Syntheses and Cellular Investigations of Di(aspartate) and ... · R. G. Waruna Jinadasa, Zehua...

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SynthesesandCellularInvestigationsofDi(aspartate)andAspartate-lysineChlorine6Conjugates

R.G.WarunaJinadasa,ZehuaZhou,M.GraçaH.VicenteandKevinM.Smith*

ELECTRONICSUPPLEMENTARYINFORMATION

Copiesof1HNMRspectra S2–S14Copiesof13CNMRspectra S15–S18Darkandphototoxicitycurves S19-S20Subcellularlocalizationforchlorine6 S21

Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2015

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1HNMRspectrumofmethylpheophorbidea(21)inacetone-d6at400MHz

N

NH N

HN

MeO2C O

CO2Me

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1HNMRspectrumof152,173-di(Asp)Ce6tert-butylmethylester(11)inacetone-d6at400MHz

S4

1HNMRspectrumof152,173-di(Asp)Ce6methylester(12)inacetone-d6at400MHz

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1HNMRspectrumof131,173-di(Asp)Ce6tert-butylmethylester(20)inacetone-d6at400MHz

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1HNMRspectrumof131,173-diaspartylchlorine6methylester(13)inMeODat400MHz

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1HNMRspectrumofethylenediaminyl(boc)pheophorbidea(25)inacetone-d6at400MHz

*Contain10%isocyclicringopenedproduct24

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1HNMRspectrumof131-ED152-ED(boc)Ce6methylester(26)inacetone-d6at400MHz

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1HNMRspectrumof131,152-ED-AspCe6tert-butylbocmethylester(28)inchloroform-dat400MHz

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1HNMRspectrumof131,152-ED-AspCe6methylester(29)inmethanol-d4at400MHz

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1HNMRspectrumofβ-alanylpheoporbidetert-butylmethylester(30)inacetone-d6at400MHz

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1HNMRspectrumofβ-alanylaspartylpheoporbidedi-tertbutylmethylester32inacetone-d6at400MHz

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1HNMRspectrumof132-EDLys-152-β-AlaAspCe6tert-butylbocmethylesterinacetone-d6at400MHz

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1HNMRspectrumof132-EDLys-152-β-AlaAspCe6methylester(34)inacetone-d6at400MHz

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1HNMRspectrumof131,173-diaspartylchlorine6methylester(13)inMeODat400MHz

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13CNMRspectrumof131,152-ED-AspCe6methylester(29)inmethanol-d4at400MHz

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13CNMRspectrumof152,173-di(Asp)Ce6methylester(12)inmethanol-d4at100MHz

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13CNMRspectrumof132-EDLys-152-β-AlaAspCe6methylester(34)inmethanol-d4at100MHz

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Darktoxicityofchlorine6(1,green)anditsderivatives152,173-di(Asp)Ce6MME(12,blue),131,173-di(Asp)Ce6MME(13,red),131,152-di(EDAsp)Ce6MME(28,purple)and131-EDLys-152-β-AlaAspCe6MME(33,black),towardHEp2cellsusingtheCellTiterBlueassay.

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Phototoxicityofchlorine6(1,green)anditsderivatives152,173-di(Asp)Ce6MME(12,blue),131,173-di(Asp)Ce6MME(13,red),131,152-di(EDAsp)Ce6MME(28,purple)and131-EDLys-152-β-AlaAspCe6MME(33,black),towardHEp2cellsusing1J/cm2lightdoseandtheCellTiterBlueassay.

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Subcellularlocalizationofchlorine6inHEp2cellsat10µMfor6h,(a)phasecontrast,(b)overlayofchlorine6andphasecontrast,(c)ERTrackerBlue,(d)overlayofchlorine6andERTrackerBlue,(e)BODIPYCeramide,(f)overlayofchlorine6andBODIPYCeramide,(g)MitoTrackerGreen,(h)overlayofchlorine6andMitoTrackerGreen,(i)LysoSensorGreen,(j)overlayofchlorine6andLysoSensorGreen.Scalebar:10µm