Synonym:CE6;ChlorinE6;chlorine6.

IUPAC/ChemicalName:2-[(7S,8S)-3-Carboxy-7-(2-carboxyethyl)-13-ethenyl-18-ethyl-7,8-dihydro-2,8,12,17-tetramethyl-21H,23H-porphin-5-yl]aceticacid

InChiKey:VAJLRIOJDADNAT-HHGNVTQFSA-N

InChiCode:InChI=1S/C34H36N4O6/c1-7-19-15(3)23-12-25-17(5)21(9-10-29(39)40)32(37-25)22(11-30(41)42)33-31(34(43)44)18(6)26(38-33)14-28-20(8-2)16(4)24(36-28)13-27(19)35-23/h7,12-14,17,21,35,38H,1,8-11H2,2-6H3,(H,39,40)(H,41,42)(H,43,44)/b23-12-,24-13-,25-12-,26-14-,27-13-,28-14-,32-22-,33-22-/t17-,21-/m0/s1

SMILESCode:O=C(O)C/C1=C2[C@@H](CCC(O)=O)[C@H](C)C(/C=C3C(C)=C(C=C)/C(N/3)=C/C(C(C)=C/4CC)=NC4=C/C5=C(C)C(C(O)=O)=C1N5)=N2

TechnicalData

AdditionalInformation

Photodynamictherapy(PDT)iscurrentlybeingusedasanalternativetherapeuticmodalityforavarietyofmalignanttumors.Thisstudywasperformedtoshowanefficientpreparationofsecondgenerationofphotosensitizerchlorine6(Ce6)withhighyieldandpurity,andtotestantitumoractivityofCe6-inducedPDT(Ce6-PDT)bothinvitroandinvivousingarattumormodel.Three-week-oldmaleSprague-Dawley(SD)ratswereinoculateds.c.ontherightflankwith5x106RK3E-rascells.Theanimalswereadmin-isteredi.v.withCe6(10mg/kg)and24hlater,PDTwasperformedusingalaserdiodeatalightdoseof100J/cm2.Ce6-PDTgeneratedreactiveoxygenspeciesandledtosignificantgrowthinhibitioninRK3E-rascell.Inaddition,Ce6-PDTinducedapoptosisthroughtheactivationofcaspase-3anditsdownstreamtarget,PARPcleavage.Theproteinlevelofanti-apoptoticbcl-2wasalsoreducedbyCe6-PDTinRK3E-rascells.Ininvivoexperiments,applicationofCe6-PDTledtoasignificantreductionoftumorsize.PCNAimmunostainingandTUNELassayrevealedthatCe6-PDTinhibitedtumorcellproliferationandincreasedapoptosis.ThesefindingssuggestthatthenewlypurifiedCe6-PDTcaneffectivelyarresttumorgrowthbyinhibitingcellproliferationandinducingapoptosis. (source:OncologyReports,2009,22(5).1085-1091.Doi:10.3892/or_00000540.Efficientpreparationofhighlypurechlorine6anditsphotodynamicanti-canceractivityinarattumormodel.Authors:Yeon-HeeMoon,Seong-MinKwon,Hyo-JunKim,Kwan-YoungJung,Jong-HwanPark,Soo-AKim,Yong-ChulKim,Sang-GunAhn,Jung-HoonYoon.Affiliations:DepartmentofPathology,SchoolofDentistry,ChosunUniversity,Gwangju500-759,Korea,http://www.spandidos-publications.com/or/22/5/1085).
Photodynamictherapy(PDT)iscurrentlybeingusedasanalternativetherapeuticmodalityforavarietyofmalignanttumors.Thisstudywasperformedtoshowanefficientpreparationofsecondgenerationofphotosensitizerchlorine6(Ce6)withhighyieldandpurity,andtotestantitumoractivityofCe6-inducedPDT(Ce6-PDT)bothinvitroandinvivousingarattumormodel.Three-week-oldmaleSprague-Dawley(SD)ratswereinoculateds.c.ontherightflankwith5x106RK3E-rascells.Theanimalswereadmin-isteredi.v.withCe6(10mg/kg)and24hlater,PDTwasperformedusingalaserdiodeatalightdoseof100J/cm2.Ce6-PDTgeneratedreactiveoxygenspeciesandledtosignificantgrowthinhibitioninRK3E-rascell.Inaddition,Ce6-PDTinducedapoptosisthroughtheactivationofcaspase-3anditsdownstreamtarget,PARPcleavage.Theproteinlevelofanti-apoptoticbcl-2wasalsoreducedbyCe6-PDTinRK3E-rascells.Ininvivoexperiments,applicationofCe6-PDTledtoasignificantreductionoftumorsize.PCNAimmunostainingandTUNELassayrevealedthatCe6-PDTinhibitedtumorcellproliferationandincreasedapoptosis.ThesefindingssuggestthatthenewlypurifiedCe6-PDTcaneffectivelyarresttumorgrowthbyinhibitingcellproliferationandinducingapoptosis. (source:OncologyReports,2009,22(5).1085-1091.
Doi:10.3892/or_00000540.
Efficientpreparationofhighlypurechlorine6anditsphotodynamicanti-canceractivityinarattumormodel.Authors:Yeon-HeeMoon,Seong-MinKwon,Hyo-JunKim,Kwan-YoungJung,Jong-HwanPark,Soo-AKim,Yong-ChulKim,Sang-GunAhn,Jung-HoonYoon.Affiliations:DepartmentofPathology,SchoolofDentistry,ChosunUniversity,Gwangju500-759,Korea,http://www.spandidos-publications.com/or/22/5/1085).
    
    
MethodstomakeChlorinE6
makeChlorinE6
ChlorinE6isanaturalproduct,andusuallycanbemadefromLivechlorella(Chlorellaellipsoidea,seethefollowingpictures)
ChlorinE6isanaturalproduct,andusuallycanbemadefromLivechlorella(Chlorellaellipsoidea,seethefollowingpictures)
 
 
    
    
  
  
ThefollowingprocedurewasreportedinOncologyReports,2009,22(5).1085-1091byKoreanscientistsYeon-HeeMoonetal. Livechlorella(Chlorellaellipsoidea)100g(driedweight)wassequentiallywashedwith500mlofwaterand300mlof50%ethanolinwatertoremovepolarmaterialsandtheresiduewasextractedtwicewith500mlof100%ethanoltoobtainchlorophyllarichfraction(extractionyield4.3%).Stirringthecombinedethanolsolutionofchlorophyllain1NHCl(pH2.5)for3hatroomtemperatureaffordedpheophytinintheformofprecipitates.Theprecipitatewasdissolvedindichloromethanewashedwithdistilledwater,driedwithanhydroussodiumsulfate,androtary-evaporatedtodryness.Theresiduewaspurifiedbyachromatographyusingneutralalumina(Aldrich,Brockmann,~150mesh)withagrADIentelutionfrom30%dichloromethaneinn-hexaneto100%dichloromethane.Themaingreenbandwascollectedandevaporatedtodryness.Thecrystallinepowderwasdissolvedinacetone,adjustedpH12.0with1NNaOH,andstirredfor12h.TheprecipitatedCe6wasfiltered,washedwithacetoneanddissolvedin100mlofwater,andfilteredtoremoveinsolubleimpurity.Afterlyophilizationofthefilteredwatersolution,afineblackpowderofCe6wasobtained.ThepurityofCe6is93-98%(yieldofCe6:1%fromdriedweightofchlorella).
Thefollowingprocedurewasreportedin
 
Livechlorella(Chlorellaellipsoidea)100g(driedweight)wassequentiallywashedwith500mlofwaterand300mlof50%ethanolinwatertoremovepolarmaterialsandtheresiduewasextractedtwicewith500mlof100%ethanoltoobtainchlorophyllarichfraction(extractionyield4.3%).Stirringthecombinedethanolsolutionofchlorophyllain1NHCl(pH2.5)for3hatroomtemperatureaffordedpheophytinintheformofprecipitates.Theprecipitatewasdissolvedindichloromethanewashedwithdistilledwater,driedwithanhydroussodiumsulfate,androtary-evaporatedtodryness.Theresiduewaspurifiedbyachromatographyusingneutralalumina(Aldrich,Brockmann,~150mesh)withagradientelutionfrom30%dichloromethaneinn-hexaneto100%dichloromethane.Themaingreenbandwascollectedandevaporatedtodryness.Thecrystallinepowderwasdissolvedinacetone,adjustedpH12.0with1NNaOH,andstirredfor12h.TheprecipitatedCe6wasfiltered,washedwithacetoneanddissolvedin100mlofwater,andfilteredtoremoveinsolubleimpurity.Afterlyophilizationofthefilteredwatersolution,afineblackpowderofCe6wasobtained.ThepurityofCe6is93-98%(yieldofCe6:1%fromdriedweightofchlorella).
 
 
 
 

References

1:LiQ,WangX,WangP,ZhangK,WangH,FengX,LiuQ.EfficacyofChlorine6-MediatedSono-PhotodynamicTherapyon4T1Cells.CancerBiotherRadiopharm.2013Nov9.[Epubaheadofprint]PubMedPMID:24206161.

2:LiZ,WangC,ChengL,GongH,YinS,GongQ,LiY,LiuZ.PEG-functionalizedironoxidenanoclustersloadedwithchlorine6fortargeted,NIRlightinduced,photodynamictherapy.Biomaterials.2013Dec;34(36):9160-70.doi:10.1016/j.biomaterials.2013.08.041.Epub2013Sep3.PubMedPMID:24008045.

3:SkripkaA,ValanciunaiteJ,DauderisG,PoderysV,KubiliuteR,RotomskisR.Two-photonexcitedquantumdotsasenergydonorsforphotosensitizerchlorine6.JBiomedOpt.2013Jul;18(7):078002.doi:10.1117/1.JBO.18.7.078002.PubMedPMID:23864017.

4:WangH,WangX,WangP,ZhangK,YangS,LiuQ.UltrasoundenhancestheefficacyofchlorinE6-mediatedphotodynamictherapyinMDA-MB-231cells.UltrasoundMedBiol.2013Sep;39(9):1713-24.doi:10.1016/j.ultrasmedbio.2013.03.017.Epub2013Jul3.PubMedPMID:23830103.

5:KimaniS,GhoshG,GhogareA,RudshteynB,BartusikD,HasanT,GreerA.Synthesisandcharacterizationofmono-,di-,andtri-poly(ethyleneglycol)chlorine6conjugatesforthephotokillingofhumanovariancancercells.JOrgChem.2012Dec7;77(23):10638-47.doi:10.1021/jo301889s.Epub2012Nov14.PubMedPMID:23126407;PubMedCentralPMCID:PMC3815657.

6:ChenB,ZhengR,LiuD,LiB,LinJ,ZhangW.Thetumoraffinityofchlorine6anditssonodynamiceffectsonnon-smallcelllungcancer.UltrasonSonochem.2013Mar;20(2):667-73.doi:10.1016/j.ultsonch.2012.09.008.Epub2012Oct2.PubMedPMID:23073382.

7:SaboktakinMR,TabatabaieRM,AminiFS,MaharramovA,RamazanovMA.Synthesisandin-vitrophotodynamicstudiesofthesuperparamagneticchitosanhydrogel/chlorinE6nanocarriers.MedChem.2013Feb;9(1):112-7.PubMedPMID:22762166.

8:LiP,ZhouG,ZhuX,LiG,YanP,ShenL,XuQ,HamblinMR.Photodynamictherapywithhyperbranchedpoly(ether-ester)chlorin(e6)nanoparticlesonhumantonguecarcinomaCAL-27cells.PhotodiagnosisPhotodynTher.2012Mar;9(1):76-82.doi:10.1016/j.pdpdt.2011.08.001.Epub2011Oct4.PubMedPMID:22369732;PubMedCentralPMCID:PMC3292741.

9:ShimG,LeeS,KimYB,KimCW,OhYK.Enhancedtumorlocalizationandretentionofchlorine6incationicnanolipoplexespotentiatethetumorablationeffectsofphotodynamictherapy.Nanotechnology.2011Sep7;22(36):365101.doi:10.1088/0957-4484/22/36/365101.Epub2011Aug12.PubMedPMID:21841215.

10:ShiH,LiuQ,QinX,WangP,WangX.Pharmacokineticstudyofanovelsonosensitizerchlorin-e6anditssonodynamicanti-canceractivityinhepatoma-22tumor-bearingmice.BiopharmDrugDispos.2011Sep;32(6):319-32.doi:10.1002/BDd.761.Epub2011Aug3.PubMedPMID:21815170.