ABSTRACT
DFKIRU,QWHJUDWHG7HVWLQJ G7HVW%DWWHU\ XUH3HUVSHFWLYHV WKH1HXURWR[LFLW\7HVW%DWWHU\ VXUH7LPHV
INTRODUCTION
JLFDO ULVNDVVHVVPHQW LVEDVHGRQTXDQWLÀFDWLRQRIDFXWHVXE [LFGRVHVWRDQLPDOVZKLFKFDQEHH[SUHVVHGDVIRUH[DPSOH OHWKDOLW\LQDSRSXODWLRQ/'ORZHVWREVHUYHGHIIHFWGRVH UYHGHIIHFWGRVH 12('$QXPEHURI VDIHW\ IDFWRUVDUH WKHQ SRODWHWKHVHGRVHVWRWKHKXPDQVLWXDWLRQDQGWRHVWDEOLVKVDIHW\
H[SRVXUH+RZHYHUWKHLQFUHDVLQJQXPEHURIFKHPLFDOVDQGWKH UG WKH XVH RI DQLPDO H[SHULPHQWDWLRQ KDYH FRQWULEXWHG WR WKH HVWV\VWHPVIRUWR[LFRORJLFDOULVNDVVHVVPHQW7KHGUDZEDFNVRI UHVXOWVREWDLQHGDUHQRWHDVLO\H[WUDSRODWHGWRWKHLQWDFWRUJDQLVP WHUPLQHGQRWGRVHDQGJHQHUDOO\WKHUHLVDODFNRIELRWUDQVIRU
PDWLRQDQGNLQHWLFF WRPHDVXUHJHQHUDOF LVWKHUHIRUHQHFHVVDU PLQHGLQYLWURFDQE DSSURDFKLVWRLQWHJU
THE STEP
7REHDEOHWRFR SDUDPHWHUV RQ SK\V FKDUDFWHULVWLFVRIWKH EHSHUIRUPHGLQDVW
,Q WKH (X (5*$77LQLWLDWHG IRU /DERUDWRU\$QLP FDOOHGWKH(5*$77 ZDV WR LQYHVWLJDWH W GDWDZKLFKZHUHLQW $ SUHYDOLGDW VHFWLRQVRIEXLOGLQJ DV F\WRWR[LFLW\ DQG EDVHGELRNLQHWLFVWD WR[LFLW\ZHUHPRGHO DQGLQYLYRWR[LFLW\
RQVLGHUDWLRQV)XUWKHUPRUHLQYLWURDVVD\VKDYHEHHQGHYHORSHG \WRWR[LFLW\UDWKHUWKDQRQPHFKDQLVPVRILPSRUWDQFHLQYLYR,W \WRÀQGDVWUDWHJ\E\ZKLFKFHOOXODUWR[LFFRQFHQWUDWLRQVGHWHU H´FRQYHUWHGµWRGRVHVWKDWDUHUHOHYDQWIRUULVNDVVHVVPHQW2QH DWHWKHLQYLWURGDWDZLWKFRPSXWHUEDVHGELRNLQHWLFPRGHOV
WISE APPROACH FOR INTEGRATED TESTING
QVWUXFWDELRNLQHWLFPRGHOLWLVQHFHVVDU\WRGHWHUPLQHUHOHYDQW LRORJLFDO FKDUDFWHULVWLFV RI DQ RUJDQLVP DQG SK\VLFRFKHPLFDO FRPSRXQGV7KHLQWHJUDWHGDSSURDFKIRUDVVHVVLQJWR[LFLW\FDQ HSZLVHSURFHGXUH
THE ECITTS PROJECT
URSHDQ 5HVHDUFK *URXS IRU $OWHUQDWLYHV LQ 7R[LFLW\ 7HVWLQJ DQLQWHUODERUDWRU\SURMHFWLQDVVRFLDWLRQZLWKWKH6ZHGLVK%RDUG DOV &HQWUDOD )|UV|NVGMXUVQlPQGHQ &)1 7KH SURMHFW ZDV &)1,QWHJUDWHG7R[LFLW\7HVWLQJ6FKHPH(&,776DQGLWVDLP KH SRVVLELOLW\ RI HVWLPDWLQJ WR[LF GRVHV IURP LQ YLWUR WR[LFLW\ HJUDWHGZLWKFRPSXWHUEDVHGELRNLQHWLFPRGHOV:DOXPHWDO LRQ VWXG\ ZDV FRQVWUXFWHG RQ WKH EDVLV RI WKUHH IXQGDPHQWDO
EORFNV)LUVWH[SHULPHQWDOGDWDIRUELRNLQHWLFPRGHOVDVZHOO WDUJHW WR[LFLW\ GDWDZHUH GHWHUPLQHG VHFRQG SK\VLRORJLFDOO\ UJHWWLVVXHFRQFHQWUDWLRQVWR[LFRORJLFDOUHVSRQVHDQGV\VWHPLF HGDQGWKLUGWKHPRGHOZDVYDOLGDWHGDJDLQVWLQYLYRNLQHWLFV SDUDPHWHUV)LJXUH
Compounds and
(LJKWZHOOFKDUD RU SHULSKHUDO QHUYRX GLD]HSDPQKH[DQH WROXHQH $Q LQ YLWUR GHVLJQHGLQRUGHUWR DQGQHXURFKHPLVWU\
7KHKXPDQQHXU PRGHOIRUV\PSDWKHW VWXGLHV7KHFHOOVFDQ ZLWKUHWLQRLFDFLG1 RI QHXULWHV 3nKOPDQ
Test Battery
FWHUL]HG WHVWFRPSRXQGVZKLFKSRVVHVVDFWLYLW\ LQ WKHFHQWUDO V V\VWHPV ZHUH VHOHFWHG 7KHVH ZHUH DFU\ODPLGH FDIIHLQH KH[DQHGLRQH OLQGDQHSDUDWKLRQSDUDR[RQSKHQ\WRLQ DQG WHVW EDWWHU\ ZLWK JHQHUDO DQG QHXURVSHFLÀF HQGSRLQWV ZDV FRYHUHQGSRLQWVIRUEDVDOF\WRWR[LFLW\PRUSKRORJ\SK\VLRORJ\ :DOXPHWDO)RUVE\HWDO REODVWRPDFHOO OLQH6+6<<KDVEHHQXVHGH[WHQVLYHO\ DV D LFJDQJOLRQFHOOV5RVVHWDODQGZDVFKRVHQIRUWKHVH EHIXUWKHUGLIIHUHQWLDWHGZKHQFXOWXUHGLQVHUXPIUHHPHGLXP 5$ZKLFKLQGXFHVWKHFHOOVWRGHYHORSDQH[WHQVLYHQHWZRUN HW DO )LJXUH )XUWKHUPRUH WKH H[FLWDELOLW\ DQG
UHFHSWRUIXQFWLRQVDU EHIRUH WKHKURI RQ VHYHUDO HQGSRLQWV SRLQWVDQGWKHDVVD\
&ULWLFDOQHXURWR[ PLQHGLQWKHPRVWV HVL]HGWRPLPLFWKH OHYHOV/2(/V7KH ELRNLQHWLFPRGHOV IR DFU\ODPLGHWKHVWXG\ EDVDOF\WRWR[LFLW\DV RUGHUWRHVWLPDWHWKH EHHQ VXJJHVWHG WKDW H[SRVXUHDUHJRRGV H[SRVXUH WRDGRVH 7LPHGHSHQGHQWQHX PDWLRQRIVXEFKURQL 7R HVWLPDWH OLQGDQH FRQVLGHUDEO\ KLJKHU DGGLWLRQDOGDWD (& FHOOVDIWHUKURIH LQ RUGHU WR VWXG\ WK WR[LFLW\ LQYLYR /' DVWKHEDVLVIRUWKHH IRU F\WRWR[LFLW\ LH VHYHUDO FHOO F\FOHV L FRQFHQWUDWLRQVLQKX
Results
7KHFHOOXODUQHXU PLQHGDIWHUKURI
HHQKDQFHG+HQFHWKHFHOOVZHUHFXOWXUHGLQ15$IRUKU H[SRVXUHZLWK WKH WHVWFRPSRXQGV$IWHUH[SRVXUH WKHHIIHFWV ZHUH LQYHVWLJDWHGDQG(&YDOXHVZHUHHVWLPDWHG7KHHQG VXVHGIRUWKHVWXG\DUHOLVWHGLQ7DEOH LFFRQFHQWUDWLRQV&1&VZHUHGHÀQHGDVWKH(&YDOXHGHWHU HQVLWLYHHQGSRLQWIRUHDFKFRPSRXQG7KH&1&VZHUHK\SRWK WR[LFWDUJHWWLVVXHFRQFHQWUDWLRQVDWWKHORZHVWREVHUYHGHIIHFW &1&VIRUHDFKFRPSRXQGZHUHLQWHJUDWHGZLWKFRPSXWHUEDVHG U UDWV DQGDFXWHDQG VXEFKURQLF/2('VZHUHHVWLPDWHG)RU ZDVH[WHQGHGE\LQWHJUDWLQJWKH(&YDOXHUHÁHFWLQJORZHVW GHWHUPLQHGE\WRWDOFHOOXODUSURWHLQFRQWHQWVHH7DEOHLQ /2('IRUDFXWHQRQOHWKDOV\VWHPLFWR[LFLW\,WKDVSUHYLRXVO\ VOLJKWO\ F\WRWR[LF FRQFHQWUDWLRQV GHWHUPLQHG DIWHU KU RI XUURJDWHVIRUDFXWHWR[LFEORRGFRQFHQWUDWLRQLQPDQKUDIWHU WKDWJLYHVPLOG WRPRGHUDWH LQWR[LFDWLRQ:DOXPHWDO ULWHGHJHQHUDWLRQZDVDOVRGHWHUPLQHGIRUDFU\ODPLGHIRUHVWL F/2('VDIWHUGLIIHUHQWH[SRVXUHWLPHV'H-RQJKHWDO LQGXFHG VXEFKURQLF /2(' IRU FRQYXOVLRQV ZKLFK RFFXU DW GRVHV WKDQ WKH /2(' IRU OHDUQLQJ GHÀFLHQFLHV 'HVL
RQ LQFUHDVHG LQWUDFHOOXODU IUHHFDOFLXPFRQFHQWUDWLRQ LQ WKH [SRVXUHZHUHLQWHJUDWHGLQWKHPRGHOIRUOLQGDQH)XUWKHUPRUH H FRUUHODWLRQ EHWZHHQ EDVDO F\WRWR[LFLW\ GDWD DQG DFXWH OHWKDO
IRU OLQGDQH(& IURP WKHSURWHLQGHWHUPLQDWLRQZDVXVHG YDOXDWLRQ(NZDOODQGFRZRUNHUVKDYHVKRZQWKDW(&YDOXHV LQKLELWLRQ RI FHOO JURZWK DQGRU FHOO GHDWK GHWHUPLQHG DIWHU Q XQDIIHFWHG FHOOV DUH JRRG VXUURJDWHV IRU OHWKDO EORRG SHDN
PDQVDIWHUDFXWHH[SRVXUH(NZDOOHWDO
RWR[LFDFWLYLWLHVIRUVL[RIWKHHLJKWWHVWFRPSRXQGVZHUHGHWHU H[SRVXUH7KH&1&VDQGWKHDGGLWLRQDO(&DQG(&YDOXHV
WKDWZHUH XVHG DV/ FRXOGQRWEHWHVWHGLQ DQG UHDFWLRQZLWK WK VLPXODWLRQRIWROXHQH
,QWHJUDWLRQRIWKH D YHU\ JRRG FRUUHODW FRPSRXQGV)LJXUH WDO/2('IRUGLD]HSD PHFKDQLVP*$%$$ LQYLYRHIIHFWWLPHWR ZDVQRWLQFOXGHGLQ GDWDRQGLD]HSDPDF NLQHWLF PRGHO DQ DF H[SHULPHQWDO/2('
CONC
7KHSUHYDOLGDWLRQ E\LQWHJUDWLQJLQYLWU LQJ+RZHYHUWKHUH
Refinement of the
7KHUHVXOWVRIGL UHOHYDQW LQYLWURHQG SRXQGV 7KXV WKH Q VWHSZLVH WHVW VWUDWHJ V\VWHPLFWR[LFLW\ED DQGEDVDOLQWUDFHOOXOD HUDWLRQ 7KH VHFRQ
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
LUSIONS AND FUTURE PERSPECTIVES
VWXG\VKRZHGWKDWLWLVSRVVLEOHWRHVWLPDWH/2('VDQG/'V RGHULYHGWR[LFLW\GDWDZLWKFRPSXWHUEDVHGELRNLQHWLFPRGHO DUHVRPHUHPDUNVWKDWKDYHWREHFRQVLGHUHG
Neurotoxicity Test Battery
D]HSDPLQGLFDWHG WKDW WKH LQYLYRHQGSRLQWVDUHGHSHQGHQWRQ SRLQWVZKLFKGHVFULEHWKHPROHFXODUPHFKDQLVPVRIWKHFRP
HXURWR[LFLW\ WHVW EDWWHU\ ZKLFK FDQ EH GHVLJQHG DV D WLHUHG \ VKRXOG FRQVLVW RI D À[HG SDUW IRU HVWLPDWLRQ RI D VSHFLÀF VDOF\WRWR[LFLW\SK\VLRORJ\IRULQVWDQFHHQHUJ\PHWDEROLVP U&DFRQFHQWUDWLRQVDQGSRVVLEO\D[RQRSDWK\QHXULWHGHJHQ G SDUW QHXURFKHPLVWU\ VKRXOG EH ÁH[LEOH DQG LQGLYLGXDOO\
GHVLJQHGIRUGLIIHUHQ EDVHGRQNQRZOHGJH GDWDKXPDQGDWDDQ
Different Exposu
7KHQHXURWR[LFD YLWUR'HWHUPLQDWLRQ HGO\LPSURYHGWKHH HW DO )XUWKH WHUPPLQXWHVDVFR
WFODVVHVRIFRPSRXQGV7KHVSHFLÀFHQGSRLQWVVKRXOGEHFKRVHQ DERXWWR[LFRORJLFDOPHFKDQLVPVRIUHODWHGFRPSRXQGV,QYLYR GNQRZOHGJHDERXWVWUXFWXUHDFWLYLW\UHODWLRQVKLSVZLOOEHXVHIXO
re Times
FWLYLWLHVVKRXOGEHGHWHUPLQHGDIWHUYDULRXVH[SRVXUHWLPHVLQ RIDFU\ODPLGHLQGXFHGGHJHQHUDWLRQRIQHXULWHVRYHUWLPHPDUN VWLPDWLRQRIQHXURWR[LFLW\DIWHUVXEFKURQLFH[SRVXUH'H-RQJK UPRUH QHXURFKHPLFDOSURSHUWLHVPD\EHGLIIHUHQW DIWHU VKRUW PSDUHGWRORQJWHUPGD\VH[SRVXUH
,Q FRQFOXVLRQ WK XVLQJFHOOXODULQYLWUR FDQ EH D SRZHUIXO W WR[LFRORJLFDOPHFKDQ UHVXOWVRI WKHSURMHF NLQHWLFPRGHOVRQLQ PRGHOVRIKXPDQVF KXPDQPHWDEROLVPE JHQHUDOSUREOHPRIY ZKLFKPDNHVLWGLIÀF 1HYHUWKHOHVVRXUVWX WKHVDPHTXDOLW\DVW
7KLVLQYHVWLJDWLRQ WLRQRI$OWHUQDWLYH0 PDOV&)1:HWKDQ WLRQWRWKHELRNLQHWLF
H (&,776 SUHYDOLGDWLRQ VWXG\ LQGLFDWHG WKDW WKH VWUDWHJ\ RI PHWKRGVLQWHJUDWHGZLWKFRPSXWHUEDVHGELRNLQHWLFPRGHOLQJ RRO IRU ULVN DVVHVVPHQW+RZHYHU JHQHUDO DVZHOO DV VSHFLÀF LVPVPXVWEHFRQVLGHUHGIRU WKHGHVLJQRI WHVWEDWWHULHV7KH WDOVRUHYHDOHGWKDW LW LVSRVVLEOH WREXLOGFRPSXWHUEDVHGELR YLWURGDWDIRUWKHUDW,QH[WHQVLRQFRPSXWHUEDVHGELRNLQHWLF DQEHFRQVWUXFWHGZKHQ LQYLWURPHWKRGVIRUGHWHUPLQDWLRQRI ORRG²EUDLQEDUULHUSDUWLWLRQFRHIÀFLHQWVHWFDUHDYDLODEOH2QH DOLGDWLRQLVWKHOLPLWHGDPRXQWRIUHOLDEOHKXPDQWR[LFLW\GDWD XOWWRYDOLGDWHDOWHUQDWLYHWHVWVIRUDODUJHUVHWRIWHVWFRPSRXQGV GLHVKDYHVKRZQWKDWDOWHUQDWLYHVFDQHVWLPDWHWR[LFGRVHVZLWK UDGLWLRQDODQLPDOWHVWV
ACKNOWLEDGMENTS
ZDVÀQDQFHGE\JUDQWVIURPWKH(XURSHDQ&HQWHUIRU9DOLGD HWKRGV(&9$0DQGWKH6ZHGLVK%RDUGIRU/DERUDWRU\$QL NDOOSDUWLFLSDQWVLQWKH(&,776SURMHFWIRUYDOXDEOHFRQWULEX PRGHODQGVFLHQWLÀFGLVFXVVLRQV
REFERENCES