毒胡萝卜素(TG)说明书

ThapsigarginCatalogNumberT9033StorageTemperature-20°CCASRN:67526-95-8Synonyms:Octanoicacidderivativeofazuleno[4,5-b]furan1DescriptionAppearance:SolidorclearfilminsidevialMolecularformula:C34H50O12Molecularweight:650.75AnHPLCmethodfortheseparationanddeterminationofthemainthapsigarginsfoundintheplanthasbeenreported.3Thestereochemistry,13CNMRspectraldataandtheX-rayanalysisfortheepoxidederivativehavebeenreported.4,5PrecautionsandDisclaimerThisproductisforR&Duseonly,notfordrug,household,orotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.MethodOfPreparationThapsigargin(TG)isextractedfromtheplantThapsiagarganicaL.withmethanolandpurifiedbyHPLC.6Amethodofisolationfromtheplantextracthasbeenreported.7Storage/StabilityTheproductshouldbestableforatleastoneyearwhenstoredat–20°C.6Solubility/SolutionStability:TGissolubleinDMSO8andinethanol9.NoinitialconcentrationsofTGweregivenforeithersolventbutitwasnotedthatthefinalconcentrationswere:inDMSO,lessthan1%(v/v)8;ethanol,didnotexceed1%ofthetotalvolume(workingconcentrationsinethanolwerefrom70-140μM).9Neitheramountofsolventhadanyadverseeffectsinthedescribedsystems.Preparationofa3%(w/v)solutionindeuteratedDMSOwasdescribed.4TGhasbeensolubilizedinacetonitrileat10mg/mlproducingaclear,colorlesssolution.2TGisalsosolubleinmethanol,diethylether,andmethylenechloride.6Solutionsarerecommendedtobepreparedfresh.6However,ifnecessary,solutionscanbestoredat–20°Cunderargonornitrogen.6A1mMsolutioninDMSOwasreportedlystoredat–20°C(nostoragetimegiven).10Usage/Applications:TGisaneffectiveinhibitoroftheCa2+ionpumpproteinsofintracellularmembraneslocatedinsarcoplasmicreticulum(SR)andendoplasmicreticulum(ER)ofskeletal,cardiac,muscleandbrainmicrosomes.9,11Aconcentrationof100nMwaseffectiveininhibitingtheSRCa2+-ATPasefromcardiacandskeletalmuscles.TheactionofTGisrestrictedtoenzymesintheSRpumpinintactcardiacmyocytes.12TGinteractsstronglywiththeSRofCa2+-transportATPasetoforma1:1stoichiometriccomplex.TheproposedmechanismofinhibitionofCa2+-ATPaseactivityincludingtheinhibitionofCa2+ionbindingandenzymephosphorylationhavebeenreported.13,14,15TGreleasesintracellularCa2+inrathepatocytesbyspecificinhibitionofamicrosomal(ER)isoformofCa2+-ATPaseenzyme.16TG,acellpermeantagent,activatesplatelets(at1μM)andthisismediatedbyanincreaseinthecytoplasmicfreecalciumionlevelprobablythroughstimulationofthepassivecalciumtransportthroughspecificchannels(asshownbyproteinphosphorylation).8,17,18TGisapotentactivatorofdifferentcelltypesinvolvedintheinflammatoryprocess.19,20Thecompoundhasalsobeenshowntopromotesecond-stagecarcinogenesisinmouseskinwhichdoesnotinvolvedirectactivationofproteinOOCOCCH3CCH3HH3COHOOHCH3CCH3OCH3OCOCH2CH2CH3OCH2CH2CH2CH2CH2CH2H3COOHkinaseC(asTPAdoes*)thissuggeststhatitisanon-TPAtypetumorpromoter.21Theincubationofisolatedratadipocyteswith100nMTGincreasestheintracellularcytosolicCa2+ioncontent2-3-fold.TGreducedtheinsulin-stimulatedglucosetransportbyabout40%.22Proteinsynthesisincellswasinhibitedupto10nMofTG.23TG,upto300nM,activatesanon-selectiveunivalentandbivalentSK&F96365-andGd3+-sensitivecationentrypathwayinhumanneutrophils.10At200nM,TGincreasesthesodium-dependentphosphateuptakeinHeLacellsbymobilizationofintracellularcalciumindependentofproteinkinaseC.24Additionalstudiesreportthatinositoltriphosphate(IP3)andTGdiscriminateERstoresofCa2+ionsinratbrainandthatCa2+ioninfluxinhumanTlymphocytesisinducedindependentlyofIP3bymobilizationofintracellularCa2+ionstores.25,26At10ng/ml,TGstimulatesarachidonicacidmetabolisminratperitonealmacrophagesbyactingasaselectiveCa2+ionmobilizerwithoutactivatingproteinkinaseC.27TGmobilizesintracellularCa2+byanIP3-independentmechanisminparotidacinarcellsat2μM.28TGat1.7μMhasbeenshowntoreleaseCa2+ionsfromanintracellularIP3-sensitiveCa2+ionstore,inaneuronalcelllinebyamechanismindependentofthehydrolysisofphosphoinositidesandactivationofproteinkinaseC.29StudiesontheeffectofTGonvascularcontractilityconcludedthatTGinhibitstheabilityofcAMPtostimulateCa2+ionuptakeintothestorebyblockingitsCa2+ionpump.30GeneralNotes:TGisatumorpromotingplantsesquiterpenelactoneextractwithauniquebiologicalactivityasCa2+-ATPaseinhibitorsinanimalcells.9,11,12,15,16,18Itisaskinirritant,aplateletactivating17,inflammatory19,20andtumorpromoting21agent.TGistoxicandapossiblecarcinogen.References1.ChemicalAbstractsRegistrydata,AmericanChemicalSociety.2.SigmaQualityControlData3.Jager,A.K.etal.,J.Chromatogr.,634,135(1993).4.Christensen,S.B.,andSchaumburg,K.,J.Org.Chem.,48,396,(1983).5.Christensen,S.B.,etal.,J.Org.Chem.,47,649,(1982).6.Supplierinformation7.Christensen,S.B.,etal.,TetrahedronLett.,21,3829,(1980).8.Thastrup,O.,etal.,Biochem.Biophys.Res.Commun.,142,654,(1987).9.Kijima,Y.,etal.,J.Biol.Chem.,266,22912,(1991).10.Wenzel-Seifert,K.,etal.,Biochem.J.,314,679,(1996).11.Lytton,J.,etal.,J.Biol.Chem.,266,17067,(1991)12.RogersT.B.,etal.,Biosc.,Rep.,15,341,(1995).13.Sagara,Y.,etal.,J.Biol.Chem.,267,12606,(1992).14.Sagara,Y.,etal.,J.Biol.Chem.,267,1286,(1992).15.Inesi,G.,andSagara,Y