-2-oxo-1h-pyridin-3-yl)methyl]-3-[ethyl(oxan-4-yl)amino]-2-methyl-5-[4-(morpholin-4-ylmethyl)phenyl]benzamide Chemical compound C=1C(C=2C=CC(CN3CCOCC3)=CC=2)=CC(C(=O)NCC=2C(NC(C)=CC=2C)=O)=C(C)C=1N(CC)C1CCOCC1 NSQSAUGJQHDYNO-UHFFFAOYSA-N 0.000 description 1
- HPODOLXTMDHLLC-QGZVFWFLSA-N n-[(4-methoxy-6-methyl-2-oxo-1h-pyridin-3-yl)methyl]-2-methyl-1-[(1r)-1-[1-(2,2,2-trifluoroethyl)piperidin-4-yl]ethyl]indole-3-carboxamide Chemical compound C1=C(C)NC(=O)C(CNC(=O)C=2C3=CC=CC=C3N([C@H](C)C3CCN(CC(F)(F)F)CC3)C=2C)=C1OC HPODOLXTMDHLLC-QGZVFWFLSA-N 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000002611 ovarian Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000003757 phosphotransferase inhibitor Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000007420 reactivation Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 210000003289 regulatory T cell Anatomy 0.000 description 1
- 230000008672 reprogramming Effects 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 102000009076 src-Family Kinases Human genes 0.000 description 1
- 108010087686 src-Family Kinases Proteins 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 210000004981 tumor-associated macrophage Anatomy 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 210000005166 vasculature Anatomy 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
- A61K31/381—Heterocyclic compounds having sulfur as a ring hetero atom having five-membered rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
- A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4535—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a heterocyclic ring having sulfur as a ring hetero atom, e.g. pizotifen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
- A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
- A61K31/7076—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/463—Cellular immunotherapy characterised by recombinant expression
- A61K39/4631—Chimeric Antigen Receptors [CAR]
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
- A61K39/464402—Receptors, cell surface antigens or cell surface determinants
- A61K39/464411—Immunoglobulin superfamily
- A61K39/464412—CD19 or B4
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
- A61K47/551—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0041—Xanthene dyes, used in vivo, e.g. administered to a mice, e.g. rhodamines, rose Bengal
- A61K49/0043—Fluorescein, used in vivo
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0052—Small organic molecules
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70503—Immunoglobulin superfamily
- C07K14/7051—T-cell receptor (TcR)-CD3 complex
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
- A61K2239/46—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the cancer treated
- A61K2239/48—Blood cells, e.g. leukemia or lymphoma
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
Abstract
本文中提供了与靶向配体缀合的药物载荷,其被专门设计成递送至耗竭的CAR T细胞以使这些CAR T细胞恢复活力。被靶向的CAR T细胞以融合受体进行修饰,所述融合受体可与所述靶向配体结合并且使所缀合的药物载荷内化,以执行其对耗竭的CAR T细胞的调节功能。
Description
技术领域
本公开内容提供了使癌抗原耗竭的嵌合抗原受体T(chimeric antigen receptorT,CAR T)细胞恢复活力(rejuvenate)的系统。具体地,所述系统在经典CAR构建体(construct)中包含融合受体,其中所述融合受体提供了配体结合模块(ligand bindingmodule),其识别高亲和力配体-载荷药物缀合物(ligand-payload drug conjugate)以递送药物载荷,所述药物载荷被设计成阻断耗竭的CAR T中的抑制性信号传导或通过抗原非依赖性途径重新激活CAR T。
背景技术
在过去的二十年中,嵌合抗原受体(CAR)T细胞治疗领域取得了巨大进步。CAR构建体由四个部分组成:(1)针对肿瘤特异性抗原的胞外结合部分,(2)铰链结构域,(3)跨膜结构域,以及(4)多种激活结构域(例如,CD28、4-1BB和CD3ζ链)的组合。在针对CD19阳性B细胞白血病的CAR T治疗中取得了最令人印象深刻的成功,其中在多项临床试验中已达到超过80%的完全缓解率。
相比之下,迄今为止已证明通过CAR T治疗实体瘤更具挑战性。在临床前研究中,在免疫缺陷小鼠中使用同基因小鼠模型或异种移植肿瘤模型已经取得了巨大的成功。然而,涉及实体瘤(例如,乳腺癌、卵巢癌或肺癌)的临床试验均没有显示出与对照相比的改善。通常来说,其具有与其他过继细胞治疗所面临的相同局限性,包括(1)肿瘤穿透性差;(2)低氧压;(3)免疫抑制性肿瘤微环境,其包括肿瘤相关的巨噬细胞、成纤维细胞和抑制性细胞因子1。CAR T细胞与肿瘤浸润淋巴细胞(TIL)一样,可以通过这种抑制性微环境再培养(re-educated)并转变为“功能低下”状态,其特征在于共抑制分子(即,PD-1、Tim-3、LAG-3等)的过表达、降低的INFγ分泌和杀伤能力。来自卵巢癌患者样品的数据显示,大多数PD-1+CD8+T细胞缺乏CD 127的表达,已知CD 127对于T细胞中的效应子-记忆的转变是重要的。LAG-3的过表达也与TCR特异性CD8+TIL的效应子功能负相关。此外,PD-1+LAG-3+双阳性T细胞表现出较低的INFγ产生2。类似地,NY-ESO-1 TCR特异人T细胞在小鼠实体瘤模型中功能低下,表明由于微环境和通过持续暴露于抗原对T细胞的持续激活二者造成的共抑制分子的高表达和低效的抗肿瘤效果3。因此,为了更好的实体瘤治疗,非常期望逆转抑制性微环境,更重要的是,使耗竭的CAR T细胞恢复活力。
发明概述
本公开内容提供了使耗竭的经典CAR T细胞恢复活力的系统。所述系统包含至少两种组分:第一组分是包含与药物载荷共价连接的靶向配体的缀合物;并且第二组分是与膜锚定模块(membrane-anchoring module)连接的靶向配体结合模块。第二组分的靶向配体结合模块以高亲和力识别第一组分中的靶向配体以形成复合物,并且载荷药物阻断耗竭的CAR T的抑制性信号传导,或者通过抗原非依赖性途径重新激活所述CAR T。膜锚定模块介导该双组分复合物内化到耗竭的CAR T细胞中。
在一些优选的实施方案中,前述第一组分的靶向配体是叶酸(folate)、FITC或FK506。
在一些优选的实施方案中,前述第二组分的靶向配体结合模块包含叶酸受体、抗FITC抗体片段或FKBP。
在一些优选的实施方案中,前述膜锚定模块是叶酸受体。
在一些优选的实施方案中,前述第一组分包含在靶向配体与载荷药物之间的可释放的接头(linker)。
在一些优选的实施方案中,前述第一组分包含在靶向配体与载荷药物之间的不可释放的接头。
在一些优选的实施方案中,前述靶向配体与靶向配体结合模块之间的结合亲和力在亚纳摩范围(sub-nanomolar range)内。
在一些优选的实施方案中,前述药物载荷是Toll样受体7(Toll Like Receptor7,TLR7)激动剂或干扰素基因刺激物(Simulator of interferon genes,STING)激动剂。
在一些优选的实施方案中,前述药物载荷是以下蛋白质的抑制剂:SHP1/2、TC-PTP或DGKα、TGFβ。
在一些优选的实施方案中,前述TLR7激动剂具有以下结构:
在一些优选的实施方案中,前述第一组分是具有以下结构的荧光素-TLR7激动剂:
在一些优选的实施方案中,前述第一组分是具有以下结构的FK506-TLR7激动剂:
在一些优选的实施方式中,前述第一组分是以下之一:
在一些优选的实施方案中,前述第一组分包含选自以下TC-PTP磷酸酶抑制剂的载荷药物:
在一些优选的实施方案中,前述磷酸酶抑制剂与荧光素或FK506(他克莫司)连接以形成以下结构:
在一些优选的实施方案中,前述第一组分中的载荷药物包含以下结构之一的STING激动剂:
在一些优选的实施方案中,前述第一组分在靶向配体与载荷药物之间包含间隔基(spacer),所述间隔基选自以下结构:
本公开内容还提供了使耗竭的CAR T细胞恢复活力的方法。所述方法包括以下步骤:
a.向耗竭的CAR T细胞提供包含缀合物的第一组分,其中所述缀合物包含通过可释放或不可释放的接头与药物载荷共价连接的靶向配体;
b.向所述耗竭的CAR T细胞提供包含与耗竭的CAR构建体连接的融合受体的第二组分,其中所述融合受体包含靶向配体结合模块和膜锚定模块;
c.使第二组分的靶向配体结合模块与第一组分中的靶向配体结合以形成复合物;
d.使膜锚定模块介导复合物向耗竭的CAR T细胞的内化;
e.使载荷药物阻断耗竭的CAR T的抑制性信号传导或通过抗原非依赖性途径重新激活所述CAR T。
在一些优选的实施方案中,前述方法携带在耗竭的CAR T的内体中执行其功能的载荷药物,并且靶向配体与载荷药物通过不可释放的接头连接。
在一些优选的实施方案中,前述方法携带作为耗竭的CAR T的胞质溶胶中的游离药物执行其功能的载荷药物,并且靶向配体与载荷药物通过可释放的接头连接。
在一些优选的实施方案中,前述方法中第一组分的靶向配体是叶酸、FITC或FK506。
在一些优选的实施方案中,前述方法中第二组分的靶向配体结合模块是抗FITC、叶酸受体或FKBP。
在一些优选的实施方案中,前述方法中第二组分的靶向配体结合模块是叶酸受体α(FRa)。
在一些优选的实施方案中,前述方法中第一组分的药物载荷是Toll样受体7(TLR7)激动剂或干扰素基因刺激物(STING)激动剂。
在一些优选的实施方案中,前述方法中第一组分的药物载荷是以下蛋白质的抑制剂:SHP1/2、TC-PTP或DGKα、TGFβ。
在一些优选的实施方案中,前述方法中TLR7激动剂具有以下结构:
在一些优选的实施方案中,前述方法中第一组分是具有以下结构的荧光素-TLR7激动剂:
在一些优选的实施方案中,前述方法中第一组分是具有以下结构的FK506-TLR7激动剂:
在一些优选的实施方案中,前述方法中第一组分是:
在一些优选的实施方案中,前述方法中第一组分包含选自以下TC-PTP磷酸酶抑制剂的载荷药物:
在一些优选的实施方案中,前述方法中第一组分包含磷酸酶抑制剂,其与荧光素或FK506(他克莫司)连接以形成以下结构:
在一些优选的实施方案中,前述方法中第一组分包含以下结构的STING激动剂的载荷药物:
在一些优选的实施方案中,前述方法中的第一组分包含在靶向配体与载荷药物之间的间隔基,所述间隔基选自以下结构:
参考以下附图、相关说明和权利要求书,将更好地理解本发明的这些和其他特征、方面和优点。
附图简述
图1. 1a,耗竭模型的图示说明。1b,在体外新鲜Raji细胞刺激3次之后抗CD19 CART细胞变得耗竭,如通过降低的杀伤作用显示的。
图2.秘密通道(secret passageway)的图示说明。FKBP或抗FITC与FRa连接作为融合受体,其不断内化并且将FK506或FITC连接的载荷递送到细胞中。
图3.FK506和FITC与相应融合受体的结合亲和力。FK506-罗丹明对FKBP-FR融合受体显示Kd=3.39nM,而FITC-AF647对抗FITC-FR显示Kd=8.03nM。
图4. 4a,TLR7激动剂的结构。4b,TLR7激动剂对耗竭的抗CD19 CAR T细胞的恢复活力作用,表现为提高的杀伤作用、INFr水平和降低的共抑制分子水平。
图5.TLR7激动剂的化学结构。
图6.用于诱导CAR T细胞耗竭的体外模型。(A)将CD19+Raji与抗CD19CAR T细胞以1∶1的比例共培养,每12小时添加新鲜的Raji细胞。(B)随着刺激次数(添加Raji细胞的次数)增加,CAR T细胞的裂解作用逐渐降低。CD19-K562细胞用作对照。(C)CD4和CD8阳性CART细胞中stim1和stim3的共抑制分子PD-1、LAG3和Tim3的表达水平变化。
图7.TLR7激动剂和PTP1b抑制剂使耗竭的CAR T细胞恢复活力的作用的评价。(A)TLR7激动剂和PTP1b抑制剂的化学结构。(B至C)将耗竭的CAR T细胞与不同浓度的TLR7激动剂和PTP1b抑制剂一起孵育,通过裂解作用和INFγ(B)以及孵育之后PD-1LAG-3和Tim3的表达水平(C)进行监测。*表示p值<0.05,**<0.01,ns=不显著。
图8.TLR7激动剂用于不可释放的配体靶向递送的潜在衍生位点的评价。(A)TLR7激动剂类似物的化学结构。(B)将耗竭的CAR T细胞与不同浓度的TLR7类似物一起孵育,测量裂解作用并且与未处理组进行比较。
图9.使用FITC作为靶向配体的TLR7激动剂的可释放和不可释放靶向递送的设计和评价。(A)FITC-TLR7激动剂的化学结构的图示。(B)将耗竭的CAR T细胞与不同浓度的可释放的FITC-TLR7和不可释放的FITC-JTLR7激动剂一起孵育,测量裂解作用并且与未处理组进行比较。*表示p值<0.05,**<0.01,ns=不显著。(C)左侧,与FITC scFv(灰色)结合的FITC(绿色)的晶体结构(PDB:1X9Q,左侧),FITC到FTIC scFv边缘之间的距离测量为约右侧,类似地,显示了与TLR7(灰色)结合的R-848(红色)的晶体结构(PDB:5GMF),R-848到TLR7边缘之间的距离为约(D)图示出了用于不可释放的FITC-TLR7激动剂的两种可能的工作机制,“到达(Reaching)”或“跳跃(Jumping)”模式。
图10.可用于靶向配体-载荷缀合物设计的可变刚性和疏水性的化学接头。
发明详述
尽管在本文的附图和说明中详细示出和描述了本公开内容的概念,但是附图及其说明中的结果在特征上应被认为是示例性的而不是限制性的;应当理解,仅示出和描述了示例性实施方案,并且希望保护落入本公开内容精神内的所有改变和修改。
除非另有定义,否则科学和技术术语具有与本公开内容所属领域的普通技术人员的通常理解相同的含义。
嵌合抗原受体(CAR)T细胞治疗最近在治疗多种类型的造血癌症中获得了实质性的成功。同时,人们还应该认识到,一些淋巴瘤和大多数实体瘤病例在CAR T细胞治疗的情况下仍然具有非常低的响应率或高的复发率。这主要是由于以下三种原因之一或组合:1.在CAR T细胞的选择压力下出现了抗原阴性癌细胞集落,如在用抗CD 19 CAR T细胞治疗的CD 19阴性ALL复发的情况中看到的;2.由于异常的肿瘤脉管系统、密集的基质屏障和抑制性微环境,实体瘤中CAR T细胞的初始归巢和增殖受阻;3.持续肿瘤抗原暴露之后,CAR T细胞逐渐耗竭并且裂解作用降低。假设对于给定的实体瘤患者不存在抗原丢失(通过活检采样验证),则CAR T细胞治疗潜在失败的原因很可能是由后两种原因引起的。因此,为了提高CAR T细胞在实体瘤中的效力,非常期望用于CAR T细胞的体内评价和恢复活力的实用方法。
在这里,我们描述了CAR T细胞中私有通道融合受体的新设计,作为实现这两个目标的通用平台。该FITC-FR融合受体由两部分构成,位于N端的作为配体结合结构域的针对FITC的scFv,以及位于C端的作为GPI锚定和内化结构域的FRα。当在CAR T细胞上独立表达时,FITC-FR融合受体可以被FITC-免疫激动剂特异性靶向,以克服抑制性肿瘤微环境中CART细胞的耗竭状态。这些免疫激动剂通常会引起强烈的自身免疫副作用,并且现在可以以FITC靶向形式全身给药并安全地递送至FITC-FR阳性CAR T细胞。在过去的几十年中,该领域在细胞类型、递送方法和合适的疾病模型方面取得了巨大的进步。就细胞类型而言,目前的细胞治疗可大致分为嵌合抗原受体(CAR)、用于肿瘤模型的细胞和基于干细胞的再生医学。
CAR T也称为嵌合T细胞受体、嵌合免疫受体或人工T细胞受体,其能够使免疫效应细胞(通常为T细胞或NK细胞)识别具有相应抗原的靶细胞并发挥其细胞毒活性。CAR-T技术的出现和发展为某些类型的癌症带来了希望,这继而使CAR-T成为生物医学研究和临床研究两个领域的超级明星。在美国申请No.15/296,666(其内容通过引用完整并入)中公开了一些传统的和改进的CAR T细胞设计。在‘666申请中,通过提供表达CAR的细胞毒性淋巴细胞来产生CAR系统,所述CAR靶向不被所治疗的对象的细胞产生或表达的部分。因此,该CAR系统允许将细胞毒性淋巴细胞集中靶向至靶细胞,例如癌细胞。被靶向的部分是还包含肿瘤细胞受体的配体的小缀合分子(SCM)的一部分。与表达CAR的细胞毒性淋巴细胞一起施用SCM导致将细胞毒性淋巴细胞应答仅靶向表达与SCM结合的肿瘤受体的那些细胞。
尽管CAR T细胞治疗在研究和临床应用领域二者都取得了飞速的发展,但是存在着伴随CAR T治疗的担忧。一种致命的副作用是因肿瘤细胞的快速裂解产生细胞因子风暴,以及其杀伤带有CAR的正常细胞。为了克服这些副作用,在PCT/US2018/018557(其内容通过引用完整并入)中开发了将具有特定药物载荷的CAR T细胞靶向递送至靶肿瘤细胞以控制这样的副作用的方法。简单地说,工程化蛋白与高亲和力靶向配体偶联,其中该靶向配体携带至少一种药物载荷,所述药物载荷待通过工程化蛋白被CAR T细胞内化以调节移植的细胞的治疗效果。
CAR T治疗的另一个局限性是其在癌抗原的反复刺激之后变得耗竭的趋势。证明了T细胞的耗竭表型的可逆性,因为从实体瘤组织分离的T细胞如果在重新刺激之前保持远离抗原(“静止”)过夜,则表现出较高INFγ分泌和杀伤作用4。然而,如果可以使用药物以更临床相关的方式实现恢复活力,则将更具吸引力:阻断抑制性信号传导或通过其他途径激活T细胞。在临床上,靶向检查点抑制剂(即PD-1、CTLA-4等)的抗体在实体瘤中已显示出一定的成功,然而,经常发现需要组合的两种或更多种靶标56。此外,抗体治疗在实体瘤中具有差的穿透性。因此,较少看到关于实体瘤中CAR T与检查点抑制剂抗体的联合治疗的报道。抑制介导TCR失活的磷酸酶(例如SHP1/27和TC-PTP8)是阻断抑制途径的另一种方法。这些磷酸酶的敲除实验和小分子抑制剂二者均显示出降低TCR阈值和提高T细胞活性的强效作用,但它们均未用于CAR T治疗。DGKα是TCR信号传导的另一种生理抑制剂并且其在耗竭的TIL中过表达。DGKα将DAG分解代谢为PA,从而降低DAG水平,其导致Ras和MARK ERK信号传导减弱。DGKα抑制剂恢复脱粒并提高TIL和CAR T的杀伤作用39。使T细胞恢复活力的另一种方法是通过抗原非依赖性途径激活它。已知某些病原体模式识别(PPR)受体(包括Toll样受体(TLR))确实在包括T细胞在内的非髓样细胞群上表达,并且可以以类似的方式激活。研究还表明,TLR210、411和7/812 13激动剂可以激活CD8 T细胞并且提高INFγ分泌。然而,由于TLR激动剂的全身给药的强烈副作用14,这些激动剂均未用于CAR T治疗中以重新激活T细胞或改变免疫抑制性微环境。TLR激动剂对肿瘤细胞本身的争议性作用也阻碍了这一过程15。干扰素基因刺激物(STING)是在外周淋巴组织中的造血细胞(包括T细胞、髓样细胞和单核细胞)中广泛表达的胞质溶胶DNA传感器(CDS)。STING激动剂已被用作许多免疫治疗的免疫刺激剂,并且在CAR T治疗中也可能具有显著的作用。然而,尽管前述抑制剂和激动剂可能对CART细胞具有显著的恢复活力作用,但由于其高度强效的促炎功能,如果全身给药,则很可能可引起严重的副作用。因此,非常期望将潜在载荷靶向递送至CAR T细胞。
为了解决特异性递送问题,我们设计了秘密通道平台,其可以在T细胞中与CAR构建体一起表达,使得某些载荷可以全身给药并且仅在CAR T细胞内特异性积累,而不接触其他细胞。该系统由通过T2A自剪切序列连接的融合受体和经典CAR构建体组成。融合受体包含两部分:(1)配体结合模块,其可以识别高亲和力配体-载荷缀合物;(2)膜结合受体模块,其可以介导受体/缀合物复合物内化到细胞中。针对部分1选择了两个蛋白质/配体对:FKBP/FK506以及针对FITC的scFv(4M5.3)/FITC,这是因为以下原因:(1)在天然细胞膜上不存在FKBP或4M5.3确保了将载荷特异性递送至融合受体阳性CAR T细胞,从而降低了对其他细胞的副作用;(2)蛋白质/配体对之间的亚纳摩结合亲和力促进了靶细胞内足够的载荷积累。对于部分2中的膜结合受体,选择了叶酸受体α(FRa),因为其具有固有内化特性而与叶酸的结合无关16。我们还设计了靶标-载荷连接系统,其中根据靶标,载荷可通过不可释放的接头或二硫键可释放接头与靶向配体连接。更具体地,由于TLR7位于内体中,所以秘密通道递送的TLR7激动剂在通过受体介导的内化进入内体后就可以尽快发挥其功能。因此,在这种情况下,不需要可释放的接头。而对于位于胞质溶胶而不是内体中的其他靶标,例如SHP1/2、TC-PTP、DGK、TGFβ和STING,从靶标-载荷缀合物中释放游离药物对于从内体逃逸是必要的。总之,这个秘密通道系统提供了用于将多种载荷特异性体内递送至耗竭的CAR T细胞的通用平台。
通过以下实验实施例将更好地理解本发明的这些和其他特征、方面和优点。
方法:
抗CD19 CAR T细胞的耗竭和药物处理:
将抗CD19 CAR T细胞与Raji以1∶1的比例在6孔板中共培养,同时每12小时向相同孔添加新鲜Raji细胞。通过流式细胞术计数来定量杀伤作用和共抑制标志物。对于药物处理,在用Raji细胞刺激4轮之后,将耗竭的抗CD19 CAR T与不同浓度的药物一起再孵育12小时,然后类似地进行定量。
靶向配体结合测定
将融合受体阳性细胞与不同浓度不同的某些配体-染料分子在4度下一起孵育30分钟。孵育之后,将细胞用PBS清洗两次,然后进行流式细胞术。MFI或位移百分比用于结合曲线和Kd的计算。
材料
细胞系和人T细胞
含有10%热灭活的胎牛血清和1%青霉素-链霉素的DMEM(Gibco)用于培养MDAMB-231和MDA-MB-231 CD19+细胞。通过Ficoll密度梯度离心(GE Healthcare Lifesciences,#17-5442-02)从获自健康志愿者的人全血中分离外周血单核细胞(PBMC)。使用EasySepTM人T细胞分离试剂盒(STEM CELL technologies,#17951)从PBMC中富集纯CD3+T细胞。
用于耗竭的CAR T细胞的体外恢复活力的潜在载荷的评价
将抗CD19 CAR T细胞与CD19+Raji细胞以1∶1的比例在12孔板中以每孔2×106个CAR T和2×106个Raji细胞的密度共培养,每12小时添加新的Raji细胞,添加三次,然后测试Raji细胞群、裂解作用和共抑制受体以确认CAR T细胞的耗竭。流式细胞术和基于萤光素酶的测定二者用于定量裂解作用。为了测试潜在载荷的恢复活力效力,然后将这种细胞混合物转移到96孔板,每孔约2×105个细胞,并且添加不同浓度的药物。12小时之后,再次测试Raji细胞群、裂解作用和共抑制受体并且与PBS处理组进行比较。
实施例
实施例1.体外抗CD19 CAR T细胞的耗竭
在该实施例中,我们说明了耗竭的CAR T细胞的模型。简单地说,图1a示出了如方法部分中所述将Raji细胞(一种B淋巴瘤细胞)与FMC63 CAR T细胞(一种抗CD19 CAR T细胞)共培养,并且连续三天每12小时向共培养物添加新鲜Raji细胞。图1b示出了这些CAR T细胞在体外通过新鲜Raji细胞刺激3次之后变得耗竭,如通过降低的杀伤作用指示的。共培养的K562细胞用作阴性对照。
实施例2.秘密通道的设计
在该实施例中,我们表明将载荷药物递送至靶细胞类型的秘密通道的图示说明。FKBP或抗FITC与FRa连接作为融合受体,其能够结合与特定药物载荷连接的靶向配体FK506或FITC。由于FRa的性质,其不断内化并且将FK506或FITC连接的载荷递送到CAR T细胞中。因此,在该实施例中,当CAR T通过CAR与其靶细胞结合时,CAR T表面上的抗CD 19分子结合癌细胞的CD-19,CAR T内部的递送的载荷药物可以执行其功能,即基于载荷功能来调节CART活性。例如,一些载荷药物可以在必要时被引导对PD-1、CTLA4或LAG3 T细胞功能调节分子发挥作用以使CAR T恢复活力。
实施例3.通过TLR7激动剂逆转耗竭的抗CD19 CAR T细胞
在该实施例中,我们表明靶向配体FK506或FITC成功地结合其各自的融合受体(通过G4S连接至叶酸受体的FKBP或FITC-AF647)。在该实施例中,载荷药物是成像剂罗丹明,以显示载荷在融合受体转染的Jurkat细胞中的分布。FK506和FITC的结合亲和力通过竞争性结合来计算,并且FK506-罗丹明对FKBP-FR融合受体显示Kd=3.39nM,而FITC-AF647对抗FITC-FR显示Kd=8.03nM。
实施例4.潜在的可释放和不可释放的FK506-TLR7激动剂和FITC-TLR7激动剂的设计
在该实施例中,我们提供了具有以下结构的与Toll样受体7激动剂缀合的靶向配体FK506以处理耗竭的CAR T细胞。当TLR7激动剂载荷药物靶向耗竭的CAR T细胞时,观察到增强的杀伤作用剂量依赖性模式。因此,CAR T活性的指示因子IFNγ表达水平相对于载荷药物浓度以剂量依赖性方式升高;并且T细胞效应抑制分子Tim 3的表达水平呈反剂量依赖性方式,即Tim 3的表达随载荷药物浓度的增加而降低。
FK506-TLR7激动剂缀合物应具有以下给出的结构之一:
类似地,荧光素-TLR7激动剂缀合物应具有以下给出的结构之一,并且如果功能性CAR T具有抗FITC融合受体,则使耗竭的CAR T恢复活力:
实施例5.用于使耗竭的CAR T细胞恢复活力的其他潜在载荷的设计
在该实施例中,我们提供了可以以较低的纳摩范围效价逆转CAR T耗竭的其他潜在载荷的结构列表。
1)TC-PTP磷酸酶抑制剂应具有以下结构:
上述磷酸酶抑制剂可以通过以下方式与荧光素或FK506(他克莫司)连接:
2)STING激动剂应具有以下结构:
实施例6.靶向配体与潜在载荷之间的间隔基的设计
以下是可用于连接靶向配体和任何潜在载荷的间隔基的列表:
实施例7.磷酸酶抑制剂和TLR7激动剂使耗竭的CAR T细胞恢复活力的能力的评价
CAR T细胞治疗在实体瘤中的一个主要局限性是其在用癌抗原重复刺激之后变得耗竭的趋势。然而,这种现象并非CAR T细胞特有的,在慢性病毒感染4和肿瘤浸润淋巴细胞5二者中均有描述。T细胞的耗竭表型的可逆性已在研究中得到证实,其中从实体瘤组织分离的T细胞如果在重新刺激之前保持远离抗原(“静止”)过夜,则表现出较高INFγ分泌和杀伤作用57。然而,如果可以使用市售治疗剂以临床上更相关的方式实现恢复活力,则将更具吸引力:阻断抑制性信号传导或通过其他途径激活T细胞。靶向检查点抑制剂(即PD-1、CTLA-4等)的抗体在临床的实体瘤中已显示出一些成功151,然而,经常发现需要组合的两种或更多种靶标。此外,抗体治疗在实体瘤中还具有差的穿透性,并且这可能导致针对实体瘤中CART细胞与检查点阻断(ICB)的联合治疗的报道较少。
抑制介导TCR失活的磷酸酶(例如SHP1/2和TC-PTP)是阻断递补CAR T信号传导(tonic CAR T signaling)的潜在途径。SHP1/2磷酸酶负责介导来自PD-1和其他耗竭标志物的信号。数据表明,SHP1/2磷酸酶抑制剂或沉默可以提高T细胞和CAR T细胞的活性6-9。已知TC-PTP是T细胞活性信号传导的重要参与者。具有T细胞特异性TC-PTP缺陷的小鼠由于提高的抗原驱动的T细胞活化而具有提高的对炎症和自身免疫的易感性。TC-PTP使TCR下游的Src家族激酶失活,从而导致TCR激活的阈值11。尽管这些磷酸酶的敲除实验和小分子抑制剂二者均显示出降低TCR阈值和提高T细胞活性的强效作用,但它们均未用于CAR T治疗。代表性的SHP1/2抑制剂具有以下结构:
使T细胞恢复活力的另一种方法是通过抗原非依赖性先天免疫受体的参与来增强其活性。已知某些病原体模式识别(PPR)受体C包括Toll样受体(TLR))确实在包括T细胞在内的非髓样细胞群上表达,并且可以以类似的方式激活。研究还表明,TLR7/8激动剂和TCR信号传导的共刺激可以激活CD8 T细胞并且提高INFγ分泌13。然而,由于TLR激动剂的全身给药的强烈副作用,这些激动剂均未用于CAR T治疗中以重新激活T细胞或改变免疫抑制性微环境。TLR激动剂对肿瘤细胞的争议性作用也阻碍了TLR激动剂用于癌症免疫治疗的使用。因此,非常期望将潜在的载荷靶向递送至CAR T细胞。在文献中发现了一种强效的TLR7激动剂(图7A),其比FDA批准的咪喹莫特强约40倍。
为了建立如图6所示的体外筛选模型,将抗CD19 CAR T细胞暴露于4轮CD19阳性Raji细胞的添加,并且变得耗竭,如通过体外共培养模型中逐渐降低的裂解活性以及提高的共抑制标志物标记的。值得注意的是,培养基对于引入CAR T耗竭是重要的,并且在整个过程中都需要保持相同,而无需新的补充或更换。这表明由癌细胞和/或CAR T细胞释放到培养基中的可溶性组分(最可能是免疫抑制性细胞因子和调节剂(腺苷等))在此过程中发挥关键作用。这也表明由这种体外模型产生的CAR T细胞的耗竭处于相当柔韧而不是不可逆的状态。
与未处理组相比,用选择的TLR7激动剂和PTP1b(与TC-PTP26高度同源)抑制剂27处理已经耗竭的CAR T细胞显示能够重新激活它们(图7)。然而,在除Tim3外的共抑制标志物的表达水平中未见明显变化。如图7C所示,PTP1b抑制剂一般性地没有表现出与TLR7激动剂一样强的再活化作用。不受任何理论的束缚,该结果可能是由于目前的体外筛选模型所致,其中研究的是耗竭的“逆转”而不是“阻止”,并且在耗竭状态下磷酸酶已经“沉默”,因此其抑制将几乎没有效果。用于未来研究的经修改的筛选模型将通过在所有培养开始时添加药物并保持其余设置不变来测试磷酸酶抑制剂和其他药物的效果,重点是耗竭的“阻止”。以这种方式,可以看到磷酸酶的抑制是否可以降低CAR T的递补信号传导,同时仍保持功能性杀伤作用。在下面的研究中,我们将主要关注TLR7激动剂。
由于TLR7是存在于内体中的4个TLR家族成员之一,因此推测TLR7激动剂与我们的秘密通道靶向配体之间的不可释放的接头将保留其TLR7激动剂功能28。为此,制备了多种TLR7激动剂类似物并进行了测试,以找到合适的用于连接的衍生位点。如图8所示,在哌啶环上具有CH2OH延伸的TLR7激动剂与母体药物相比具有甚至更高的活性。因此,该衍生位点将用于不可释放的缀合物。还已经合成了二硫键连接的自牺牲形式(self-immolativeform)。为了理解该TLR7激动剂到达其自身靶标所需的距离,为不可释放的FITC-TLR7制备了FITC与TLR7激动剂之间的三种不同长度的接头(PEG3、6、16)。如图9C所示,所有不可释放的形式都有一定作用,而PEG6化合物显示出最佳的剂量依赖性响应。这些结果表明,TLR7激动剂可以通过在与FITC-FR结合时伸出或在TLR与FITC-FR之间跳跃而与TLR7对接,在这种情况下,两者之间的接头长度不是关键因素(图9D至E)。由于不可释放的FITC-TLR7被捕获在内体内部,并且每个内体的体积比胞质溶胶小得多,因此内体内的TLR7可以更快更迅速达到其功能浓度,导致较小的IC50。
实施例8.用于逆转/阻止CAR T细胞耗竭的其他潜在载荷
除TLR7激动剂外,还有如下所述多种其他潜在的载荷可以逆转/阻止CAR T细胞的耗竭。一些靶标目前可能尚无具有适合我们靶向药物递送方法的IC50的激动剂或抑制剂,但仍值得注意,可以通过其他抑制机制进行探索,例如CRISPR或靶向微RNA(microRNA)递送方法。
·STING激动剂
IFN基因刺激物(STING)是参与胞质溶胶DNA传感和随后IFN-β产生的主要衔接子之一。STING与sdDNA弱缔合,但是与通过cGMP-AMP合酶(sGAS)合成的内源环状二核苷酸GMP-AMP(cGAMP)强烈结合。其主要在巨噬细胞、T细胞、多种DC、内皮细胞以及选择的成纤维细胞和上皮细胞中表达。STING的研究主要集中于其在巨噬细胞和树突状细胞中的功能,并且最近一些小组注意到STING激活在T细胞中的直接作用40。STING激动剂可能对T细胞具有类似的促炎作用。ADU-S100是临床上探索的许多STING激动剂之一。
·DGK-α抑制剂
二酰基甘油激酶-α(DGK-α)将TCR信号传导中的第二信使二酰基甘油(DAG)与IP3一起转换为磷脂酸(PA)。DGK在CD8TIL中比在CD8-NIL中更高表达,并且其抑制促进了ERK磷酸化和裂解脱粒41。其还恢复了从体内分离的CAR TIL的裂解功能5。一些DGK抑制剂结构如下:
·TGFβRI(ALK5)抑制剂
TGFβ以其在许多免疫细胞(例如T细胞、B细胞和巨噬细胞)中的免疫抑制功能而闻名。T细胞中I型TGFβ受体(TGFβRI,也称为ALK5)的阻断逆转了肿瘤的免疫抑制环境42。在临床中利用测试的加尼斯替(galunisertib)(一水合LY2157299)和EW-7197探索了小分子抑制剂43 44。
TGFβ抑制剂的结构如下:
TGFβ抑制剂(LY2157299)。
·EZH2抑制剂
Zeste同源物增强子2(EZH2)是组蛋白H3K27甲基转移酶,其具有与Treg功能的强相关性。EZH2的遗传或药理破坏驱动肿瘤浸润Treg的促炎功能的获得45。由于慢性病毒感染中耗竭的CTL还以独特的表观遗传学变化为特征174,因此EZH2抑制剂可能能够逆转这种耗竭状态。已经开发了EZH2抑制剂的多种小分子,包括CPI1205、EPZ6438和GSK126。
因此,FITC-FR融合受体和相应的FITC靶向免疫激动剂载荷为监测和控制CAR T细胞在实体瘤中的归巢和持久性提供了通用平台。由于FITC-FR融合受体独立于CAR构建体表达,因此该方法可容易引入到CAR T细胞中以用于任何抗原。靶向配体-载荷缀合物的模块化设计还使其更易于切换和修饰。这种方法结合了细胞治疗和基于小分子的靶向药物递送的优势,并且可能需要对工程化细胞和相应的配体二者进行额外的表征。
CAR T细胞在实体瘤中的成功最可能还需要靶向微环境中其他参与者的多种方法的组合,例如通过PI3K激酶抑制剂破坏胞外基质,将抗炎M2巨噬细胞重编程为促炎M1表型,以及癌细胞上降低的MHC分子水平的上调。因此,将在临床前和临床上进行越来越多的联合治疗研究。然而,与此同时,不能忽略对CAR T细胞本身的仔细检查和控制,并且在临床上应该首先使用简单而稳健的系统(如FITC-FR融合受体)进行优化,然后再将其应用于人。
参考文献:
1.Park,J.H.;Riviere,I.;Goncn,M.;Wang,X.;Senechal,B.;Curran,K.J.;Sauter,C.;Wang,Y.;Santomasso,B.;Mead,E.;Roshal,M.;Maslak,P.;Davila,M.;Brentiens,R.l.;Sadelain,M.,Long-Tern Follow-up of CD19 CAR Thcrapy in AcuteLymphoblastic Leukemia.The New EngIand journal of medicine 2018.378(5).449-459.
2.Jiang,Y.;Li,Y.;Zhu,B.,T-cell exhaustion in the tumormicroenvironment.Cell Death Dis 2015,6,e1792.
3.Matsuzaki,J.;Gniatic,S.;Mhawech-Fauceglia,P.;Beck,A.;Miller,A.;Tsuji,T.;Eppolito,C.;Qian,F.;Lele,S.;Shrikant,P.;Old,L.J.;Odunsi,K.,Tumor-infiltrating NY-ESO-1-specific CD8+T cells are negatively regulated by LAG-3and PD-1 in human ovarian cancer.Proceedings of the National Academy ofSciences of the Unitcd Srates of America 2010,107(17),7875-80.
4.Moon,E.K.;Ranganathan,R.;Eruslanov,E.;Kim,S.;Newick,K.;O’Brien,S.;Lo,A.;Liu,X.;Zhao,Y.;Albelda,S.M.,Blockade of Programmed Death 1 Augments theAbility of Human T Cells Engineered to Target NY-ESO-1 to Control TumorGrowth after Adoptive Transfer.Clinical cancer research:an official journalof the American Association for Cancer Research 2016,22(2),436-47.
5.Moon,E.K.;Wang,L.C.;Dolfi,D.V.;Wilson,C.B.;Ranganathan,R.;Sun,J.;Kapoor,V.;Scholler,J.;Pure,E.;Milone,M.C.;June,C.H.;Riley,J.L.;Wherry,E.J.;Albelda,S.M.,Multifactorial T-cell hypofunction that is reversible can limitthe efficacy of chimeric antigen receptor-transduced human T cells in solidtumors.Clinical cancer research;an official journal of the AmericanAssociation for Cancer Research 2014,20(16),4262-73.
6.Chae,Y.K.;Arya,A.;lams,W.;Cruz,M.R.;Chandra,S.;Choi,J.;Giles,F.,Current landscape an future of dual anti-CTLA4 and PD-1/PD-L1 blockadeimmunotherapy in cancer;lessons learned from clinical trials with melanomaand non-small cell lung cancer (NSCLC).J Immunother Cancer 2018,6(1),39.
7.Ott,P.A.;Hodi,F.S.;Kaufman,H.L.;Wigginton,J.M.;Wolchok,J.D.,Combination immunotherapy:a road map.J Immunother Cancer 2017,5,16.
8.Watson,H.A.;Dolton,G.;Ohme,J.;Ladell,K.;Vigar,M.;Wehenkel,S.;Hindley,J.;Mohammed,R.N.;Miners,K.;Luckwell,R.A.;Price,D.A.;Matthews,R.J.;Ager,A.,Purity of transferred CD8(+)T cells is crucial for safety andefficacy of combinatorial tumor immunotherapy in the absence of SHP-1.Immunology and cell biology 2016,94(8),802-8.
9.Wiede,F.;Shields,B,J.;Chew,S.H.;Kyparissoudis,K.;van Vliet,C.;Galic,S.;Tremblay,M.L.;Russell,S.M.;Godfrey,D.I.;Tiganis,T.,T cell proteintyrosine phosphatase attenuates T cell signaling to maintain tolerance inmice.The Journal of clinical investigation 2011,121(12),4758-74.
10.Prinz,P.U.;Mendler,A.N.;Masouris,I.;Durner,L.;Obeerneder,R.;Noessner,E.High DGK-alpha and disabled MAPK pathways cause dysfunction ofhuman tumor-infiltrating CD8+T cells that is reversible by pharmacologicintervention.Journal of immunology 2012,188(12),5990-6000.
11.Chua,B.Y.;Olson,M.R.;Bedoui,S.;Sekiya,T.;Wong,C.Y;Turner,S.J.;Jackson,D.C.,The use of a TLR2 agonist-based adjuvant for enhancing effectorand menmory CD8 T-cell responses.Immunology and cell biology 2014,92(4),377-83.
12.Rhee,E.G.;Kelley,R.P.;Agarwal,I.;Lynch,D.M.;LaPorte,A.;Simmons,N.L.;Clark,S.L;Barouch,D.H.,TLR4 ligands augment antigen-specific CD8+Tlymphocyte responses elicited by a viral vaccine vector.Journal of virology2010,g4(19).10413-9.
13.Wiedemann,G.M.;Jacobi,S.J.;Chaloupka,M.;Krachan,A.;Hamm,S.;Strobl,S.;Baumgartner,R.;Rothenfusser,S.;Duewell,P.;Endres,S.;Kobold,S.,A novel TLR7agonist reverses NK cell anergy and cures RMA-S lymphoma-bearingmice.Oncoimmunology 2016,5(7),e1189051.
14.Cheadle.E.J.;Lipowska-Bhalla,G.;Dovedi,S.J.;Fagnano,E.;Klein,C.;Honeychurch,J.;Illidge,T.M.,A TLR7 agonist enhances the antitumor efficacy ofobinutuzumab in murine lymphoma models via NK cells and CD4 T cells.Leukemia2017,31(10),2278.
15.Hengge,U.R.;Ruzicka,T.,Topical immunomodulation in dermatology:potential of toll-like roceptor agonists.Dermatol Surg 2004,30(8),1101-12.
16.Kaczanowska,S.;Joseph,A.M.;Davila,E.,TLRagonists:our best frenemyin cancer immunotherapy.Journal of leukocyte biology 2013,93(6),847-63.
17,Bandara,N.A.;Hansen,M.J.;Low,P.S.,Effect of receptor occupancy onfolate receptor internalization.Molecular pharmaceutics 2014,11(3),1007-13.
18.Kniess,T.;Laube,M.;Wust,F.;Pietzsch,J.,Technetium-99m based smallmolecule radiopharmaceuticals and radiotracers targeting inflammation andinfection.Dalton transactions 2017,46(42),14435-14451.
19.Kularatne.S.A.;Zhou,Z.;Yang.J.;Post,C.B.;Low.P.S.,Design,synthesis,and preclinical evaluation of prostate-specific membrane antigentargeted(99m)Tc-radioimaging agents.Molecular pharmaceutics 2009,6(3),790-800.
20.Henne,W.A.;Rothenbuhler,R.;Ayala-Lopez,W.;Xia,W.;Varghese,B.;Low,P.S.,Imaging sites of infection using a 99mTc-labeled folate conjugatetargeted to folate receptor positiye macrophages.Molecular pharmaceutics2012,9(5),1435-40.
21.Kahan,S.M.;Wherry,E.J.;Zajac,A.J.,T cell exhaustion duringpersistent viral in fections.Virology 2015,479-480,180-93.
22.Stefanova,I.;Hemmer,B.;Vergelli,M.;Martin,R.;Biddison,W.E.;Germain,R.N.,TCR ligand discrimination is enforccd by competing ERK positiyeand SHP-1 negative feedback pathways.Nature immumology 2003,4(3),248-54.
23.Lorenz,U.,SHP-1and SHP-2 in T cells:two phosphatases functioningat many levels.Immunological reviews 2009,228(1),342-59.
24.Watson,H.A.;Wehenkel,S.;Matthews;,J.;Ager,A.,SHP-1:the nextcheckpoint target for cancer immunotherapy?Biochemical Society transactions2016,44(2),356-62.
25.Hebeisen,M.;Baitsch,L.;Presotto,D.;Baumgaertner,P.;Romero,P.;Michielin,O.;Speiser,D.E.;Rufer,N.,SHP-1 phosphatase activity counteractsincreased T cell receptor affinity.The Journal of clinicl in vestigation2013,123(3),1044-56.
26.Wiede,F.;Shields,B.J.;Chew,S.H.;Kyparissoudis,K.;van Vliet,C.;Galic,S.;Tremblay,M.L.;Russell,S.M.;Godfrey,D.I.;Tiganis,T.,T cell proteintyrosine phosphatase attenuates T cell signaling to maintain tolerance inmice.The Journal of clinical investigation 2011,121(12),4758-74.
27.Pike,K.A.;Hatzihristidis,T.;Bussieres-Marmen,S.;Robert,F.;Desai,N.;Miranda-Saavedra.D.;Pelletier,J.;Tremblay,M.I.,TC-PTP regulates the IL-7transcriptional response during murine early T cell development.Scientificreports 2017,7(1),13275.
28.Wiedemann,G.M.;Jacobi,S.J.;Chaloupka.M.;Krachan,A.;Hamm,S.;Strobl,S.;Baumgartner,R.;Rothenfusser,S.;Duewell,P.;Endres,S.;Kobold,S.,A novel TLR7agonist reverses NK cell anergy and cures RMA-S lymphoma-bearingmice.Oncoimmunology 2016,5(7),e1189051.
29.Caron,G.;Duluc,D.;Fremaux,I.;Jeannin,P.;David,C.;Gascan,H.;Delneste,Y.,Direct stimulation of human T cells via TLR5 and TLR7/8:flagellinand R-848 up-regulate proliferation and IFN-gamma production by memory CD4+Tcells.Journal of immunology 2005,175(3).1551-7.
30.Wille-Reece,U.;Flynn,B.J.;Lore,K.;Koup,R.A.;Miles,A.P.;Saul,A.;Kedl,R.M.;Mattapallil,J.J.;Weiss,W.R.;Roederer,M.;Seder,R.A.,Toll-likereceptor agonists influence the magnitude and quality of memory T cellresponses after prime-boost immunization in nonhuman primates.The Journal ofexperimental medicine 2006,203(5),1249-58.
31.Zarember,K.A.;Godowski,P.J.,Tissue expression of human Toll-likereceptors and differential regulation of Toll-like receptor mRNAs inleukocytes in response to microbes,their products,and cytokines.Journal ofimmunology 2002,168(2),554-61.
32.Hornung,V.;Rothenfusser,S.;Britsch,S.;Krug,A.;Jahrsdorfer,B.;Giese,T.;Endres,S.;Hartmann,G.,Quantitative expression of toll-like receptor1-10 mRNA in cellular subsets of human peripheral blood mononuclear cells andsensitivity to CpG oligodeoxynucleotides.Journal of immunology 2002,168(9),4531-7.
33.Strominger,N.L.;Brady,R.;Gullikson,G.;Carpenter,D.O.,Imiquimod-elicited emesis is mediated by the area postrema,but not by direct neuronalactivation.Brain Res Bull 2001,55(3),445-51.
34.Harrison,L.I.;Astry,C.;Kumar,S.;Yunis,C.,Pharmacokinetics of 852A,an imidazoqutinoline Toll-like receptor 7-specific agonist,followingintravenous,subcutaneous,and oral administrations in humans.Journal ofclinical pharmacology 2007,47(8),962-9.
35.Dudek,A.Z.;Yunis,C.;Harrison,L.I.;Kumar,S.;Hawkinson,R.;Cooley,S.;Vasilakos,J.P.;Gorski,K.S.;Miller,J.S.,First in human phase I trial of 852A,anovel systemic toll-like receptor 7 agonist,to actiyate innate immuneresponses in patients with advanced cancer.Clinical cancer research:anofficial journal of the American Association for Cancer Research 2007,13(23),7119-25.
36.Dummer,R.;Hauschild,A.;Becker,J.C.;Grob,J.J.;Schadendorf,D.;Tebbs,V.;Skalsky,J.;Kaehler,K.C.;Moosbauer,S.;Clark,R.;Meng,T.C.;Urosevic,M.,Anexploratory study of systemic administration of the toll-like receptor-7agonist 852A in patients with refractory metastatic melano ma.Clinical cancerresearch :an official journal of the American Association for Cancer Research2008,14(3),856-64.
37.Perkins,H.;Khodai,T.;Mechiche,H.;Colman,P.;Burden,F.;Laxton,C.;Horscroft,N.;Corey,T.;Rodrigues.D.;Rawal,J.;Heyen,J.;Fidock,M.;Westby,M.;Bright,H.,Therapy with TLR7 agonists induces lymphopenia:correlatingPharmacology to mechanism in a mouse model.J Clin Immunol 2012,32(5),1082-92.
38.Hasham,M.G.;Baxan,N.;Stuckey,D.J.;Branca,J.;Perkins,B.;Dent,O.;Duffy,T.;Hameed,T.S.;Stella,S.E.;Bellahcene,M.;Schneider,M,D.;Harding,S.E.;Rosenthal,N.;Sattler,S.,Systemic autoimmunity induced by the TLR7/8 agonistResiquimod causes myocarditis and dilated cardiomyopathy in a new mouse modelof autoimmune heart disease.Dis Model Mech 2017,10(3),259-270.
39.Katie,J.;David,N.,The discovery of a novel prototype smallmolecule TLR7 agonist for the treatment of hepatitis C virusinfection.MedChemComm 2011,2(3),185-189.
40Jones,P.;Pryde,D.C.;Tran,T.D.;Adam,F.M.;Bish,G.;Calo,F.;Ciaramella,G.;Dixon,R.;Duckworth,J.;Fox,D.N.;Hav,D.A.;Hitchin,J.;Horscroft,N.;Howard,M.;Laxton,C.;Parkinson,T.;Parsons,G.;Proctor,K.;Smith,M.C.;Smith.N.;Thomas,A.,Discovery of a highly potent series of TLR7 agonists.Bioorganic &medicinalchemistry letters 2011,21(19),5939-43.
41.Hemmi,H.;Kaisho,T.;Takeuchi,O.;Sato,S.;Sanio,H.;Hoshino,K.;Horiuchi,T.;Tomizawa,H.;Takeda,K.;Akira,S.,Small anti-viral compoundsactivate immune cells via the TLR7 MyD88-dependent signaling pathway.Natureimmunology 2002,3(2),196-200.
42.lversen,L.F.;Moller,K.B.;Pedersen,A.K.;Peters,G.H.;Petersen,A.S.;Andersen,H.S.;Branner,S.;Mortensen,S.B.;Moller,N.P.,Structure determinationof T cell protein-tvrosine phosphatase.The Journal of biological chemistry2002,277(22),19982-90.
43.Wilson,D.P.;Wan,Z.K.;Xu,W.X.;Kirincich,S.J.;Follows,B.C.;Joseph-McCarthy,D.;Foreman,K.;Moretto,A.;Wu,J.;Zhu,M.;Binrun,E.;Zhang,Y.L.;Tam,M.;Erbe,D.V.;Tobin,J.;Xu,X.;Leung,L.;Shilling,A.;Tam,S.Y.;Mansour,T.S.;Lee,J.,Structure-based optimization of protein tyrosine phosphatase 1B inhibitors:from the active site to the second phosphotyrosine binding site.Journal ofmedicinal chemistry 2007,50(19),4681-98.
44.Ignacio,B.J.;Albin,T.J.;Esser-Kahn,A.P.;Verdoes,M.,Toll-likeReceptor Agonist Conjugation:A Chemical Perspective.Bioconjugate chemistry2018,29(3),587-603.
45.Parente-Pereira,A.C.;Burnet,J.;Ellison,D.;Foster,J.;Dayies,D.M.;van der Stegen,S.;Burbridge,S.;Chiapero-Stanke,L.;Wilkie,S.;Mather,S.;Maher.J.,Trafficking of CAR-engineered human T cells following regional orsystemic adoptive transfer in SCID beige mice.J Clin Immunol 2011,31(4),710-8.
46.Dobrenkov,K.;Olszewska,M.;Likar,Y.;Shenker,L.;Gunset,G.;Cai,S.;Pillarsetty,N.;Hricak,H.;Sadelain,M.;Ponomarey,V.,Monitoring the efficacy ofadoptively transferred prostate cancer-targeted human T lymphocytes with PETand bioluminescence imaging.Journal of nuclear medicine:official publication,Society of Nuclear Medicine 2008,49(7),1162-70.
47.Emami-Shahri,N.;Foster,J.;Kashani,R.;Gazinska,P.;Cook,C.;Sosabowski,J.;Maher,J.;Papa,S.,Clinically compliantspatial and temporalimaging of chimeric antigen receptor T-cells.Nature communications 2018,9(1),1081.
48.Bruno,R.;Giannasio,P.;Ronga,G.;Baudin,E.;Travagli,J.P.;Russo,D.;Filetti.S.;Schlumberger,M.,Sodiumiodide symporter expression and radioiodinedistribution in extrathyroidal tissues.J Endocrinol Invest 2004,27(11),1010-4.
49.Vedvyas,Y.;Shevlin,E.;Zaman,M.;Min,I.M.;Amor-Coarasa,A.;Park,S.;Park,S.;Kwon.K.W.;Smith,T.;Luo,Y.;Kim,D.;Kim,Y.;Law,B.;Ting,R.;Babich,J.;Jin,M.M.,Longitudinal PET imagingdemonstrates biphasic CAR T cell responses insurvivors.JCI insight 2016,1(19).e90064.
50.Zhang,H.;Moroz,M.A.;Serganova,I.;Ku,T.;Huang,R.;Vider,J.;Maecke,H.R.;Larson,S.M.;Blasberg,R.;Smith-Jones,P.M.,Imaging expression of the humansomatostatin receptor subtype-2 reporter gene wit 68Ga-DOTATOC.Journal ofnuclear medicine:official publication,Society of Nuclear Medicine 2011,52(1),123-31.
51.Bhattacharyya,S.;Dixit,M.,Metallic radionuclides in thedevelopment of diagnostic and therapeutic radiopharmaceuticals.Daltontransactions 2011,40(23),6112-28.
52.Khan,O.;Giles.J.R.;McDonald.S.;Manne,S.;Ngiow,S.F.;Patel,K.P.;Werner,M.T.;Huang,A.C.;Alexander,K.A.;Wu,J.E.; Attanasio,J.;Yan,P.;George,S.M.;Bengsch,B.;Staupc,R.P.;Donahuc.G.;Xu.W.;Amaravadi,R.K.;Xu,X.;Karakousis,G.C.;Mitchell,T.C.;Schuchter,L.M.;Kaye,J.;Berger,S.L.;Wherry,E.J.,TOXtranscriptionally and epigenetically programs CD8(+)T cell exhaust ion.Nature2019.
53.Alfei,F.;Kanev,K.;Hofmann,M.;Wu,M.;Ghoneim,H.E.;Roelli,P.;Utzschneider,D.T.;von Hoesslin,M.;Cullen,J.G.;Fan,Y.;Eisenberg,V.;Wohlleber,D.;Steiger,K.;Merkler,D.;Delorenzi,M.;Knolle,P.A.;Cohen,C.J.;Thimme,R.;Youngblood,B.;Zehn,D.,TOX reinforces the phenotype and longevity of exhaustedT cells in chronic viral infection.Nature 2019.
54.Kawai,T.;Akira,S.,Signaling to NF-kappaB by Toll-likereceptors.Trends in molecular medicine 2007,13(11),460-9.
55.Chen,J.;Lopez-Moyado,I.F.;Seo,H.;Lio,C.J.;Hempleman,L.J.;Sekiya,T.;Yoshimura,A.;Scott-Browne,J.P.;Rao,A.,NR4Atranscription factors limit CART cell function in solid tumours.Nature 2019,567(7749),530-534.
56.Larkin,B.;Ilyukha,V.;Sorokin,M.;Buzdin,A.;Vannier,E.;Poltorak,A.,Cutting Edge:Activation of STING in T Cells Induces Type I IFN Responses andCell Death.Journal of immunology 2017,199(2),397-402.
57.Prinz,P.U.;Mendler,A.N.;Masouris,I.;Durner,L.;Oberneder,R.;Noessner,E.,High DGK-alpha and disabled MAPK pathways cause dysfunction ofhuman tumor-infiltrating CD8+T cells that is reversible by pharmacologicintervention.Journal of immunology 2012,l88(12),5990-6000.
58.Gorelik,L.;Flavell,R.A.,Immune-mediated eradication of tumorsthrough the blockade of transforming growth factor-βsignaling in Tcells.Nature medicine 2001,7(10),1118.
59.Herbertz,S.;Sawyer,J.S.;Stauber,A.J.;Gueorguieva,I.;Driscoll,K.F.;Estrem,S.T.;Cleverly,A.L.;Desaiah,D.;Guba,S.C.;Benhadji,K.A.;Slapak,C.A.;Lahn,M.M.,Clinical development of galunisertib(LY2157299 monohydrate),a smallmolecule inhibitor of transforming growth factor-beta signaling pathway.Drugdesign,develoment and therapy 2015,9,4479-99.
60.Jin,C.H.;Krishnaiah,M.;Sreenu,D.;Subrahmanyam,V.B.;Rao,K.S.;Lee,H.J.;Park,S.J.;Park,H.J.;Lee,K.;Sheen,Y.Y.;Kim,D.K.,Discovery of N-((4-([1,2,4]triazolo[1,5-aJpyridin-6-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-yl)methyl)-2-fluoroaniline(EW-7197):ahighly potent,selective,and orallybioavailable inhibitor of TGF-beta type I
receptor kinase as cancer immunotherapeutic/antifibroticagent.Journal of medicinal chemtistry 2014,57(10),4213-38.
61.Wang,D.;Quiros,J.;Mahuron,K.;Pai,C.C.;Ranzani,V.;Young,A.;Silveria,S.;Harwin,T.;Abnousian,A.;Pagani,M;Rosenblum,M.D.;Van Gool,F.;Fong,L.;Bluestone,J.A.;DuPage,M.,TargetingEZH2 Reprograms Intratumoral RegulatoryT Cells to Enhance Cancer Immunity.Cell reports 2018,23(11),3262-3274
Claims (33)
1.使耗竭的经典CAR T细胞恢复活力的系统,其包含至少两种组分:第一组分是包含与载荷药物共价连接的靶向配体的缀合物;并且第二组分是与膜锚定模块连接的靶向配体结合模块,其中所述第二组分的所述靶向配体结合模块以高亲和力识别所述第一组分中的所述靶向配体以形成复合物,所述载荷药物阻断所述耗竭的经典CAR T的抑制性信号传导,或者通过抗原非依赖性途径重新激活所述CAR T,并且其中所述膜锚定模块介导该双组分复合物内化到所述耗竭的CAR T细胞中。
2.根据权利要求1所述的系统,其中所述第一组分的所述靶向配体是叶酸、FITC或FK506。
3.根据权利要求1所述的系统,其中所述第二组分的所述靶向配体结合模块包含抗FITC抗体片段或FKBP或叶酸受体。
4.根据权利要求1所述的系统,其中所述膜锚定模块是叶酸受体。
5.根据权利要求1所述的系统,其中所述第一组分包含在所述靶向配体与所述载荷药物之间的可释放的接头。
6.根据权利要求1所述的系统,其中所述第一组分包含在所述靶向配体与所述载荷药物之间的不可释放的接头。
7.根据权利要求1所述的系统,其中所述靶向配体与所述配体结合模块之间的结合亲和力在亚纳摩范围内。
8.根据权利要求1所述的系统,其中所述药物载荷是Toll样受体7(TLR7)激动剂或干扰素基因刺激物(STING)激动剂。
9.根据权利要求1所述的系统,其中所述药物载荷是以下蛋白质的抑制剂:SHP1/2、TC-PTP或DGKα、TGFβ。
18.使耗竭的CAR T细胞恢复活力的方法,其包括:
a.向所述耗竭的CAR T细胞提供包含缀合物的第一组分,其中所述缀合物包含通过可释放或不可释放的接头与药物载荷共价连接的靶向配体;
b.向所述耗竭的CAR T细胞提供包含与耗竭的CAR构建体连接的融合受体的第二组分,其中所述融合受体包含靶向配体结合模块和膜结合受体模块;
c.使所述第二组分的所述靶向配体结合模块与所述第一组分中的所述靶向配体结合以形成复合物;
d.使该膜结合CAR模块介导所述复合物向所述耗竭的CAR T细胞的内化;
e.使所述载荷药物阻断所述耗竭的CAR T的抑制性信号传导或通过抗原非依赖性途径重新激活所述CAR T。
19.根据权利要求18所述的方法,其中所述载荷药物在所述耗竭的CAR T的内体中执行其功能,并且所述靶向配体与所述载荷药物通过不可释放的接头连接。
20.根据权利要求18所述的方法,其中所述载荷药物作为所述耗竭的CAR T的胞质溶胶中的游离药物执行其功能,并且所述靶向配体与所述载荷药物通过可释放的接头连接。
21.根据权利要求18所述的方法,其中所述第一组分的所述靶向配体是叶酸、FITC或FK506。
22.根据权利要求18所述的方法,其中所述第二组分的所述靶向配体结合模块是抗FITC、叶酸受体或FKBP。
23.根据权利要求18所述的方法,其中所述配体结合模块是叶酸受体α(FRa)。
24.根据权利要求18所述的方法,其中所述药物载荷是Toll样受体7(TLR7)激动剂或干扰素基因刺激物(STING)激动剂。
25.根据权利要求18所述的方法,其中所述药物载荷是以下蛋白质的抑制剂:SHP1/2、TC-PTP或DGKα、TGFβ。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310525133.XA CN116763943A (zh) | 2018-08-07 | 2019-07-21 | 使car t细胞恢复活力 |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201862715666P | 2018-08-07 | 2018-08-07 | |
US62/715,666 | 2018-08-07 | ||
PCT/US2019/042726 WO2020033129A1 (en) | 2018-08-07 | 2019-07-21 | Rejuvenation of car t cell |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310525133.XA Division CN116763943A (zh) | 2018-08-07 | 2019-07-21 | 使car t细胞恢复活力 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112543651A true CN112543651A (zh) | 2021-03-23 |
CN112543651B CN112543651B (zh) | 2023-06-02 |
Family
ID=69415080
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201980052586.7A Active CN112543651B (zh) | 2018-08-07 | 2019-07-21 | 使car t细胞恢复活力 |
CN202310525133.XA Pending CN116763943A (zh) | 2018-08-07 | 2019-07-21 | 使car t细胞恢复活力 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310525133.XA Pending CN116763943A (zh) | 2018-08-07 | 2019-07-21 | 使car t细胞恢复活力 |
Country Status (9)
Country | Link |
---|---|
US (2) | US20210308267A1 (zh) |
EP (1) | EP3833400A4 (zh) |
JP (1) | JP2021533166A (zh) |
KR (1) | KR20210042125A (zh) |
CN (2) | CN112543651B (zh) |
AU (1) | AU2019317278A1 (zh) |
CA (1) | CA3108710A1 (zh) |
SG (1) | SG11202100616VA (zh) |
WO (1) | WO2020033129A1 (zh) |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3589295A4 (en) | 2017-02-28 | 2020-11-04 | Endocyte, Inc. | COMPOSITIONS AND METHODS OF T CAR LYMPHOCYTE THERAPY |
WO2019144095A1 (en) | 2018-01-22 | 2019-07-25 | Seattle Children's Hospital (dba Seattle Children's Research Institute) | Methods of use for car t cells |
CN115551555A (zh) * | 2020-03-06 | 2022-12-30 | 普渡研究基金会 | 用于修饰car-t细胞活性的方法、化合物和组合物 |
CA3200076A1 (en) * | 2020-11-30 | 2022-06-02 | Purdue Research Foundation | Combinations of small molecule drug conjugate and car-expressing cytotoxic lymphocytes and methods of treating cancer using the same |
WO2023034341A1 (en) * | 2021-08-30 | 2023-03-09 | Purdue Research Foundation | Conjugates, compositions, and methods for rejuvenation of car t-cells |
WO2023156510A1 (en) * | 2022-02-18 | 2023-08-24 | F. Hoffmann-La Roche Ag | Method for detecting an analyte of interest in a sample |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090092582A1 (en) * | 2007-08-15 | 2009-04-09 | Oren Bogin | Compositions and methods for modifying properties of biologically active polypeptides |
CN105814083A (zh) * | 2013-10-15 | 2016-07-27 | 加州生物医学研究所 | 嵌合抗原受体t细胞开关和其用途 |
CN105829349A (zh) * | 2013-10-15 | 2016-08-03 | 加州生物医学研究所 | 肽嵌合抗原受体t细胞开关和其用途 |
CN106163547A (zh) * | 2014-03-15 | 2016-11-23 | 诺华股份有限公司 | 使用嵌合抗原受体治疗癌症 |
CN106459989A (zh) * | 2013-12-19 | 2017-02-22 | 诺华股份有限公司 | 人间皮素嵌合抗原受体及其用途 |
WO2017177137A1 (en) * | 2016-04-07 | 2017-10-12 | Bluebird Bio, Inc. | Chimeric antigen receptor t cell compositions |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2017504601A (ja) * | 2013-12-20 | 2017-02-09 | セレクティスCellectis | 免疫療法のためにマルチインプットシグナル感受性t細胞を操作する方法 |
WO2017035117A1 (en) * | 2015-08-24 | 2017-03-02 | University Of Houston System | Combination therpay combining car + t cells with appropriately timed immunodulatory antibodies |
WO2017177149A2 (en) * | 2016-04-08 | 2017-10-12 | Purdue Research Foundation | Methods and compositions for car t cell therapy |
CN109475558A (zh) * | 2016-05-25 | 2019-03-15 | 普渡研究基金会 | 通过靶向骨髓衍生的抑制细胞而治疗癌症的方法 |
EP3555627B1 (en) * | 2016-12-14 | 2023-11-22 | Purdue Research Foundation | Fibroblast activation protein (fap)-targeted imaging and therapy |
CA3053534A1 (en) * | 2017-02-17 | 2018-08-23 | Purdue Research Foundation | Targeted ligand-payload based drug delivery for cell therapy |
-
2019
- 2019-07-21 US US17/266,509 patent/US20210308267A1/en active Pending
- 2019-07-21 CA CA3108710A patent/CA3108710A1/en active Pending
- 2019-07-21 SG SG11202100616VA patent/SG11202100616VA/en unknown
- 2019-07-21 AU AU2019317278A patent/AU2019317278A1/en active Pending
- 2019-07-21 CN CN201980052586.7A patent/CN112543651B/zh active Active
- 2019-07-21 KR KR1020217006445A patent/KR20210042125A/ko unknown
- 2019-07-21 WO PCT/US2019/042726 patent/WO2020033129A1/en unknown
- 2019-07-21 EP EP19847190.6A patent/EP3833400A4/en active Pending
- 2019-07-21 CN CN202310525133.XA patent/CN116763943A/zh active Pending
- 2019-07-21 JP JP2021506492A patent/JP2021533166A/ja active Pending
-
2023
- 2023-12-21 US US18/392,939 patent/US20240148880A1/en active Pending
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090092582A1 (en) * | 2007-08-15 | 2009-04-09 | Oren Bogin | Compositions and methods for modifying properties of biologically active polypeptides |
CN105814083A (zh) * | 2013-10-15 | 2016-07-27 | 加州生物医学研究所 | 嵌合抗原受体t细胞开关和其用途 |
CN105829349A (zh) * | 2013-10-15 | 2016-08-03 | 加州生物医学研究所 | 肽嵌合抗原受体t细胞开关和其用途 |
CN106459989A (zh) * | 2013-12-19 | 2017-02-22 | 诺华股份有限公司 | 人间皮素嵌合抗原受体及其用途 |
CN106163547A (zh) * | 2014-03-15 | 2016-11-23 | 诺华股份有限公司 | 使用嵌合抗原受体治疗癌症 |
WO2017177137A1 (en) * | 2016-04-07 | 2017-10-12 | Bluebird Bio, Inc. | Chimeric antigen receptor t cell compositions |
Non-Patent Citations (3)
Title |
---|
JOHN VICTOR NAPOLEON,ET AL.: "Design, Synthesis, and Targeted Delivery of an Immune Stimulant that Selectively Reactivates Exhausted CAR T Cells", 《ANGEW. CHEM. INT. ED.》 * |
MIN SOO KIM,ET AL.: "Redirection of Genetically Engineered CAR‑T Cells Using Bifunctional Small Molecules", 《J. AM. CHEM. SOC.》 * |
李萍,等: "嵌合抗原受体T细胞在血液恶性肿瘤", 《内科急危重症杂志》 * |
Also Published As
Publication number | Publication date |
---|---|
JP2021533166A (ja) | 2021-12-02 |
EP3833400A1 (en) | 2021-06-16 |
CN112543651B (zh) | 2023-06-02 |
CN116763943A (zh) | 2023-09-19 |
AU2019317278A1 (en) | 2021-03-18 |
KR20210042125A (ko) | 2021-04-16 |
US20240148880A1 (en) | 2024-05-09 |
CA3108710A1 (en) | 2020-02-13 |
EP3833400A4 (en) | 2022-06-15 |
US20210308267A1 (en) | 2021-10-07 |
SG11202100616VA (en) | 2021-03-30 |
WO2020033129A1 (en) | 2020-02-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN112543651B (zh) | 使car t细胞恢复活力 | |
Siriwon et al. | CAR-T cells surface-engineered with drug-encapsulated nanoparticles can ameliorate intratumoral T-cell hypofunction | |
Taefehshokr et al. | Cancer immunotherapy: Challenges and limitations | |
ES2918501T3 (es) | Receptores de antígenos quiméricos de mesotelina humana y usos de los mismos | |
Hegde et al. | Tandem CAR T cells targeting HER2 and IL13Rα2 mitigate tumor antigen escape | |
CN107750167B (zh) | 用于治疗癌症和感染的免疫检查点调节剂的抑制剂 | |
Ampie et al. | Immunotherapeutic advancements for glioblastoma | |
Heymann et al. | Bone sarcomas in the immunotherapy era | |
Nel et al. | Multifunctional lipid bilayer nanocarriers for cancer immunotherapy in heterogeneous tumor microenvironments, combining immunogenic cell death stimuli with immune modulatory drugs | |
EP3833363A1 (en) | Methods and compositions for stimulation of chimeric antigen receptor t cells with hapten labelled cells | |
JP2022518207A (ja) | 酸化コレステロールを含有する薬物送達システム | |
Ju et al. | A carrier-free multiplexed gene editing system applicable for suspension cells | |
Riether et al. | From" magic bullets" to specific cancer immunotherapy | |
Jin et al. | Enhance anti-lung tumor efficacy of chimeric antigen receptor-T cells by ectopic expression of C–C motif chemokine receptor 6 | |
KR20190124268A (ko) | Nk 세포의 골수 호밍을 개선시키는 pm21 입자 | |
WO2021158534A1 (en) | Methods and compositions for stimulation of chimeric antigen receptor t cells with hapten labelled cells | |
Sun et al. | Downregulation of exosomal MHC-I promotes glioma cells escaping from systemic immunosurveillance | |
Grzybowski et al. | Novel dual arginase 1/2 inhibitor OATD-02 (OAT-1746) improves the efficacy of immune checkpoint inhibitors | |
San Miguel et al. | Engineering anti-myeloma responses using affinity-enhanced TCR-engineered T cells | |
CN116554357A (zh) | 一种双特异性嵌合抗原受体(car)及其在制备抗肿瘤药物中的应用 | |
Atrash et al. | Chimeric antigen receptor T-cell therapy for multiple myeloma | |
Chou | Developing Immunotherapy Targeting Immune Suppression Using Spherical Nucleic Acids (SNAS) | |
靳利远 et al. | Enhance anti-lung tumor efficacy of chimeric antigen receptor-T cells by ectopic expression of C–C motif chemokine receptor 6 | |
Zhang | Design of a Private Passageway Fusion Receptor for Sensitive Control of Adoptive Cell Therapies | |
WO2022087424A1 (en) | Monoamine oxidase blockade therapy for treating cancer through regulating tumor associated macrophages (tams) |
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 | ||
REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 40049545 Country of ref document: HK |
|
GR01 | Patent grant | ||
GR01 | Patent grant |