JP4782927B2 - 代理細胞ベースシステム及びｃ型肝炎ウイルスｎｓ３プロテアーゼの活性を評価する方法 - Google Patents
代理細胞ベースシステム及びｃ型肝炎ウイルスｎｓ３プロテアーゼの活性を評価する方法 Download PDFInfo
- Publication number
- JP4782927B2 JP4782927B2 JP2000615652A JP2000615652A JP4782927B2 JP 4782927 B2 JP4782927 B2 JP 4782927B2 JP 2000615652 A JP2000615652 A JP 2000615652A JP 2000615652 A JP2000615652 A JP 2000615652A JP 4782927 B2 JP4782927 B2 JP 4782927B2
- Authority
- JP
- Japan
- Prior art keywords
- protease
- hcv
- activity
- expression
- protein
- 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.)
- Expired - Fee Related
Links
- 230000000694 effects Effects 0.000 title claims abstract description 111
- 238000000034 method Methods 0.000 title claims description 57
- 108700012707 hepatitis C virus NS3 Proteins 0.000 title claims description 4
- 238000003776 cleavage reaction Methods 0.000 claims abstract description 121
- 230000007017 scission Effects 0.000 claims abstract description 120
- 101800001838 Serine protease/helicase NS3 Proteins 0.000 claims abstract description 94
- 108091023040 Transcription factor Proteins 0.000 claims abstract description 84
- 102000040945 Transcription factor Human genes 0.000 claims abstract description 80
- 108010076039 Polyproteins Proteins 0.000 claims abstract description 64
- 230000014509 gene expression Effects 0.000 claims abstract description 61
- 108700008625 Reporter Genes Proteins 0.000 claims abstract description 35
- 239000012634 fragment Substances 0.000 claims abstract description 34
- 101800001554 RNA-directed RNA polymerase Proteins 0.000 claims abstract description 30
- 150000001875 compounds Chemical class 0.000 claims abstract description 23
- 101800001014 Non-structural protein 5A Proteins 0.000 claims abstract 4
- 210000004027 cell Anatomy 0.000 claims description 108
- 108090000623 proteins and genes Proteins 0.000 claims description 105
- 108020004414 DNA Proteins 0.000 claims description 54
- 108091005804 Peptidases Proteins 0.000 claims description 37
- 239000004365 Protease Substances 0.000 claims description 37
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 32
- 102000053602 DNA Human genes 0.000 claims description 27
- 210000004962 mammalian cell Anatomy 0.000 claims description 22
- 238000001890 transfection Methods 0.000 claims description 20
- 108020004511 Recombinant DNA Proteins 0.000 claims description 16
- 210000003494 hepatocyte Anatomy 0.000 claims description 16
- 239000003112 inhibitor Substances 0.000 claims description 16
- 239000012528 membrane Substances 0.000 claims description 13
- 150000007523 nucleic acids Chemical class 0.000 claims description 13
- 230000004927 fusion Effects 0.000 claims description 11
- 102000039446 nucleic acids Human genes 0.000 claims description 10
- 108020004707 nucleic acids Proteins 0.000 claims description 10
- 239000013613 expression plasmid Substances 0.000 claims description 9
- 241000714474 Rous sarcoma virus Species 0.000 claims description 6
- 230000005945 translocation Effects 0.000 claims description 6
- 210000002472 endoplasmic reticulum Anatomy 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 210000003292 kidney cell Anatomy 0.000 claims description 4
- 238000013519 translation Methods 0.000 claims description 4
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 claims description 3
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims description 3
- 230000000977 initiatory effect Effects 0.000 claims description 3
- 238000011156 evaluation Methods 0.000 claims description 2
- 210000002950 fibroblast Anatomy 0.000 claims description 2
- 210000004698 lymphocyte Anatomy 0.000 claims description 2
- 230000001939 inductive effect Effects 0.000 claims 1
- 238000003556 assay Methods 0.000 abstract description 37
- 101800001019 Non-structural protein 4B Proteins 0.000 abstract description 29
- 230000003612 virological effect Effects 0.000 abstract description 12
- 238000012216 screening Methods 0.000 abstract description 11
- 230000000120 cytopathologic effect Effects 0.000 abstract description 10
- 230000027455 binding Effects 0.000 abstract description 8
- 238000000423 cell based assay Methods 0.000 abstract description 7
- 238000012360 testing method Methods 0.000 abstract description 7
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 3
- 238000013518 transcription Methods 0.000 abstract description 3
- 230000035897 transcription Effects 0.000 abstract description 3
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 101800001020 Non-structural protein 4A Proteins 0.000 abstract 1
- 241000711549 Hepacivirus C Species 0.000 description 101
- 102000004169 proteins and genes Human genes 0.000 description 91
- 235000018102 proteins Nutrition 0.000 description 90
- 239000005089 Luciferase Substances 0.000 description 63
- 108060001084 Luciferase Proteins 0.000 description 63
- 239000013612 plasmid Substances 0.000 description 45
- 150000001413 amino acids Chemical class 0.000 description 35
- 235000001014 amino acid Nutrition 0.000 description 27
- 230000035772 mutation Effects 0.000 description 24
- 239000004098 Tetracycline Substances 0.000 description 21
- 229960002180 tetracycline Drugs 0.000 description 21
- 229930101283 tetracycline Natural products 0.000 description 21
- 235000019364 tetracycline Nutrition 0.000 description 21
- 150000003522 tetracyclines Chemical class 0.000 description 21
- 239000000047 product Substances 0.000 description 20
- 239000013598 vector Substances 0.000 description 18
- 230000003321 amplification Effects 0.000 description 17
- 238000003199 nucleic acid amplification method Methods 0.000 description 17
- 238000012545 processing Methods 0.000 description 16
- 230000001419 dependent effect Effects 0.000 description 13
- 108090000765 processed proteins & peptides Proteins 0.000 description 13
- 230000004913 activation Effects 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 12
- 102000004196 processed proteins & peptides Human genes 0.000 description 12
- 241000700605 Viruses Species 0.000 description 11
- 108091008146 restriction endonucleases Proteins 0.000 description 11
- 239000000758 substrate Substances 0.000 description 11
- 238000001262 western blot Methods 0.000 description 11
- 108020001507 fusion proteins Proteins 0.000 description 10
- 102000037865 fusion proteins Human genes 0.000 description 10
- 229920001184 polypeptide Polymers 0.000 description 10
- 239000002243 precursor Substances 0.000 description 10
- 208000015181 infectious disease Diseases 0.000 description 9
- 239000002773 nucleotide Substances 0.000 description 9
- 125000003729 nucleotide group Chemical group 0.000 description 9
- 101710118188 DNA-binding protein HU-alpha Proteins 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 101710144128 Non-structural protein 2 Proteins 0.000 description 8
- 101710199667 Nuclear export protein Proteins 0.000 description 8
- 241000700618 Vaccinia virus Species 0.000 description 8
- 239000002299 complementary DNA Substances 0.000 description 8
- 238000003670 luciferase enzyme activity assay Methods 0.000 description 8
- 108091028043 Nucleic acid sequence Proteins 0.000 description 7
- 108091034117 Oligonucleotide Proteins 0.000 description 7
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 7
- 239000012190 activator Substances 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 6
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 6
- 235000004279 alanine Nutrition 0.000 description 6
- 238000004113 cell culture Methods 0.000 description 6
- 238000010367 cloning Methods 0.000 description 6
- 230000002950 deficient Effects 0.000 description 6
- 239000000499 gel Substances 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 230000002441 reversible effect Effects 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 5
- 108060004795 Methyltransferase Proteins 0.000 description 5
- 102000035195 Peptidases Human genes 0.000 description 5
- 230000004071 biological effect Effects 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 230000007246 mechanism Effects 0.000 description 5
- 238000005457 optimization Methods 0.000 description 5
- 230000036961 partial effect Effects 0.000 description 5
- 238000003752 polymerase chain reaction Methods 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- 102100039556 Galectin-4 Human genes 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 4
- 101710137500 T7 RNA polymerase Proteins 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- 238000012761 co-transfection Methods 0.000 description 4
- 210000003527 eukaryotic cell Anatomy 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 230000000670 limiting effect Effects 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 230000010076 replication Effects 0.000 description 4
- 239000004475 Arginine Substances 0.000 description 3
- 108020004705 Codon Proteins 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 241000238631 Hexapoda Species 0.000 description 3
- 101000608765 Homo sapiens Galectin-4 Proteins 0.000 description 3
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 3
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 3
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 3
- 108010006035 Metalloproteases Proteins 0.000 description 3
- 102000005741 Metalloproteases Human genes 0.000 description 3
- -1 NS5A Proteins 0.000 description 3
- 101710152005 Non-structural polyprotein Proteins 0.000 description 3
- 101710114167 Polyprotein P1234 Proteins 0.000 description 3
- 101710124590 Polyprotein nsP1234 Proteins 0.000 description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 3
- 101710172711 Structural protein Proteins 0.000 description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 108010005774 beta-Galactosidase Proteins 0.000 description 3
- 102000005936 beta-Galactosidase Human genes 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 238000010276 construction Methods 0.000 description 3
- 235000018417 cysteine Nutrition 0.000 description 3
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 230000005714 functional activity Effects 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 208000019423 liver disease Diseases 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 210000004940 nucleus Anatomy 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 230000023603 positive regulation of transcription initiation, DNA-dependent Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 238000003757 reverse transcription PCR Methods 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
- 230000001052 transient effect Effects 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- 208000006154 Chronic hepatitis C Diseases 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 208000005176 Hepatitis C Diseases 0.000 description 2
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- 101710150451 Protein Bel-1 Proteins 0.000 description 2
- 108010022999 Serine Proteases Proteins 0.000 description 2
- 102000012479 Serine Proteases Human genes 0.000 description 2
- 108090000340 Transaminases Proteins 0.000 description 2
- 102000003929 Transaminases Human genes 0.000 description 2
- 108091061763 Triple-stranded DNA Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- 210000004899 c-terminal region Anatomy 0.000 description 2
- 229910000389 calcium phosphate Inorganic materials 0.000 description 2
- 239000001506 calcium phosphate Substances 0.000 description 2
- 235000011010 calcium phosphates Nutrition 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229960005091 chloramphenicol Drugs 0.000 description 2
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 239000005547 deoxyribonucleotide Substances 0.000 description 2
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000001952 enzyme assay Methods 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- 208000010710 hepatitis C virus infection Diseases 0.000 description 2
- 238000012203 high throughput assay Methods 0.000 description 2
- 238000013537 high throughput screening Methods 0.000 description 2
- 210000005260 human cell Anatomy 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 239000013600 plasmid vector Substances 0.000 description 2
- 230000020978 protein processing Effects 0.000 description 2
- 230000002797 proteolythic effect Effects 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- 230000002103 transcriptional effect Effects 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 2
- 238000010200 validation analysis Methods 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- 230000029812 viral genome replication Effects 0.000 description 2
- 238000012800 visualization Methods 0.000 description 2
- 210000005253 yeast cell Anatomy 0.000 description 2
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 description 1
- HBZBAMXERPYTFS-SECBINFHSA-N (4S)-2-(6,7-dihydro-5H-pyrrolo[3,2-f][1,3]benzothiazol-2-yl)-4,5-dihydro-1,3-thiazole-4-carboxylic acid Chemical compound OC(=O)[C@H]1CSC(=N1)c1nc2cc3CCNc3cc2s1 HBZBAMXERPYTFS-SECBINFHSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 108700028369 Alleles Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 101710132601 Capsid protein Proteins 0.000 description 1
- 206010057573 Chronic hepatic failure Diseases 0.000 description 1
- RGJOEKWQDUBAIZ-IBOSZNHHSA-N CoASH Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCS)O[C@H]1N1C2=NC=NC(N)=C2N=C1 RGJOEKWQDUBAIZ-IBOSZNHHSA-N 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 208000003322 Coinfection Diseases 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 238000007399 DNA isolation Methods 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 230000004568 DNA-binding Effects 0.000 description 1
- 208000010334 End Stage Liver Disease Diseases 0.000 description 1
- 241000991587 Enterovirus C Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 241000710781 Flaviviridae Species 0.000 description 1
- 108010001515 Galectin 4 Proteins 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 206010019786 Hepatitis non-A non-B Diseases 0.000 description 1
- 101000979342 Homo sapiens Nuclear factor NF-kappa-B p105 subunit Proteins 0.000 description 1
- 108010058683 Immobilized Proteins Proteins 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 101800000021 N-terminal protease Proteins 0.000 description 1
- 101800001292 Non-structural protein 2-3 Proteins 0.000 description 1
- 101150038760 Ns3 gene Proteins 0.000 description 1
- 102100023050 Nuclear factor NF-kappa-B p105 subunit Human genes 0.000 description 1
- 102000008021 Nucleoside-Triphosphatase Human genes 0.000 description 1
- 108010075285 Nucleoside-Triphosphatase Proteins 0.000 description 1
- 101800000212 P1A protein Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 101800000862 Protease cofactor Proteins 0.000 description 1
- 108090000944 RNA Helicases Proteins 0.000 description 1
- 102000004409 RNA Helicases Human genes 0.000 description 1
- 108091027981 Response element Proteins 0.000 description 1
- IWUCXVSUMQZMFG-AFCXAGJDSA-N Ribavirin Chemical compound N1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 IWUCXVSUMQZMFG-AFCXAGJDSA-N 0.000 description 1
- 108091028664 Ribonucleotide Proteins 0.000 description 1
- 241000700584 Simplexvirus Species 0.000 description 1
- 241000710960 Sindbis virus Species 0.000 description 1
- 230000024932 T cell mediated immunity Effects 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 206010046865 Vaccinia virus infection Diseases 0.000 description 1
- 108020005202 Viral DNA Proteins 0.000 description 1
- 108700022715 Viral Proteases Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- QWXOJIDBSHLIFI-UHFFFAOYSA-N [3-(1-chloro-3'-methoxyspiro[adamantane-4,4'-dioxetane]-3'-yl)phenyl] dihydrogen phosphate Chemical compound O1OC2(C3CC4CC2CC(Cl)(C4)C3)C1(OC)C1=CC=CC(OP(O)(O)=O)=C1 QWXOJIDBSHLIFI-UHFFFAOYSA-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
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 101150010487 are gene Proteins 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000013611 chromosomal DNA Substances 0.000 description 1
- 208000011444 chronic liver failure Diseases 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- RGJOEKWQDUBAIZ-UHFFFAOYSA-N coenzime A Natural products OC1C(OP(O)(O)=O)C(COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCS)OC1N1C2=NC=NC(N)=C2N=C1 RGJOEKWQDUBAIZ-UHFFFAOYSA-N 0.000 description 1
- 239000005516 coenzyme A Substances 0.000 description 1
- 229940093530 coenzyme a Drugs 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- ZTOGYGRJDIWKAQ-UHFFFAOYSA-N cyclohexene-1,2-diamine Chemical compound NC1=C(N)CCCC1 ZTOGYGRJDIWKAQ-UHFFFAOYSA-N 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- KDTSHFARGAKYJN-UHFFFAOYSA-N dephosphocoenzyme A Natural products OC1C(O)C(COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCS)OC1N1C2=NC=NC(N)=C2N=C1 KDTSHFARGAKYJN-UHFFFAOYSA-N 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 238000011143 downstream manufacturing Methods 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 238000009585 enzyme analysis Methods 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 238000012188 high-throughput screening assay Methods 0.000 description 1
- 230000028996 humoral immune response Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 238000003367 kinetic assay Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- ADKOXSOCTOWDOP-UHFFFAOYSA-L magnesium;aluminum;dihydroxide;trihydrate Chemical compound O.O.O.[OH-].[OH-].[Mg+2].[Al] ADKOXSOCTOWDOP-UHFFFAOYSA-L 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000004660 morphological change Effects 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 238000007899 nucleic acid hybridization Methods 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229960000329 ribavirin Drugs 0.000 description 1
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 description 1
- 239000002336 ribonucleotide Substances 0.000 description 1
- 125000002652 ribonucleotide group Chemical group 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 238000007423 screening assay Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 108700020534 tetracycline resistance-encoding transposon repressor Proteins 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000003151 transfection method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- JLEXUIVKURIPFI-UHFFFAOYSA-N tris phosphate Chemical compound OP(O)(O)=O.OCC(N)(CO)CO JLEXUIVKURIPFI-UHFFFAOYSA-N 0.000 description 1
- 108010036927 trypsin-like serine protease Proteins 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 208000007089 vaccinia Diseases 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 210000000605 viral structure Anatomy 0.000 description 1
- 210000002845 virion Anatomy 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- 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
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6897—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids involving reporter genes operably linked to promoters
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/04—Fusion polypeptide containing a localisation/targetting motif containing an ER retention signal such as a C-terminal HDEL motif
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/50—Fusion polypeptide containing protease site
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
- C07K2319/71—Fusion polypeptide containing domain for protein-protein interaction containing domain for transcriptional activaation, e.g. VP16
-
- 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
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/24011—Poxviridae
- C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
- C12N2710/24141—Use of virus, viral particle or viral elements as a vector
- C12N2710/24143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- 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
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/24011—Flaviviridae
- C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
- C12N2770/24222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Gastroenterology & Hepatology (AREA)
- Analytical Chemistry (AREA)
- Virology (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US13236099P | 1999-05-04 | 1999-05-04 | |
| US60/132,360 | 1999-05-04 | ||
| PCT/CA2000/000496 WO2000066623A2 (en) | 1999-05-04 | 2000-05-01 | Surrogate cell-based system and method for assaying the activity of hepatitis c virus ns3 protease |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2002542812A JP2002542812A (ja) | 2002-12-17 |
| JP2002542812A5 JP2002542812A5 (https=) | 2007-06-21 |
| JP4782927B2 true JP4782927B2 (ja) | 2011-09-28 |
Family
ID=22453660
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2000615652A Expired - Fee Related JP4782927B2 (ja) | 1999-05-04 | 2000-05-01 | 代理細胞ベースシステム及びｃ型肝炎ウイルスｎｓ３プロテアーゼの活性を評価する方法 |
Country Status (14)
| Country | Link |
|---|---|
| US (3) | US7276373B2 (https=) |
| EP (1) | EP1177211B1 (https=) |
| JP (1) | JP4782927B2 (https=) |
| AT (1) | ATE386746T1 (https=) |
| AU (1) | AU772386B2 (https=) |
| CA (1) | CA2365262C (https=) |
| DE (1) | DE60038113T2 (https=) |
| DK (1) | DK1177211T3 (https=) |
| ES (1) | ES2300262T3 (https=) |
| HU (1) | HUP0200945A3 (https=) |
| IL (2) | IL145376A0 (https=) |
| MX (1) | MXPA01011045A (https=) |
| NZ (1) | NZ515861A (https=) |
| WO (1) | WO2000066623A2 (https=) |
Families Citing this family (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2365262C (en) * | 1999-05-04 | 2008-06-10 | Boehringer Ingelheim (Canada) Ltd. | Surrogate cell-based system and method for assaying the activity of hepatitis c virus ns3 protease |
| AU5920400A (en) * | 1999-07-07 | 2001-01-22 | Du Pont Pharmaceuticals Company | Cell-based assay systems for examining hcv ns3 protease activity |
| US6986892B1 (en) | 1999-11-24 | 2006-01-17 | Chiron Corporation | Immunogenic Hepatitis C virus non-structural polypeptides |
| WO2001038360A2 (en) * | 1999-11-24 | 2001-05-31 | Chiron Corporation | Novel hcv non-structural polypeptide |
| US6921634B2 (en) * | 1999-12-23 | 2005-07-26 | Board Of Regents, The University Of Texas System | Replication competent hepatitus C virus and methods of use |
| CA2370803A1 (en) * | 2000-02-25 | 2001-08-30 | Cambridge Drug Discovery, Ltd. | Methods for identifying a protease inhibitor |
| MXPA03005638A (es) | 2000-12-22 | 2004-04-05 | Boehringer Ingelheim Ca Ltd | Auto replicacion de la molecula de arn de virus de la hepatitis c. |
| DE10234418A1 (de) * | 2002-07-29 | 2004-02-12 | Bayer Ag | Verfahren zur Untersuchung und Beurteilung von Virusenzymen im HTS |
| EP1670415A4 (en) * | 2003-09-12 | 2007-12-05 | Vertex Pharma | ANIMAL MODEL FOR STUDYING PROTEASES ACTIVITY AND HEPATIC DISORDERS |
| WO2005037214A2 (en) | 2003-10-14 | 2005-04-28 | Intermune, Inc. | Macrocyclic carboxylic acids and acylsulfonamides as inhibitors of hcv replication |
| MX2008001166A (es) | 2005-07-25 | 2008-03-18 | Intermune Inc | Nuevos inhibidores macrociclicos de la replicacion del virus de hepatitis c. |
| AU2006301966A1 (en) | 2005-10-11 | 2007-04-19 | Array Biopharma, Inc. | Compounds and methods for inhibiting hepatitis C viral replication |
| EP2392589A3 (en) | 2005-11-11 | 2012-06-20 | Vertex Pharmaceuticals Incorporated | Hepatitis C virus variants |
| US7705138B2 (en) | 2005-11-11 | 2010-04-27 | Vertex Pharmaceuticals Incorporated | Hepatitis C virus variants |
| WO2013027217A1 (en) * | 2011-08-22 | 2013-02-28 | Ramot At Tel-Aviv University Ltd. | Zymoxins and methods of using the same |
| US9353423B2 (en) | 2012-11-07 | 2016-05-31 | Gilead Sciences, Inc. | HCV genotype 6 replicons |
| US20210087548A1 (en) * | 2017-07-24 | 2021-03-25 | The Board Of Trustees Of The Leland Stanford Junior University | Compositions and Methods for Inducing Protein Function |
| WO2022094463A1 (en) * | 2020-11-02 | 2022-05-05 | Regents Of The University Of Minnesota | Live cell assay for protease inhibition |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06303980A (ja) * | 1990-06-25 | 1994-11-01 | Handai Biseibutsubiyou Kenkyukai | 非Ａ非Ｂ型肝炎ウイルス遺伝子ｃＤＮＡおよび抗原ポリペプチド |
| JP2002523102A (ja) * | 1998-08-31 | 2002-07-30 | バーテックス ファーマシューティカルズ インコーポレイテッド | プロテアーゼ活性を測定するために有用な融合タンパク質、ｄｎａ分子、ベクター、および宿主細胞 |
| JP2003523717A (ja) * | 1998-08-05 | 2003-08-12 | アゴーロン・ファーマシューティカルズ・インコーポレイテッド | Ｃ型肝炎ウイルスプロテアーゼ阻害物質の細胞ベースの評価で使用するレポーター遺伝子系 |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU652941B2 (en) * | 1990-06-25 | 1994-09-15 | Research Foundation For Microbial Diseases Of Osaka University, The | Non-A, Non-B hepatitis virus particles |
| US5861267A (en) * | 1995-05-01 | 1999-01-19 | Vertex Pharmaceuticals Incorporated | Methods, nucleotide sequences and host cells for assaying exogenous and endogenous protease activity |
| IT1277914B1 (it) * | 1995-08-22 | 1997-11-12 | Angeletti P Ist Richerche Bio | Procedimento per produrre - in forma pura e in quantita' elevate - polipeptidi con l'attivita' proteolitica della proteasi ns3 di hcv, e |
| KR100209095B1 (ko) * | 1996-06-28 | 1999-07-15 | 성재갑 | C형 간염 바이러스의 프로테아제의 활성을 측정할 수 있는 c형 간염 대체 바이러스, 그 재조합 유전자 및 그 용도 |
| CA2365262C (en) * | 1999-05-04 | 2008-06-10 | Boehringer Ingelheim (Canada) Ltd. | Surrogate cell-based system and method for assaying the activity of hepatitis c virus ns3 protease |
-
2000
- 2000-05-01 CA CA002365262A patent/CA2365262C/en not_active Expired - Fee Related
- 2000-05-01 MX MXPA01011045A patent/MXPA01011045A/es active IP Right Grant
- 2000-05-01 DE DE60038113T patent/DE60038113T2/de not_active Expired - Lifetime
- 2000-05-01 NZ NZ515861A patent/NZ515861A/en not_active IP Right Cessation
- 2000-05-01 AT AT00922396T patent/ATE386746T1/de active
- 2000-05-01 ES ES00922396T patent/ES2300262T3/es not_active Expired - Lifetime
- 2000-05-01 IL IL14537600A patent/IL145376A0/xx unknown
- 2000-05-01 DK DK00922396T patent/DK1177211T3/da active
- 2000-05-01 EP EP00922396A patent/EP1177211B1/en not_active Expired - Lifetime
- 2000-05-01 HU HU0200945A patent/HUP0200945A3/hu unknown
- 2000-05-01 WO PCT/CA2000/000496 patent/WO2000066623A2/en not_active Ceased
- 2000-05-01 JP JP2000615652A patent/JP4782927B2/ja not_active Expired - Fee Related
- 2000-05-01 AU AU42825/00A patent/AU772386B2/en not_active Ceased
-
2001
- 2001-09-11 IL IL145376A patent/IL145376A/en not_active IP Right Cessation
-
2002
- 2002-12-23 US US10/328,206 patent/US7276373B2/en not_active Expired - Lifetime
- 2002-12-23 US US10/328,127 patent/US20030162168A1/en not_active Abandoned
-
2005
- 2005-05-27 US US11/140,379 patent/US7838207B2/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06303980A (ja) * | 1990-06-25 | 1994-11-01 | Handai Biseibutsubiyou Kenkyukai | 非Ａ非Ｂ型肝炎ウイルス遺伝子ｃＤＮＡおよび抗原ポリペプチド |
| JP2003523717A (ja) * | 1998-08-05 | 2003-08-12 | アゴーロン・ファーマシューティカルズ・インコーポレイテッド | Ｃ型肝炎ウイルスプロテアーゼ阻害物質の細胞ベースの評価で使用するレポーター遺伝子系 |
| JP2002523102A (ja) * | 1998-08-31 | 2002-07-30 | バーテックス ファーマシューティカルズ インコーポレイテッド | プロテアーゼ活性を測定するために有用な融合タンパク質、ｄｎａ分子、ベクター、および宿主細胞 |
Also Published As
| Publication number | Publication date |
|---|---|
| MXPA01011045A (es) | 2002-07-22 |
| ES2300262T3 (es) | 2008-06-16 |
| DE60038113D1 (de) | 2008-04-03 |
| IL145376A (en) | 2009-09-01 |
| DE60038113T2 (de) | 2009-02-12 |
| EP1177211A2 (en) | 2002-02-06 |
| HUP0200945A2 (en) | 2002-08-28 |
| IL145376A0 (en) | 2002-06-30 |
| EP1177211B1 (en) | 2008-02-20 |
| NZ515861A (en) | 2004-08-27 |
| WO2000066623A3 (en) | 2001-03-22 |
| US20050227226A1 (en) | 2005-10-13 |
| WO2000066623A2 (en) | 2000-11-09 |
| CA2365262C (en) | 2008-06-10 |
| US7276373B2 (en) | 2007-10-02 |
| US7838207B2 (en) | 2010-11-23 |
| DK1177211T3 (da) | 2008-06-02 |
| CA2365262A1 (en) | 2000-11-09 |
| US20030162168A1 (en) | 2003-08-28 |
| HUP0200945A3 (en) | 2004-07-28 |
| US20030162169A1 (en) | 2003-08-28 |
| AU4282500A (en) | 2000-11-17 |
| ATE386746T1 (de) | 2008-03-15 |
| AU772386B2 (en) | 2004-04-29 |
| JP2002542812A (ja) | 2002-12-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4782927B2 (ja) | 代理細胞ベースシステム及びｃ型肝炎ウイルスｎｓ３プロテアーゼの活性を評価する方法 | |
| Flajolet et al. | A genomic approach of the hepatitis C virus generates a protein interaction map | |
| JP4223068B2 (ja) | Ｃ型肝炎ウイルス（ｈｃｖ）用の機能性ｄｎａクローン及びその使用 | |
| Satoh et al. | Cleavage of hepatitis C virus nonstructural protein 5A by a caspase-like protease (s) in mammalian cells | |
| JP2001017187A (ja) | Ｃ型肝炎ウイルス細胞培養系、ｃ型肝炎ウイルス−ｒｎａ−構築物、細胞培養系または構築物の使用、ｃ型肝炎ウイルス−ｒｎａ−構築物の細胞培養に適合した突然変異体を獲得する方法、ｃ型肝炎ウイルス−全長ゲノム、ｃ型肝炎ウイルス−部分ゲノム、または任意のｃ型肝炎ウイルス−構築物の突然変異体の製法、細胞培養に適合したｃ型肝炎ウイルス−構築物、その突然変異体、ｃ型肝炎ウイルス−全長ゲノムの突然変異体、ｃ型肝炎ウイルス粒子またはウイルス様粒子、およびこれで感染した細胞 | |
| AU759425B2 (en) | Reporter gene system for use in cell-based assessment of inhibitors of the hepatitis C virus protease | |
| HUP0302593A2 (hu) | Önállóan replikálódó RNS molekula hepatitis C vírusból | |
| Shimoike et al. | Down-regulation of the internal ribosome entry site (IRES)-mediated translation of the hepatitis C virus: critical role of binding of the stem-loop IIId domain of IRES and the viral core protein | |
| JP2002542812A5 (https=) | ||
| CA2543846C (en) | Hcv replicons containing ns5b from genotype 2b | |
| Huang et al. | A human hepatoma cell line expressing hepatitis C virus nonstructural proteins tightly regulated by tetracycline | |
| JP3878132B2 (ja) | 精製活性型ｈｃｖｎｓ２／３プロテアーゼ | |
| Pacini et al. | Reporter substrates for assessing the activity of the hepatitis C virus NS3-4A serine protease in living cells | |
| Song et al. | Development of an in vivo assay system suitable for screening inhibitors of hepatitis C viral protease | |
| US20040229213A1 (en) | Exhaustive analysis of viral protein interactions by two-hybrid screens and selection of correctly folded viral interacting polypeptides | |
| US20010034019A1 (en) | Chimeric HCV/GBV-B viruses | |
| CA2437072A1 (en) | In vitro system for replication of rna-dependent rna polymerase (rdrp) viruses | |
| CA2331786A1 (en) | Exhaustive analysis of viral protein interactions by two-hybrid screens and selection of correctly folded viral interacting polypeptides | |
| CA2483513A1 (en) | Reporter-selectable hepatitis c virus replicon | |
| Appel | Structural and functional analysis of the hepatitis C virus non-structural protein 5A | |
| AU2002240214A1 (en) | In vitro system for replication of RNA-dependent RNA polymerase (RDRP) viruses |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20070425 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20070425 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20100311 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20100611 |
|
| A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20100618 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20100910 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20101118 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20110216 |
|
| A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20110223 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110518 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20110609 |
|
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20110708 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20140715 Year of fee payment: 3 |
|
| R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| LAPS | Cancellation because of no payment of annual fees |