Design and in Vitro Characterization of Tricyclic Benzodiazepine Derivatives as Potent and Selective Antileukemic Agents

Authors:

• Adam Mieczkowski,
• T. Frączyk,
• Mateusz Psurski,
• Patrycja Wińska,
• Paweł Siedlecki,
• Monika Dziełak,
• Damian Trzybiński,
• Marcin Wilczek,
• Maciej Bagiński,
• Bartosz Bieszczad,
• Krzysztof Woźniak

Abstract

DesignandinVitroCharacterizationofTricyclicBenzodiazepineDerivativesasPotentandSelectiveAntileukemicAgentsAdamMieczkowski,*aTomaszFrączyk,a, bMateuszPsurski,cPatrycjaWińska,dPawełSiedlecki,aMonikaDziełak,aDamianTrzybiński,eMarcinWilczek,fMaciejBagiński,a, fBartoszBieszczad,aandKrzysztofWoźniakeaInstituteof Biochemistryand Biophysics,PolishAcademyof Sciences,Pawinskiego5a, 02-106Warsaw,Poland,e-mail:amiecz@ibb.waw.plbMedicalUniversityof Warsaw,Nowogrodzka59, 02-006Warsaw,PolandcHirszfeldInstituteof Immunologyand ExperimentalTherapy,PolishAcademyof Sciences,R. Weigla12,53-114Wroclaw,PolanddFacultyof Chemistry,WarsawUniversityof Technology,Noakowskiego3, 00-664Warsaw,PolandeBiologicaland ChemicalResearchCentre,Universityof Warsaw,Żwirkii Wigury101, 02-089Warsaw,PolandfFacultyof Chemistry,Universityof Warsaw,Pasteura1, 02-093Warsaw,PolandCurrentlyavailablechemotherapeutictreatmentsfor bloodcancers(leukemia)usuallyhave strongsideeffects.Moreselective,efficient,and less toxic anticanceragentsare needed.We synthesizedseven,new,opticallypure (12aS)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dionederivativesand examinedtheircytotoxicitytowardseight cancercell lines, includingurinarybladder(TCC-SUP,UM-UC-3,KU-19-9),colon (LoVo),and breast(MCF-7,MDA-MB-231)cancerrepresentatives,as well as two leukemiccell lines (MV-4-11,CCRF-CEM)and normalmurinefibroblasts(Balb/3T3)as referencecell line. Threeof the sevennewly-obtainedcompounds((12aS)-8-bromo-2-(3-phenylbenzoyl)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dione,(12aS)-8,9-dimethoxy-2-(4-phenylbenzoyl)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dioneand (12aS)-8-nitro-2-(4-phenylbenzoyl)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dione,showedenhancedactivityand selectiv-ity towardthe leukemicMV-4-11cell lines whencomparedto our previouslyreportedcompounds,with IC50valuesin the rangeof 2.9–5.6μM. Additionally,(12aS)-9-nitro-2-(4-phenylbenzoyl)-1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dioneexhibiteda strongcytotoxiceffectagainstthe leukemicCCRF-CEM(IC50=6.1μM)and MV-4-11(IC50=11.0μM) cell lines, a moderatecytotoxiceffecttowardothertumorlines (IC50=31.8–55.0μM) and very weak cytotoxiceffecttowardthe Balb/3T3referencecell lines. Selectedcompoundswerefurtherevaluatedfor their potentialto induceapoptoticcell deathin MV-4-11cells by measuringcaspase-3activity.We also establishedthe crystalstructureof three productsand investigatedthe effectof 22 derivativesof 1,3,4,12a-tetrahydropyrazino[2,1-c][1,4],12(2H,11H)-dioneon the activityof the cancer-associatedenzymeautotaxin.All compoundsprovedto be weak inhibitorsof autotaxin,althoughsome (R) and (S) enantiomershadKivaluesof 10–19μM. The obtainedresultsshowedthat the testedcompoundsexhibiteda selectiveantileukemiceffect,whichappearednot to be relateddirectlyto autotaxin.Moleculartargetsresponsiblefor thiseffectremainto be identified.The newlyobtainedcompoundscan be used in the searchfor new, selectiveanticancertherapies.