WO2011036883A1 - Anti-influenza virus agent - Google Patents
Anti-influenza virus agent Download PDFInfo
- Publication number
- WO2011036883A1 WO2011036883A1 PCT/JP2010/005762 JP2010005762W WO2011036883A1 WO 2011036883 A1 WO2011036883 A1 WO 2011036883A1 JP 2010005762 W JP2010005762 W JP 2010005762W WO 2011036883 A1 WO2011036883 A1 WO 2011036883A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- influenza virus
- extract
- inhibitor
- karin
- viral
- Prior art date
Links
- 241000700605 Viruses Species 0.000 title claims abstract description 44
- 206010022000 influenza Diseases 0.000 title claims abstract description 37
- 239000000284 extract Substances 0.000 claims abstract description 47
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 31
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 31
- 239000003112 inhibitor Substances 0.000 claims abstract description 29
- 241000712461 unidentified influenza virus Species 0.000 claims abstract description 29
- 238000012360 testing method Methods 0.000 claims abstract description 18
- 230000003612 virological effect Effects 0.000 claims abstract description 18
- 108091034135 Vault RNA Proteins 0.000 claims abstract description 17
- 230000000694 effects Effects 0.000 claims abstract description 16
- 108020004999 messenger RNA Proteins 0.000 claims abstract description 15
- 238000001179 sorption measurement Methods 0.000 claims abstract description 15
- 239000004480 active ingredient Substances 0.000 claims abstract description 12
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 10
- 230000034217 membrane fusion Effects 0.000 claims abstract description 9
- 238000005194 fractionation Methods 0.000 claims description 7
- 108010003533 Viral Envelope Proteins Proteins 0.000 claims description 6
- 235000013361 beverage Nutrition 0.000 claims description 5
- 235000013305 food Nutrition 0.000 claims description 5
- 235000017831 Pseudocydonia sinensis Nutrition 0.000 claims description 4
- 230000002949 hemolytic effect Effects 0.000 claims description 2
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims 7
- 241001507760 Chaenomeles sinensis Species 0.000 claims 2
- 241000972773 Aulopiformes Species 0.000 claims 1
- VKYKSIONXSXAKP-UHFFFAOYSA-N hexamethylenetetramine Chemical compound C1N(C2)CN3CN1CN2C3 VKYKSIONXSXAKP-UHFFFAOYSA-N 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 235000019515 salmon Nutrition 0.000 claims 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 45
- 239000000419 plant extract Substances 0.000 abstract description 4
- 235000017788 Cydonia oblonga Nutrition 0.000 abstract 1
- 230000002587 anti-hemolytic effect Effects 0.000 abstract 1
- 239000003219 hemolytic agent Substances 0.000 abstract 1
- 101000894525 Homo sapiens Transforming growth factor-beta-induced protein ig-h3 Proteins 0.000 description 34
- 102100021398 Transforming growth factor-beta-induced protein ig-h3 Human genes 0.000 description 34
- 210000004027 cell Anatomy 0.000 description 23
- 230000002401 inhibitory effect Effects 0.000 description 15
- 230000005764 inhibitory process Effects 0.000 description 12
- 235000009508 confectionery Nutrition 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 9
- 208000015181 infectious disease Diseases 0.000 description 9
- 206010018910 Haemolysis Diseases 0.000 description 8
- 101710154606 Hemagglutinin Proteins 0.000 description 8
- 101710093908 Outer capsid protein VP4 Proteins 0.000 description 8
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 description 8
- 101710176177 Protein A56 Proteins 0.000 description 8
- 230000008588 hemolysis Effects 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 239000000185 hemagglutinin Substances 0.000 description 7
- 239000003826 tablet Substances 0.000 description 7
- 239000002299 complementary DNA Substances 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- 102000005348 Neuraminidase Human genes 0.000 description 5
- 108010006232 Neuraminidase Proteins 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 239000003205 fragrance Substances 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 4
- 102100034343 Integrase Human genes 0.000 description 4
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 4
- 235000015218 chewing gum Nutrition 0.000 description 4
- 229940112822 chewing gum Drugs 0.000 description 4
- 239000002324 mouth wash Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000007921 spray Substances 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 238000013518 transcription Methods 0.000 description 4
- 230000035897 transcription Effects 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 208000035473 Communicable disease Diseases 0.000 description 3
- 239000003155 DNA primer Substances 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 101150118742 NP gene Proteins 0.000 description 3
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 108010006785 Taq Polymerase Proteins 0.000 description 3
- 244000269722 Thea sinensis Species 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 235000015110 jellies Nutrition 0.000 description 3
- 239000008274 jelly Substances 0.000 description 3
- 239000013615 primer Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 230000029812 viral genome replication Effects 0.000 description 3
- 230000009385 viral infection Effects 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- WJLVQTJZDCGNJN-UHFFFAOYSA-N Chlorhexidine hydrochloride Chemical compound Cl.Cl.C=1C=C(Cl)C=CC=1NC(N)=NC(N)=NCCCCCCN=C(N)N=C(N)NC1=CC=C(Cl)C=C1 WJLVQTJZDCGNJN-UHFFFAOYSA-N 0.000 description 2
- 206010011224 Cough Diseases 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 244000251905 Pseudocydonia sinensis Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 235000006468 Thea sinensis Nutrition 0.000 description 2
- 208000036142 Viral infection Diseases 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000003443 antiviral agent Substances 0.000 description 2
- 235000015895 biscuits Nutrition 0.000 description 2
- 235000020279 black tea Nutrition 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 229960004504 chlorhexidine hydrochloride Drugs 0.000 description 2
- 235000019219 chocolate Nutrition 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229940051866 mouthwash Drugs 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 239000002304 perfume Substances 0.000 description 2
- 235000013824 polyphenols Nutrition 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 2
- 229940081974 saccharin Drugs 0.000 description 2
- 235000019204 saccharin Nutrition 0.000 description 2
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 235000014347 soups Nutrition 0.000 description 2
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-3',4',5,7-Tetrahydroxy-2,3-trans-flavan-3-ol Natural products C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 description 1
- PFTAWBLQPZVEMU-UKRRQHHQSA-N (-)-epicatechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-UKRRQHHQSA-N 0.000 description 1
- 229930013783 (-)-epicatechin Natural products 0.000 description 1
- 235000007355 (-)-epicatechin Nutrition 0.000 description 1
- UBCHPRBFMUDMNC-UHFFFAOYSA-N 1-(1-adamantyl)ethanamine Chemical compound C1C(C2)CC3CC2CC1(C(N)C)C3 UBCHPRBFMUDMNC-UHFFFAOYSA-N 0.000 description 1
- XGRSAFKZAGGXJV-UHFFFAOYSA-N 3-azaniumyl-3-cyclohexylpropanoate Chemical compound OC(=O)CC(N)C1CCCCC1 XGRSAFKZAGGXJV-UHFFFAOYSA-N 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 101100313763 Arabidopsis thaliana TIM22-2 gene Proteins 0.000 description 1
- 208000006820 Arthralgia Diseases 0.000 description 1
- 241000282465 Canis Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 235000012766 Cannabis sativa ssp. sativa var. sativa Nutrition 0.000 description 1
- 235000012765 Cannabis sativa ssp. sativa var. spontanea Nutrition 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 240000006766 Cornus mas Species 0.000 description 1
- 235000003363 Cornus mas Nutrition 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 241000508269 Psidium Species 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 230000006819 RNA synthesis Effects 0.000 description 1
- 238000010240 RT-PCR analysis Methods 0.000 description 1
- 208000036071 Rhinorrhea Diseases 0.000 description 1
- 206010039101 Rhinorrhoea Diseases 0.000 description 1
- 235000001537 Ribes X gardonianum Nutrition 0.000 description 1
- 235000001535 Ribes X utile Nutrition 0.000 description 1
- 235000016919 Ribes petraeum Nutrition 0.000 description 1
- 244000281247 Ribes rubrum Species 0.000 description 1
- 235000002355 Ribes spicatum Nutrition 0.000 description 1
- 241000220222 Rosaceae Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- -1 Sucrose fatty acid ester Chemical class 0.000 description 1
- 108700005077 Viral Genes Proteins 0.000 description 1
- 108020000999 Viral RNA Proteins 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- ABUBSBSOTTXVPV-UHFFFAOYSA-H [U+6].CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O Chemical compound [U+6].CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O.CC([O-])=O ABUBSBSOTTXVPV-UHFFFAOYSA-H 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- DKNWSYNQZKUICI-UHFFFAOYSA-N amantadine Chemical compound C1C(C2)CC3CC2CC1(N)C3 DKNWSYNQZKUICI-UHFFFAOYSA-N 0.000 description 1
- 229960003805 amantadine Drugs 0.000 description 1
- 235000010208 anthocyanin Nutrition 0.000 description 1
- 229930002877 anthocyanin Natural products 0.000 description 1
- 239000004410 anthocyanin Substances 0.000 description 1
- 150000004636 anthocyanins Chemical class 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 210000003403 autonomic nervous system Anatomy 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 238000010804 cDNA synthesis Methods 0.000 description 1
- 235000009120 camo Nutrition 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000005607 chanvre indien Nutrition 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 208000024829 digestive system symptom Diseases 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 210000001163 endosome Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 238000012869 ethanol precipitation Methods 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- ZJYYHGLJYGJLLN-UHFFFAOYSA-N guanidinium thiocyanate Chemical compound SC#N.NC(N)=N ZJYYHGLJYGJLLN-UHFFFAOYSA-N 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 239000011487 hemp Substances 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N hydrochloric acid Substances Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 230000009610 hypersensitivity Effects 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007937 lozenge Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000004779 membrane envelope Anatomy 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 208000020470 nervous system symptom Diseases 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000015205 orange juice Nutrition 0.000 description 1
- 239000010330 ougon Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 229960000888 rimantadine Drugs 0.000 description 1
- 239000012898 sample dilution Substances 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 229960004711 sodium monofluorophosphate Drugs 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 235000019871 vegetable fat Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- ARAIBEBZBOPLMB-UFGQHTETSA-N zanamivir Chemical compound CC(=O)N[C@@H]1[C@@H](N=C(N)N)C=C(C(O)=O)O[C@H]1[C@H](O)[C@H](O)CO ARAIBEBZBOPLMB-UFGQHTETSA-N 0.000 description 1
- 229960001028 zanamivir Drugs 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
Definitions
- the present invention relates to an anti-influenza virus agent, an influenza virus adsorption inhibitor, a virus mRNA synthesis inhibitor in influenza virus-infected cells, and a virus vRNA in influenza virus-infected cells, which contains a plant-derived extract having an influenza virus infection-suppressing action as an active ingredient.
- the present invention relates to a synthesis inhibitor, a viral cRNA synthesis inhibitor in influenza virus-infected cells, a hemolysis inhibitor in a viral membrane fusion activity test, and an influenza virus envelope disruptor.
- the influenza virus that causes the influenza epidemic is an RNA virus having an envelope membrane with a diameter of about 1 / 10,000 mm. Although it is classified into three types, A, B, and C, due to the difference in antigenicity, it is A type and B type that show a trendy spread. Two types of glycoproteins, hemagglutinin (HA) and neuraminidase (NA), protrude in the form of spikes on the surface of these virus particles, and gene RNA segmented into 8 segments exists inside. HA and NA on the surface of the virus frequently mutate within the same subtype, and new antigenic variants appear every year.
- HA hemagglutinin
- NA neuraminidase
- Influenza virus released by cough droplets enters from the human nose and mouth, adsorbs to the mucosal epithelial cells of the upper respiratory tract by the spike-like glycoprotein HA on the surface of the virus, and begins to proliferate after entering the cells.
- Recent research has revealed the mechanism of viral infection.
- the virus binds to a receptor consisting of a sugar chain on the surface layer of human target cells, is taken into the endosome, enters the cell by fusion of the viral membrane and endosomal membrane, undergoes shelling and translocation, and the expression of the viral gene Replication occurs, and finally progeny virus particles are formed and propagated by budding from the host cell membrane.
- influenza virus causes symptoms such as sudden fever, headache, joint pain, general malaise in a few days, followed by respiratory symptoms such as cough, sore throat, runny nose and stuffy nose. Unlike the so-called cold, it is highly infectious and causes an explosive epidemic in a short period of time.
- the structure of the HA protein of influenza virus repeats mutations every year, and the fact that antibodies made by past infections are not very useful is also a factor that spreads infection.
- antiviral drugs such as amantadine, rimantadine, and zanamivir have been developed, but side effects such as hypersensitivity, neuropsychiatric symptoms, digestive system symptoms, and autonomic nervous system symptoms have been reported. Caution must be taken.
- influenza viruses are thought to be difficult to suppress by vaccination because influenza viruses infect and proliferate in the airway mucosal epithelium and the epidemic type of the year cannot be accurately predicted. Frequent gargling, avoiding throat dryness, and adequate nutrition and rest are currently considered the most effective preventive measures. Development of an anti-influenza virus agent that is highly effective in suppressing infection and has no safety problems and can be used on a daily basis is desired.
- Non-patent Documents 1 and 2 polyphenol components of tea and black tea have been reported as anti-influenza materials derived from natural products (Non-patent Documents 1 and 2), and it has become clear that gargle of black tea suppresses actual viral infections through tests using humans. (Non-Patent Document 3). In addition, it has been reported that ougon-derived flavonoid components exhibit an influenza infection suppression effect due to viral sialidase inhibitory activity (Non-patent Document 4).
- Patent Document 1 Kameda Futoshi Koshiketsu
- Patent Document 2 Kuroburi Currant Extract
- Patent Document 3 Potato Anthocyanin Pigment
- Patent Document 4 Guava Leaf Extract
- an anti-influenza agent containing an extract of the flower bud or petal as an active ingredient is disclosed (Patent Document 6), an extract of Karin (Patent Document 8), and a column fraction of Karin.
- the anti-influenza virus effect of is reported.
- influenza virus adsorption inhibitory effect of the column fraction of Karin shown in the patent of the present invention, the viral mRNA synthesis inhibitory effect in influenza virus-infected cells, the virus vRNA synthesis inhibitory effect in influenza virus-infected cells, and the virus cRNA in influenza virus-infected cells
- the synthesis inhibitory effect the hemolysis inhibitory effect in the virus membrane fusion activity test, and the envelope destruction effect of influenza virus, which have been revealed for the first time by the present invention.
- An object of the present invention is to use a highly safe plant extract that can be used with peace of mind on a daily basis, an influenza virus adsorption inhibitor, a virus mRNA synthesis inhibitor in influenza virus-infected cells, and a virus vRNA synthesis inhibitor in influenza virus-infected cells. It is intended to provide an agent, a virus replication inhibitor in influenza virus-infected cells, a hemolysis inhibitor in a virus membrane fusion activity test, and an influenza virus envelope disruptor.
- the present inventors focused on highly safe kalin without side effects, and used RT-PCR analysis of mRNA, vRNA and cRNA in MDCK cells added with influenza virus, and chicken erythrocytes. The effect of the extract of karin on hemolysis and observation of virus particles with an electron microscope was found, and the product of the present invention was completed.
- the present invention relates to an influenza virus adsorption inhibitor characterized by comprising an extract of karin (Chaenomeles sinensis, Pseudocydonia sinensis), a viral mRNA synthesis inhibitor in influenza virus-infected cells, and a virus in influenza virus-infected cells.
- karin Choenomeles sinensis, Pseudocydonia sinensis
- a viral mRNA synthesis inhibitor in influenza virus-infected cells and a virus in influenza virus-infected cells.
- They are a vRNA synthesis inhibitor, a viral cRNA synthesis inhibitor in influenza virus-infected cells, a hemolysis inhibitor in a viral membrane fusion activity test, and an influenza virus envelope disruptor.
- this invention is the food-drinks which have an anti-influenza virus effect
- the present invention comprises a highly safe extract of karin as an active ingredient, an influenza virus adsorption inhibitor strong against influenza virus, a virus mRNA synthesis inhibitor in influenza virus-infected cells, a virus vRNA synthesis inhibitor in influenza virus-infected cells,
- the present invention provides a virus replication inhibitor in influenza virus-infected cells, a hemolysis inhibitor in a virus membrane fusion activity test, and an envelope disruptor for influenza virus.
- the extract of Karin which is an active ingredient of the present invention, has high safety, it can be adsorbed, impregnated and added to a mask, an air conditioner filter, clothing, wet tissue, a spray solution, etc. Can be widely used in daily life.
- the product of the present invention is effective in preventing influenza virus infection and treating diseases caused by influenza virus.
- the destruction activity to the viral envelope membrane of a karin extract is shown.
- the adsorption inhibitory activity, vRNA synthesis inhibitory activity, mRNA synthesis inhibitory activity, and cRNA synthesis inhibitory activity of the karin extract are shown.
- the hemolysis inhibitory activity of a karin extract is shown.
- ⁇ It is desirable to use the fruits of karin as the raw material for the product of the present invention.
- the method for obtaining the extract of the present invention from the pulverized plant is not particularly limited, but water, methanol, ethanol, n-propanol, and lower alcohols such as n-butanol, ether, ethyl acetate, acetone, glycerin, propylene glycol.
- One or two or more mixed solvents of organic solvents such as these are added, and extraction is performed by a conventional extraction method.
- the extraction condition is not particularly limited, but it is preferably about 1 to 5 hours at 50 to 90 ° C.
- the extract can be filtered and the extraction solvent can be distilled off, followed by concentration or lyophilization under reduced pressure. Further, those obtained by fractionating and purifying these extracts by organic solvent fractionation, column chromatography or the like can also be used.
- the form of use of the product of the present invention is not particularly limited, and powders, tablets, troches can be obtained by adding a solvent, a dispersant, a pharmaceutical carrier, an emulsifier, a diluent, a stabilizer, etc. to the plant extract exemplified as an active ingredient.
- a solvent e.g., ethanol
- emulsifier e.g., a sulfate
- a diluent e.g., diluent
- a diluent emulsifier
- a diluent emulsifier
- a diluent emulsifier
- a diluent e.g., diluent
- a stabilizer e.g., a stabilizer, etc.
- the plant extract exemplified as an active ingredient.
- an infection control product that can contribute to influenza prevention can be provided.
- the amount of adsorbed and added plant extract in these applications varies depending on the form of the infection control product, and cannot be specified in general, but is added so that the proportion is 0.001 to 5% by weight. Is preferred.
- the present invention is highly safe, for example, it is blended into confectionery such as chewing gum, candy, tablet confectionery, gummy jelly, chocolate and biscuits, ice cream, sorbet and other frozen confectionery, beverages, soups, jams and other food and drink, Can be used.
- confectionery such as chewing gum, candy, tablet confectionery, gummy jelly, chocolate and biscuits, ice cream, sorbet and other frozen confectionery, beverages, soups, jams and other food and drink, Can be used.
- the amount to be added varies depending on the form of use and the taste of the extract, but it is added in an amount of 0.001 to 5% by weight, preferably about 0.01 to 1% by weight based on the food or drink. Is preferred.
- a reflux condenser is attached to a flask to which 300 g of dried Karin fruit and 3000 ml of 50% ethanol are added, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered, the solvent was removed, and freeze-dried to obtain 69 g of extract.
- the Karin 50% ethanol extract was applied to a Diaion HP20 (Mitsubishi Kasei) column. Elution was carried out in the order of water, 20% ethanol, 40% ethanol, and 60% ethanol. The component (CSD3) eluted with this ethanol concentration 40% aqueous solution was collected. The yield of this fraction was about 8.6 g as dry weight.
- the phenol content of this product by the vanillin-hydrochloric acid method was 54% [based on ( ⁇ )-epicatechin], and it was confirmed that it mainly contained phenolic substances.
- Viral adsorption inhibitory effect, vRNA synthesis inhibitory effect, mRNA synthesis inhibitory effect, and cRNA synthesis inhibitory effect by CSD3 were extracted from viral RNA and analyzed by RT-PCR.
- a sample stock solution was prepared by dissolving CSD3 in 50% ethanol to 50 mg / ml.
- Sample stock solutions are appropriately diluted with Tris-Glucose-Saline (TGS) (from 10 2 to 10 6- fold dilution), and 50 ⁇ l of these sample dilutions and virus solution (4 ⁇ 10 4 to 4 ⁇ 10 6 PFU (place forming unit) ) / Ml) 50 ⁇ l was mixed and allowed to react at room temperature for 10 minutes.
- TGS Tris-Glucose-Saline
- 0.1 ml of this was inoculated into a single-layer culture (6-well plate) of Madin-Darby canine kidney (MDCK) cells to which 0.4 ml of TGS was added, and the virus was adsorbed at room temperature for 30 minutes. After removing 0.5 ml of the culture solution and culturing at 37 ° C. for 0, 1, 2, 3, 4, 6, 8, 12 hours, the viral genomic RNA in the infected cells was dissolved in guanidine thiocyanate 25% It was recovered by ethanol precipitation. Details of the cDNA synthesis method and the PCR amplification method from RNA in each test method are shown below.
- RNA a reverse transcriptase reaction was carried out using Super Script III (Invitrogen) reverse transcriptase and T7 cRNA (1-12) primer according to the attached instructions to synthesize vRNA cDNA.
- PCR amplification was performed with a set of oligonucleotide primers specific to the virus (Udorn) NP gene. PCR was performed for 45 cycles (5 seconds, 95 ° C., 34 seconds, 64 ° C.) using SYBR Premix taq polymerase II (Invitrogen).
- the number of vRNA copies per hole obtained is shown in FIG.
- the adsorption copy number of CSD3 untreated virus at 0 hour was 6.7 ⁇ 10 8 / hole, but it was 2.2 ⁇ 10 8 / hole by 1 ⁇ g / ml CSD3 treatment, and adsorption was suppressed to about 1/3.
- the amount of vRNA synthesized by the CSD3 untreated virus was 2.2 ⁇ 10 10 / well (after 12 hours of culture), whereas that of CSD3 treated at 1 ⁇ g / ml was 3.2 ⁇ 10 8 / well, and the amount of vRNA synthesized was about Reduced to 1/70.
- RNA synthesis inhibition test In the virus primary / secondary transcription inhibition test by CSD3, reverse transcription was performed using reverse transcriptase and T7T (18) VN primer to synthesize mRNA cDNA. PCR was amplified with a set of oligonucleotide primers specific to the NP gene using the synthesized cDNA as a template. PCR was performed for 45 cycles (5 seconds, 95 ° C., 34 seconds, 64 ° C.) using SYBR Premix taq polymerase II (Invitrogen).
- the number of mRNA copies per hole obtained is shown in FIG.
- the copy number of mRNA of CSD3 untreated virus was 2.6 ⁇ 10 10 / well (after 12 hours of culture), whereas that of CSD3 treatment at 1 ⁇ g / ml was 4.3 ⁇ 10 7 / well, and the amount of mRNA synthesis was about Reduced to 1/600.
- CRNA synthesis inhibition test In the viral replication stage suppression test using CSD3, reverse transcription was performed using reverse transcriptase and T7 vRNA (1-13) primer to synthesize cRNA cDNA. PCR was amplified with a set of oligonucleotide primers specific to the NP gene using the synthesized cDNA as a template. PCR was performed for 45 cycles (5 seconds, 95 ° C., 34 seconds, 64 ° C.) using SYBR Premix taq polymerase II (Invitrogen). The number of copies of cRNA obtained for each well is shown in FIG.
- CSD3 untreated virus cRNA copy number was 9.7 ⁇ 10 6 / well (after 8 hours of culture), whereas 1 ⁇ g / ml CSD3 treatment was 2.5 ⁇ 10 4 / well, and the amount of cRNA synthesis was about Reduced to 1/400.
- CSD3 was added to the virus (16000HA) and kept at room temperature for 1 hour. Observation of the virus particles with an electron microscope was performed by a negative staining method with 2% uranium acetate.
- FIG. 1 is an electron microscopic image showing the disrupting activity of a karin extract (CSD3) on the virus envelope membrane, (a) shows the case of adding the karin extract, and (b) shows no addition of the karin extract. Show the case.
- Prescription ethanol of spray liquid 25.0% by weight Citric acid 1.5% by weight Trisodium citrate 1.0 wt% CSD3 0.5 wt% Water remaining 100.0% by weight
- Prescription sugar for tablet candy 76.1% by weight Glucose 19.0% by weight Sucrose fatty acid ester 0.2% by weight Fragrance 0.2 wt% CSD3 0.5 wt% Water remaining 100.0% by weight
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- Botany (AREA)
- Engineering & Computer Science (AREA)
- Mycology (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Virology (AREA)
- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
- Medical Informatics (AREA)
- Alternative & Traditional Medicine (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Pulmonology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Description
カリンの乾燥粉砕物を50%EtOH抽出した後ダイヤイオンHP-20 を用いて分画し、5つの分画物(CSD1~5)を得た。これらの分画物のうち,最も活性の高かったCSD3(40%EtOH溶出画分)を試料とした。 (Preparation of Karin extract)
The dry pulverized product of Karin was extracted with 50% EtOH and then fractionated using Diaion HP-20 to obtain five fractions (
吸着抑制試験およびvRNA合成抑制試験では、回収した0.5~5
μgのRNAから、添付説明書に従って、Super Script III(インビトロジェン社)逆転写酵素およびT7 cRNA(1-12)プライマーを用いて逆転写反応を行い、vRNAのcDNAを合成した。合成したcDNAを鋳型として、ウイルス(Udorn)NP遺伝子特異的な1組のオリゴヌクレオチドプライマーによりPCRの増幅を行った。PCRは、SYBR Premix taq polymeraseII(インビトロジェン社)を用いて45サイクル(5秒、95℃、34秒、64℃)実施した。 (Influenza virus adsorption inhibition test and vRNA synthesis inhibition test)
In the adsorption inhibition test and the vRNA synthesis inhibition test, the collected 0.5 to 5
From the μg of RNA, a reverse transcriptase reaction was carried out using Super Script III (Invitrogen) reverse transcriptase and T7 cRNA (1-12) primer according to the attached instructions to synthesize vRNA cDNA. Using the synthesized cDNA as a template, PCR amplification was performed with a set of oligonucleotide primers specific to the virus (Udorn) NP gene. PCR was performed for 45 cycles (5 seconds, 95 ° C., 34 seconds, 64 ° C.) using SYBR Premix taq polymerase II (Invitrogen).
CSD3によるウイルスの一次/二次転写抑制試験では、逆転写酵素およびT7T(18)VNプライマーを用いて逆転写反応を行い、mRNAのcDNAを合成した。合成したcDNAを鋳型として、NP遺伝子特異的な1組のオリゴヌクレオチドプライマーによりPCRの増幅を行った。PCRは、SYBR Premix taq polymeraseII(インビトロジェン社)を用いて45サイクル(5秒、95℃、34秒、64℃)実施した。 (MRNA synthesis inhibition test)
In the virus primary / secondary transcription inhibition test by CSD3, reverse transcription was performed using reverse transcriptase and T7T (18) VN primer to synthesize mRNA cDNA. PCR was amplified with a set of oligonucleotide primers specific to the NP gene using the synthesized cDNA as a template. PCR was performed for 45 cycles (5 seconds, 95 ° C., 34 seconds, 64 ° C.) using SYBR Premix taq polymerase II (Invitrogen).
CSD3によるウイルスの複製段階抑制試験では、逆転写酵素およびT7 vRNA(1-13)プライマーを用いて逆転写反応を行い、cRNAのcDNAを合成した。合成したcDNAを鋳型として、NP遺伝子特異的な1組のオリゴヌクレオチドプライマーによりPCRの増幅を行った。PCRは、SYBR Premix taq polymeraseII(インビトロジェン社)を用いて45サイクル(5秒、95℃、34秒、64℃)実施した。得られた各穴あたりのcRNAのコピー数を図2に示す。CSD3未処理ウイルスのcRNAのコピー数は9.7×106/穴(培養8時間後)であったのに対し、1μg/mlのCSD3処理では2.5×104/穴であり、cRNA合成量は約1/400に減少した。 (CRNA synthesis inhibition test)
In the viral replication stage suppression test using CSD3, reverse transcription was performed using reverse transcriptase and T7 vRNA (1-13) primer to synthesize cRNA cDNA. PCR was amplified with a set of oligonucleotide primers specific to the NP gene using the synthesized cDNA as a template. PCR was performed for 45 cycles (5 seconds, 95 ° C., 34 seconds, 64 ° C.) using SYBR Premix taq polymerase II (Invitrogen). The number of copies of cRNA obtained for each well is shown in FIG. CSD3 untreated virus cRNA copy number was 9.7 × 10 6 / well (after 8 hours of culture), whereas 1 μg / ml CSD3 treatment was 2.5 × 10 4 / well, and the amount of cRNA synthesis was about Reduced to 1/400.
CSD3を5%エタノールに溶解(5 mg/ml)したものを試料原液とした。インフルエンザウイルス(10HAから320HA)とCSD3(試料原液をTGSで102から106倍希釈)を、10分間室温下でインキュベートした。反応後、CSD3で処理したインフルエンザウイルスと10%ニワトリ赤血球100μlを混合し、30分間室温で保持した。赤血球を遠心分離して集め、沈殿をpH5.3の緩衝液(136mM NaCl、2.68mM KCl、10mM CH3COONa)に懸濁させた。懸濁液を37℃で30分間インキュベート後、遠心分離した。上清の409nmの吸光度を測定し、カリン未処理ウイルスの吸光度を1としカリン添加による溶血活性を評価した。 (Hemolysis suppression test)
CSD3 dissolved in 5% ethanol (5 mg / ml) was used as a sample stock solution. Influenza virus (10 HA to 320 HA) and CSD3 (sample stock solution diluted 10 2 to 10 6 times with TGS) were incubated for 10 minutes at room temperature. After the reaction, influenza virus treated with CSD3 and 100 μl of 10% chicken erythrocytes were mixed and kept at room temperature for 30 minutes. Red blood cells were collected by centrifugation, and the precipitate was suspended in pH 5.3 buffer (136 mM NaCl, 2.68 mM KCl, 10 mM CH 3 COONa). The suspension was incubated at 37 ° C. for 30 minutes and then centrifuged. The absorbance at 409 nm of the supernatant was measured, and the hemolytic activity by adding karin was evaluated with the absorbance of the untreated virus as 1.
カリン抽出物(CSD3)処理ウイルスでは一次転写より前段階でウイルス増殖が停止することが示された。ウイルスの吸着・膜融合段階での失活も示されたが、それ以上にRNAの合成が抑制されており、膜融合から一次転写までの過程に関わる何らかの欠陥があることが示唆された。電子顕微鏡で示されたエンベロープの損傷がこの欠陥の原因となっている可能性がある。 (Conclusion)
It was shown that the virus growth was stopped at the stage before primary transcription in the kalin extract (CSD3) -treated virus. Inactivation at the stage of virus adsorption and membrane fusion was also shown, but RNA synthesis was further suppressed, suggesting that there was some defect related to the process from membrane fusion to primary transcription. Damage to the envelope shown in the electron microscope may be responsible for this defect.
エタノール 2.0 重量%
香料 1.0 重量%
サッカリン 0.05 重量%
塩酸クロルヘキシジン 0.01 重量%
CSD3 0.5 重量%
水 残
100.0 重量% Prescription ethanol for mouthwash 2.0% by weight
Fragrance 1.0% by weight
Saccharin 0.05 wt%
Chlorhexidine hydrochloride 0.01% by weight
CSD3 0.5 wt%
Water remaining
100.0% by weight
エタノール 5.0 重量%
CSD3 1.0 重量%
水 残
100.0 重量% Prescription ethanol for inhalation 5.0 wt%
CSD3 1.0 wt%
Water remaining 100.0% by weight
ブドウ糖 72.3 重量%
乳糖 19.0 重量%
アラビアゴム 6.0 重量%
香料 1.0 重量%
モノフルオロリン酸ナトリウム 0.7 重量%
CSD3 1.0 重量%
100.0 重量% Lozenge prescription glucose 72.3% by weight
Lactose 19.0% by weight
Gum arabic 6.0% by weight
Fragrance 1.0% by weight
Sodium monofluorophosphate 0.7% by weight
CSD3 1.0 wt%
100.0% by weight
エタノール 25.0 重量%
クエン酸 1.5 重量%
クエン酸三ナトリウム 1.0 重量%
CSD3 0.5 重量%
水 残
100.0 重量% Prescription ethanol of spray liquid 25.0% by weight
Citric acid 1.5% by weight
Trisodium citrate 1.0 wt%
CSD3 0.5 wt%
Water remaining 100.0% by weight
ガムベース 20.0 重量%
砂糖 54.7 重量%
グルコース 15.0 重量%
水飴 9.3 重量%
香料 0.5 重量%
CSD3 0.2 重量%
100.0 重量% Chewing gum formula gum base 20.0% by weight
54.7% by weight sugar
Glucose 15.0% by weight
Minamata 9.3 wt%
Fragrance 0.5% by weight
CSD3 0.2 wt%
100.0% by weight
砂糖 50.0 重量%
水飴 34.0 重量%
クエン酸 1.0 重量%
香料 0.2 重量%
CSD3 0.4 重量%
水 残
100.0 重量% Candy prescription sugar 50.0% by weight
Minamata 34.0 wt%
Citric acid 1.0% by weight
Fragrance 0.2 wt%
CSD3 0.4 wt%
Water remaining 100.0% by weight
砂糖 76.1 重量%
グルコース 19.0 重量%
ショ糖脂肪酸エステル 0.2 重量%
香料 0.2 重量%
CSD3 0.5 重量%
水 残
100.0 重量% Prescription sugar for tablet candy 76.1% by weight
Glucose 19.0% by weight
Sucrose fatty acid ester 0.2% by weight
Fragrance 0.2 wt%
CSD3 0.5 wt%
Water remaining 100.0% by weight
オレンジ果汁 30.00 重量%
異性化糖 15.24 重量%
クエン酸 0.10 重量%
ビタミンC 0.04 重量%
香料 0.10 重量%
CSD3 0.10 重量%
水 残
100.00 重量% Beverage prescription orange juice 30.00% by weight
Isomerized sugar 15.24% by weight
Citric acid 0.10% by weight
Vitamin C 0.04% by weight
Perfume 0.10% by weight
CSD3 0.10 wt%
Water remaining 100.00% by weight
トウモロコシ澱粉 55.0 重量%
カルボキシセルロース 40.0 重量%
CSD3 5.0 重量%
100.0 重量% Formulation of powdered corn starch 55.0% by weight
Carboxycellulose 40.0% by weight
CSD3 5.0 wt%
100.0% by weight
ラクトース 70.0 重量%
結晶性セルロース 15.0 重量%
ステアリン酸マグネシウム 5.0 重量%
CSD3 10.0 重量%
100.0 重量% Tablet prescription lactose 70.0% by weight
Crystalline cellulose 15.0% by weight
Magnesium stearate 5.0 wt%
CSD3 10.0 wt%
100.0% by weight
エタノール 2.00 重量%
香料 1.00 重量%
サッカリン 0.05 重量%
塩酸クロルヘキシジン 0.01 重量%
CSD3 0.50 重量%
水 残
100.00 重量% Prescription of gargle ethanol 2.00% by weight
Perfume 1.00% by weight
Saccharin 0.05 wt%
Chlorhexidine hydrochloride 0.01% by weight
CSD3 0.50 wt%
Water remaining 100.00% by weight
ゼラチン 60.00 重量%
水飴 23.00 重量%
砂糖 8.50 重量%
植物油脂 4.50 重量%
マンニトール 2.95 重量%
レモン果汁 1.00 重量%
CSD3 0.05 重量%
100.00 重量% Gummy jelly prescription gelatin 60.00 wt%
Minamata 23.00 wt%
Sugar 8.50% by weight
Vegetable oil and fat 4.50% by weight
Mannitol 2.95% by weight
Lemon juice 1.00% by weight
CSD3 0.05 wt%
100.00% by weight
This application claims priority from Japanese Patent Application No. 2009-218643 filed on September 24, 2009 and Japanese Patent Application No. 2009-278462 filed on December 8, 2009. The contents of which are incorporated herein by reference.
Claims (9)
- カリン(Chaenomeles sinensis, Pseudocydonia sinensis)の抽出物を有効成分とすることを特徴とする抗インフルエンザウイルス剤。 An anti-influenza virus agent characterized by comprising an extract of Chaenomeles sinensis, Pseudocydonia sinensis as an active ingredient.
- カリンを50%エタノール水を用いて、抽出し、その抽出液をカラム分画して精製することにより得た抽出物を有効成分とすることを特徴とする抗インフルエンザウイルス剤。 An anti-influenza virus agent comprising, as an active ingredient, an extract obtained by extracting salmon carin with 50% ethanol water, and subjecting the extract to column fractionation and purification.
- カリンを50%エタノール水を用いて、抽出し、その抽出液をカラム分画して精製することにより得た抽出物を有効成分とすることを特徴とするインフルエンザウイルス吸着抑制剤。 An influenza virus adsorption inhibitor characterized by extracting karin with 50% ethanol water and using an extract obtained by fractionating and purifying the extract as a column.
- カリンを50%エタノール水を用いて、抽出し、その抽出液をカラム分画して精製することにより得た抽出物を有効成分とすることを特徴とするインフルエンザウイルス感染細胞におけるウイルスmRNA合成抑制剤。 Inhibitor of viral mRNA synthesis in influenza virus-infected cells, characterized in that an extract obtained by extracting karin with 50% ethanol water and purifying the extract by column fractionation is used as an active ingredient .
- カリンを50%エタノール水を用いて、抽出し、その抽出液をカラム分画して精製することにより得た抽出物を有効成分とすることを特徴とするインフルエンザウイルス感染細胞におけるウイルスvRNA合成抑制剤。 Inhibitor of viral vRNA synthesis in influenza virus-infected cells, characterized in that an extract obtained by extracting karin with 50% ethanol water and purifying the extract by column fractionation is used as an active ingredient .
- カリンを50%エタノール水を用いて、抽出し、その抽出液をカラム分画して精製することにより得た抽出物を有効成分とすることを特徴とするインフルエンザウイルス感染細胞におけるウイルスcRNA合成抑制剤。 An inhibitor of viral cRNA synthesis in influenza virus-infected cells, characterized in that an extract obtained by extracting karin with 50% ethanol water and purifying the extract by column fractionation is used as an active ingredient .
- カリンを50%エタノール水を用いて、抽出し、その抽出液をカラム分画して精製することにより得た抽出物を有効成分とすることを特徴とするウイルス膜融合活性試験における溶血抑制剤。 A hemolytic inhibitor in a viral membrane fusion activity test, characterized in that an extract obtained by extracting karin with 50% ethanol water and purifying the extract by column fractionation is used as an active ingredient.
- カリンを50%エタノール水を用いて、抽出し、その抽出液をカラム分画して精製することにより得た抽出物を有効成分とすることを特徴とするインフルエンザウイルスのエンベロープ破壊剤。 An influenza virus envelope disrupting agent comprising an extract obtained by extracting karin with 50% ethanol water and purifying the extract by column fractionation.
- カリン抽出物を含有する抗インフルエンザウイルス作用を有する飲食品。 Food / beverage products having anti-influenza virus action, containing karin extract.
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2010800432467A CN102573866A (en) | 2009-09-24 | 2010-09-24 | Anti-influenza virus agent |
JP2011532910A JPWO2011036883A1 (en) | 2009-09-24 | 2010-09-24 | Anti-influenza virus agent |
KR1020127010366A KR20120087928A (en) | 2009-09-24 | 2010-09-24 | Anti-influenza virus agent |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2009-218643 | 2009-09-24 | ||
JP2009218643 | 2009-09-24 | ||
JP2009278462 | 2009-12-08 | ||
JP2009-278462 | 2009-12-08 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2011036883A1 true WO2011036883A1 (en) | 2011-03-31 |
Family
ID=43795649
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2010/005762 WO2011036883A1 (en) | 2009-09-24 | 2010-09-24 | Anti-influenza virus agent |
Country Status (5)
Country | Link |
---|---|
JP (1) | JPWO2011036883A1 (en) |
KR (1) | KR20120087928A (en) |
CN (1) | CN102573866A (en) |
TW (1) | TW201125571A (en) |
WO (1) | WO2011036883A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103501798A (en) * | 2011-04-26 | 2014-01-08 | 罗蒂株式会社 | Pandemic influenza antiviral agent |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005343836A (en) * | 2004-06-04 | 2005-12-15 | Lotte Co Ltd | Anti-influenza virus agent, and infection inhibiting article containing the same and food and drink |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN100544750C (en) * | 2006-10-13 | 2009-09-30 | 解会元 | Medicated wine composition |
-
2010
- 2010-09-24 JP JP2011532910A patent/JPWO2011036883A1/en active Pending
- 2010-09-24 CN CN2010800432467A patent/CN102573866A/en active Pending
- 2010-09-24 TW TW099132393A patent/TW201125571A/en unknown
- 2010-09-24 WO PCT/JP2010/005762 patent/WO2011036883A1/en active Application Filing
- 2010-09-24 KR KR1020127010366A patent/KR20120087928A/en not_active Application Discontinuation
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005343836A (en) * | 2004-06-04 | 2005-12-15 | Lotte Co Ltd | Anti-influenza virus agent, and infection inhibiting article containing the same and food and drink |
Non-Patent Citations (1)
Title |
---|
J. ETHNOPHARMACOL., vol. 118, 2008, pages 108 - 112 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103501798A (en) * | 2011-04-26 | 2014-01-08 | 罗蒂株式会社 | Pandemic influenza antiviral agent |
Also Published As
Publication number | Publication date |
---|---|
CN102573866A (en) | 2012-07-11 |
TW201125571A (en) | 2011-08-01 |
JPWO2011036883A1 (en) | 2013-02-14 |
KR20120087928A (en) | 2012-08-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
KR101139376B1 (en) | Anti influenza viral drug and, viral-infection suppression products and foods comprising the same | |
JP4216013B2 (en) | Anti-influenza virus agent | |
CA2721040C (en) | Procyanidin extracts of cinnamon, litchi and arachis useful in the treatment of retroviral infections | |
JP2011126834A (en) | Antiviral agent | |
WO2011036883A1 (en) | Anti-influenza virus agent | |
KR20210002378A (en) | Hepatoprotective composition containing pini pollen extract as effective component | |
US10136663B2 (en) | Curcuminoid-based compound/stevioside-containing complex for the prevention and treatment of an influenza virus infection | |
JP2009269861A (en) | Prophylactic and therapeutic agent for viral infection derived from cassis fruit | |
KR20220018953A (en) | Antivirus composition comprising Pteridium aquilinum extract or fraction thereof | |
WO2012147327A1 (en) | Pandemic influenza antiviral agent | |
US8603548B2 (en) | Anti-avian influenza virus agent, and product containing anti-avian influenza virus agent | |
JP5303482B2 (en) | Anti-influenza virus agent and influenza infection suppression product obtained by adsorbing, impregnating and adding the same | |
JP5343016B2 (en) | Anti-influenza virus agent and influenza infection suppression product obtained by adsorbing, impregnating and adding the same | |
JP2004002361A (en) | Anti-influenza virus agent | |
KR102676865B1 (en) | Composition for preventing, ameliorating or treating coronavirus infectious disease comprising herbal medicine extract as effective component | |
KR102578871B1 (en) | Composition for preventing, ameliorating or treating coronavirus infectious disease comprising herbal medicine extract as effective component | |
KR20240059530A (en) | Composition for preventing, ameliorating or treating coronavirus infectious disease comprising herbal medicine extract as effective component | |
TWI389700B (en) | A novel standardized composition, method of manufacture and use in the resolution of rna virus infection | |
JP4847712B2 (en) | Antioxidant composition containing acacia bark | |
KR101376206B1 (en) | Composition having antibacterial or antiviral activity containing sorghum natural colors extract or polyphenol compound separated from it | |
JP2007223970A (en) | Avian influenza virus infection inhibitor |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
WWE | Wipo information: entry into national phase |
Ref document number: 201080043246.7 Country of ref document: CN |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 10818560 Country of ref document: EP Kind code of ref document: A1 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2011532910 Country of ref document: JP Ref document number: 12012500575 Country of ref document: PH |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1201001291 Country of ref document: TH |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 20127010366 Country of ref document: KR Kind code of ref document: A |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 10818560 Country of ref document: EP Kind code of ref document: A1 |