Hotline:400-820-3792Inhibitors ? ScreeningLibraries ? Proteinswww.MedChemEIspinesibmesylateCat.No.:HY-50759ACASNo.:514820-03-2Synonyms:SB-715992mesylate;CK-0238273mesylate分子式:C??H??ClN?O?S分子量:613.17作用靶點:Kinesin;Apoptosis作用通路:CellCycle/DNADamage;Cytoskeleton;Apoptosis儲存方式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY生物活性Ispinesib(SB-715992)mesylate是一種特異性的紡錘體驅(qū)動蛋白KSP抑制劑，Kiapp值為1.7nM。IC50&TargetKSP1.7nM(Kiapp)體外研究Ispinesib(150nM)mesylateinhibitsBT-474andMDA-MB-468celllines,withGI50sof45and19nM,respectively[2].Ispinesib(SB715992,15and30nM)mesylatesuppressestheproliferationofPC-3prostatecancercellby48.65%and52.16%,andinducesapoptosisofprostatecancercellby1094.88%and1516.70%,respectively.Ispinesibmesylateupregulatesgenesresponsibleforapoptosisandcellcyclearrest,anddownregulatesgenesresponsibleforcellproliferationandsurvival.Theanti-proliferationandpro-apoptoticactivitiesofIspinesibmesylatecanbeenhancedbygenistein[3].體內(nèi)研究Ispinesib(SCID,8mg/kg;nude,10mg/kg,q4d×3)mesylatereducestumorvolumeinmicebearingtumorxenograftsofER-positive(MCF7),HER2-positive(KPL4,HCC1954,andBT-474),andtriple-negative(MDA-MB-468)breastcancercellsviai.p.onedoseevery4daysrepeatedthreetimes[2].PROTOCOLKinaseAssay[1]Kinesinspecificityanalysisiscarriedoutusingapyruvatekinase?lactatedehydrogenasedetectionsystemthatcouplestheproductionofADPtooxidationofNADH.Absorbancechangesaremonitoredat340nm.1/3 MasterofBioactiveMolecules—您身邊的抑制劑大師www.MedChemESteady-statestudiesusingnanomolarconcentrationsofKSPareperformedusingasensitivefluorescence-basedassayutilizingapyruvatekinase,pyruvateoxidase,andhorseradishperoxidasecoupleddetectionsystemthatcouplesthegenerationofADPtooxidationofAmplexRedtofluorescentresorufin.Generationofresorufinismonitoredbyfluorescence(λexcitation=520nmandλemission=580nm).Steady-statebiochemicalexperimentsareperformedinPEM25buffer[25mMPipes-K+(pH6.8),2mMMgCl2,1mMEGTA]supplementedwith10μMpaclitaxelforexperimentsinvolvingmicrotubules.TheIC50forsteady-stateinhibitionisdeterminedat500μMATP,5μMMTs,and1nMKSPinPEM25buffer.Kiapp(apparentinhibitordissociationconstant)estimatesofIspinesibareextractedfromtheconcentration-responsecurves,withexplicitcorrectionforenzymeconcentration[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[3]PC-3prostatecancercellsareseededin96wellplatesatadensityof4×103cells/well.PC-3cellsareincubatedfor24hourstoallowattachmenttothesurfaceofeachwellofthetissuecultureplate.Then,thecellsaretreatedwithvaryingconcentrationofreagentsandincubatedfor1to3days.First,PC-3cellsaretreatedwith15and30nMofIspinesib,respectively.Second,PC-3cellsaresubjectedtocombinationaltreatmentswith7.5or10nMofIspinesibplus30μMofgenistein.Finally,PC-3cellsarepre-treatedwith30μMofgenisteinfor24hoursfollowedbytreatmentwith15nMofIspinesib.Controlcellsaretreatedwith0.3mMNa2CO3(vehiclecontrol).Aftertreatment,PC3cellsareincubatedat37°CwithMTT(0.5mg/mL)for2hoursandisopropylalcoholatroomtemperaturefor1hour.ThespectrophotometricabsorbanceofeachsampleisthendeterminedbyusingULTRAMultifunctionalMicroPlateReaderat595nm[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMicewithatumorvolumeof-250mm3receiveasingledoseofIspinesib(10mg/kg).Tumorsaredissected,Administration[2]fixedin10%bufferedformalin,andembeddedinparaffin,and5-μmtissuesectionsareprepared.Antigenretrievalisdonebyboilingin50mMcitratebuffer(pH5.5),andsectionsarethenincubatedin3%hydrogenperoxide,washedinPBS-0.1%Tween,andblockedin10%goatserum.Phospho-histoneH3(PH3)antibodyisdetectedusingAlexaFluor488secondaryantibody.Imagesaretakenwithamicroscopeat×10magnificationandcapturedusingMetaMorphsoftwaretoquantifyPH3expressionbycomputingthearearatioofPH3-positivecellspertotalcells.Ki67/cleavedcaspase-3stainingisdone.NonfluorescentimagesaretakenonanOlympusBX41microscopeat×20magnification[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.客戶使用本產(chǎn)品發(fā)表的科研文獻RedoxBiol.2019Feb:21:101112.CancerLett.2021May28:506:1-10.Cancers(Basel).2024Nov5;16(22):3732.Gene.2025Apr3:955:149458.UrolOncol.2023May;41(5):253.e11-253.e20.Seemorecustomervalidationsonwww.MedChemEREFERENCES2/3 MasterofBioactiveMolecules—您身邊的抑制劑大師www.MedChemELadL,etal.MechanismofinhibitionofhumanKSPbyispinesib.Biochemistry.2008Mar18;47(11):3576-85.PurcellJW,etal.Activityofthekinesinspindleproteininhibitorispinesib(SB-715992)inmodelsofbreastcancer.ClinCancerRes.2010Jan15;16(2):566-76.DavisDA,etal.Increasedtherapeuticpotentialofanexperimentalanti-mitoticinhibitorSB715992bygenisteininPC-3human