CN108064153A - 用于调节gst-pi基因的rna剂 - Google Patents
用于调节gst-pi基因的rna剂 Download PDFInfo
- Publication number
- CN108064153A CN108064153A CN201580071210.2A CN201580071210A CN108064153A CN 108064153 A CN108064153 A CN 108064153A CN 201580071210 A CN201580071210 A CN 201580071210A CN 108064153 A CN108064153 A CN 108064153A
- Authority
- CN
- China
- Prior art keywords
- gst
- nucleic acid
- acid molecules
- cancer
- molecule
- 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
- 108090000623 proteins and genes Proteins 0.000 title description 22
- 230000009368 gene silencing by RNA Effects 0.000 claims abstract description 78
- 239000002773 nucleotide Substances 0.000 claims abstract description 71
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 68
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 67
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 66
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 66
- 230000000692 anti-sense effect Effects 0.000 claims abstract description 56
- 201000011510 cancer Diseases 0.000 claims abstract description 49
- 230000014509 gene expression Effects 0.000 claims abstract description 41
- 239000000203 mixture Substances 0.000 claims abstract description 40
- 108091081021 Sense strand Proteins 0.000 claims abstract description 38
- 238000000034 method Methods 0.000 claims abstract description 33
- 108020004999 messenger RNA Proteins 0.000 claims abstract description 30
- 230000000295 complement effect Effects 0.000 claims abstract description 22
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 19
- 201000010099 disease Diseases 0.000 claims abstract description 14
- 102000040430 polynucleotide Human genes 0.000 claims abstract description 10
- 108091033319 polynucleotide Proteins 0.000 claims abstract description 10
- 239000002157 polynucleotide Substances 0.000 claims abstract description 10
- 108091030071 RNAI Proteins 0.000 claims abstract description 6
- 108020004459 Small interfering RNA Proteins 0.000 claims description 108
- 206010028980 Neoplasm Diseases 0.000 claims description 60
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 51
- 102000040650 (ribonucleotides)n+m Human genes 0.000 claims description 33
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 8
- 230000030279 gene silencing Effects 0.000 claims description 8
- 238000012226 gene silencing method Methods 0.000 claims description 7
- 201000005202 lung cancer Diseases 0.000 claims description 7
- 208000020816 lung neoplasm Diseases 0.000 claims description 7
- 239000004055 small Interfering RNA Substances 0.000 claims description 6
- 201000009030 Carcinoma Diseases 0.000 claims description 5
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 5
- 238000007385 chemical modification Methods 0.000 claims description 5
- 239000002502 liposome Substances 0.000 claims description 5
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 5
- 206010009944 Colon cancer Diseases 0.000 claims description 4
- 108091027967 Small hairpin RNA Proteins 0.000 claims description 4
- 230000002401 inhibitory effect Effects 0.000 claims description 4
- 230000036210 malignancy Effects 0.000 claims description 4
- 108091070501 miRNA Proteins 0.000 claims description 4
- 239000002679 microRNA Substances 0.000 claims description 4
- 201000003076 Angiosarcoma Diseases 0.000 claims description 3
- 208000003174 Brain Neoplasms Diseases 0.000 claims description 3
- 206010006187 Breast cancer Diseases 0.000 claims description 3
- 208000026310 Breast neoplasm Diseases 0.000 claims description 3
- 208000017897 Carcinoma of esophagus Diseases 0.000 claims description 3
- 208000005243 Chondrosarcoma Diseases 0.000 claims description 3
- 208000001333 Colorectal Neoplasms Diseases 0.000 claims description 3
- 206010061825 Duodenal neoplasm Diseases 0.000 claims description 3
- 208000000461 Esophageal Neoplasms Diseases 0.000 claims description 3
- 201000008808 Fibrosarcoma Diseases 0.000 claims description 3
- 208000022072 Gallbladder Neoplasms Diseases 0.000 claims description 3
- 208000001258 Hemangiosarcoma Diseases 0.000 claims description 3
- 208000018142 Leiomyosarcoma Diseases 0.000 claims description 3
- 206010024612 Lipoma Diseases 0.000 claims description 3
- 206010025323 Lymphomas Diseases 0.000 claims description 3
- 206010039491 Sarcoma Diseases 0.000 claims description 3
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 3
- 208000002495 Uterine Neoplasms Diseases 0.000 claims description 3
- 201000001531 bladder carcinoma Diseases 0.000 claims description 3
- 208000022033 carcinoma of urethra Diseases 0.000 claims description 3
- 208000006990 cholangiocarcinoma Diseases 0.000 claims description 3
- 201000000312 duodenum cancer Diseases 0.000 claims description 3
- 201000010175 gallbladder cancer Diseases 0.000 claims description 3
- 206010017758 gastric cancer Diseases 0.000 claims description 3
- 201000010536 head and neck cancer Diseases 0.000 claims description 3
- 208000014829 head and neck neoplasm Diseases 0.000 claims description 3
- 208000032839 leukemia Diseases 0.000 claims description 3
- 201000007270 liver cancer Diseases 0.000 claims description 3
- 208000014018 liver neoplasm Diseases 0.000 claims description 3
- 235000013372 meat Nutrition 0.000 claims description 3
- 201000008968 osteosarcoma Diseases 0.000 claims description 3
- 201000009410 rhabdomyosarcoma Diseases 0.000 claims description 3
- 201000011549 stomach cancer Diseases 0.000 claims description 3
- 206010042863 synovial sarcoma Diseases 0.000 claims description 3
- 210000001550 testis Anatomy 0.000 claims description 3
- 208000010570 urinary bladder carcinoma Diseases 0.000 claims description 3
- 206010046766 uterine cancer Diseases 0.000 claims description 3
- 206010060862 Prostate cancer Diseases 0.000 claims description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 2
- 108091036066 Three prime untranslated region Proteins 0.000 claims description 2
- 210000003734 kidney Anatomy 0.000 claims description 2
- 230000000505 pernicious effect Effects 0.000 claims 2
- 206010053717 Fibrous histiocytoma Diseases 0.000 claims 1
- 208000007766 Kaposi sarcoma Diseases 0.000 claims 1
- 208000001119 benign fibrous histiocytoma Diseases 0.000 claims 1
- 208000029742 colonic neoplasm Diseases 0.000 claims 1
- 125000003473 lipid group Chemical group 0.000 claims 1
- 210000001365 lymphatic vessel Anatomy 0.000 claims 1
- 208000030940 penile carcinoma Diseases 0.000 claims 1
- 201000008174 penis carcinoma Diseases 0.000 claims 1
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 abstract description 77
- 150000001875 compounds Chemical class 0.000 abstract description 18
- 210000004027 cell Anatomy 0.000 description 48
- 101000907904 Homo sapiens Endoribonuclease Dicer Proteins 0.000 description 29
- 102100023387 Endoribonuclease Dicer Human genes 0.000 description 28
- 239000000758 substrate Substances 0.000 description 26
- 239000000725 suspension Substances 0.000 description 22
- 241000699666 Mus <mouse, genus> Species 0.000 description 21
- 230000000694 effects Effects 0.000 description 16
- 238000002360 preparation method Methods 0.000 description 15
- 238000001727 in vivo Methods 0.000 description 13
- 150000002632 lipids Chemical group 0.000 description 10
- 210000001519 tissue Anatomy 0.000 description 10
- 108091028664 Ribonucleotide Proteins 0.000 description 9
- 238000011081 inoculation Methods 0.000 description 9
- 238000012545 processing Methods 0.000 description 9
- 239000002336 ribonucleotide Substances 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 8
- 238000009472 formulation Methods 0.000 description 8
- 125000002652 ribonucleotide group Chemical group 0.000 description 8
- 210000003711 chorioallantoic membrane Anatomy 0.000 description 7
- 231100000673 dose–response relationship Toxicity 0.000 description 7
- 230000005764 inhibitory process Effects 0.000 description 7
- 238000002347 injection Methods 0.000 description 7
- 239000007924 injection Substances 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 7
- 230000002829 reductive effect Effects 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 238000001890 transfection Methods 0.000 description 7
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 6
- 230000003993 interaction Effects 0.000 description 6
- 230000002265 prevention Effects 0.000 description 6
- 230000001225 therapeutic effect Effects 0.000 description 6
- 108091023037 Aptamer Proteins 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 239000005547 deoxyribonucleotide Substances 0.000 description 5
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 235000013399 edible fruits Nutrition 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 238000011065 in-situ storage Methods 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 101150082998 pi gene Proteins 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000012124 Opti-MEM Substances 0.000 description 4
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 210000000038 chest Anatomy 0.000 description 4
- 239000012228 culture supernatant Substances 0.000 description 4
- 208000035475 disorder Diseases 0.000 description 4
- 230000002349 favourable effect Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- 230000001875 tumorinhibitory effect Effects 0.000 description 4
- SNKAWJBJQDLSFF-NVKMUCNASA-N 1,2-dioleoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC SNKAWJBJQDLSFF-NVKMUCNASA-N 0.000 description 3
- 108010024636 Glutathione Proteins 0.000 description 3
- 206010025066 Lung carcinoma cell type unspecified stage 0 Diseases 0.000 description 3
- 101710190786 PI protein Proteins 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 239000012298 atmosphere Substances 0.000 description 3
- 235000012000 cholesterol Nutrition 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 229960003180 glutathione Drugs 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 210000004072 lung Anatomy 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 238000011580 nude mouse model Methods 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 210000002381 plasma Anatomy 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 229940124597 therapeutic agent Drugs 0.000 description 3
- 238000002054 transplantation Methods 0.000 description 3
- 230000004614 tumor growth Effects 0.000 description 3
- 230000005760 tumorsuppression Effects 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 208000006644 Malignant Fibrous Histiocytoma Diseases 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 206010054184 Small intestine carcinoma Diseases 0.000 description 2
- 208000015778 Undifferentiated pleomorphic sarcoma Diseases 0.000 description 2
- 210000001015 abdomen Anatomy 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000012520 frozen sample Substances 0.000 description 2
- 229930195733 hydrocarbon Natural products 0.000 description 2
- 150000002430 hydrocarbons Chemical class 0.000 description 2
- 206010020718 hyperplasia Diseases 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 239000006210 lotion Substances 0.000 description 2
- 208000012804 lymphangiosarcoma Diseases 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 108010082117 matrigel Proteins 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000002777 nucleoside Substances 0.000 description 2
- 125000003835 nucleoside group Chemical group 0.000 description 2
- 201000002528 pancreatic cancer Diseases 0.000 description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 102000016914 ras Proteins Human genes 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000002103 transcriptional effect Effects 0.000 description 2
- 239000003981 vehicle Substances 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- 239000013603 viral vector Substances 0.000 description 2
- 230000003442 weekly effect Effects 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 208000035657 Abasia Diseases 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 208000007860 Anus Neoplasms Diseases 0.000 description 1
- 206010073360 Appendix cancer Diseases 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- 108020004635 Complementary DNA Proteins 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical group OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 108010007355 Glutathione S-Transferase pi Proteins 0.000 description 1
- 102000007648 Glutathione S-Transferase pi Human genes 0.000 description 1
- 102100030943 Glutathione S-transferase P Human genes 0.000 description 1
- 101001010139 Homo sapiens Glutathione S-transferase P Proteins 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- 206010069755 K-ras gene mutation Diseases 0.000 description 1
- 208000008839 Kidney Neoplasms Diseases 0.000 description 1
- 101150105104 Kras gene Proteins 0.000 description 1
- 108091026898 Leader sequence (mRNA) Proteins 0.000 description 1
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- 239000012098 Lipofectamine RNAiMAX Substances 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 108091007412 Piwi-interacting RNA Proteins 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000012979 RPMI medium Substances 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 206010038389 Renal cancer Diseases 0.000 description 1
- 108010057163 Ribonuclease III Proteins 0.000 description 1
- 102000003661 Ribonuclease III Human genes 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- JCAQMQLAHNGVPY-UUOKFMHZSA-N [(2r,3s,4r,5r)-3,4-dihydroxy-5-(2,2,4-trioxo-1h-imidazo[4,5-c][1,2,6]thiadiazin-7-yl)oxolan-2-yl]methyl dihydrogen phosphate Chemical compound O[C@@H]1[C@H](O)[C@@H](COP(O)(O)=O)O[C@H]1N1C(NS(=O)(=O)NC2=O)=C2N=C1 JCAQMQLAHNGVPY-UUOKFMHZSA-N 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 230000001594 aberrant effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 201000011165 anus cancer Diseases 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 208000021780 appendiceal neoplasm Diseases 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000011717 athymic nude mouse Methods 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 238000011953 bioanalysis Methods 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- -1 cationic lipid Chemical class 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000003837 chick embryo Anatomy 0.000 description 1
- 230000001010 compromised effect Effects 0.000 description 1
- 239000006184 cosolvent Substances 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 230000006651 lactation Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 125000005647 linker group Chemical group 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 201000005296 lung carcinoma Diseases 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 239000004530 micro-emulsion Substances 0.000 description 1
- 238000007431 microscopic evaluation Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000003961 penetration enhancing agent Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920000768 polyamine Polymers 0.000 description 1
- 229920000570 polyether Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000002271 resection Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 231100000161 signs of toxicity Toxicity 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 229940098465 tincture Drugs 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000002463 transducing effect Effects 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000006648 viral gene expression Effects 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0071—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by measuring fluorescence emission
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/6527—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having nitrogen and oxygen atoms as the only ring hetero atoms
- C07F9/6533—Six-membered rings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1135—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against oncogenes or tumor suppressor genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
-
- 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
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/32—Chemical structure of the sugar
- C12N2310/322—2'-R Modification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/34—Spatial arrangement of the modifications
- C12N2310/344—Position-specific modifications, e.g. on every purine, at the 3'-end
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/35—Nature of the modification
- C12N2310/351—Conjugate
- C12N2310/3515—Lipophilic moiety, e.g. cholesterol
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/50—Physical structure
- C12N2310/53—Physical structure partially self-complementary or closed
- C12N2310/531—Stem-loop; Hairpin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/30—Special therapeutic applications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/30—Special therapeutic applications
- C12N2320/31—Combination therapy
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/30—Special therapeutic applications
- C12N2320/32—Special delivery means, e.g. tissue-specific
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/30—Special therapeutic applications
- C12N2320/35—Special therapeutic applications based on a specific dosage / administration regimen
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/50—Methods for regulating/modulating their activity
- C12N2320/53—Methods for regulating/modulating their activity reducing unwanted side-effects
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Plant Pathology (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Nanotechnology (AREA)
- Pathology (AREA)
- Virology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Medical Informatics (AREA)
- Optics & Photonics (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
Abstract
本发明提供利用RNA干扰来调节人GST‑π表达的化合物、组合物和方法。所述RNA干扰分子可用于预防或治疗例如恶性肿瘤等疾病的方法。核酸分子具有以下特征:a)多核苷酸有义链和多核苷酸反义链;b)所述分子的每条链长度为15~30个核苷酸;c)反义链中,由15~30个核苷酸构成的连续区域与编码GST‑π的mRNA序列互补;及d)有义链的至少一部分与反义链的至少一部分互补,并且,所述分子具有长度为15~30个核苷酸的双链区。
Description
技术领域
本发明涉及由基于核酸的分子构成的生物药物和治疗剂的领域。具体而言,本发明涉及利用RNA干扰(RNAi)来调节人GST-π表达的化合物和组合物。
序列表
本申请包括以电子版形式提交的序列表ASCII文件,该文件创建于2015年12月23日,文件名为ND5123202WO_SL.txt,大小为442,955字节,其全部内容通过参考并入本说明书中。
背景技术
已经发现,各种人癌组织与呈现KRAS基因突变有关。某些情况下,所述组织还呈现升高的谷胱甘肽S-转移酶-π(GST-π)表达水平(Miyanishi et al.,Gastroenterology,2001,Vol.121:865-874,摘要)例如,在患有各种胃肠道恶性肿瘤的患者中,观察到了升高的血清GST-π水平(Niitsu et al.,Cancer,1989,Vol.63,No.2,pp.317-323,摘要)。
GST-π是隶属GST家族的酶之一,其通过催化疏水性和亲电子性的化合物与还原型谷胱甘肽缀合而参与解毒。利用siRNA可在体外使GST-π表达降低(Niitsu et al.,US2014/0315975 Al)。
用于抑制GST-π表达的治疗剂要求高效的siRNA序列和结构。
因此,需要用于抑制GST-π表达的siRNA序列、化合物和结构。
发明内容
本发明涉及利用RNA干扰来调节人GST-π表达的化合物、组合物和方法。
在一些实施方式中,本发明提供用于GST-π的RNA干扰基因沉默的分子。
在另一些实施方式中,本发明的结构、分子和组合物可用于预防或治疗与GST-π相关的疾病、或改善恶性肿瘤等与GST-π相关的病症或紊乱的症状的方法。
本发明包括以下实施方式:
核酸分子,其中:
a)所述分子具有多核苷酸有义链和多核苷酸反义链;
b)所述分子的每条链长度为15~30个核苷酸;
c)所述反义链中的15~30个核苷酸的连续区域与编码GST-π的mRNA序列互补;
d)所述有义链的至少一部分与所述反义链的至少一部分互补,并且,所述分子具有长度为15~30个核苷酸的双链区。
在一些实施方式中,核酸分子可在反义链具有15~30个核苷酸的连续区域,所述区域与编码GST-π的mRNA序列互补,并且,所述区域位于分子的双链区。
在其他实施方式中,核酸分子可在反义链具有由15~30个核苷酸构成的连续区域,所述区域与编码GST-π的mRNA序列互补。
本发明的化合物可含有编码GST-π的mRNA序列,所述序列选自由序列号l的5’UTR第1~249位、序列号l的CDS第250~882位、序列号l的3’UTR第883~986位组成的组。
在某些实施方式中,核酸分子的每条链的长度可以为18~22个核苷酸。核酸分子的双链区长度可以为19个核苷酸。
在一些替代形式中,核酸分子可具有多核苷酸有义链和多核苷酸反义链,所述有义链和反义链以单链形式连接,并且形成一端以环(loop)连接的双链区。
一些实施方式中,本申请公开的核酸分子可具有平末端。在某些实施方式中,核酸分子可具有一个或多个3’悬挂。
本发明提供一系列核酸分子,其是具有基因沉默活性的RNAi分子。本发明的核酸分子可以是dsRNA、siRNA、micro-RNA、或具有基因沉默活性的shRNA、及DNA指导的RNA(ddRNA)、Piwi-相互作用RNA(piRNA)、或重复相关siRNA(rasiRNA)。核酸分子可具有GST-π表达抑制活性。
本发明的实施方式还提供GST-π敲低的IC50小于100pM的核酸分子。
本发明还包括含有一种或多种本发明的核酸分子、和药学上允许的担体(carrier)的组合物。在某些实施方式中,所述担体可以是脂质分子或脂质体。
对于本发明的化合物和组合物而言,通过将其施予至有需要的受试者,所述化合物或组合物对于预防或治疗GST-π相关疾病的的方法有用。
本发明的方法可使用本发明的化合物来对恶性肿瘤进行预防或治疗。该恶性肿瘤呈现于各种疾病,例如,与GST-π表达相关的癌症、由表达KRAS突变的细胞导致的癌症、肉瘤、纤维肉瘤、恶性纤维性组织细胞瘤、脂肪肉瘤、横纹肌肉瘤、平滑肌肉瘤、血管肉瘤、卡波西肉瘤、淋巴管肉瘤、滑膜肉瘤、软骨肉瘤、骨肉瘤、癌、脑瘤、头颈癌、乳腺癌、肺癌、食道癌、胃癌、十二指肠癌、阑尾癌、结肠直肠癌、直肠癌、肝癌、胰腺癌、胆囊癌、胆管癌、肛门癌、肾癌、尿道癌、膀胱癌、前列腺癌、睾丸癌、子宫癌、卵巢癌、皮肤癌、骨癌、白血病、恶性淋巴瘤、上皮性恶性肿瘤、和非上皮性恶性肿瘤。
附图说明
图1显示了序列号1(其为作为靶标的人谷胱甘肽S-转移酶-π(GST-π)mRNA的核酸序列),其被公开于GenBank登录号NM000852.3(hGSTPl),长度为986个核苷酸。
图2显示了GST-πsiRNA的体内敲低效果。如图2所示,在体内观察到了以GST-π作为靶标的siRNA对于GST-π的剂量依赖性敲低。
图3显示了GST-πsiRNA对于增殖的抑制。如图3所示,在A549细胞系(体外)中,观察到了以GST-π作为靶标的siRNA对于增殖的剂量依赖性抑制。
图4显示了GST-πsiRNA的抑制效果。利用的胰腺癌异种移植模型被施予了相对较低的量的以GST-π作为靶标的siRNA(0.75mg/kg)。GST-πsiRNA展示出显著的肿瘤抑制效果。
具体实施方式
本发明涉及用于基于核酸的治疗剂的化合物、组合物和方法,所述治疗对GST-π的表达进行调节。
在一些实施方式中,本发明提供具有RNA干扰活性的分子,以及能够使GST-π表达沉默的结构和组合物
本申请公开的结构和组合物可用于预防或治疗各种疾病,例如恶性肿瘤。
在另一些实施方式中,本发明提供用于递送或摄取一种或多种本发明的治疗性RNAi分子的组合物,以及使用所述组合物的方法。本发明的基于RNA的组合物可用于预防或治疗例如癌症等恶性肿瘤的方法。
本发明的治疗性组合物包括具有RNA干扰活性的核酸分子。所述治疗性核酸分子可以GSTP1(GST-π)作为靶标,实现基因沉默。
在各种实施方式中,本发明提供一系列能够作为小干扰RNA(siRNA)发挥作用的分子,并且,这些分子能够对GST-π基因表达加以调节或使其沉默。
本发明的siRNA可用于预防或治疗恶性肿瘤。
本发明的实施方式还提供用于将本发明的siRNA递送至需要预防或治疗恶性肿瘤的受试者的介质(vehicle)、制剂(formulation)、或脂质纳米粒制剂。本发明还包括向哺乳动物施予作为治疗剂的siRNA的方法。
对于本发明的治疗性分子和组合物而言,通过将其施予至有需要的受试者,所述分子和组合物可用于RNA干扰介导的GST-π相关疾病的预防或治疗。
本发明的方法可使用本发明的化合物来对恶性肿瘤进行预防或治疗。该恶性肿瘤呈现于各种疾病,例如:高水平表达GST-π的癌症、由表达KRAS突变的细胞导致的癌症、肉瘤、纤维肉瘤、恶性纤维性组织细胞瘤、脂肪肉瘤、横纹肌肉瘤、平滑肌肉瘤、血管肉瘤、卡波西肉瘤、淋巴管肉瘤、滑膜肉瘤、软骨肉瘤、骨肉瘤、癌、脑瘤、头颈癌、乳腺癌、肺癌、食道癌、胃癌、十二指肠癌、结肠直肠癌、肝癌、胰腺癌、胆囊癌、胆管癌、肾癌、尿道癌、膀胱癌、前列腺癌、睾丸癌、子宫癌、卵巢癌、皮肤癌、白血病、恶性淋巴瘤、上皮性恶性肿瘤、和非上皮性恶性肿瘤。
在某些实施方式中,本发明的治疗性分子的组合可用于GST-π基因表达的沉默或抑制。
本发明提供一系列核酸分子,所述分子具有多核苷酸有义链和多核苷酸反义链;所述分子的每条链长度为15~30个核苷酸;反义链中的15~30个核苷酸的连续区域与编码GST-π的mRNA序列互补;有义链的至少一部分与反义链的至少一部分互补,并且,分子具有长度为15~30个核苷酸的双链区。
本发明的RNAi分子可在反义链具有15~30个核苷酸的、与编码GST-π的mRNA序列互补的连续区域,所述区域位于分子的双链区。
在一些实施方式中,RNAi分子可在反义链具有15~30个核苷酸的连续区域,所述区域与序列编码GST-π的mRNA序列互补。
本发明的实施方式还可提供下述方法:在有需要的哺乳动物中,预防、治疗或改善恶性肿瘤的一种或多种症状的方法、或降低发展恶性肿瘤的风险的方法、或延迟恶性肿瘤发病的方法。
GST-π和RNAi分子
图1显示了作为靶标的人谷胱甘肽S-转移酶-π(GST-π)mRNA的核酸序列的一个例子,其公开于GenBank登录号NM 000852.3(hGSTPl),长度为986个核苷酸。
本领域技术人员可以理解,已提交的序列可随时间而变更,以引入核酸分子中任何需要的改变。
本发明的实施方式可提供使用小核酸分子的组合物和方法,其用于GST-π基因表达沉默。核酸分子的例子包括具有RNA干扰活性的分子(RNAi分子)、短干扰RNA(siRNA)、双链RNA(dsRNA)、micro-RNA(miRNA)、和短发夹RNA(shRNA)分子、以及DNA指导的RNA(ddRNA)、Piwi-相互作用RNA(piRNA),和重复相关siRNA(rasiRNA)。这些分子能够介导对于GST-π基因表达的RNA干扰。
本申请公开的组合物和方法也可用于治疗受试者的各种恶性肿瘤。
本发明的核酸分子和方法可用于下调GST-π编码基因的表达。
本发明的组合物和方法可包含一种或多种核酸分子,所述核酸分子能够单独或通过组合来调节或调控GST-π蛋白及/或GST-π蛋白编码基因的表达、和与恶性肿瘤等GST-π相关疾病、病症或紊乱的维持及/或发展相关的蛋白及/或GST-π蛋白编码基因的表达。
参考GST-π的示例性序列,对本发明的组合物和方法进行说明。本领域技术人员可以理解,本发明的各方面和实施方式可对应任何相关的GST-π基因、序列、或变体(例如,同源基因和转录变体)、以及多态性、例如与任何GST-π基因相关的单核苷酸多态性(SNP)。
在一些实施方式中,本发明的组合物和方法可提供下调GST-π基因(例如,人GST-π)表达的双链短干扰核酸(siRNA)分子。
本发明的RNAi分子以GST-π和任何能够提供其他靶序列的同源序列作为靶标,例如使用互补序列或通过并入非典型(canonical)碱基对例如错配和/或摆动(wobble)碱基对(其可提供额外的靶序列)。
在鉴定错配的情况下,可以使用非典型碱基对,例如错配和/或摆动碱基来产生靶向多于一个基因序列的核酸分子。例如,UU和CC碱基对等非常规碱基对可用于生成能以具有序列同源性的不同的GST-π靶序列作为靶标的核酸分子。由此,RNAi分子可将同源基因的保守核苷酸序列作为靶标,从而可使用1种RNAi分子来抑制多个基因的表达。
在一些方面,本发明的组合物和方法包含对于GST-πmRNA具有活性的RNAi分子,所述RNAi分子含有与任意的编码GST-π的mRNA互补的序列。
在一些实施方式中,本申请公开的RNAi分子可具有对于GST-πRNA的活性,所述RNAi分子含有与具有GST-π变体编码序列(例如,本领域已知的与恶性肿瘤相关的GST-π基因突变)的RNA互补的序列。
在另一些实施方式中,本发明的RNAi分子可含有能够与GST-π基因的核苷酸序列相互作用、且能够介导GST-π基因表达沉默的核苷酸序列。
本发明的以GST-πmRNA作为靶标的RNAi分子的例子示于表1和表2。
表1:GST-πRNAi分子序列
表1备注:大写字母A、G、C、U分别表示ribo-A、ribo-G、ribo-C和ribo-U。小写字母a、g、c、t分别表示2’-脱氧-A、2’-脱氧-G、2’-脱氧-C和脱氧胸苷。
表2:GST-πRNAi分子序列
表2备注:大写字母A、G、C、U分别表示ribo-A、ribo-G、ribo-C和ribo-U。小写字母a、g、c、t分别表示2’-脱氧-A、2’-脱氧-G、2’-脱氧-C和脱氧胸苷。
例如,本发明的siRNA可具有序列号1341的反义链、和序列号1276的有义链,或经过化学修饰的上述链。
例如,本发明的siRNA可具有反义链序列号1305的反义链、和序列号1240的有义链,或经过化学修饰的上述链。
对于化学修饰而言,可在链中任意位置的任意氨基酸含有2’-OMe取代基,以及本领域已知的其他修饰。
调节GST-π和治疗恶性肿瘤的方法
本发明的实施方式可提供用于下调或抑制GST-π及/或GST-π蛋白的表达的RNAi分子。
在一些实施方式中,本发明的RNAi分子可用于下调或抑制GST-π及/或GST-π蛋白的表达,所述GST-π及/或GST-π蛋白源于可能与例如恶性肿瘤等疾病或病症相关的GST-π单倍型多态性。
对于GST-π的蛋白或mRNA的水平的监控可用于表征基因沉默,以及测定本发明的化合物和组合物的效果。
本申请公开的RNAi分子可单独使用,也可与其他siRNA组合使用来调节一个或多个基因的表达。
本申请公开的RNAi分子可单独使用,也可与其他已知的药物组合或缀合使用,由此预防或治疗与GST-π相关的疾病、或改善例如恶性肿瘤等与GST-π相关的病症或紊乱的症状。
本发明的RNAi分子可用于以序列特异性方式调节或抑制GST-π表达。
本申请公开的RNAi分子可包含引导链(guide strand),其是一系列连续的核苷酸,且至少部分与GST-πmRNA互补。
在特定的方面,可使用本发明的RNAi分子通过RNA干扰对恶性肿瘤进行治疗。
对于恶性肿瘤的治疗而言,可利用合适的以细胞为基础的模型、动物模型(体外或体内)进行表征。
对于恶性肿瘤的治疗而言,可通过测定受影响组织的细胞中的GST-πmRNA水平或GST-π蛋白水平进行表征。
对于恶性肿瘤的治疗而言,可通过对受影响的器官或组织进行非侵入性的医学扫描进行表征。
本发明的实施方式可包括对有需要的受试者的GST-π相关疾病或病症的症状进行预防、治疗、或改善的方法。
在一些实施方式中,对受试者的恶性肿瘤的症状进行预防、治疗、或改善的方法可包括向受试者施予本发明的RNAi分子,由此调节受试者或生物体中的GST-π基因的表达。
在一些实施方式中,本发明包括通过使细胞或生物体与本发明的RNAi分子接触来下调细胞或生物体中的GST-π基因表达的方法。
RNA干扰
RNA干扰(RNAi)是指由短干扰RNA(siRNA)介导的动物中的序列特异性转录后基因沉默。例如,参见Zamore et al.,Cell,2000,Vol.101,pp.25-33;Fire et al.,Nature,1998,Vol.391,pp.806811;Sharp,Genes&Development,1999,Vol.13,pp.139-141。
细胞中的RNAi反应可以由双链RNA(dsRNA)引发,但其机理尚未彻底研明。在细胞中,作为RNase III酶的Dicer酶可作用于特定的dsRNA。例如,参见Zamore et al.,Cell,2000,Vol.101,pp.25-33;Hammond et al.,Nature,2000,Vol.404,pp.293-296。Dicer可将dsRNA加工成更短的dsRNA片段,即siRNA。
一般而言,siRNA的长度可以为21~约23个核苷酸,并且包含长度为约19个核苷酸的碱基对双链区。
RNAi涉及被称为RNA诱导沉默复合物(RISC)的核酸内切酶复合物。siRNA具有的反义链或引导链进入RISC复合物,并介导单链靶RNA的切割,所述单链靶RNA含有与siRNA双链体的反义链互补的序列。siRNA的另一条链为随从链(passenger strand)。靶RNA的切割在与siRNA双链体的反义链互补的区域中部发生。例如,参见Elbashir et al.,Genes&Development,2001,Vol.15,pp.188-200。
本文中,用语“有义链”表示siRNA分子的下述核苷酸序列,所述序列与该siRNA分子的反义链的至少一部分完全互补或部分互补。siRNA分子的有义链可含有与靶核酸序列具有同源性的核酸序列。
本文中,用语“反义链”表示与靶核酸序列的至少一部分完全互补或部分互补的siRNA分子的核苷酸序列。siRNA分子的反义链可含有与该siRNA分子的有义链的至少一部分互补的核酸序列。
RNAi分子可通过以序列特异性方式介导RNA干扰来下调或敲低基因表达。例如,参见Zamore et al.,Cell,2000,Vol.101,pp.25-33;Elbashir et al.,Nature,2001,Vol.411,pp.494-498;Kreutzer et al.,WO2000/044895;Zernicka-Goetz et al.,WO2001/36646;Fire et al.,WO1999/032619;Plaetinck et al.,WO2000/01846;Mello etal.,WO2001/029058。
本文中,基因表达方面的用语“抑制”、“下调”或“降低”是指,观察到基因表达、或编码一种或多种蛋白的mRNA分子的水平、或一种或多种编码蛋白的活性低于不存在本发明的RNAi分子或siRNA的情况。例如,可观察到较之不存在本发明的RNAi分子或siRNA的情况而言,表达水平、mRNA水平、或编码蛋白的活性水平降低至少1%、或至少10%、或至少20%、或至少50%、或至少>90%、或更多。
RNAi分子还可用于敲低病毒基因表达,从而能够影响病毒复制。
RNAi分子可以由独立的多核苷酸链制得:有义链或随从链、和反义链或引导链。引导链与随从链至少部分互补。引导链和随从链可形成具有约15~49个碱基对的双链区。
在一些实施方式中,siRNA双链区可具有碱基对。
在某些实施方式中,RNAi分子的长度包括具有RISC活性的双链区,由此可在RISC复合物中发挥作用。
在另一些实施方式中,RNAi分子可以作为Dicer底物发挥作用,由此被转换为能够在RISC复合物中发挥作用的RNAi分子。
在一些方面,RNAi分子可在长链分子相反的两端具有互补的引导序列部分和随从序列部分,由此,分子能够介由互补的序列部分而形成双链区,在双链区的一端介由核苷酸接头或非核苷酸接头将链彼此连接。例如,发夹结构或茎环结构。接头与链的相互作用可以是共价键或非共价相互作用。
本申请公开的RNAi分子可包含将核酸的有义区域与反义区域接合的核苷酸接头、非核苷酸接头、或核苷酸/非核苷酸混合接头。核苷酸接头的长度可以为≥2个核苷酸,例如约3、4、5、6、7、8、9、或10个核苷酸。核苷酸接头可以是核酸适体。本文中,用语“适体”或“核酸适体”表示与靶分子特异性结合的核酸分子,所述核酸分子的序列中包含天然能够被靶分子识别的序列。或者,适体也可以是天然不会与核酸结合的靶分子结合的核酸分子。例如,适体可用于与蛋白质的配体结合区域结合,由此阻碍天然存在的配体与蛋白的相互作用。例如,参见Gold et al.,Annu Rev Biochem,1995,Vol.64,pp.763-797;Brody et al.,J.Biotechnol.,2000,Vol.74,pp.5-13;Hermann et al.,Science,2000,Vol.287,pp.820-825。
非核苷酸接头的例子包括脱碱基核苷酸、聚醚、聚胺、聚酰胺、肽、碳水化合物、脂质、聚烃、或其他聚合物,例如,具有例如2~100个乙二醇单元的聚乙二醇。一些例子记载于Seela et al.,Nucleic Acids Research,1987,Vol.15,pp.3113-3129;Cload et al.,J.Am.Chem.Soc,1991,Vol.113,pp.6324-6326;Jaeschke et al.,Tetrahedron Lett.,1993,Vol.34,pp.301;Arnold et al.,WO 1989/002439;Usman et al.,WO1995/006731;Dudycz et al.,WO 1995/011910,and Ferentz et al.,J.Am.Chem.Soc,1991,Vol.113,pp.4000-4002。
RNAi分子可在双链区具有一个或多个悬挂。悬挂是碱基未配对的单链区,长度可为1~8个核苷酸或更长。悬挂可以是3’悬挂,其中,链的3’端具有长度为1~8个核苷酸的单链区。悬挂可以是5’悬挂,其中,链的5’端具有长度为1~8个核苷酸的单链区。
RNAi分子的悬挂的长度可以相同,也可以不同。
RNAi分子可具有一个或多个平末端,即,双链区的末端没有悬挂,两条链的碱基配对直到双链区的末端。
本申请公开的RNAi分子可具有一个或多个平末端,也可具有一个或多个悬挂,或者具有平末端和悬挂的组合。
RNAi分子链的5’端可以是平末端,或者也可以是悬挂。RNAi分子链的3’端可以是平末端,或者也可以是悬挂。
RNAi分子链的3’端为悬挂时,5’端可以是平末端。RNAi分子链的5’端为悬挂时,3’端可以是平末端。
在一些实施方式中,RNAi分子的两端均为平末端。
在另一些实施方式中,RNAi分子的两端可具有悬挂。
5’端和3’端的悬挂的长度可以不同。
在某些实施方式中,RNAi分子可具有平末端,即,在反义链的5’端和有义链的3’端不具有任何悬挂的核苷酸。
在另一些实施方式中,RNAi分子可具有平末端,即,在反义链的3’端和有义链的5’端不具有任何悬挂的核苷酸。
RNAi分子可在双链区具有碱基错配。
RNAi分子的悬挂中的任意的核苷酸可以是脱氧核糖核苷酸,也可以是核糖核苷酸。
RNAi分子可在5’端具有一个或多个脱氧核糖核苷酸,另一条链的3’端可不具有悬挂,或不具有脱氧核糖核苷酸悬挂。
RNAi分子可在3’端具有一个或多个脱氧核糖核苷酸,另一条链的5’端可不具有悬挂,或不具有脱氧核糖核苷酸悬挂。
在一些实施方式中,RNAi分子的一个或多个、或者全部悬挂的核苷酸可以为2’-脱氧核糖核苷酸。
Dicer底物RNAi分子
在一些方面,RNAi分子可具有适合作为Dizcer底物的长度,其能够被加工成具有RISC活性的RNAi分子。例如,参见Rossi et al.,US2005/0244858。
作为Dicer底物的双链RNA(dsRNA)具有充分的长度,使其能够被Dicer加工成活性RNAi分子,并且,具有以下一种或多种性质:(i)作为Dicer底物的dsRNA可以是不对称的,例如,可在反义链具有3’悬挂;及(ii)Dicer底物dsRNA可在有义链的3’末端具有修饰,由此引导Dicer结合和加工的方向,将dsRNA加工成活性RNAi分子。
在某些实施方式中,作为Dicer底物的dsRNA中最长的链可以24~30个核苷酸。
Dicer底物dsRNA可以是对称的或不对称的。
在一些实施方式中,Dicer底物dsRNA可具有长度为22~28个核苷酸的有义链、和长度为24~30个核苷酸的反义链。
在某些实施方式中,Dicer底物dsRNA可在反义链的3’端具有悬挂。
在另一些实施方式中,Dicer底物dsRNA可含有长度为25个核苷酸的有义链、和长度为27个核苷酸的反义链,所述反义链具有2个碱基的3’悬挂。悬挂的长度可以为1、2或3个核苷酸。有义链也可具有5’磷酸基团(phosphate)。
不对称的Dicer底物dsRNA可在有义链的3’端代替2个核糖核苷酸而含有2个脱氧核糖核苷酸。
Dicer底物dsRNA的有义链的长度可以为约22~约30;或约22~约28;或约24~约30;或约25~约30;或约26~约30;或约26~29;或约27~约28个核苷酸。
Dicer底物dsRNA的有义链的长度可以为22、23、24、25、26、27、28、29或30个核苷酸。
在某些实施方式中,Dicer底物dsRNA可具有有义链和反义链,它们的长度至少为25个核苷酸,且不多于约30个核苷酸。
在某些实施方式中,Dicer底物dsRNA可具有长度为26~29个核苷酸的有义链和反义链。
在某些实施方式中,Dicer底物dsRNA可具有长度为27个核苷酸的有义链和反义链。
Dicer底物dsRNA的有义链和反义链的长度可以相同(例如,具有平末端的情况),也可以不同(例如,具有悬挂的情况),或者可含有1个平末端和1个悬挂。
Dicer底物dsRNA可具有长度为19、20、21、22、23、24、25、26或27个核苷酸的双链区。
Dicer底物的反义链dsRNA可含有在生物环境中(真核细胞的细胞质中)与有义链序列的至少一部分退火(anneal)的任意序列。
具有有义链和反义链的Dicer底物可被其他的结构连接,例如接头基团或接头寡核苷酸。例如,接头将dsRNA的两条链连接,由此通过退火形成发夹结构。
Dicer底物的有义链和反义链通常是互补的,但可含有碱基对错配。
在一些实施方式中,Dicer底物dsRNA可以是不对称的,例如,有义链具有22~28个核苷酸,反义链具有24~30个核苷酸。
Dicer底物dsRNA链、尤其是反义链中的区域的序列长度可以为至少19个核苷酸,这些核苷酸位于接近反义链3’端的长度为21个核苷酸的区域,并且与由靶基因产生的RNA的核苷酸序列充分互补。
Dicer底物dsRNA的反义链可在5’端具有1~9个核糖核苷酸,从而使其长度成为22~28个核苷酸。若反义链的长度为21个核苷酸,则可在3’端添加1~7个核糖核苷酸、或2~5个核糖核苷酸、或4个核糖核苷酸。添加的核糖核苷酸可以为任意的序列。
Dicer底物dsRNA的有义链可含有24~30个核苷酸。为了在生物环境下与反义链结合,有义链可实质上与反义链互补。
RNAi分子的使用方法
本发明的核酸分子和RNAi分子可通过直接应用而递送至细胞或组织,也可与担体或稀释剂组合。
对于本发明的核酸分子和RNAi分子而言,可使用担体或稀释剂、或任何其他发挥辅助、促进或加快进入细胞的作用的递送介质(vehicle)(例如,病毒序列、病毒物质、或者脂质或脂质体制剂),由此将分子直接用于递送或施予至细胞、组织、器官、或受试者。
本发明的核酸分子和RNAi分子可以与阳离子性脂质形成复合物、被脂质体包封、或以其它方式递送至靶细胞或靶组织。核酸或核酸复合物可通过直接经皮施用、透皮施用、或注射而局部施予至体外或体内的有关组织。
递送体系可包括:例如,水性和非水性的凝胶、乳霜(cream)、乳剂(emulsion)、微乳剂、脂质体、软膏、水性和非水性的溶液、乳液(lotion)、气溶胶(aerosols)、烃类基剂和粉末,并且可含有例如助溶剂、渗透促进剂等赋形剂。
本申请公开的组合物和方法可包含表达载体,所述载体以能够使核酸分子表达的方式而含有编码至少1种本发明的RNAi分子的核酸序列。
本发明的核酸分子和RNAi分子可利用插入DNA或RNA载体的转录单元表达。重组载体可以是DNA质粒或病毒载体。病毒载体可用于使核酸分子短暂表达。
例如,载体可含有编码RNAi分子双链体的2条链、或自身互补从而能形成RNAi分子的单个核酸分子的序列。表达载体可含有编码2个或更多核酸分子的核酸序列。
核酸分子可利用真核生物启动子在细胞中表达。本领域技术人员熟知,核酸可利用合适的DNA/RNA载体而在真核细胞中表达。
在一些方面,病毒构建体可用于将表达构建体导入细胞,由此转录该表达构建体编码的dsRNA构建体。
脂质制剂可通过静脉注射、肌肉内注射、腹腔内注射、口服、吸入或本领域已知的其他方法施予至动物。
可使用已知能够用于施予寡核苷酸的药学上允许的制剂。
实施例
实施例1:为了测定siRNA敲低效果,利用A549细胞系实施了体外转染。如表3所示,观察到了siRNA对于GST-πmRNA的剂量依赖性敲低。
表3:A549细胞系中GST-πmRNA的剂量依赖性敲低
siRNA | IC50(pM) |
A9 | 27、29 |
B2 | 121 |
B3 | 235 |
B4 | 229 |
B13 | 23、34 |
BU02 | 21、25、34 |
实施例2:体外敲低的方案
在转染前1天,向96孔板中注入细胞(2 x103细胞/孔,100μl含有10%FBS的DMEM(HyClone Cat.#SH30243.01)),在37℃的孵育器(湿润气氛,空气中CO2浓度为5%)中培养。转染前,将培养基置换为90μl含有2%FBS的Opti-MEM I减血清培养基(Life TechnologiesCat.#31985-070)。将0.2μl的Lipofectamine RNAiMax(Life Technologies Cat.#13778-100)与4.8μl的Opti-MEM I于室温混合5分钟。将1μl的siRNA与4μl的Opti-MEM I混合,进而加入LF2000溶液,轻轻混合,无需振荡(vortex)。于室温等待5分钟。将混合物于室温孵育10分钟,形成RNA-RNAiMax复合物。向孔中加入10μl的RNA-RNAiMax复合物,用手轻轻摇动平板。将细胞在37℃的孵育器(湿润气氛,空气中CO2浓度为5%)中孵育2小时。将培养基置换为新鲜的含有2%FBS的Opti-MEM I减血清培养基(Life Technologies Cat.#31985-070)。转染24小时后,用冰冷的PBS洗涤1次细胞。用50μl的Cell-to-Ct裂解液(LifeTechnologies Cat.#4391851C)裂解细胞,于室温处理5~30分钟。加入5μl的终止溶液,并于室温孵育2分钟。立刻使用TAQMAN通过RT-qPCR测定mRNA水平。或者,也可将试样于-80℃冷冻,在稍后进行分析。
实施例3:图2显示了GST-πsiRNA的体内敲低效果。如图2所示,在体内观察到了以GST-π作为靶标的BU02siRNA对于GST-π的剂量依赖性敲低。
实施例4:图3显示了以GST-π作为靶标的siRNA对于细胞增殖的抑制。如图3所示,在体外A549细胞系中观察到了以GST-π作为靶标的siRNA对于增殖的剂量依赖性抑制。
实施例5:图4显示了GST-πsiRNA(BU02)的肿瘤抑制效果。利用的胰腺癌异种移植模型被施予了相对较低的量的以GST-π作为靶标的siRNA(0.75mg/kg)。在第28天,GST-πsiRNA展示出显著且出乎意料的有利的肿瘤抑制效果。
该实验中,从ATCC获得了A549细胞系和PANC-1细胞系。将细胞悬浮液与冰上融化的BD matrigel以1:1的比例充分混合,用于注射。使用25G注射针和针筒,在每只小鼠(雌性无胸腺裸鼠,6~8周,Charles River)的右侧腹皮下接种0.1ml含有2.5x106个(A549)或2.5x106个(PANC-1)细胞的接种液(每只小鼠接种1剂接种液)。小鼠在接种时被麻醉。在培养的肿瘤达到约250~350mm3(A549)或约150~250mm3(PANC-1)当天,经由动物的尾部血管进行弹丸注射(bolus injection)。在给药后不同的时间点利用高浓度CO2杀死动物并切除肿瘤。先称取肿瘤的湿重,然后将其分成3部分,用于测量GST-π敲低、siRNA生物分布、和生物标志物分析。用液氮快速冷冻试样,并保存于-80℃,直到准备进行生物分析为止。
实施例6:原位A549肺癌小鼠模型。本发明的GST-πsiRNA能够导致原位肺癌肿瘤在体内显著缩小。该实施例中,在以脂质体制剂的形式施予至无胸腺裸鼠的原位肺癌肿瘤的情况下,GST-πsiRNA在体内发挥了基因敲低效能。
一般而言,原位肿瘤模型可呈现药物效果、效能和提高的预估性能的直观临床意义。原位肿瘤模型中,肿瘤细胞被直接植入与细胞来源种类相同的器官。
将处理组和介质对照组剖检时的最终原发瘤重量进行比较,由此对siRNA制剂对于人A549肺癌的抗肿瘤效果进行评价。
在体内观察到了由基于BU2结构的GST-πsiRNA(序列号1276、1341)对于原位肺癌肿瘤的抑制。利用的原位A549肺癌小鼠模型被施予了相对较低的量(2mg/kg)的以GST-π作为靶标的siRNA。
在为期6周的该研究中,GST-πsiRNA呈现了显著且出乎意料的有利的肿瘤抑制效果。43天后,GST-πsiRNA呈现了明显有利的肿瘤抑制,较之对照而言,肿瘤的平均体积显著缩小2.8倍。
该研究中,使用了5~6周龄的雄性NCr nu/nu小鼠。在实验期间,将实验动物饲育在经过HEPA过滤的环境中。siRNA制剂在使用前于4℃保存,并在注射进小鼠10分钟前使其升温至室温。
对于该A549人肺癌原位模型而言,在外科原位移植(SOI)当天,从接受A549肿瘤异种移植的动物皮下部位采集备份肿瘤(stock tumor),置于RPMI-1640培养基中。摘除坏死组织,将活体组织切割成1.5~2mm3的小片。通过异氟烷吸入将动物麻醉,用碘酒和酒精消毒手术区。使用一对手术剪在小鼠左胸壁切开约1.5cm长的横向切口。切开第三、四肋骨之间,暴露左肺。使用8-0外科缝线(尼龙),将一片A549肿瘤移植至肺表面。使用6-0缝线(丝)将胸壁缝合。使用装有25G X 11/2针头的3cc注射器进行胸内穿刺,使肺重新膨胀,由此排出胸腔中残余的空气。使用6-0缝线(丝)将胸壁缝合。上述所有手术步骤均在HEPA过滤层流操作台上使用7x放大倍数的显微镜实施。
肿瘤移植3天后,将荷瘤模型小鼠随机分组,每组10只小鼠。对于处理组而言,在肿瘤移植3天后开始对10只小鼠进行处理。
对于处理组而言,制剂为脂质体组合物(可离子化脂质:胆固醇:DOPE:DOPC:DPPE-PEG-2K:DSPE-PEG-2K)。脂质体包封GST-πsiRNA。
对于研究终点而言,在处理开始42天后杀死实验小鼠。切除原发瘤,并用电子天平称重,用于后续分析。
对于化合物毒性的估测而言,在整个实验期间,处理组和对照组的小鼠的平均体重维持在正常范围内。在小鼠中未观察到其他毒性症状。
实施例7:以GST-π作为靶标的小干扰RNA(siRNA)对于小鼠中A549细胞生长和绒毛尿囊膜(CAM)上血管形成的效果的分析。构建3对GST-πsiRNA质粒和非沉默质粒(non-silencing-plasmid),利用LIPOFECTAMINE 2000将它们分别转染至A549细胞。通过ELISA和实时RT-PCR选出最有效的一对GST-πsiRNA质粒。将使用选出的GST-πsiRNA质粒转染的A549细胞、使用非沉默质粒转染的A549细胞、和未经过转染的A549细胞分别接种至裸鼠。将雏鸡胚胎随机地分成4组,用不同的溶液处理CAM 48小时:以DMEM培养基作为阴性对照组,以未经转染的A549细胞培养上清液作为阳性对照组,以GST-πsiRNA A549细胞培养上清液作为GST-πsiRNA组,以非沉默siRNA A54细胞培养上清液作为非沉默siRNA组。在第12天收集CAM,供于显微镜分析。
较之对照组而言,GST-πsiRNA质粒在使GST-πmRNA降低的同时诱导了由A549细胞分泌的GST-π的减少。较之非沉默siRNA组而言,GST-πsiRNA组中,小鼠异种移植肿瘤的平均体积减小;异种移植瘤生长至50mm3的时间被延迟。异种移植瘤中的GST-π成分减少。CAM分析中,阴性对照组的GST-π成分为0,而GST-πsiRNA组的GST-π成分较之非沉默siRNA组或阳性对照组而言降低了20~70%;较之阴性对照组而言,GST-πsiRNA组或非沉默siRNA组或阳性对照组中,CAM的血管分支点增加;较之阴性对照组而言,GST-πsiRNA组中CAM的总血管长度增加,而在非沉默siRNA组或阳性对照组中也增加。向GST-πsiRNA组添加含有GST-π的细胞培养上清液时,较之阴性对照组而言,微血管的增生增加,而在非沉默siRNA组或阳性对照组中,观察到了明显增生的血管。
实施例8:细胞培养。在添加有10%FBS(FBS,Invitrogen)的F-12K培养基(ATCC)中培养人非小细胞肺癌细胞系A549(37℃,湿润气氛,空气中CO2浓度为5%)。使用各种慢病毒转导颗粒,按照各制造商(Sigma-Aldrich)的说明分别转导A549TR细胞,制备稳定表达对照、或GST siRNA的细胞。
使用2.5μg/mL嘌呤霉素(Invivogen)进行筛选,12天后,用克隆量筒分离具有抗性的克隆,然后利用含有嘌呤霉素的培养基进行扩增和维持。
实施例9:以GST-π作为靶标的siRNA导致体内肿瘤体积的显著缩小。
使用了脂质制剂,由此将siRNA包封在纳米粒子中,并递送至严重联合免疫缺陷小鼠中的人A549肺癌细胞异种移植瘤。对异种移植瘤进行了检测,由此鉴定KRAS突变的存在、或较之正常细胞而言的异常表达水平。成功培育肿瘤(>100mm3)后,用以GST-π作为靶标的siRNA或对照(非特异性)siRNA对小鼠进行处理,每天2次,持续2周。在必须将对照组安乐死时终止实验。
结果:利用以GST-π作为靶标的siRNA的处理可防止肿瘤增大,并导致肿瘤体积急剧减小。
将回收的肿瘤切割,利用TUNEL染色使其直观化。经过以GST-π作为靶标的siRNA处理的肿瘤呈现了明显更高的细胞凋亡水平。从肿瘤中提取RNA,并实施实时PCR,对GST-π的特异性敲低进行检测。
结果:利用以GST-π作为靶标的siRNA的处理使GST-π的体内表达急剧降低。
实施例10:本发明的GST-πsiRNA呈现出提高的血清稳定性。
在人血清中孵育GST-πsiRNA,在各时间点利用HPLS/LCMS检测残留的siRNA。GST-πsiRNA(序列号1276、1341)的有义链和反义链在血清中的半衰期(t1/2)均为约100分钟。
实施例11:本发明的GST-πsiRNA的制剂在血浆中呈现了提高的稳定性。
在血浆中孵育制剂形式的GST-πsiRNA,在各时间点检测残留的siRNA。GST-πsiRNA(序列号1276、1341)制剂在血浆中的半衰期(t1/2)明显长于100小时。
将GST-πsiRNA制成包含组合物(电离脂质:胆固醇:DOPE:DOPC:DPPE-PEG-2K)(25:30:20:20:5)的脂质体制剂。脂质体纳米粒子的Z均大小为40.0nm,并且包封有91%的siRNA。
将制剂在含有50%人血清的PBS中孵育40分钟、1.5小时、3小时、24小时、和96小时。利用基于ELISA的分析测定GST-πsiRNA的量。
实施例12:本发明的GST-πsiRNA能够使癌异种移植肿瘤在体内显著减小。以脂质体制剂的形式施予至癌异种移植肿瘤的情况下,GST-πsiRNA能够在体内发挥基因敲低效能。
观察到了GST-πsiRNA(序列号1276、1341)的肿瘤抑制效果。在体内观察到了以GST-π作为靶标的siRNA对于GST-πmRNA的剂量依赖性敲低。利用的癌异种移植模型使用了以GST-π作为靶标的siRNA。
GST-π的siRNA制剂在施予后数天内呈现了显著且出乎意料的有利的肿瘤抑制效果。在以脂质制剂的形式注射4天后,利用GST-πsiRNA的处理导致GST-πmRNA表达显著降低。以更高的剂量即4mg/kg注射,24小时后,观测到GST-πmRNA表达显著下降约40%。
以包含组合物(可离子化脂质:胆固醇:DOPE:DOPC:DPPE-PEG-2K)(25:30:20:20:5)的脂质体制剂的形式单次注射施予GST-πsiRNA,施予量为10mL/kg。
对于癌异种移植模型而言,从ATCC获得了A549细胞系。在添加有10%胎牛血清、100U/ml盘尼西林和100μg/ml链霉素的RPMI-1640中维持细胞。在接种前48小时稀释(split)细胞,从而使细胞在被收集时处于对数生长期。使用胰蛋白酶-EDTA轻度消化细胞后,从组织培养物中收集细胞。在台盼蓝的存在下,用血细胞计数器计数并测定活细胞的数量(仅计数活细胞)。在无血清RPMI培养基中重新悬浮细胞,使浓度为4x107/ml。然后将细胞悬浮液与冰上融化的BD matrigel以1:1的比例充分混合,用于注射。
小鼠为Charles River Laboratory的雌性无胸腺裸鼠(nu/nu),经过免疫妥协,6~8周龄,每组3只小鼠。
对于肿瘤模型的制备而言,使用25G注射针和针筒,在每只小鼠的右侧腹皮下接种0.1ml含有2x106个A549细胞的接种液,每只小鼠接种1剂接种液。小鼠在接种时未被麻醉。
对于肿瘤体积的测量和随机化而言,肿瘤大小的测量结果取至0.1mm。肿瘤体积利用下式算出:肿瘤体积=长x宽2/2。每周检测2次肿瘤体积。当培养的肿瘤生长至约350~600mm3时,将小鼠按照各时间点分组。按照给药方案,在同一天施予试验制剂。
对于施予量而言,在培养的肿瘤生长至约350~600mm3当天,从4℃冰箱中取出试验制剂。为了形成均质的溶液,在装入注射器之前,手动将装在瓶中的制剂颠倒数次。
对于体重而言,小鼠称重至0.1g。在接下来的数周,每周检测并记录体重2次,包括研究结束当天。
对于肿瘤的采集而言,在给药后0、24、48、72、96(可选)、168小时利用高浓度CO2杀死动物,并切除肿瘤。先称取肿瘤的湿重,然后将其分成3部分,用于分析KD、分配和生物标志物。用液氮快速冷冻试样,并保存于-80℃,直到供于处理为止。
本文描述的实施方案不是限制性的,并且本领域技术人员可以容易地理解,可以不进行过度实验而测试本文所述修饰的特定组合以鉴定具有改进的RNAi活性的核酸分子。
出于所有目的,本文特别提及的所有出版物、专利和文献通过引用整体并入本文。
应当理解,本发明不限于所述特定方法、方案、材料和试剂,因为它们可能变化。还应当理解,本文使用的术语仅用于描述特定实施方案的目的,并不意图限制本发明的范围。对于本领域技术人员将显而易见的是,在不背离本说明书的范围和宗旨的情况下,可以对本文所公开的描述进行各种替换和修改,并且这些实施方案在本说明书和所附权利要求的范围内。
必须指出,本文和所附权利要求中所使用的单数形式包括复数指称,上下文另有明确说明除外。同样地,术语“一个”(或“一种”)、“一个或更多个(一种或更多种)”和“至少一个/种”在本文可以互换使用。另外要注意,术语“包括”、“包含”、“含有”、“含”和“具有”可以互换使用,并且应该被广义、无限制地解释。
除非本文另有说明,否则本文中对数值范围的记载仅仅意在作为对单独提到落入该范围的每个单独的值的速记法,每个单独的值均被并入说明书中,如同它们被单独记载于本文中一样。对于马库什组,本领域技术人员将认识到,该描述包括个体成员以及马库什组成员的亚组。
即使没有进一步的阐述,本领域技术人员亦可基于上述描述最大程度地利用本发明。因此,所附的具体实施方案应被解释为仅是说明性的,而不是以任何方式限制本公开的其余部分。
本说明书中公开的所有特征可以以任何组合而联用。本说明书中公开的每个特征可以由服务于相同、等同或相似目的的替代特征来代替。
Claims (23)
1.核酸分子,其中:
a)所述分子具有多核苷酸有义链和多核苷酸反义链;
b)所述分子的每条链长度为15~30个核苷酸;
c)所述反义链中的15~30个核苷酸的连续区域与编码GST-π的mRNA序列互补;
d)所述有义链的至少一部分与所述反义链的至少一部分互补,并且,所述分子具有长度为15~30个核苷酸的双链区。
2.如权利要求1所述的核酸分子,其中,所述反义链为序列号1341,所述有义链为序列号1276,或是它们的经化学修饰的链。
3.如权利要求1所述的核酸分子,其中,所述反义链为序列号1305,所述有义链为序列号1240,或是它们的经化学修饰的链。
4.如权利要求1所述的核酸分子,其中,所述与编码GST-π的mRNA序列互补的反义链的15~30个核苷酸的连续区域位于所述分子的双链区。
5.如权利要求1所述的核酸分子,其中,与编码GST-π的mRNA序列互补的反义链的15~30个核苷酸的连续区域选自人GSTPl序列,并且,人GSTPl mRNA为序列号l。
6.如权利要求1所述的核酸分子,其中,所述编码GST-π的mRNA序列选自由序列号l的5’UTR第1~249位、序列号l的CDS第250~882位、序列号l的3’UTR第883~986位组成的组。
7.如权利要求1所述的核酸分子,其中,所述反义链含有选自序列号609~1215中的任一序列。
8.如权利要求1所述的核酸分子,其中,所述反义链含有选自序列号1281~1345中的任一序列。
9.如权利要求1所述的核酸分子,其中,所述分子由选自由序列号1240和1305、序列号1265和1330、序列号1267和1332、序列号1269和1334、及序列号1276和341组成的组中的反义链和有义链的对构成。
10.如权利要求1所述的核酸分子,其中,所述分子的每条链长度为18~22个核苷酸。
11.如权利要求1所述的核酸分子,其中,所述双链区的长度为19个核苷酸。
12.如权利要求1所述的核酸分子,其中,所述多核苷酸有义链和所述多核苷酸反义链以单链形式连接,并且形成一端以环连接的双链区。
13.如权利要求1所述的核酸分子,其中,所述分子具有平末端。
14.如权利要求1所述的核酸分子,其中,所述分子具有一个或多个3’悬挂。
15.如权利要求1所述的核酸分子,其中,所述分子为具有基因沉默活性的RNAi分子。
16.如权利要求1所述的核酸分子,其中,所述分子为具有基因沉默活性的dsRNA、siRNA、micro-RNA、或shRNA。
17.如权利要求1所述的核酸分子,其中,所述分子具有GST-π表达抑制活性。
18.如权利要求1所述的核酸分子,其中,所述分子的GST-π敲低的IC50小于100pM。
19.组合物,其含有一种或多种权利要求1~18中任一项所述的核酸分子、和药学上允许的担体。
20.如权利要求19所述的组合物,其中,所述担体为脂质分子或脂质体。
21.治疗GST-π表达相关疾病的方法,其中,所述方法包括向有需要的受试者施予权利要求19所述的组合物。
22.如权利要求21所述的方法,其中,所述疾病为恶性肿瘤。
23.如权利要求22所述的方法,其中,所述恶性肿呈现为选自由下述疾病组成的组中的疾病:与GST-π表达相关的癌症、由表达KRAS突变的细胞导致的癌症、肉瘤、纤维肉瘤、恶性纤维性组织细胞瘤、脂肪肉瘤、横纹肌肉瘤、平滑肌肉瘤、血管肉瘤、卡波西肉瘤、淋巴管肉瘤、滑膜肉瘤、软骨肉瘤、骨肉瘤、癌、脑瘤、头颈癌、乳腺癌、肺癌、食道癌、胃癌、十二指肠癌、结肠直肠癌、结肠癌、肝癌、胰腺癌、胆囊癌、胆管癌、肾癌、尿道癌、膀胱癌、前列腺癌、睾丸癌、阴茎癌、子宫癌、卵巢癌、皮肤癌、骨癌、白血病、恶性淋巴瘤、上皮性恶性肿瘤、和非上皮性恶性肿瘤。
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2014266198 | 2014-12-26 | ||
JP2014-266198 | 2014-12-26 | ||
US201562184239P | 2015-06-24 | 2015-06-24 | |
US62/184,239 | 2015-06-24 | ||
US201562266664P | 2015-12-13 | 2015-12-13 | |
US62/266,664 | 2015-12-13 | ||
PCT/US2015/067561 WO2016106406A2 (en) | 2014-12-26 | 2015-12-28 | Rna agents for gst-pi gene modulation |
Publications (1)
Publication Number | Publication Date |
---|---|
CN108064153A true CN108064153A (zh) | 2018-05-22 |
Family
ID=56151549
Family Applications (7)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201580071233.3A Active CN107108686B (zh) | 2014-12-26 | 2015-12-28 | 用于恶性肿瘤的rna干扰组合物和方法 |
CN201580071081.7A Active CN108064155B (zh) | 2014-12-26 | 2015-12-28 | 用于p21基因调节的rna剂 |
CN201580071234.8A Pending CN108024961A (zh) | 2014-12-26 | 2015-12-28 | 使用靶向Hsp47和p21的RNAi分子的恶性肿瘤治疗组合物和方法 |
CN201580071191.3A Active CN107106591B (zh) | 2014-12-26 | 2015-12-28 | 用于gst-pi基因调节的rna干扰剂 |
CN201580071209.XA Active CN107106592B (zh) | 2014-12-26 | 2015-12-28 | 用于p21基因调节的rna干扰剂 |
CN201580071210.2A Pending CN108064153A (zh) | 2014-12-26 | 2015-12-28 | 用于调节gst-pi基因的rna剂 |
CN201580071232.9A Pending CN107106564A (zh) | 2014-12-26 | 2015-12-28 | 用于治疗与kras突变相关的恶性肿瘤的方法和组合物 |
Family Applications Before (5)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201580071233.3A Active CN107108686B (zh) | 2014-12-26 | 2015-12-28 | 用于恶性肿瘤的rna干扰组合物和方法 |
CN201580071081.7A Active CN108064155B (zh) | 2014-12-26 | 2015-12-28 | 用于p21基因调节的rna剂 |
CN201580071234.8A Pending CN108024961A (zh) | 2014-12-26 | 2015-12-28 | 使用靶向Hsp47和p21的RNAi分子的恶性肿瘤治疗组合物和方法 |
CN201580071191.3A Active CN107106591B (zh) | 2014-12-26 | 2015-12-28 | 用于gst-pi基因调节的rna干扰剂 |
CN201580071209.XA Active CN107106592B (zh) | 2014-12-26 | 2015-12-28 | 用于p21基因调节的rna干扰剂 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201580071232.9A Pending CN107106564A (zh) | 2014-12-26 | 2015-12-28 | 用于治疗与kras突变相关的恶性肿瘤的方法和组合物 |
Country Status (10)
Country | Link |
---|---|
US (14) | US10264976B2 (zh) |
EP (9) | EP3236969A4 (zh) |
JP (9) | JP6457645B2 (zh) |
KR (4) | KR20170098929A (zh) |
CN (7) | CN107108686B (zh) |
AU (4) | AU2015369596A1 (zh) |
BR (2) | BR112017013599A2 (zh) |
CA (4) | CA2971881C (zh) |
RU (4) | RU2719185C2 (zh) |
WO (7) | WO2016106405A1 (zh) |
Families Citing this family (34)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10792299B2 (en) | 2014-12-26 | 2020-10-06 | Nitto Denko Corporation | Methods and compositions for treating malignant tumors associated with kras mutation |
ES2789049T3 (es) * | 2014-12-26 | 2020-10-23 | Nitto Denko Corp | Agentes de interferencia de ARN para modulación génica de GST-pi |
US11045488B2 (en) | 2014-12-26 | 2021-06-29 | Nitto Denko Corporation | RNA interference agents for GST-π gene modulation |
US10264976B2 (en) | 2014-12-26 | 2019-04-23 | The University Of Akron | Biocompatible flavonoid compounds for organelle and cell imaging |
US20180002702A1 (en) * | 2014-12-26 | 2018-01-04 | Nitto Denko Corporation | Methods and compositions for treating malignant tumors associated with kras mutation |
WO2016210190A1 (en) * | 2015-06-24 | 2016-12-29 | Nitto Denko Corporation | Ionizable compounds and compositions and uses thereof |
KR20180086260A (ko) | 2015-12-13 | 2018-07-30 | 닛토덴코 가부시키가이샤 | 높은 활성 및 감소한 오프 타겟을 위한 sirna 구조 |
CA3023487A1 (en) | 2016-05-10 | 2017-11-16 | National University Corporation Tokyo Medical And Dental University | Expression inhibitor of inflammation promoting factor, screening method for active ingredient thereof, expression cassette useful for said method, diagnostic agent and diagnosis method |
WO2018151840A2 (en) * | 2017-02-16 | 2018-08-23 | Nitto Denko Corporation | Methods and compositions for treating malignant tumors |
AU2018359904B2 (en) * | 2017-11-06 | 2023-02-23 | Nitto Denko Corporation | Fusogenic compounds for delivery of biologically active molecules |
CA3082006A1 (en) * | 2017-11-09 | 2019-05-16 | National University Corporation Tokyo Medical And Dental University | Inhibitor of the expression of cancer-promoting factors, screening method for active ingredient thereof, expression cassette useful in said method, diagnostic drug, and diagnosticmethod |
CN109777798A (zh) * | 2017-11-13 | 2019-05-21 | 深圳华大生命科学研究院 | 一种基于CRISPR技术治疗KRAS突变恶性肿瘤的sgRNA及其应用 |
JP6952594B2 (ja) * | 2017-12-15 | 2021-10-20 | 洋司郎 新津 | 細胞増殖抑制剤及びそれを含むがんの治療若しくは予防用医薬組成物 |
CN108486011B (zh) * | 2018-03-27 | 2020-05-05 | 山东大学 | 一种三联苯化合物、制备方法及其应用 |
JP7432929B2 (ja) * | 2018-05-31 | 2024-02-19 | コリア ユニバーシティ リサーチ アンド ビジネス ファウンデーション | マイクロrnaの非正規標的を抑制するrna干渉誘導核酸およびその用途 |
WO2020009189A1 (ja) * | 2018-07-05 | 2020-01-09 | 洋司郎 新津 | Braf阻害剤によるがん細胞の逆説的増殖を抑制する薬剤 |
US20210260095A1 (en) * | 2018-08-22 | 2021-08-26 | Nitto Denko Corporation | Medicine using hsp47 inhibitor to enhance sensitivity to chemotherapeutic agent |
CN112739381A (zh) * | 2018-08-22 | 2021-04-30 | 日东电工株式会社 | 使用了hsp47的抑制物质的癌转移抑制 |
JP2022512077A (ja) * | 2018-11-16 | 2022-02-02 | 日東電工株式会社 | Rna干渉送達製剤及び悪性腫瘍のための方法 |
CN113038956A (zh) * | 2018-12-05 | 2021-06-25 | 日东电工株式会社 | 癌处置用RNAi分子 |
EP3909588A4 (en) | 2019-01-10 | 2023-01-04 | Osaka University | IMMUNO-STIMULATING COMPOSITION |
CN113677373A (zh) * | 2019-03-28 | 2021-11-19 | 日东电工株式会社 | RNAi分子 |
JP2019116507A (ja) * | 2019-04-25 | 2019-07-18 | 有限会社オービット | Hsp47の発現促進剤、脱毛抑制方法、Hsp47の発現促進剤の製造方法及び飲食物の製造方法 |
EP3965781A2 (en) * | 2019-07-02 | 2022-03-16 | Argonaute Rna Limited | Apolipoprotein b antagonist |
US20220267778A1 (en) * | 2019-07-30 | 2022-08-25 | Shionogi & Co., Ltd. | Nucleic acid drug targeting murf1 |
CN112280800B (zh) * | 2020-10-19 | 2022-06-07 | 上海市东方医院(同济大学附属东方医院) | 一种构建体及其在制备动物衰老细胞示踪和衰老细胞清除药物中的应用 |
EP4267741A2 (en) * | 2020-12-28 | 2023-11-01 | 1E Therapeutics, Ltd. | P21 mrna target areas for silencing |
KR20230133859A (ko) | 2020-12-28 | 2023-09-19 | 1이 테라퓨틱스 엘티디. | p21 mRNA 표적화 DNA자임 |
KR20230101285A (ko) * | 2021-12-29 | 2023-07-06 | 의료법인 명지의료재단 | K-ras 특이적 활성화 T 세포를 포함하는 유방암의 예방 및 치료용 약제학적 조성물 및 이의 제조방법 |
KR20230101287A (ko) * | 2021-12-29 | 2023-07-06 | 의료법인 명지의료재단 | K-ras 특이적 활성화 T 세포를 포함하는 흑색종의 예방 및 치료용 약제학적 조성물 및 이의 제조방법 |
KR20230101286A (ko) * | 2021-12-29 | 2023-07-06 | 의료법인 명지의료재단 | K-ras 특이적 활성화 T 세포를 포함하는 폐 선암종의 예방 및 치료용 약제학적 조성물 및 이의 제조방법 |
KR20230101283A (ko) * | 2021-12-29 | 2023-07-06 | 의료법인 명지의료재단 | K-ras 특이적 활성화 T 세포를 포함하는 폐 유두상 선암종의 예방 및 치료용 약제학적 조성물 및 이의 제조방법 |
KR20230101284A (ko) * | 2021-12-29 | 2023-07-06 | 의료법인 명지의료재단 | K-ras 특이적 활성화 T 세포를 포함하는 대장암의 예방 및 치료용 약제학적 조성물 및 이의 제조방법 |
WO2023210713A1 (ja) * | 2022-04-27 | 2023-11-02 | 国立大学法人京都大学 | 心外膜細胞再生促進剤および心外膜細胞の再生促進方法 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007061922A2 (en) * | 2005-11-17 | 2007-05-31 | Children's Medical Center Corporation | Methods to predict and prevent resistance to taxoid compounds |
CN103619355A (zh) * | 2011-06-21 | 2014-03-05 | 日东电工株式会社 | 细胞凋亡诱导剂 |
Family Cites Families (85)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU630076B2 (en) | 1987-09-21 | 1992-10-22 | Gen-Probe Incorporated | Non-nucleotide linking reagents for nucleotide probes |
US5204241A (en) | 1990-10-22 | 1993-04-20 | Oxi-Gene Inc. | Glutathione-S-transferase mu as a measure of drug resistance |
US5786336A (en) | 1991-04-29 | 1998-07-28 | Terrapin Technologies, Inc. | Target-selective protocols based on mimics |
US5658780A (en) | 1992-12-07 | 1997-08-19 | Ribozyme Pharmaceuticals, Inc. | Rel a targeted ribozymes |
ES2186690T3 (es) | 1993-09-02 | 2003-05-16 | Ribozyme Pharm Inc | Acido nucleico enzimatico que contiene no-nucleotidos. |
WO1995011910A1 (en) | 1993-10-27 | 1995-05-04 | Ribozyme Pharmaceuticals, Inc. | 2'-amido and 2'-peptido modified oligonucleotides |
AU715524B2 (en) | 1995-06-07 | 2000-02-03 | Telik, Inc. | Metabolic effects of certain glutathione analogs |
US5968737A (en) | 1996-11-12 | 1999-10-19 | The University Of Mississippi | Method of identifying inhibitors of glutathione S-transferase (GST) gene expression |
US6506559B1 (en) | 1997-12-23 | 2003-01-14 | Carnegie Institute Of Washington | Genetic inhibition by double-stranded RNA |
CA2328731A1 (en) | 1998-04-16 | 1999-10-28 | Atsushi Imaizumi | Glutathione derivatives and dosage forms thereof |
GB9827152D0 (en) | 1998-07-03 | 1999-02-03 | Devgen Nv | Characterisation of gene function using double stranded rna inhibition |
DE19956568A1 (de) | 1999-01-30 | 2000-08-17 | Roland Kreutzer | Verfahren und Medikament zur Hemmung der Expression eines vorgegebenen Gens |
WO2001029058A1 (en) | 1999-10-15 | 2001-04-26 | University Of Massachusetts | Rna interference pathway genes as tools for targeted genetic interference |
GB9927444D0 (en) | 1999-11-19 | 2000-01-19 | Cancer Res Campaign Tech | Inhibiting gene expression |
US20070083945A1 (en) | 2000-03-10 | 2007-04-12 | Byrum Joseph R | Nucleic acid molecules and other molecules associated with plants |
US20030144236A1 (en) | 2000-03-29 | 2003-07-31 | Weiss Robert H | Novel specific inhibitor of the cyclin kinase inhibitor p21 (wafl/cip1) |
EP1305450A2 (en) * | 2000-07-28 | 2003-05-02 | Compugen Inc. | Oligonucleotide library for detecting rna transcripts and splice variants that populate a transcriptome |
PL218876B1 (pl) | 2000-12-01 | 2015-02-27 | Europaisches Lab Für Molekularbiologie Embl | Wyizolowana cząsteczka dwuniciowego RNA, sposób wytwarzania cząsteczki dwuniciowego RNA, zastosowanie cząsteczki dwuniciowego RNA do wytwarzania leku do modulowania działania genu związanego z patogenem, genu związanego z nowotworem, oraz genu związanego z chorobą autoimmunologiczną, sposób in vitro kierowania miejscowo specyficznymi interferencjami RNA w komórce, zastosowanie sposobu in vitro do ustalania działania genu w komórce oraz do modulowania działania genu w komórce, środek farmaceutyczny, komórka eukariotyczna transfekowana cząsteczką RNA lub cząsteczką DNA kodującą tę cząsteczkę RNA, oraz zastosowanie komórki eukariotycznej w procedurach analitycznych oraz w procedurach preparatywnych |
AU2004266311B2 (en) * | 2001-05-18 | 2009-07-23 | Sirna Therapeutics, Inc. | RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA) |
US20030099974A1 (en) | 2001-07-18 | 2003-05-29 | Millennium Pharmaceuticals, Inc. | Novel genes, compositions, kits and methods for identification, assessment, prevention, and therapy of breast cancer |
US20040142325A1 (en) * | 2001-09-14 | 2004-07-22 | Liat Mintz | Methods and systems for annotating biomolecular sequences |
US20040029275A1 (en) * | 2002-08-10 | 2004-02-12 | David Brown | Methods and compositions for reducing target gene expression using cocktails of siRNAs or constructs expressing siRNAs |
PT2284266E (pt) * | 2002-11-14 | 2013-12-17 | Thermo Fisher Scient Biosciences Inc | Siarn contra tp53 |
US20040219600A1 (en) | 2002-12-13 | 2004-11-04 | Williams Robert Wood | Method for determining sensitivity to environmental toxins and susceptibility to parkinson's disease |
AU2003224132A1 (en) | 2003-04-24 | 2004-11-19 | Galapagos Genomics N.V. | Effective sirna knock-down constructs |
US20050142596A1 (en) * | 2003-11-14 | 2005-06-30 | Krolewski Andrzej S. | Methods of diagnosing renal and cardiovascular disease |
JP5243789B2 (ja) | 2004-03-15 | 2013-07-24 | シティ・オブ・ホープ | 二本鎖rnaによる遺伝子発現の特異的阻害のための方法及び組成物 |
EP2322650A1 (en) | 2004-05-14 | 2011-05-18 | Rosetta Genomics Ltd | MicroRNAs and uses thereof |
MX2007002294A (es) * | 2004-08-26 | 2007-10-19 | Engeneic Molecular Delivery Pty Ltd | Suministro de acidos nucleicos funcionales a celulas mamiferas via minicelulas intactas, derivadas bacterialmente. |
US9393315B2 (en) * | 2011-06-08 | 2016-07-19 | Nitto Denko Corporation | Compounds for targeting drug delivery and enhancing siRNA activity |
ES2443229T3 (es) | 2004-12-22 | 2014-02-18 | Nitto Denko Corporation | Portador de fármacos y kit portador de fármacos para la inhibición de la fibrosis |
EP1874793A4 (en) | 2005-04-15 | 2008-12-24 | Univ Texas | DISTRIBUTION OF SIRNA BY NEUTRAL LIPID COMPOSITIONS |
US20070134687A1 (en) * | 2005-09-12 | 2007-06-14 | Aurelium Biopharma Inc. | Focused microarray and methods of diagnosing cancer |
ES2324128A1 (es) | 2005-09-29 | 2009-07-30 | Proyecto De Biomedicina Cima, S.L. | Metodo para el diagnostico de carcinoma hepatocelular mediante el empleo de marcadores moleculares. |
US20090220956A1 (en) | 2005-10-25 | 2009-09-03 | Dimitry Serge Antoine Nuyten | Prediction of Local Recurrence of Breast Cancer |
US20070197460A1 (en) | 2005-11-01 | 2007-08-23 | Alnylam Pharmaceuticals, Inc. | Rnai inhibition of influenza virus replication |
RU2448974C2 (ru) * | 2005-11-01 | 2012-04-27 | Элнилэм Фармасьютикалз, Инк. | РНКи-ИНГИБИРОВАНИЕ РЕПЛИКАЦИИ ВИРУСА ГРИППА |
JP5336853B2 (ja) * | 2005-11-02 | 2013-11-06 | プロチバ バイオセラピューティクス インコーポレイティッド | 修飾siRNA分子およびその使用法 |
US7729737B2 (en) | 2005-11-22 | 2010-06-01 | Isense Corporation | Method and apparatus for background current arrangements for a biosensor |
PL1957044T3 (pl) | 2005-12-01 | 2013-11-29 | Pronai Therapeutics Inc | Amfoteryczny preparat liposomowy |
US9572886B2 (en) * | 2005-12-22 | 2017-02-21 | Nitto Denko Corporation | Agent for treating myelofibrosis |
JP5342834B2 (ja) * | 2008-09-05 | 2013-11-13 | 日東電工株式会社 | 骨髄線維症処置剤 |
WO2007130604A2 (en) * | 2006-05-04 | 2007-11-15 | Baylor Research Institute | Anti-tumor activity of an oncolytic adenovirus-delivered oncogene sirna |
KR101036126B1 (ko) * | 2006-05-11 | 2011-05-23 | 알닐람 파마슈티칼스 인코포레이티드 | Pcsk9 유전자의 발현을 억제하기 위한 조성물 및 방법 |
WO2008109432A2 (en) | 2007-03-02 | 2008-09-12 | The Board Of Regents Of The University Of Texas System | Therapeutic targeting of interleukins using sirna in neutral liposomes |
TWI407971B (zh) | 2007-03-30 | 2013-09-11 | Nitto Denko Corp | Cancer cells and tumor-related fibroblasts |
WO2008124634A1 (en) | 2007-04-04 | 2008-10-16 | Massachusetts Institute Of Technology | Polymer-encapsulated reverse micelles |
EP2164966A2 (en) | 2007-07-05 | 2010-03-24 | Novartis Ag | Dsrna for treating viral infection |
ES2873350T3 (es) | 2007-08-27 | 2021-11-03 | 1Globe Health Inst Llc | Composiciones de ARN interferente asimétrico y usos de las mismas |
WO2009033284A1 (en) | 2007-09-14 | 2009-03-19 | Mcmaster University | Inhibitors of collagen biosynthesis as anti-tumor agents |
JP2011513342A (ja) * | 2008-03-06 | 2011-04-28 | ロッタファルム・ソシエタ・ペル・アチオニ | 関節炎、癌および関連疼痛の治療のための2−アリールおよび2−ヘテロアリール4h−1−ベンゾピラン−4−オン−6−アミジノ誘導体 |
CA2732229C (en) | 2008-07-25 | 2023-10-17 | Alnylam Pharmaceuticals, Inc. | Enhancement of sirna silencing activity using universal bases or mismatches in the sense strand |
CN102159247A (zh) * | 2008-07-30 | 2011-08-17 | 日东电工株式会社 | 药物载体 |
EP2496700B1 (en) * | 2009-11-04 | 2017-03-01 | The University Of British Columbia | Nucleic acid-containing lipid particles and related methods |
DK2509991T3 (en) * | 2009-12-09 | 2015-12-21 | Nitto Denko Corp | MODULATION OF HSP47 EXPRESSION |
AU2011220749B2 (en) | 2010-02-24 | 2017-02-02 | Bodysync, Inc. | Methods for determining gene-nutrient interactions |
US20130004494A1 (en) | 2010-03-12 | 2013-01-03 | Ellen Heber-Katz | Inhibition of P21 and Use Thereof for Inducing Tissue Regeneration |
US8372819B2 (en) * | 2010-04-11 | 2013-02-12 | Salk Institute For Biological Studies | Methods and compositions for targeting skip |
US8828944B2 (en) | 2010-04-22 | 2014-09-09 | Institut Gustave Roussy | Compounds and uses thereof to induce an immunogenic cancer cell death in a subject |
EP2566955A4 (en) | 2010-05-06 | 2014-10-29 | Stem Cell Medicine Ltd | STEM CELL BANK FOR PERSONALIZED MEDICINE |
JP5950428B2 (ja) * | 2010-08-05 | 2016-07-13 | 日東電工株式会社 | 線維化組織から正常組織を再生するための組成物 |
CA2810825A1 (en) * | 2010-09-30 | 2012-04-05 | Yoshiro Niitsu | Modulation of timp1 and timp2 expression |
US9339513B2 (en) | 2010-11-09 | 2016-05-17 | Alnylam Pharmaceuticals, Inc. | Lipid formulated compositions and methods for inhibiting expression of Eg5 and VEGF genes |
US9011903B2 (en) * | 2011-06-08 | 2015-04-21 | Nitto Denko Corporation | Cationic lipids for therapeutic agent delivery formulations |
DK2998289T3 (da) * | 2011-06-08 | 2019-09-16 | Nitto Denko Corp | Forbindelser til at målrette lægemiddellevering og fremme sirna-aktivitet |
TWI658830B (zh) | 2011-06-08 | 2019-05-11 | 日東電工股份有限公司 | Hsp47表現調控強化用類視色素脂質體 |
US20140351961A1 (en) * | 2011-08-31 | 2014-11-27 | Alexzander A. Asea | Compositions and methods for treatment of metastatic cancer |
US9579338B2 (en) | 2011-11-04 | 2017-02-28 | Nitto Denko Corporation | Method of producing lipid nanoparticles for drug delivery |
US9631192B2 (en) | 2011-11-17 | 2017-04-25 | The United States Of America, As Represented By The Secretary, Department Of Health & Human Services | Auto-recognizing therapeutic RNA/DNA chimeric nanoparticles (NP) |
EP2849762A4 (en) * | 2012-05-16 | 2016-02-24 | Aadigen Llc | MULTI-TARGET MODULATION FOR TREATING FIBROSIS AND INFLAMMATORY DISORDERS |
US20140134158A1 (en) | 2012-05-22 | 2014-05-15 | Alberto Bardelli | Kras mutations and resistance to anti-egfr treatment |
KR102102801B1 (ko) | 2012-06-08 | 2020-04-22 | 닛토덴코 가부시키가이샤 | 치료제 전달 제형용 지질 |
WO2013192364A1 (en) | 2012-06-22 | 2013-12-27 | The University Of Vermont And State Agricultural College | Treatments of oxidative stress conditions |
CN104684584A (zh) * | 2012-07-02 | 2015-06-03 | 费卜卢斯坦丁公司 | Gpbp-1的抑制及其治疗用途 |
WO2014022739A2 (en) | 2012-08-03 | 2014-02-06 | Alnylam Pharmaceuticals, Inc. | Modified rnai agents |
JP6340162B2 (ja) * | 2012-12-20 | 2018-06-06 | 日東電工株式会社 | アポトーシス誘導剤 |
JP6352950B2 (ja) | 2013-03-08 | 2018-07-04 | ノバルティス アーゲー | 活性薬物の送達のための脂質と脂質組成物 |
CN103695421B (zh) * | 2013-12-09 | 2016-06-15 | 浙江大学 | 一种特异抑制p21基因表达的siRNA及其应用 |
US10792299B2 (en) * | 2014-12-26 | 2020-10-06 | Nitto Denko Corporation | Methods and compositions for treating malignant tumors associated with kras mutation |
US10264976B2 (en) | 2014-12-26 | 2019-04-23 | The University Of Akron | Biocompatible flavonoid compounds for organelle and cell imaging |
US20180002702A1 (en) | 2014-12-26 | 2018-01-04 | Nitto Denko Corporation | Methods and compositions for treating malignant tumors associated with kras mutation |
CN113559267B (zh) * | 2014-12-26 | 2024-01-12 | 日东电工株式会社 | 细胞死亡诱导试剂、细胞增殖抑制试剂及用于治疗由细胞增殖异常导致的疾病的医药组合物 |
US20160187319A1 (en) | 2014-12-26 | 2016-06-30 | Nitto Denko Corporation | Cell death-inducing agent, cell growth-inhibiting agent, and pharmaceutical composition for treatment of disease caused by abnormal cell growth |
KR20180086260A (ko) * | 2015-12-13 | 2018-07-30 | 닛토덴코 가부시키가이샤 | 높은 활성 및 감소한 오프 타겟을 위한 sirna 구조 |
JP6899201B2 (ja) | 2016-06-23 | 2021-07-07 | 日東電工株式会社 | 細胞死誘導剤、細胞増殖抑制剤及び細胞の増殖異常に起因する疾患の治療用医薬組成物 |
-
2015
- 2015-11-06 US US14/934,665 patent/US10264976B2/en not_active Expired - Fee Related
- 2015-12-28 BR BR112017013599-0A patent/BR112017013599A2/pt not_active Application Discontinuation
- 2015-12-28 JP JP2017534297A patent/JP6457645B2/ja active Active
- 2015-12-28 CN CN201580071233.3A patent/CN107108686B/zh active Active
- 2015-12-28 AU AU2015369596A patent/AU2015369596A1/en not_active Abandoned
- 2015-12-28 RU RU2017126613A patent/RU2719185C2/ru active
- 2015-12-28 JP JP2017534286A patent/JP6793649B2/ja active Active
- 2015-12-28 CA CA2971881A patent/CA2971881C/en active Active
- 2015-12-28 WO PCT/US2015/067560 patent/WO2016106405A1/en active Application Filing
- 2015-12-28 JP JP2017534279A patent/JP6865169B2/ja active Active
- 2015-12-28 EP EP15874365.8A patent/EP3236969A4/en not_active Withdrawn
- 2015-12-28 EP EP15874361.7A patent/EP3236974B9/en active Active
- 2015-12-28 BR BR112017013597-3A patent/BR112017013597B1/pt active IP Right Grant
- 2015-12-28 JP JP2017534275A patent/JP2018512373A/ja active Pending
- 2015-12-28 CA CA2972270A patent/CA2972270A1/en not_active Abandoned
- 2015-12-28 JP JP2017534274A patent/JP2018513104A/ja active Pending
- 2015-12-28 CN CN201580071081.7A patent/CN108064155B/zh active Active
- 2015-12-28 WO PCT/US2015/067561 patent/WO2016106406A2/en active Application Filing
- 2015-12-28 US US14/979,574 patent/US10047111B2/en not_active Ceased
- 2015-12-28 AU AU2015369592A patent/AU2015369592B2/en active Active
- 2015-12-28 AU AU2015369595A patent/AU2015369595A1/en not_active Abandoned
- 2015-12-28 EP EP15874363.3A patent/EP3236976B1/en active Active
- 2015-12-28 WO PCT/US2015/067559 patent/WO2016106404A2/en active Application Filing
- 2015-12-28 EP EP15874367.4A patent/EP3236973A4/en not_active Withdrawn
- 2015-12-28 CN CN201580071234.8A patent/CN108024961A/zh active Pending
- 2015-12-28 KR KR1020177020894A patent/KR20170098929A/ko unknown
- 2015-12-28 EP EP15874362.5A patent/EP3236975A4/en not_active Ceased
- 2015-12-28 CN CN201580071191.3A patent/CN107106591B/zh active Active
- 2015-12-28 EP EP15874366.6A patent/EP3240796B1/en active Active
- 2015-12-28 US US14/979,573 patent/US9580710B2/en not_active Ceased
- 2015-12-28 JP JP2017534287A patent/JP2018513669A/ja active Pending
- 2015-12-28 RU RU2017126610A patent/RU2756253C2/ru active
- 2015-12-28 CN CN201580071209.XA patent/CN107106592B/zh active Active
- 2015-12-28 EP EP15874364.1A patent/EP3236945B1/en active Active
- 2015-12-28 KR KR1020177020901A patent/KR20170093988A/ko unknown
- 2015-12-28 RU RU2017126598A patent/RU2017126598A/ru not_active Application Discontinuation
- 2015-12-28 WO PCT/US2015/067556 patent/WO2016106401A2/en active Application Filing
- 2015-12-28 AU AU2015369598A patent/AU2015369598A1/en not_active Abandoned
- 2015-12-28 EP EP20156350.9A patent/EP3683309A1/en active Pending
- 2015-12-28 WO PCT/US2015/067558 patent/WO2016106403A2/en active Application Filing
- 2015-12-28 US US14/979,568 patent/US10405749B2/en active Active
- 2015-12-28 CA CA2972265A patent/CA2972265A1/en not_active Abandoned
- 2015-12-28 JP JP2017534294A patent/JP6730285B2/ja active Active
- 2015-12-28 WO PCT/US2015/067553 patent/WO2016106400A2/en active Application Filing
- 2015-12-28 WO PCT/US2015/067557 patent/WO2016106402A1/en active Application Filing
- 2015-12-28 CA CA2972268A patent/CA2972268A1/en not_active Abandoned
- 2015-12-28 US US14/979,571 patent/US9695206B2/en active Active
- 2015-12-28 EP EP20207751.7A patent/EP3798308A1/en active Pending
- 2015-12-28 US US14/979,567 patent/US10047110B2/en active Active
- 2015-12-28 CN CN201580071210.2A patent/CN108064153A/zh active Pending
- 2015-12-28 RU RU2017126601A patent/RU2017126601A/ru unknown
- 2015-12-28 US US14/979,566 patent/US9771582B2/en not_active Ceased
- 2015-12-28 KR KR1020177020898A patent/KR102527430B1/ko active IP Right Grant
- 2015-12-28 KR KR1020177020900A patent/KR20170096199A/ko unknown
- 2015-12-28 US US14/979,577 patent/US20160215286A1/en not_active Abandoned
- 2015-12-28 CN CN201580071232.9A patent/CN107106564A/zh active Pending
-
2017
- 2017-04-17 US US15/489,650 patent/US20170218365A1/en not_active Abandoned
- 2017-06-27 US US15/634,771 patent/US10023597B2/en active Active
-
2019
- 2019-09-19 US US16/576,102 patent/USRE48887E1/en active Active
-
2020
- 2020-02-28 US US16/804,696 patent/USRE49229E1/en active Active
- 2020-08-11 US US16/990,860 patent/USRE49431E1/en active Active
-
2021
- 2021-11-11 JP JP2021183788A patent/JP7307137B2/ja active Active
- 2021-11-12 US US17/454,757 patent/US20220087531A1/en not_active Abandoned
-
2023
- 2023-06-29 JP JP2023106970A patent/JP2023123743A/ja active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007061922A2 (en) * | 2005-11-17 | 2007-05-31 | Children's Medical Center Corporation | Methods to predict and prevent resistance to taxoid compounds |
CN103619355A (zh) * | 2011-06-21 | 2014-03-05 | 日东电工株式会社 | 细胞凋亡诱导剂 |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108064153A (zh) | 用于调节gst-pi基因的rna剂 | |
US11926831B2 (en) | SiRNA structures for high activity and reduced off target | |
JP6978561B2 (ja) | GST−π遺伝子を調節するためのRNA干渉剤 | |
US20220047619A1 (en) | Rna interference agents for gst-pi gene modulation | |
US11045488B2 (en) | RNA interference agents for GST-π gene modulation | |
TWI715594B (zh) | 用於麩胱甘肽S轉移酶Pi(GST-π)基因調控之RNA干擾劑 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
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: 20180522 |