VOLUM E - O FO FSUBMISSIO N 

# syngenta

Norflurazo n Reques t fo r Waive r o f Endocrin e Disruptio n Screenin g Progra mAssay s Assessmen t
DAT A REQUIREMENT(S) : 

### No t Applicabl e AUTHOR(S) : 

### W . Thoma s Beidler , Ph.D . Jame s T . Stevens , Ph.D . Jonatha n M . Akins , Ph.D. , DAB T COMPLETIO N DATE : 

### February 4 , 201 0 PERFORMIN G LABORATORY : 

### Syngent aCro p Protection , Inc . 41 0 Swin g Roa d Greensboro , N C 2740 9 US A LABORATOR Y PROJEC T ID : 

### Report Number : TK002342 5 Tas k Number : TK002342 5 SUBMITTER/SPONSOR : 

### Syngent aCro p Protection , Inc . I J 41 0 Swin g Roa d" a<> < 

### Pos t Offic e Bo x 18300 * * ' Greensboro , N C 27419-830 0 VOLUM E 1 O F1 O FSTUD Y PAG E 1 O F4 3

### STATEMEN T O FDAT A CONFIDENTIALIT Y CLAIM S

STATEMENT O FN O DAT A CONFIDENTIALIT Y CLAIM S UNDE RSPECIFIED FIFR A PROVISION S N o clai mo f

confidentialit

y i s made for

any informatio n containedi n thi s study o n th e basi s o f it s fallin g withi n th e scop e o f FIFR A Sectio n 10(d)(l)(A ) (disclose s manufacturin g o r qualit y contro l processes) , (B ) (disclose s th e detail s o f method s fo r

testing , detectin g o r measurin g th e quantit

y o f an y deliberatel y adde diner t ingredien t

o f apesticide) , o r

(C ) (disclose s th e identity o r

percentag e quantit

y o f an y deliberatel y adde diner t ingredien t

o f a pesticide) . Company : Syngent aCro p Protection , Inc . Compan y Agent : Jerry Wells , Ph.D . U . S . Produc t

Registratio n Manager

Date Syngent ai s th e owne r

o f thi s study . Syngent aha s submitte dthi s material t o th e Unite d State s Environmental Protectio n Agenc y specificall y unde r th e provision s containe di n FIFRA a s amende dand , hereby , consent s to us e an ddisclosure o f thi s materia l b y EP A accordin g t o FIFRA . Notwithstandin g th e wordin g o f our markin g TRAD E SECRET , thi s marking , b y itself, convey s n o supplemental claim s o f confidentiality unde r

FIFRA Section s 10(a

) o r 10(b ) (addressin g protectio n o f trad e secret s an dcommercia l an dfinancia l information) . I n submittin g thi s material t o EP A accordin g t o metho dan dforma t requirement s containe di n P RNotic e 86-5 , w e d o no t waiv e any protectio n o r

right involvin g thi s material tha t

woul dhav e bee n claime db y th e compan y i f thi s materia l ha dno t

bee n . J . submitte dt o th e EPA , no r d o w e waiv e an y protectio n o r righ t provide dunde r

FIFR A * Sectio n 3 (concernin g dat aexclusivit y an ddat acompensation ) o r FIFR A Sectipn.lO(g ) ; » 0 * (prohibitin g disclosure t o foreig n an dmultinational pesticid e companie s o r tjieirlagents) . .

Repor t Number : TK002342 5 Pag e 2 o f 4 3

### GOO D LABORATOR Y PRACTIC E COMPLIANC E STATEMENT

A s thi s volum e contain s comment s an dassessment s o f informatio n an ddat ai n report s submitte dseparately , aGoo dLaborator y Practic e Complianc e Statement i s no t appropriate . Stud y Director : There i s n o GLP Study Directo r fo r thi s volume .

W . Thoma s Beidler , Ph.D . Dat e / Autho r Performin g Laboratory : Syngent aCrop Protection , Inc . 41 0 Swin g Roa d Greensboro , N C 2740 9 US A

, 2 0 f O JoWha n M . Akins , Ph.^J&B T Date Representativ e o f

Submitter/Sponso r Submitter/Sponsor : SyngentaCrop Protection , Inc . 41 0 Swin g Roa d Pos t Offic e Bo x 18300 Greensboro , N C 27419-830 0 US A ' • •

• > • • • *

Report Number : TK002342 5 Pag e 3 o f 4 3

### TABL E O FCONTENT S

STATEMEN T O FDAT A CONFIDENTIALIT Y CLAIM S 1 GOO D LABORATOR Y PRACTIC E COMPLIANC E STATEMEN T 2 TABLE O FCONTENT S 3 1. 0 EXECUTIV E SUMMARY 5 2. 0 INTRODUCTIO N 6 3. 0 OTHE RSCIENTIFICALL Y RELEVAN T INFORMATIO N (OSRI) :

EXISTIN G NORFLURAZO N STUDIE S SATISFY EDS P TIE R1 ASSA Y REQUIREMENT S 8 3. 1 Reproductio n Studie s ........................................................................................ . 9 3. 2 Developmenta l Toxicity Studie s ..................................................................... . 1 0 3. 3 Roden t

Subchroni c an dChroni c Toxicity/Carcinogenicity Studie s ............... . 1 1 3. 4 Fis han dAmphibia n Studie s ........................................................................... . 1 2 4. 0 RATIONAL E FO RACCEPTANC E O FNORFLURAZO N OSR I FO RSPECIFI C TIE R1 ASSAY S 1 2 4. 1 Estroge n Recepto r Bindin g Assay s (OPPT S 890.1250 ; OPPT S 890.1300 ) ... . 1 2 4. 2 Androge n Recepto r Bindin g (OPPT S 890.1150 ) ............................................ . 1 3 4. 3 Aromatas e -Huma n Recombinan t

(OPPT S 890.1200 ) ................................... . 1 5 4. 4 Steroidogenesis - Huma n Cel l Lin e H295 R(OPPT S 890.1550 ) ................... . 1 6 4. 5 Hershberge r - Rat (OPPT S 890.1400 ) ............................................................ . 1 7 4. 6 Mal e Puberta l - Rat (OPPT S 890.1500 ) .......................................................... . 1 8 4. 7 Femal e Puberta l - Rat (OPPT S 890.1450 ) ...................................................... . 2 0 4. 8 Uterotrophi c - Rat (OPPT S 890.1600 ) ........................................................... . 2 2 4. 9 Fis hShor t Ter mReproductio n (OPPT S 890.1350 ) ................................... . .....2 3 4.1 0 Amphibia n Metamorphosi s (OPPT S 890 . 1 1 00 ) .................................... . ..;.,... . 2 3 5. 0 CONCLUSIO N ../. ^ 2 3 6. 0 REFERENCE S I'T l * " " 2 5 APPENDICE S SECTIO N ••;•• • •••" • 2 7 

### APPENDI X 1 Titl e Page s o f Cite dStudie s ................................................ . ".?• " ....... . :.V..V.2 8 * ~ ^ ••*•&.._ - • •
•



### Report Number : TK002342 5 Pag e 4 o f 4 3

### LIS T O FTABLE S

Tabl e 1 Norflurazo n OSRI Studie s 8 Tabl e 2 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte d withNorflurazo n whic har e Relevan t t o i n vitr o E RBindin g an dt o th e Estrogeni c Mod e o f

Actio n 1 3 Tabl e 3 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte d withNorflurazo n whic har e Relevan t t o i n vitr o A RBindin g an dt o th e Androgeni c Mod e o f

Actio n 1 4 Tabl e 4 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte d withNorflurazo n whic har e Relevan t t o Aromatas e Inhibitio n 1 6 Tabl e 5 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte d wit hNorflurazo n whic har e Relevan t t o th e Steroid s Alteration s that migh t b e Detecte di n th e Steroidogenesi s Scree n 1 7 Tabl e 6 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte d wit hNorflurazo n Compare dt o thos e Measure dh i th e Hershberge r Screen... . 1 8 Tabl e 7 Functionall y Equivalen t Endpoint s i n FIFR A Guidelin e Studie s Conducte d wit hNorflurazo n Compare dt o thos e Measure di n th e Puberta l Mal e Screen . 1 9

••••• • * *,, *



### •••• • I *,. *

Report Number : TK002342 5 Pag e 5 o f 4 3

### 1. 0 EXECUTIV E SUMMARY

Thi s documen t

provide s th e scientifi c justificatio n toward s awaive r fo r th e majorit y o f

th e Endocrin e Disruptio n Screenin g Progra mTie r 1 assay s i n th e norflurazo n tes t orde r base do n th e existenc e o f

Othe r Scientificall y Relevan t

Informatio n (OSRI) . EP A develope dan drecentl y publishe dth e guideline s fo r th e Endocrin e Disruptio n Screenin g Progra m(EDSP ) Tier

1 Screenin g (T1S ) assay s t o identif y potentia l endocrine-modulatin g chemicals , an dEP A sen t tes t order s t o registran t

o f norflurazon . However , EP A als o indicate dtha t

"othe r scientificall y relevan t

informatio n ("OSRI" ) that i s functionall y equivalen t

t o th e EDS P Tie r

1 assay s may b e utilize di n lie u o f conductin g th e Tier

1 assays . Th e objectiv e o f

thi s documen t

i s t o provid e th e OSRI that i s availabl e fo r

norflurazo n an dt o obtai n waiver s fo r

th e followin g EDS P Tie r 1 assay s o f

th e tes t order : 890.115 0 - Androge n Recepto r Bindin g (Ra

t Prostate ) 890.120 0 - Aromatas e (Huma n Recombinant ) 890.125 0 - Estroge n Recepto r Bindin g 890.130 0 - Estroge n Recepto r Transcriptiona l Activatio n (Huma n Cel l Lin e HeLa-9903 ) 890.140 0 - Hershberge r

(Rat ) 890.145 0 - Femal e Puberta l (Rat ) 890.150 0 - Mal e Puberta l (Rat ) 890.155 0 - Steroidogenesi s (Huma n Cel l Lin e - H295R ) 890.160 0 - Uterotrophi c (Rat ) Th e OSR I fo r

norflurazo n i s base do n tw o differen t sources : th e EPA' s ToxCast TM progra m result s an dth e U S EPA' s 40CF RPart 15 8 guidelin e studies . Norflurazo n wa s teste da s par t o f th e EPA' s ToxCast TM Phas e 1 screen , wher e th e NI H Chemica l Genomic s Cente r conducte destroge n an dandroge n recepto r bindin g assay s an daNovascree n assay fo r aromatas e inhibition . I n addition , endocrine-relate dendpoint s hav e alread y bee n assesse di n existin g EPA-require dguidelin e studie s fo r norflurazon , whic h provid e furthe r weigh t o f evidenc e tha t

th e OSR I provid e "functionall y equivalent " informatio n t o tha t obtaine di n th e propose dT1 S battery . I n conclusion , norflurazo n alread y ha s bee n thoroughl y evaluate dvi acritica l apica l endpoint s tha t correspon dt o th e EDS P Tie r 1 screens . Therefore , Syngent aCro p Protection , Inc. , request s that

th e Environmenta l Protectio n Agenc y waiv e th e requiremen t t o conduc t

th e majorit y o f

th e norflurazo n EDS P tes t order assay s base do n th e followin g scientifi c justification . 1 . U S EP A ha s recentl y sponsore dan dproduce dOSR I fo r th e followin g studies/endpoints , therefor e thes e studie s shoul db e waive dfro mth e EDS P Tie r 1 scree n requirements :

• 890.115 0 - Androgen Recepto r Bindin g (Rat

Prostate ) • 890.120 0 - Aromatas e (Huma n Recombinant ) • 890.125 0 - Estroge n Recepto r Bindin g • 890.130 0 - Estroge n Recepto r Transcriptiona l Activatio n (Huma n Cel l Lin e HeLa-9903 ) Report Number : TK002342 5 Pag e 6 o f 4 32 . Tie r 2 studie s hav e bee n previousl y conducte dfo r norflurazo n a s part

o f th e 40CF R Part 15 8 dat arequirement s (e.g. , rat reproductio n study , developmenta l toxicity , 90 - day studies , chroni c an doncogenicity studies) . Thes e EP A guidelin e studie s serv e a s OSRI fo r th e followin g studies/endpoints , therefor e thes e studie s shoul db e waive d fro mth e EDS P Tier

1 scree n requirements : 890.140 0 - Hershberge r

(Rat ) 890.145 0 - Femal e Puberta l (Rat ) 890.150 0 - Mal e Puberta l (Rat ) 890.155 0 - Steroidogenesi s (Huma n Cel l Lin e - H295R ) 890.160 0 - Uterotrophi c (Rat ) 3 . Th e existin g databas e fo r norflurazo n doe s no t

contai n th e specifi c OSRI fo r th e fis h an damphibia n studies , an dSyngent ai s no t providin g arational e t o waiv e thes e specifi c assays . However , th e existin g mammalia n toxicit y databas e combine dwith th e fis hearl y lif

e stag e studie s provid e significan t informatio n tha t

ma y b e judge da s adequat e OSR I fo r

awaive r o r partia l waive r

o f th e followin g tw o studies :

• 890.110 0 - Amphibia n Metamorphosi s (Frog ) • 890.135 0 - Fis h Short-Ter mReproductio n 

### 2. 0 INTRODUCTIO N Norflurazo n i s aselectiv e pre-emergen t herbicid e use dt o contro l germinatin g annua l grasse s an dbroadlea f

weed s i n fruits , vegetables , nuts , cotton , peanuts , soybeans , an dvariou s nonagricultura l an dindustria l areas . Norflurazo n wa s firs t registere da s apesticid e i n th e U.S . i n 197 4 an dha s undergon e registration , wit h th e lates t Registratio n Eligibilit y Decisio n occurrin g i n o r

aroun d2002 , an dwa s judge dt o mee t curren t dat arequirement s an d regulatory standards . Th e Federa l Foo dDru g an dCosmeti c Ac t require dth e U . S . Environmenta l Protectio n Agenc y (EPA ) t o "develo p ascreenin g program , usin g appropriat e validate dtes t system s an d othe r scientificall y relevan t

informatio n t o determin e whethe r certai n substance s ma y hav e a n effec t i n human s tha t

i s similar

t o a n effec t

produce db y anaturall y occurrin g estrogen , o r othe r effect s a s [EPA ] may designat e [2 1 U.S.C . 346 a(p)] . Sectio n 408(p ) (3 ) specificall y require dtha t

th e Administrato r

"shal l provid e fo r

th e testin g o f

al l pesticid e chemicals. " [2 1 U.S.C . 346 a(p ) (3)] . EP A develope dan drecentl y publishe dth e guideline s fo r th e Endocrin e Disruptio n Screenin g Program(EDSP ) Tier

1 Screenin g (T1S ) assay s t o identif y potentia l endocrine-modulatin g chemicals , an dEP A sent

tes t order s fo r

norflurazo n t o th e registran t Syngent aCro p Protection . However , EP A als o indicate dtha t "other scientificall y relevan t

informatio n ("OSRI" ) tha t

i s functionall y equivalen t t o th e EDS P Tie r

1 assay s may b e utilize di n lie u o f conductin g th e Tie r

1 assays . EP A indicate dth e followin g criteri afo r OSRI . Report Number : TK002342 5 Pag e 7 o f 4 3

### 1 ) Th e OSRI studie s mus t follo w accepte dscientifi c methodolog y o r protocol , includin g bu t no t limite dt o thos e presente di n th e EPA' s harmonize dtes t guidelin e compendiu m(se e http://www.epa.gov/endo) , and : a . Adher e t o goo dlaboratory practice s b . Ar e o f hig hquality , achievin g th e objectiv e t o provid e reasonabl e assuranc e tha t achemical doe s o r

doe s no t hav e th e potentia l t o interac t wit hestrogen , androgen , o r

thyroi dsystems . c . Includ e dat ao r

areferenc e t o th e dat afo r eac htes t that i s required , alon g wit h arational e tha t

explain s ho w th e existin g study tha t i s cite d/ submitte d satisfie s par t

o r al l o f th e Order . i . Deviation s fro mth e protocol s validate dfo r th e Tie r 1 assay s mus t

b e identified , alon g wit ha n explanatio n fo r th e deviations , includin g a n explanatio n a s t o why , notwithstandin g th e deviations , th e protoco l use dshoul dstil l b e considere da s providin g an accepte dscientifi c methodolog y o r

protocol . 2 ) B e classifie db y EP A a s acceptabl e i f i t ha s bee n previousl y submitted . a . Provid e MRI D numbe r an dcop y o f

th e titl e pag e (d o no t

resubmi t astud y tha t ha s previousl y bee n submitte dt o th e Agency . b . Identif y an y actio n take n b y th e Agenc y o n th e protocol , i f i t ha dbee n previousl y reviewe db y EPA . Identif y manne r i n whic hal l Agenc y comments , concern s o r

issue s wer e addresse di n th e fina l protoco l an dstudy . c . Offe r t o pa y compensatio n t o origina l dat asubmitte r i f yo u ar e citin g astud y o f whic hyo u ar e no t

th e origina l dat asubmitter . Th e objectiv e o f

thi s documen t

i s t o provid e th e OSR I tha t i s availabl e fo r

norflurazo n an dt o obtai n waiver s fo r

th e followin g EDS P Tie r

1 assays : 890.115 0 - Androgen Recepto r Bindin g (Rat

Prostate ) 890.120 0 - Aromatas e (Huma n Recombinant ) 890.125 0 - Estroge n Recepto r Bindin g 890.130 0 - Estroge n Receptor Transcriptiona l Activatio n (Huma n Cel l Lin e HeLa-9903 ) 890.140 0 - Hershberge r

(Rat ) 890.145 0 - Femal e Puberta l (Rat ) 890.150 0 - Mal e Puberta l (Rat ) 890.155 0 - Steroidogenesi s (Huma n Cel l Lin e - H295R ) 890.160 0 - Uterotrophi c (Rat ) 

### Thi s documen t

review s th e endocrine-relate dendpoint s alread y routinel y assesse di n existin g EPA-require dFIFR A guidelin e studie s an dothe r testin g o f norflurazo n b y EP A (e.g. , EP A dat ageneratio n fo r

norflurazo n fro mth e ToxCast TM program) , whic hprovid e weigh t o f evidenc e tha t

th e OSRI provid e "functionall y equivalent " informatio n whic hfulfil s th e requiremen t

fo r th e majorit y o f

th e screen s i n th e T1 S battery .

Report Number : TK002342 5 Pag e 8 o f 4 3

### 3. 0 OTHERSCIENTIFICALL Y RELEVAN T INFORMATIO N

(OSRI) : EXISTIN G NORFLURAZO N STUDIE S SATISFY EDS P TIER1 ASSA Y REQUIREMENT S Th e OSRI fo r

norflurazo n i s base do n tw o differen t sources : th e EPA' s ToxCast TM program result s o n norflurazo n an dth e U S EPA' s 40CF RPart 15 8 guidelin e studie s fo r norflurazo n registratio n (Tabl e 1) . Th e ToxCas t dat aprovide s substantiall y simila r methodolog y a s th e EDS P Tier

1 screen s wit h regar dt o estrogen , androgen , an daromatas e activity . Th e USEP A 40CF RPart

15 8 dat aprovide s th e Tier 2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmenta l toxicity , reproductio n effects , chroni c toxicity an dothe r apica l endpoint s o f

endocrin e functio n effects . Tabl e 1 Norflurazo n OSR I Studie s OSRI Dat aSourc e U S EP A ToxCas t

Th e USEP A 40CF R par t 15 8 dat a requirement s Stud y

Estroge n Bindin g Androge n Bindin g Aromatas e 90-Day Subchroni c Rat . (1971) . FIFR A §82-1 . 6 - Mont hDo g Chroni c Toxicity . (1973) . FIFR A §83- 1 Ra t Chroni c Toxicity /

Carcinogenicity . (1975) . FIFR A §83-1 . Rabbi t Developmenta l Toxicity . (1983) . FIFR A §83-3 . Rat Developmenta l Toxicity . (1972) . FIFR A §83-3 . 3-Generatio n Ra t

Reproduction . (1975) . FIFRA §83-4 . 2-Generatio n Ra t

Reproduction . (1994) . FIFR A §83- 4 MRID #

-

-

- 0009105 5 00111618 . 008201 9 an d 0011164 9 0013115 2 0006362 1 008075 0 4352230 1 OSRI Assessmen t

Substantiall y Similar methodolog y b y USEPA ; GL P Substantiall y Similar methodolog y b y USEPA ; GL P Substantiall y Similar methodolog y b y USEPA ; GL P USEP A Guidelin e study , Pre GL P USEP A Guidelin e study , Pre GL P USEP A Guidelin e study , Pre GL P USEP A Guidelin e study , GL P USEP A Guidelin e study , Pre GL P USEP A Guidelin e study , Pre GL P USEP A Guidelin e study , GL P Th e EPA' s ToxCast TM progra mha s alread y evaluate dth e norflurazo n i n assay s similar t o th e estroge n (OPPT S 890.130 0 - Estroge n Recepto r Transcriptiona l Activation) , androge n (OPPT S 890.115 0 - Androge n Recepto r Binding) , an daromatas e (OPPT S 890.120 0 - Aromatas e - Huma n Recombinant ) screen s require db y th e EDS P Tie r 1 batter y (Kavloc k

e t al. , 2009 ; Houck , 2009) . Th e protocol s fo r assay s use di n th e ToxCast TM evaluatio n ar e no t currentl y available , howeve r th e assay s wer e conducte db y an dfo r EPA . Thes e EPA supporte dassay s apparentl y mee t th e qualit y standard s o f

adherenc e t o th e spiri t

o f goo d laboratory practice s an dar e o f hig h quality , achievin g th e objectiv e t o provid e reasonabl e assuranc e that

achemica l doe s o r

doe s no t hav e th e potentia l t o interac t with estrogen , androge n an daromatas e receptor s (El i Lill y an dCompan y an dNI H Chemica l Genomic s Center , 2008) . Report Number : TK002342 5 Pag e 9 of4 3

### I n additio n th e Reregistratio n Eligibility Decisio n (RED ) Documen t fo r norflurazo n wa s issue di n 200 2 (USEPA , 2002) ; thi s Reregistratio n Eligibilit y Decisio n documen t reflect s a reassessmen t

o f al l dat awhic hwer e submitte di n respons e t o th e Registratio n Standar dan d th e subsequen t

DCIs . Th e toxicolog y databas e fo r norflurazo n wa s considere dcomplete . Norflurazo n i s liste di n th e IE H REDIPE D repor t (IEH , 2005 ) wit hth e designatio n that n o dat ai s availabl e i n regar dt o norflurazo n potentia l t o disrup t th e endocrin e system . N o othe r scientificall y relevan t

informatio n consisten t

withth e EDS P Tie r

1 assay s wa s foun dfo r norflurazo n i n th e literature . 3. 1 Reproductio n Studie s Th e followin g reproductio n studie s hav e bee n conducte dwithnorflurazon ; aone-generatio n stud y i n th e mous e (guidelin e 83-3 ; MRI D 00080751) , atwo-generatio n reproductio n stud y i n th e rat (guidelin e 83-4 ; MRI D 435223-01 ) an dathree-generatio n study i n th e rat (guidelin e 83-4 ; MRI D 00080750) . Al l o f thes e studie s were determine db y th e USEP A a s guidelin e acceptabl e excep t th e one-generatio n mous e whic hwa s foun dt o b e supplemental . A one-generatio n reproductio n study wa s conducte di n mal e an dfemal e CD- I (HaM/IC R Swiss ) mice . I n thi s study (WARF , 1975b) , norflurazo n technical wa s administere di n th e die t a t nomina l dose s o f

0 , 85 , 170 , o r

34 0 pp m(0 , 12.1 , 24.3 , o r 48. 5 mg/kg/day) . There wer e n o treatment-relate deffect s reporte di n parenta l mic e o r i n offsprin g at

an y o f th e dos e level s tested . Th e systemi c an dreproductiv e NOE Lwa s determine dt o b e 48. 5 mg/kg/day , an dth e systemi c an dreproductiv e LE Lwer e determine dt o b e > 48. 5 mg/kg/day . Thi s stud y wa s classifie da s core supplementary data . A three-generatio n reproductiv e toxicity stud y wa s conducte di n mal e an dfemal e Sprague Dawle y rat s (WARF , 11975b) . I n thi s study , norflurazo n technical (98.8 % a.i. ) wa s administere di n th e die t

a t nomina l dose s o f

0 , 125 , 375 , o r

102 5 pp m(0 , 6.25 , 18.75 , o r 51.2 5 mg/kg/day) . Th e F Q generatio n o f rat s receive dnorflurazo n technica l at nomina l dos e level s o f 0 , 125 , 250 , o r

50 0 pp m(0 , 6.25 , 12.5 , o r 25. 0 mg/kg/day ) fo r

1 8 week s prio r

to matin g t o produc e th e F] a

### an dFi b generation . Subsequen t t o this , th e Fj a

### generatio n wa s use d fo r atw o year

feedin g study , whil e th e Fi b generation s receive dnorflurazo n technica l a t 0 , 125 , 375 , o r

102 5 pp mfo r

1 3 week s an dwer e the n mate dt o produc e th e p2 a

### an dF2 b generations . Th e F2 alitte r continue dt o receiv e tes t

articl e an dwer e sacrifice djus t prio r t o delivery o f

asecon dlitte r

(Fsb) , whic hwa s examine dfo r developmenta l effects . There wer e n o dat apresente dfo r

parenta l rat s i n thi s study . Reproductiv e dat apresente dindicate dn o apparen t effect s o f norflurazo n treatmen t

o n reproductiv e performanc e at an y dos e leve l tested . Th e systemi c toxicity NOE Lwa s determine dt o b e 18.7 5 mg/kg/day , an dth e systemi c toxicit y LE Lwa s determine dt o b e 51.2 5 mg/kg/day . Th e reproductiv e toxicit y NOE Lwa s determine dt o b e 51.2 5 mg/kg/day an dth e reproductiv e toxicity LE Lwa s determine dt o b e > 51.2 5 mg/kg/day . Du e t o th e lac k o f parenta l toxicity dat ai n thi s study , th e systemi c toxicit y NOE Lan dLE Lwer e base do n th e result s o f subchroni c an dchroni c testin g withnorflurazon , i n whic ha n effec t leve l o f 2 5 mg/kg/da y wa s indicated . However , th e Healt hEffect s Divisio n RfD/Qualit y Assuranc e Committe e determine dtha t i t wa s no t Report Number : TK002342 5 Pag e 1 0 o f 4 3

### appropriat e t o extrapolat e asystemi c NOEL/LE Lt o thi s study base do n effect s observe d

fro msubchroni c an dchroni c toxicity studies . Nonetheless , th e Committe e fel t that th e conclusion s draw n fro mth e subchroni c an dchroni c toxicit y studie s coul db e use dt o justif y that th e dos e level s wer e probabl y adequate . Therefore , th e systemi c toxicity NOE Lan d LE Lwer e considere dappropriate . A 2-generatio n reproductiv e toxicity stud y wa s conducte di n mal e an dfemal e Wistar rat s (Eschbac he t

al. , 1994) . I n thi s study , norflurazo n technical wa s administere di n th e die t at dos e level s o f

0,150 , 750 , o r

150 0 pp move r

2 generation s (0 , 10.2 , 50.8 , o r 102. 5 mg/kg/day fo r F 0 

### males ; 0 , 12.1 , 62.0 , o r 129. 7 mg/kg/day fo r

F 0 

### females ; 0 , 13.2 , 67.8 , o r 138. 6 mg/kg/day fo r

F l males ; 17.1 , 81.7 , an d173. 0 mg/kg/day fo r F I females) . Th e NOE Lfo r systemi c toxicity wa s determine dt o b e 15 0 pp m(10. 2 mg/kg/day fo r male s an d12. 1 mg/kg/day fo r

females) , an dth e systemi c toxicit y LE Lwa s determine dt o b e 75 0 pp m(50. 8 mg/kg/day fo r

male s an d62. 0 mg/kg/day fo r females) , base do n significan t increase s i n live r an dkidne y weight s observe di n bot hgeneration s o f

parenta l rat s an dth e increase dincidenc e o f hepatocellular

hypertrophy i n bothgeneration s o f parenta l rats . Th e reproductiv e toxicit y NOE Lwa s determine dt o b e 75 0 pp m(50. 8 mg/kg/da y fo r male s an d62. 0 mg/kg/day fo r females) , an dth e reproductiv e toxicit y LE Lwa s determine dt o b e 150 0 pp m(102. 5 mg/kg/day fo r

male s an d129. 7 mg/kg/day fo r females) , base do n an increas e i n pu p deaths , a n increas e i n stillborn pup s i n th e Fi b an dFi alitters , an dadecrease dlactatio n inde x i n th e F 2 b 

### litter . Th e studie s examinin g developmental an dreproductiv e toxicit y o f norflurazo n wer e considere db y th e HealthEffect s Divisio n RfD/Pee r Revie w Committe e i n ameetin g hel d Marc h16 , 199 5 (USEPA , 2002) . Th e committe e conclude dtha t treatmen t wit hnorflurazo n wa s associate dwithreproductiv e toxicity i n th e for mo f increase dpu p death , increase d stillbor n pups , an dincrease dpu p death s betwee n day s 5-1 4 o f lactatio n at

ados e o f 1500 pp m[12 9 an d17 3 mg/kg/da y fo r F 0 

### an dF I females , respectively ] (USEPA , 2002) . 3. 2 Developmenta l Toxicity Studie s Developmenta l toxicity studie s wit hnorflurazo n hav e bee n conducte di n th e rat (guidelin e 83-3 ; MRI D 00063621 ) an dth e rabbi t (guidelin e 83-3 ; MRI D 0013115 1 an d00131152) . Al l o f thes e studie s wer e determine db y th e USEP A a s guidelin e acceptable . I n adevelopmenta l toxicity (teratology ) stud y (Hartman , 1972) , rat s o f th e Sprague-Dawle y strai n receive d0 , 100 , 200 , o r 40 0 mg/kg/da y norflurazo n technica l b y oral gavag e o n gestatio n day s 6 throug h1 5 inclusive. Th e materna l toxicit y NOE Lwa s determine dt o b e < 10 0 mg/kg/day , an dth e materna l toxicity LE Lwa s determine dt o b e < 10 0 mg/kg/day , base d o n reduction s i n bod y weigh t gai n fo r th e perio do f

dosin g an dfo r

th e dosin g plu s post dosin g period . Th e developmenta l toxicit y NOE Lwa s determine dt o b e > 40 0 mg/kg/day an dth e developmenta l toxicit y LE Lwa s determine dt o b e > 40 0 mg/kg/day . Developmenta l toxicit y wa s suggeste dat

th e 40 0 mg/kg/da y dos e leve l i n th e for mo f an increas e i n bipartit e thoraci c vertebra e (10th-13th ) an dan increas e i n rudimentar y 14t h ribs . However , thes e increase s wer e no t

statisticall y significan t

an dar e believe dt o b e secondary t o materna l effect s at th e hig hdos e (USEPA , 2002) . Report Number : TK002342 5 Pag e 1 1 o f 4 3A developmental toxicity stud y wa s conducte di n Ne w Zealan dWhit e rabbits , whic h

receive d0 , 10 , 30 , o r

6 0 mg/k g /da y norflurazo n technical , b y ora l gavag e o n gestatio n day s 7 throug h 1 9 inclusive . Th e materna l toxicit y NOE L wa s determine dt o b e 3 0 mg/kg/day , an dth e materna l toxicity LE L wa s determine dt o b e 6 0 mg/kg/day , base do n decrease dbod y weigh t gai n an dclinica l toxicity (abortion ) at thi s dos e level . Th e developmenta l toxicity NOE L wa s determine dt o b e 3 0 mg/kg/day , an dth e developmenta l toxicit y LE L wa s determine dt o b e 6 0 mg/kg/day , base do n statisticall y significan t increase s i n th e feta l incidenc e o f

incompletel y ossifie dfronta l bones , caudal vertebrae , unossifie dfirs t metacarpal , unossifie dmiddl e phalan x o f th e fift hdigi t

o f th e forelimb , an dunossifie d proxima l epiphysi s o f

th e tibi aat

th e 6 0 mg/kg/day dos e level . A 12 % decreas e i n mea n feta l weigh t wa s als o observe dat

th e 6 0 mg/kg/da y dos e level . Health Effect s Divisio n RfD/Pee r Revie w Committe e whic h me t i n Marc h 16 , 199 5 an d conclude dthat

developmenta l toxicity wa s no t eviden t i n treate drats , bu t

developmenta l toxicit y i n rabbit s wa s eviden t at ados e o f 6 0 mg/kg/da y i n th e for mo f decrease dmean feta l weight , sligh t

delay s i n ossificatio n o f

th e skul l an dlimbs , an dan increas e i n th e incidenc e o f 13t h ribs . Thes e developmenta l effect s occurre dat ados e tha t

wa s maternall y toxi c (USEPA , 2002) . 

### 3. 3 Roden t

Subchroni c an dChroni c Toxicity/Carcinogenicit y Studie s I n a90-day rat

feedin g stud y (Fogleman , 1971) , th e systemi c NOE L wa s determine dt o b e 12. 5 mg/kg/day i n males , an d2 5 mg/kg/day i n females . Th e 6-month do g feedin g stud y (Klotzch e e t

al . 1973 ) ha dasystemi c NOE L o f 1.5 3 mg/kg/da y fo r

male s an d1.5 8 mg/kg/day fo r

females ; th e systemi c LE L wa s determine dt o b e 5.0 2 mg/kg/day fo r male s an d4.7 7 mg/kg/da y fo r

female s base do n increase dabsolut e an drelativ e live r weigh t an d increase dcholestero l i n both sexes , h i a9-month rat feedin g stud y (Harris , 1972) , th e systemi c NOE L wa s determine dt o b e 12. 5 mg/kg/day fo r bot h sexes , an dth e systemi c LE L wa s determine dt o b e 2 5 mg/kg/da y i n both sexe s base do n th e dose-relate dincreas e i n live r weigh t i n mal e an dfemal e rat s at 3 9 weeks , th e increas e i n gona dweigh t o f females , an dth e microscopi c change s observe di n kidney s o f both sexes . Further , i n th e rat

combine dchroni c toxicity/carcinogenicity stud y (Warf, 1975c) , th e systemi c NOE L wa s determine dt o b e 18.7 5 mg/kg/da y fo r both sexes . Th e systemi c LE L wa s determine dt o b e 51.2 5 mg/kg/da y i n both sexe s (HDT) , base do n th e increase dkidne y weigh t an daccompanyin g microscopi c pathologi c changes , a s wel l a s th e increas e i n live r weigh t i n mal e an dfemal e rat s an dth e increas e i n thyroi dweigh t i n males . Ther e wa s n o evidenc e o f

carcinogenicity fo r

norflurazon . A two-year carcinogenicity stud y i n mic e (WARF , 1975d) , th e systemi c NOE L wa s determine dt o b e 12. 8 mg/kg/day fo r mal e mice , an d58. 7 mg/kg/da y fo r

femal e mice . Th e systemi c LE L wa s determine dt o b e 58. 7 mg/kg/da y fo r

mal e mice , base do n th e increase dincidenc e o f enlarge dspleen , increase d absolut e an drelativ e live r weight , an dincrease dincidenc e o f nephritis . Th e systemi c LE L wa s determine dt o b e 218. 8 mg/kg/day fo r femal e mice , base do n th e increase dincidenc e o f enlarge dlive r

an dcysti c ovaries , th e increase dabsolut e an drelativ e live r weight , an dth e increase dincidenc e o f

pyelonephritis . Base dupo n statisticall y significan t pair-wis e comparison s o f

th e incidenc e o f

live r adenoma s an dcombine dlive r adenomas/carcinoma s a s Repor t Number : TK002342 5 Pag e 1 2 o f 4 3

### wel l a s statisticall y positiv e trend s fo r thes e lesion s i n mal e CD- I mic e receivin g 218. 8 mg/kg/day norflurazo n technica l i n th e die t fo r u p t o 10 4 weeks , th e OPP/HealthEffect s Divisio n Carcinogenicity Peer Revie w Committee , norflurazo n wa s classifie da s ano n quantifiabl e Group C - possibl e huma n carcinogen . 3. 4 Fis h an dAmphibia n Studie s Norflurazo n ha s bee n teste di n th e guidelin e acut e an dchroni c toxicity test s i n fish . Fis h earl y lif e cycl e studie s wer e conducte dwit hnorflurazo n i n th e rainbo w trou t (EG& G Bionomics , 1982a) an dfathea dminno w (EG& GBionomics , 1982b ) an dther e wer e n o endocrine-mediate dchanges . T o ou r knowledge , n o norflurazo n toxicit y dat aha s bee n generate dwithamphibia n species . 

### 4. 0 RATIONAL E FO RACCEPTANC E O FNORFLURAZO N OSRI FO RSPECIFI C TIE R1 ASSAY S 4. 1 Estroge n Recepto r Bindin g Assay s (OPPT S 890.1250 ; OPPT S 890.1300) Th e norflurazo n OSRI fo r

th e Estroge n Recepto r

Bindin g Assa y i s base do n tw o differen t sources : th e EP A ToxCast TM programresult s o n norflurazo n an dth e U S EPA' s 40CF RPart 15 8 guideline s studie s supportin g th e norflurazo n registration . Th e ToxCas t dat aprovide s substantiall y similar

endpoints/methodolog y a s th e EDS P Tie r 1 scree n an dth e USEP A 40CF RPar t

15 8 dat aprovide s th e Tie r 2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmenta l toxicity , reproductio n effects , chroni c toxicit y an dother apica l endpoint s o f

endocrin e functio n effects . EPA' s ToxCast TM progra mresults : Norflurazo n ha s bee n previousl y teste dfo r estroge n bindin g i n th e EP A ToxCas t program(Kavloc k

e t al , 2009 ; Houck , 2009) . Th e protoco l utilize dwa s deeme dappropriat e b y U S EP A fo r estroge n bindin g potentia l (se e El i Lill y an d Compan y an dNI H Chemica l Genomic s Center , 2008) . Th e protocol s fo r th e assay s use di n th e ToxCast TM evaluatio n ar e no t currentl y available , howeve r th e assay s wer e conducte db y an dfo r EPA . Thes e EPA-supporte dassay s apparentl y mee t th e qualit y standard s o f adherenc e t o th e spiri t

o f goo dlaboratory practice s an dar e o f hig hquality , achievin g th e objectiv e t o provid e reasonabl e assuranc e tha t achemical doe s o r

doe s no t hav e th e potentia l t o interac t wit hestroge n receptor s (El i Lill y an dCompan y an dNI H Chemica l Genomic s Center , 2008) . Base do n th e existenc e o f

thi s data , Syngent arequest s awaive r fo r th e EDS P Tie r

1 scree n fo r estroge n binding . U S EPA' s 40CF RPart

15 8 dat arequirements : Norflurazo n ha s an extensiv e toxicity database , includin g th e 2-generatio n reproductio n stud y whic hha s bee n designate db y th e EP A a s th e Tie r

2 stud y i n th e EDS P progra mfo r mammalia n endocrin e toxicit y characterization . Repor t Number : TK002342 5 Pag e 1 3 o f 4 3I t

i s importan t

t o recogniz e i n ligh t

o f th e us e o f

othe r scientificall y relevan t

informatio n tha t ther e ar e endpoint s measure di n th e breadth o f th e EP A guidelin e studie s whic h can detec t effect s o n th e endocrin e system . Fo r example , an increas e i n uterin e weigh t usuall y reflect s a respons e t o estrogen , o r

afunctionally-equivalen t measur e o f

estroge n actin g throug h an estroge n receptor-mediate dreaction . Th e endpoint s measure di n th e EP A guidelin e studie s fo r norflurazo n whic h can detec t afunctionall y equivalen t respons e t o i n vitr o estroge n recepto r bindin g (OPPT S 890.125 0 an d890.1300) , bot h agonis man dantagonism , are presente di n Tabl e 2 . Tabl e 2 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte dwit h Norflurazo n whic h ar e Relevan t t o i n vitro E RBindin g an dt o th e Estrogeni c Mod e o f Actio n 3-Generatio n Ra t Reproduction . FIFRA §83-4 ; EP A MRI D 008075 0 2-Generatio n Ra t Reproduction . FIFRA §83-4 ; EP A MRI D 4352230 1 Organ weight : Uterus , Ovary , Pituitar y Histopathology : Uterus , Vagina , Cervix , Ovary , Pituitary , Femal e Reproductiv e indices : mating , conception , fertility , gestation , an dlactatio n Dystoci a Pu p se x rati o No . pup s "missexe d"at

birt h compar e t o se x a t

cul l an d necrops y Normalit y o f

externa l genitali a i n pup s an dF l adult s Tim e t o matin g Gros s lesion s Rabbi t Developmenta l Toxicity . FIFRA §83-3 . EP A MRI D 0013 1 15 2 Ra t Developmenta l Toxicity . FIFRA §83-3 . EP A MRI D 0006362 1 Organ weight : Uteru s (gravid ) Numbe r o f corpor alutea , implantatio n sites , fetuse s

— Feta l se x rati o Feta l reproductiv e trac t anomalie s

—

_ 90-Day Subchroni c

Rat . FIFRA §82-1 . EPA MRI D 0009105 5 Orga n weight : Adrenals , Ovary Histopathology : Uterus , Ovary , Adrenals , Pituitary , Thyroi d 

# ~

—

—

— Gros s lesion s 6 - Mont h Do g Chroni c Toxicity . FIFRA §83-1 . EP A MRI D 00 1 1 1 6 1 8 . Ra t Chroni c Toxicity / Carcinogenicity . FIFRA §83-1 . EPA MRID' s 008201 9 an d0011164 9 Organ weight : Uterus , Ovary , Pituitary , Thyroid , Adrena l Histopathology : Uterus , Ovary , Mammar y gland , Pituitary , Thyroid , Adrena l

—

—

— Gros s lesion s I f norflurazo n wer e estrogeni c o r anti-estrogenic , change s i n uterin e an dovaria n weight s shoul dmanifes t

themselve s i n no t

onl y FIFRA §83- 4 an dFIFR A §83-3 , bu t als o FIFRA §82- 1 an dFIFRA §83- 1 guidelin e studies . Likewise , change s i n th e histopatholog y o f thes e endocrin e tissue s an daccessor y se x tissue s lik e th e mammar y glan dshoul doccur . Needles s t o say , effect s o n reproductiv e indice s woul db e observe di n th e EDS P Tier I I gol dstandar d study , th e 2-generatio n reproductio n stud y i n rats . Therefore , base do n th e availabl e norflurazo n OSRI , Syngent arequest s awaive r fro mthi s EDS P Tie r

1 screen . 

### 4. 2 Androge n Recepto r Bindin g (OPPT S 890.1150) Th e norflurazo n OSRI fo r th e Androge n Recepto r

Bindin g Assa y i s base do n tw o differen t sources : th e EP A ToxCast TM progra mresult s o n norflurazo n an dth e U S EPA' s 40CF RPart 15 8 guideline s studie s supportin g th e norflurazo n registration . Th e ToxCas t dat aprovide s substantiall y simila

r methodology/endpoint s a s th e EDS P Tie r 1 scree n an dth e USEP A Repor t Number : TK002342 5 Pag e 1 4 o f 4 3

### 40CF RPart 15 8 dat aprovide s th e Tie r 2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmenta l toxicity , reproductio n effects , chroni c toxicity an dother apical endpoint s o f

endocrin e functio n effects . EPA' s ToxCast TM progra mresults : Norflurazo n ha s bee n previousl y teste dfo r androge n bindin g i n th e EP A ToxCas t program(Kavloc k

e t al , 2009 ; Houck , 2009) . Th e protoco l utilize dwa s deeme dappropriat e b y U S EP A fo r androge n bindin g potentia l (se e El i Lill y an d Compan y an dNI H Chemica l Genomic s Center , 2008) . Th e protocol s fo r assay s use di n th e ToxCast TM evaluatio n ar e no t currentl y available ; however , th e assay s wer e conducte db y an d fo r EPA . Thes e EPA-supporte dassay s apparentl y mee t th e qualit y standard s o f

adherenc e t o th e spiri t o f goo dlaboratory practice s an dar e o f hig hquality , achievin g th e objectiv e t o provid e reasonabl e assuranc e that achemica l doe s o r

doe s no t hav e th e potentia l t o interac t withandroge n receptor s (El i Lill y an dCompan y an dNI H Chemica l Genomic s Center , 2008) . Base do n th e existenc e o f thi s data , Syngent arequest s awaive r fo r th e EDS P Tie r

1 scree n fo r androge n binding . U S EPA' s 40CF RPart

15 8 dat arequirements : Norflurazo n ha s an extensiv e toxicit y database , including th e 2-generatio n reproductio n stud y whic hha s bee n designate db y th e EPA a s th e Tie

r 2 study i n th e EDS P progra mfo r mammalian endocrin e toxicit y characterization . Th e endpoint s measure di n th e existin g FIFRA guidelin e studies provid e functiona l equivalent s t o i n vitr o androge n recepto r bindin g (OPPT S 890.1150) . Thes e parameter s fo r

studie s wit hnorflurazo n ar e give n i n Tabl e 3 . Tabl e 3 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte dwit hNorflurazo n whic har e Relevan t t o i n vitro A RBindin g an dt o th e Androgeni c Mod e o f Actio n 3-Generatio n Ra t Reproduction . FIFR A §83-4 ; EP A MRI D 008075 0 2 -Generatio n Ra t Reproduction . FIFR A §83^» ; EP A MRI D 4352230 1 Organ weight : Testes , Pituitary , Thyroids , Adrenal s Histopathology : Testes , Prostate , Semina l Vesicles , Epididymide s Mal e Reproductiv e indices : Mating , Fertility , Conceptio n Tim e t o matin g Normalit y o f

externa l genitali a No . pup s "missexe d"at

birt h compar e t o se x at

cul l an d necrops y Pu p se x rati o Undescende dteste s i n F l male s Gros s lesion s Rabbi t Developmenta l Toxicity . FIFR A §83-3 . EP A MRI D 0013 1 15 2 Ra t Developmenta l Toxicity . FIFR A §83-3 . EP A MRI D 0006362 1

— Feta l Reproductiv e trac t anomalie s Feta l se x rati o

—

— 90-Day Subchroni c

Rat . FIFR A §82-1 . EP A MRI D 0009 1 05 5 Organ weight : Testes , Epididymide s Histopathology : Testes , Epididymides , Semina l vesicles , Prostat e

—

—

—

— Gros s lesion s 6 - Mont hDo g Chroni c Toxicity . FIFR A §83-1 . EP A MRI D 00 1 1 1 6 1 8 . Ra t Chroni c Toxicity / Carcinogenicity . FIFR A §83-1 . EP A MRID' s 008201 9 an d0011164 9 Organ weight : Testes , Epididymides , Prostate , Thyroid , Adrenals , Pituitar y Histopathology : Testes , Epididymides , Prostate , Adrenals , Thyroid , Pituitar y

—

—

—

— Gros s lesion s 

### Report Number : TK002342 5 Pag e 1 5 o f 4 3I f

norflurazo n wer e androgeni c o r anti-androgenic , thes e effect s woul db e note di n th e existin g FIFRA §83-4 , FIFR A §82- 1 an dFIFR A §83- 1 guidelin e studie s throug h alteration s i n endocrin e and/o r

accessor y se x tissu e weigh t

and/o r pathology , alteratio n o f

reproductio n an dfecundit y i n FIFRA §83- 4 o r se x rati o i n FIFR A §83- 3 studies.Therefore , base do n th e availabl e norflurazo n OSRI , Syngent arequest s awaive r fro mthi s EDS P Tier

1 Screen . 

### 4. 3 Aromatas e -Huma n Recombinan t (OPPT S 890.1200 ) Th e norflurazo n OSRI fo r th e aromatas e assay i s base do n tw o differen t sources : th e EP A ToxCast TM programresult s o n norflurazo n an dth e U S EPA' s 40CF RPart 15 8 guideline s studie s supportin g th e norflurazo n registration . Th e ToxCas t dat aprovide s substantiall y similar methodology/endpoint s a s th e EDS P Tie r 1 scree n an dth e USEP A 40CF RPart 15 8 dat aprovide s th e Tie r

2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmenta l toxicity , reproductio n effects , chroni c toxicity an dother apical endpoint s o f endocrin e functio n effects . EPA' s ToxCast TM progra mresults : Norflurazo n ha s bee n previousl y teste dfo r aromatas e assay i n th e EP A ToxCas t program(Kavloc k

e t al. , Houck , 2009) . Th e protoco l utilize dwa s deeme dappropriat e b y U S EP A fo r assessin g effect s o n aromatase . Th e protocols fo r assay s use di n th e ToxCast TM evaluatio n ar e no t currentl y available ; howeve r th e assay s wer e conducte db y an dfo r

EPA . Thes e EPA-supporte dassay s apparentl y mee t th e quality standard s o f

adherenc e t o th e spiri t

o f goo dlaboratory practice s an dar e o f hig h quality , achievin g th e objectiv e t o provid e reasonabl e assuranc e tha t achemica l doe s o r

doe s no t hav e th e potentia l t o interac t

wit h aromatas e activit y (Eli Lill y an dCompan y an dNI H Chemica l Genomic s Center , 2008) . Base do n th e existenc e o f thi s data , Syngent arequests awaive r fo r th e EDS P Tie r

1 scree n fo r

assessin g aromatas e effects . U S EPA' s 40CFRPart

15 8 dat arequirements : Norflurazo n ha s a n extensiv e toxicit y database , including th e 2-generatio n reproductio n stud y whic h ha s bee n designate db y th e EP A a s th e Tie r

2 stud y i n th e EDS P progra mfo r mammalia n endocrin e toxicit y characterization . Althoug h th e measuremen t o f inhibitio n o f

aromatas e [Human Recombinant ] (OPPT S 890.1200 ) i s unlik e direc t alteratio n o f

bindin g o f

asteroi dhormon e with it s receptor , th e aromatas e focuse s o n th e alteratio n o f estroge n formatio n an dtherefor e th e endpoint s tha t ar e measure di n th e FIFRA guidelin e studie s ar e functionall y equivalen t t o thos e evaluate di n OPPT S 890.125 0 and/o r

OPPT S 890.1300 . Thes e endpoint s ar e presente di n Tabl e 4 . OPPT S 890.120 0 wil l onl y detec t areductio n i n aromatas e activit y whic h wil l resul t i n a reductio n i n testosteron e o r an anti-androgeni c response . Agai n thi s woul db e detecte di n existin g EP A guidelin e studie s that wer e conducte dusin g th e FIFR A §83-4 , FIFRA §83-3 , FIFRA §82- 1 an dFIFR A §83- 1 guidelines . Therefore , base do n th e availabl e norflurazo n OSRI , Syngent arequest s awaive r fro mthi s EDS P Tie r

1 Screen .
Repor t Number : TK002342 5 Pag e 1 6 o f 4 3

### Tabl e 4 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte dwit hNorflurazo n whic har e Relevan t t o Aromatas e Inhibitio n 3-Generatio n Rat Reproduction . FIFRA §83-4 ; EP A MRI D 008075 0 2 -Generatio n Ra t Reproduction . FIFRA §83-4 ; EP A MRI D 4352230 1 Organ weight : Pituitary , Testes , Epididymides , Uterus , Ovary , Adrenal , Thyroi d Histopathology : Pituitary , Testes , Semina l Vesicle , Epididymides , Prostate , Uterus , Ovary Reproductiv e indices : Mal e an dfemal e , Mating , Fertility , Conception , Femal e Gestatio n an dLactatio n No . pup s "missexe d"at

birt h compar e t o se x a t

cul l an d necrops y Normalit y o f

externa l genitali a Gros s lesion s Rabbi t Developmenta l Toxicity . FIFRA §83-3 . EPA MRI D 0013115 2 Ra t Developmenta l Toxicity . FIFRA §83-3 . EP A MRI D 0006362 1 Organ weight : Uteru s

Numbe r o f corpor alutea , implantatio n sites , fetuse s

Feta l reproductiv e trac t anomalies

— 90-Day Subchroni c Rat . FIFRA §82-1 . EP A MRI D 0009105 5 Organ weight : Testes , Uterus , Ovary , Thyroid , Adrena l Histopathology : Testes , Epididymides , Prostate , Semina l vesicles , Uterus , Ovary , Mammar y gland , Thyroid , Adrenal , Pituitar y

— Gros s lesion s 6 - Mont hDo g Chroni c Toxicity . FIFRA §83-1 . EPA MRI D 00111618 . Ra t Chroni c Toxicity / Carcinogcnicity . FIFRA §83-1 . EPA MRID' s 008201 9 and00 1 1164

9 Organ weight : Testes , Uterus , Ovary , Thyroi d(non-rodent) , Adrena l Histopathology : Testes , Epididymides , Prostate , Semina l vesicles , Uterus , Ovary , Mammar y gland , Thyroi d, Adrenal , Pituitar y

— Gros s lesion s 

### 4. 4 Steroidogenesi s - Huma n Cell Lin e H295 R(OPPT S 890.1550) Th e norflurazo n OSRI fo r Steroidogenesi s assa

y i s base do n th e exitin g U S EPA' s 40CF R Part 15 8 guidelin e studie s that suppor t th e norflurazo n registration . Th e USEP A 40CF RPart 15 8 dat aprovide s th e Tie r 2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmental toxicity , reproductio n effects , chroni c toxicit y an dothe r apical endpoint s o f endocrin e functio n effects . A s wit haromatas e inhibition , th e endpoint s measure di n th e H295 RSteroidogenesi s scree n (OPPT S 890 . 1550 ) are alteration s o f estradio l an dtestosterone . Therefore , th e relevan t endpoint s measure di n th e FIFR A guidelin e studie s encompas s th e organ s an dtissue s tha t woul db e potentiall y affecte db y th e alteratio n o f bot hsteroid s (Tabl e 5) . I f norflurazo n ha d effect s o n Steroidogenesis , th e EP A guidelin e studies , FIFRA §83-4 , FIFRA §83-3 , FIFR A §82- 1 an dFIFR A §83-1 , woul db e capabl e o f detectin g estrogenic , anti-estrogenic , androgeni c an danti-androgeni c change s i n th e apica l endpoints . Therefore , base do n th e availabl e norflurazo n OSRI , Syngent arequest s awaive r fro mthi s EDS P Tie r

1 Screen .

Report Number : TK002342 5 Pag e 1 7 o f 4 3

### Tabl e 5 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte dwith Norflurazo n whic har e Relevan t t o th e Steroid s Alteration s tha t

migh t b e Detecte di n th e Steroidogenesi s Scree n 3-Generatio n Rat Reproduction . FIFR A §83-4 ; EP A MRI D 008075 0 2-Generatio n Ra t Reproduction . FIFR A §83-4 ; EP A MRI D 4352230 1 Organ weight : Pituitary , Testes , Epididymides , Uterus , Ovary , Adrenal , Thyroi d Histopathology : Pituitary , Testes , Semina l Vesicle , Epididymides , Prostate , Uterus , Ovary Reproductiv e indice s No . pup s "missexe d"at

birt h compar e t o se x at

cul l an d necrops y

— Corpor alute a Normalit y o f

externa l genitali a

— Gros s lesion s Rabbi t Developmenta l Toxicity . FIFRA §83-3 . EP A MRI D 00 1 3 1 15 2 Ra t Developmenta l Toxicity . FIFRA §83-3 . EP A MRI D 0006362 1 Organ weight : Uteru s

Numbe r o f corpor alutea , implantatio n sites , fetuse s

— Corpor alute a Feta l reproductiv e trac t anomalie s Numbe r o f implantatio n site s

— 90-Da y Subchroni c Rat . FIFRA §82-1 . EPA MRI D 0009105 5 Orga n weight : Pituitary , Testes , Ovary , Thyroid , Adrenal Histopathology : Testes , Epididymides , Prostate , , Uterus , Ovary , Thyroid , Adrenal , Pituitar y

— Seru mcholestero l

—

—

— Gros s lesion s 6 - Mont hDo g Chroni c Toxicity . FIFRA §83-1 . EPA MRI D 00111618 . Ra t Chroni c Toxicity / Carcinogenicity . FIFR A §83-1 . EPA MRID' s 00820 1 9 and00 1 1 1 64 9 Organ weight : Testes , Uterus , Ovary , Thyroi d(non-rodent) , Adrenal Histopathology : Testes , Epididymides , Prostate , Semina l vesicles , Uterus , Ovary , Thyroi d, Adrenal , Pituitary , Mammar y glan d

—

—

—

— Gros s lesion s 

### 4. 5 Hershberge r

- Rat (OPPT S 890.1400 ) Th e norflurazo n OSRI fo r Hershberge r

assay i s base do n th e exitin g U S EPA' s 40CF RPart 15 8 guidelin e studie s tha t support th e norflurazo n registration . Th e USEP A 40CF RPart 15 8 dat aprovide s th e Tie r

2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmenta l toxicity , reproductio n effects , chroni c toxicity an dothe r apica l endpoint s o f endocrin e functio n effects . I n viv o EDS P screen s focu s o n man y o f th e apica l endpoin t

tha t ar e measure di n FIFRA guidelin e studie s fo r

developmental , reproductive , subchroni c an dchroni c toxicity . Th e endpoint s measure di n th e Hershberge r scree n (OPPT S 890.1400 ) ar e presente di n Tabl e 6 agains t th e FIFR A guidelin e studie s showin g functiona l equivalence . Whil e th e Hershberge r

assay ha s no t

bee n specificall y conducte dwithnorflurazon , th e FIFR A guidelin e studie s conducte dwit hnorflurazo n captur e importan t parameter s suc ha s chang e i n teste s weigh t

and , i n th e cas e o f

FIFRA §83-4 , prostat e weigh t whic har e evaluate d i n th e Hershberge r

screen . However , th e FIFRA studie s als o includ e th e measuremen t o f more definitiv e endpoint s suc h a s histopatholog y o f th e teste s (whic hha s bee n remove di n th e Hershberger ) an dreproductiv e an ddevelopmenta l parameter s i n model s wit ha n intac t hypothalamic-pituitary-gonada l axis . Therefore , base do n th e availabl e norflurazo n OSRI , Syngent arequest s awaive r fro mthi s EDS P Tie r

1 Screen . Report Number : TK002342 5 Pag e 1 8 o f 4 3

### Tabl e 6 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte dwit hNorflurazo n Compare dt o thos e Measure di n th e Hershberge r

Scree n Hershberge r

Endpoint s

Cowper' s Glan dWeigh t Teste s Weigh t Epididymide s Weigh t Ventra l prostat e Weigh t Semina l Vesicl e Weigh t Levator Ani bulbocavernosu s Muscl e Weigh t Glan s Peni s Weigh t (castrate dmale )

—

—

— 3-Generatio n Ra t

Reproduction . FIFRA §83-4 ; EPA MRI D 008075 0 2-Gcneratio n Ra t Reproduction . FIFRA §83-4 ; EPA MRI D 4352230 1

—

X X X X 

# ~

— Rabbi t

Developmenta l Toxicity . FIFRA §83-3 . EPA MRI D 0013115 2 Ra t Developmenta l Toxicity . FIFRA §83-3 . EP A MRI D 0006362 1

—

—

—

—

— 

# ~

— 90-Da y

Subchroni c Rat . FIFR A §82-1 . EPA MRI D 0009105 5

— X X X

— 

# "

— 6 - Mont hDo g Chroni c

Toxicity . FIFRA §83 - 1 . EP A MRI D 00111618 . Ra t Chroni c Toxicity / Carcinogenicity . FIFR A §83-1 . EPA MRID' s 008201 9 an d 00111649

—

X X X

— 

# ~

— Additiona l Relevan t

Parameter s i n Existin g Studie s Histopathology : Testes , Epididymides , Prostat e Reproductiv e Indice s Pu p se x rati o Gros s lesion s Normalit y o f mal e genitali a Numbe r o f corpor a lutea , implantatio n sites , fetuse s Feta l se x rati o

— Histopathology :

Testes , Epididymides , Prostate , Semina l vesicle s

— Gros s lesion s Histopathology : Testes ,

Epididymides , Prostate , Semina l vesicle s

— Gros s lesion s 

### 4. 6 Mal e Puberta l - Rat (OPPT S 890.1500) Th e norflurazo n OSR I fo r mal e puberta l assay i s base do n th e exitin g U S EPA' s 40CF RPart 15 8 guidelin e studie s tha t

ar e require dfo r

th e norflurazo n registration . Th e USEP A 40CF R Part 15 8 dat aprovide s th e Tie r 2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmental toxicity , reproductio n effects , chroni c toxicity an dothe r apica l endpoint s o f endocrin e functio n effects . Despit e th e ag e o f

th e 3-generatio n rat

reproductio n stud y (WARF , 1975b) , th e stud y measure dbod y weights , foo dconsumption s an dth e weight s o f th e liver , kidneys , pituitary , testes , thyroid s an dadrenal s i n al l generation s o f males ; furthe r

ahistopathologi c evaluatio n wa s performe do n thes e organs . I n addition , amor e modern 2-generatio n reproductio n stud y conducte dwit h norflurazo n (Eschbac he t al. , 1994 ) recorde dth e bod y weigh t an dfoo d consumptio n fo r

F Q an dF I male s throughou t th e study , th e pu p se x distribution , an dth e examinatio n o f

smear s fo r

spermatozo aand/o r

appearanc e matin g period , whic hi s no t onl y an assessmen t

o f mal e fertilit y bu t

als o tha t norma l puberta l developmen t ha s occurred . I n addition , th e pituitary , prostate , semina l vesicle s wit hcoagulatin g gland , an dteste s wit h Repor t Number : TK002342 5 Pag e 1 9 o f 4 3epididymide s wer e evaluate dhistopathologicall y fo r th e F O an dF I males . Therefore , thes e FIFRA§83- 4 guidelin e studie s wit h norflurazo n cover nearl y al l th e endpoint s measure di n OPPT S 890.150 0 an dth e FIFRA §82- 1 an d§83- 1 studie s capture s th e clinica l chemistry (Table s 7) . Tabl e 7 Functionall y Equivalen t Endpoint s i n FIFR

A Guidelin e Studie s Conducte dwit h Norflurazo n Compare dt o thos e Measure di n th e Puberta l Mal e Scree n Puberta l Mal e Endpoint s

Growth(dail y bod y weight ) Ag e an dWeigh t

a t Preputia l Separatio n Semina l Vesicl e an d Coagulatin g glan d weigh t (withan d withou t fluid ) Ventra l Prostat e Weigh t Dorsolatera l Prostat e Weigh t Levato r An i plu s Bulbocavernosu s Muscl e Comple x (LABC)Weigh t Epididymi s Weigh t Teste s Weigh t Thyroi dWeigh t Adrena l Weigh t Pituitar y Weigh t Live r Weigh t Kidne y Weigh t Bloo dChemistry , standar dpane l Thyroi dHormon e Level s (T4 , TSH ) Testosteron e level s Teste s histopatholog y (1 ) Thyroi dhistopatholog y Epididymide s histopatholog y (1 ) Kidne y histopatholog y 3-Generatio n Ra t Reproduction . FIFRA §83-4 ; EPA MRI D 008075 0 2-Generatio n Ra t Reproduction . FIFRA §83-4 ; EP A MRI D 4352230 1 X

— X X

—

X X X X X X X

—

—



### — X X X X Rabbi t

Developmenta l Toxicity . FIFRA §83 - 3 . EPA MRI D 0013115 2 Ra t Developmenta l Toxicity . FIFRA §83 - 3 . EP A MRI D 0006362 1 X

—

_ .

—

X X

— ,

—

—

—

—

—



### — X

- X

— 90-Da

y Subchroni c Rat . FIFR A §82-1 . EPA MRI D 0009105 5 X

—

.. .

—

X X X X X X X X

—



### — X X X X 6 - Mont h Do g Chroni c

Toxicity . FIFRA §83-1 . EPA MRI D 00111618 . Ra t Chroni c Toxicity / Carcinogenicity . FIFR A §83-1 . EPA MRID' s 008201 9 an d 00111649 X

—

.. .

—

X X X X X X X X

—



### — X X X X

Report Number : TK002342 5 Pag e 2 0 o f 4 3

### Tabl e 7 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte dwit hNorflurazo n Compare dt o thos e Measure di n th e Puberta l Mal e Scree n (continued ) Puberta l Mal e Endpoint s 3-Generatio n Rat Reproduction . FIFRA §83-4 ; EPA MRI D 008075 0 2 -Generatio n Ra t Reproduction . FIFRA §83-4 ; EPA MRI D 4352230 1 Rabbi t

Developmenta l Toxicity . FIFRA §83-3 . EPA MRI D 0013115 2 Ra t Developmenta l Toxicity . FIFRA §83-3 . EPA MRI D 0006362 1 90-Da y Subchroni c Rat . FIFRA §82-1 . EP A MRI D 0009105 5 6 - Mont hDo g Chroni c

Toxicity . FIFRA §83-1 . EP A MRI D 00 1 1 1 6 1 8 . Ra t Chroni c Toxicity/ Carcinogenicity . FIFRA §83-1 . EPA MRID' s 008201 9 an d00111649 Additiona l Relevan t

Parameter s i n Existin g Studie s

—

.. .

—

— Reproductiv e Indice s

Numbe r o f corpor a lutea , implantatio n sites , fetuse s

No . pup s "missexe d "at birt hcompar e t o se x a t cul l an d necrops y Pu p se x rati o Normalit y o f

externa l genitali a Gros s lesion s —

Feta l se x rati o Feta l anomalie s

— —

Histopatholog y Pituitary , Adrenal , Prostate , Semina l vesicles , Liver , Kidne y

—

— Gros s lesion s —

Histopathology : Pituitary , Adrenal , Prostate , Semina l vesicles , Liver , Kidne y

—

— Gros s lesion s 

### Whil e th e mal e puberta l assa y ha s no t bee n specificall y conducte dwit hnorflurazon , th e FIFRA guidelin e studie s conducte dwit hnorflurazo n captur e importan t parameter s measure d i n OPPT S 890.1500 . Therefore , base do n th e availabl e norflurazo n OSRI , Syngent arequest s awaive r fro mthi s EDS P Tie r

1 Screen . 4. 7 Femal e Puberta l - Rat (OPPT S 890.1450 ) Th e norflurazo n OSRI fo r femal e puberta l assay i s base do n th e exitin g U S EPA' s 40CF R Part 15 8 guidelin e studie s tha t ar e require dfo r

th e norflurazo n registration . Th e USEP A 40CF RPart

15 8 dat aprovide s th e Tier 2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmenta l toxicity , reproductio n effects , chroni c toxicity an dothe r apica l endpoint s o f

endocrin e functio n effects . Fo r th e females , th e 3-generatio n rat reproductio n study (WARF , 1975b ) measure dbod y weight s an dmajo r

organ s weight s o f

kidneys , pituitary , ovaries , thyroid s an dadrenal s fo r female s a s define di n OPPT S 890.145 0 a s wel l a s histopathologi c evaluatio n o f thes e organs . I n addition , th e 2-generatio n reproductio n stud y conducte dwithnorflurazo n (Eschbac he t al. , 1994 ) recorde dth e bod y weigh t an dfoo dconsumptio n fo r F Q an dF I female s throughou t th e study , th e pu p se x distribution , an dth e assessmen t o f mating , fertility , gestation , an d Report Number : TK002342 5 Pag e 2 1 o f 4 3lactatio n indice s whic h measur e femal e development , fecundit y an dreproductiv e success . I n

addition , th e pituitary , ovaries , uterus , vagin aan dcervi x wer e evaluate dhistopathologicall y fo r th e F 0 an dF I females . Therefore , thes e FIFRA§83- 4 guidelin e studie s with norflurazo n cover nearl y al l th e endpoint s measure di n OPPT S 890.145 0 an dth e FIFRA §82- 1 an d§83- 1 studie s capture s th e clinica l chemistry (Tabl e 8) . Tabl e 8 Functionall y Equivalen t Endpoint s i n FIFR

A Guidelin e Studie s Conducte dwith Norflurazo n Compare dt o thos e Measure di n th e Puberta l Femal e Scree n Puberta l Femal e Endpoint s

Growth(dail y bod y weight ) Ag e an dWeigh t

a t Vagina l openin g Uteru s (blotted ) Weigh t Ovarie s (paired ) Weigh t Thyroi dWeigh t Pituitar y Weigh t Adrenal s (paired ) Weigh t Uteru s histopatholog y Ovary histopatholog y Thyroi d(colloi dare aan d follicula r cell height ) histopatholog y Bloo dChemistry , standar dpane l Thyroi dHormon e Level s Estrou s Cyclicit y

—

—

— 3-Generario n Ra t

Reproduction . FIFR A §83-4 ; EPA MRI D 008075 0 2-Gcncratio n Ra t Reproduction . FIFRA §83-4 ; EP A MRI D 4352230 1 X

— X X X X X X X X

—

— X Rabbi t

Developmenta l Toxicity . FIFRA §83-3 . EPA MRI D 0013115 2 Ra t Developmenta l Toxicity . FIFR A §83-3 . EP A MRI D 0006362 1

—

—

—

.. .

—

—

—

—

.. .

—

—

— 90-Da

y Subchroni c Rat . FIFRA §82-1 . EPA MRI D 0009105 5 X

— X X X

— X X X X X

—



### — 6 - Mont h Do g Chroni c

Toxicity . FIFRA §83-1 . EP A MRI D 00 1 1 1 6 1 8 . Ra t Chroni c Toxicity / Carcinogenicity . FIFR A §83-1 . EPA MRID' s 008201 9 an d00111649

X

— X X X X X X X X X

—



### — Additiona l Relevan t

Parameter s i n Existin g Studie s Reproductiv e indice s

Pu p se x rati o Normalit y o f

externa l Genitali a Gros s lesion s Numbe r o f corpor a lutea , implantatio n sites , fetuse s

Feta l se x rati o Feta l anomalie s Histopathology :

Pituitary , Adrenal , Mammary glan d Gros s lesion s Histopathology :

Pituitary , Adrenal , Mammar y glan d

Gros s lesions , masse s Whil e th e femal e puberta l assay ha s no t bee n specificall y conducte dwith norflurazon , th e FIFR A guidelin e studie s conducte dwith norflurazo n captur e th e importan t parameter s measure di n OPPT S 890.1450 . Therefore , base do n th e availabl e norflurazo n OSRI , Syngent arequest s awaive r fro mthi s EDS P Tie r

1 Screen . Report Number : TK002342 5 Pag e 2 2 o f 4 3

### 4. 8 Uterotrophi c - Rat (OPPT S 890.1600 ) Th e norflurazo n OSRI fo r Uterotrophi c assay i s base do n th e exitin g U S EPA' s 40CFRPart 15 8 guidelin e studie s tha t ar e require dfo r

th e norflurazo n registration . Th e USEP A 40CFR Part 15 8 dat aprovide s th e Tie r 2 assessmen t

fo r potentia l effect s o n subchroni c toxicity , developmenta l toxicity , reproductio n effects , chroni c toxicity an dothe r apica l endpoint s o f endocrin e functio n effects . Althoug h th e Uterotrophi c Scree n ha s onl y on e functiona l endpoint , uterin e weigh t a s a surrogat e fo r

measurin g arespons e t o achemica l whic h reflect s o n th e mod e o f actio n o f estrogen , i t

fall s shor t o f measurin g th e suite o f

endpoint s whic h coul dsignal estrogeni c effect s that

are measure di n th e existin g FIFRA§83-4 , FIFRA §82- 1 an d§83- 1 guidelin e studie s (Tabl e 9) . Tabl e 9 Functionall y Equivalen t Endpoint s i n FIFRA Guidelin e Studie s Conducte dwit h Norflurazo n Compare dt o thos e Measure di n th e Uterotrophi c Scree n Uterotrophi c Scree n Endpoint s

Uteru s Weigh t Uterin e histopatholog y (optional )

—

—

—

— 3-Generatio n Ra t

Reproduction . FIFR A §83-4 ; EPA MRI D 008075 0 2-Generatio n Ra t Reproduction . FIFR A §83-4 ; EPA MRI D 4352230 1

— X Rabbi t

Developmenta l Toxicity . FIFRA §83-3 . EPA MRI D 0013115 2 Ra t Developmenta l Toxicity . FIFRA §83-3 . EP A MRI D 0006362 1 X (with placenta )

— 90-Day

Subchroni c Rat . FIFR A §82-1 . EP A MRI D 0009105 5

— X 6 - Mont hDo g

Chroni c Toxicity . FIFR A §83-1 . EPA MRI D 00111618 . Ra t Chroni c Toxicity / Carcinogenicity . FIFR A §83-1 . EPA MRlD' s 008201 9 an d 0011164 9 X

— Additiona l Relevan t

Parameter s i n Existin g Studie s Orga n weight : Ovary Histopathology : Ovary , Uteru s Reproductiv e indice s Tim e t o Mating Pu p se x rati o Gros s lesion s —

Numbe r o f corpor alutea , implantatio n sites , fetuse s Feta l se x rati o Organ weight :

Ovary Histopathology : Ovary , Uterus , Mammary glan d

—

— Gros s lesion s Orga n weight :

Ovary Histopathology : Ovary , Uterus , Mammar y glan d

—

—

— Whil e th e Uterotrophi c assay ha s no t bee n specificall y conducte dwit h norflurazon , th e FIFRA guidelin e studie s conducte dwith norflurazo n captur e importan t measurin g endpoint s importan t i n measurin g estrogeni c effects . Therefore , base do n th e availabl e norflurazo n OSRI , Syngent arequests awaive r fro mthi s EDS P Tie r

1 Screen . Report Number : TK002342 5 Pag e 2 3 o f 4 3

### 4. 9 Fis h Short Ter mReproductio n (OPPT S 890.1350) Th e norflurazo n OSRI fo r

th e fis hshor

t termreproductio n stud y i s limite dt o th e exitin g U S EPA' s 40CF RPar

t 15 8 guidelin e studie s that are require dfo r

th e norflurazo n registration . Thes e studie s includ e acut e an dchroni c studie s i n fis han devidenc e fro mth e mammalia n toxicity database . Th e chroni c fis hstudie s availabl e fo r norflurazo n ar e limite dt o th e earl y lif e stag e tests , whic hd o no t includ e al l o f

th e stud y endpoint s require db y th e fis hshor t term reproductio n study . Therefore , Syngent ai s no t seekin g awaive r

at thi s tim e fo r

thi s study . 4.1 0 Amphibia n Metamorphosi s (OPPT S 890.1100) Th e norflurazo n OSRI fo r

th e amphibian metamorphosi s study i s limite dt o th e exitin g U S EPA' s 40CF RPart

15 8 guidelin e studie s tha t ar e require dfo r

norflurazo n registration . Thes e studie s includ e acute an dchroni c studie s i n fis han devidenc e fro mth e mammalia n toxicity database . Base do n ou r

revie w o f th e existin g database , ther e ar e n o existin g amphibia n studie s withnorflurazon , and , therefore , Syngent ai s no t seekin g awaive r

at thi s tim e fo r

thi s study . 

### 5. 0 CONCLUSIO N Th e objectiv e o f

thi s documen t

i s t o provid e th e OSRI that i s availabl e fo r

norflurazo n i n th e for mo f U S EP A testin g an dFIFR A guidelin e (Tier 2 ) studie s an dt o obtai n waiver s fo r certai n EDS P Tier

1 assay s where scientificall y justified . I n conclusion , norflurazo n alread y ha s bee n thoroughl y evaluate drelativ e t o th e critical apica l endpoint s covere db y th e EDS P Tie r 1 screens . Therefore , Syngent aCro p Protection , Inc. , request s tha t

th e Environmenta l Protectio n Agenc y waiv e th e requiremen t t o conduc t th e majorit y o f

th e norflurazo n EDS P tes t orde r assay s base do n th e followin g scientifi c justification . 1 . U S EP A ha s recentl y sponsore dan dproduce dOSR I fo r th e followin g studies/endpoints , therefor e thes e studie s shoul db e waive dfro mth e EDS P Tie r 1 scree n requirements :

• 890.115 0 - Androge n Recepto r Bindin g (Ra t

Prostate ) • 890.120 0 - Aromatas e (Huma n Recombinant ) • 890.125 0 - Estroge n Recepto r Bindin g • 890.130 0 - Estroge n Recepto r Transcriptiona l Activatio n (Huma n Cel l Lin e HeLa-9903 ) 

### 2 . Tie r 2 studie s hav e bee n previousl y conducte dfo r norflurazo n a s part

o f th e 40CF RPart 15 8 dat arequirement s (e.g. , rat reproductio n study , developmental toxicity , 90-da y studies , chroni c an doncogenicity studies) . Thes e EP A guidelin e studie s serv e a s OSRI fo r th e followin g studies/endpoints , therefor e thes e studie s shoul db e waive dfro mth e EDS P Tie r

1 scree n requirements: 890.140 0 - Hershberge r

(Rat ) 890.145 0 - Femal e Puberta l (Rat ) 890.150 0 - Mal e Puberta l (Rat ) 890.155 0 - Steroidogenesi s (Huma n Cel l Lin e - H295R ) 890.160 0 - Uterotrophi c (Rat ) 

### Repor t Number : TK002342 5 Pag e 2 4 o f 4 3Th e existin g databas e fo r norflurazo n doe s no t

contai n th e specifi c OSRI fo r th e fis han d amphibia n studies , an dSyngent ai s no t providin g arational e t o waiv e thes e specifi c assays . However , th e existin g mammalia n toxicit y databas e combine dwit h th e fis hearl y lif e stag e studie s provid e significan t informatio n tha t

may b e judge da s adequat e OSR I fo r awaive r o r partia l waive r

o f th e followin g tw o studies :

• 890.110 0 - Amphibia n Metamorphosi s (Frog ) • 890.1 3 5 0 - Fis h Short-TermReproductio n

Report Number : TK002342 5 Pag e 2 5 o f 4 3

### 6. 0 REFERENCE S

EG& GBionomic s (1982a) : Earl y Lif e Cycl e Stud y i n Rainbo w Trout . MRI D 00218047 . EG& GBionomic s (1982b) : Earl y Lif e Cycl e Study i n Fathea dMinnows . MRI D 0018048 . El i Lill y an dCompan y an dNI H Chemica l Genomic s Center . (2008) . Assa y Guidanc e Manua l Versio n 5.0.' . http://www.ncgc.nih.gov/guidance/manua l toc.html . (Las t accesse d 1/25/2010) . Eschbach , B. ; Aerni , R. ; Hertl , P. ; e t al . (1994) : Tw o Generatio n Reproductio n Study i n Rats : Norflurazon : Revise dFina l Report . Projec t No . 440R . (Unpublishe dstud y prepare db y Sando z Agro , Ltd.) . EPA MRID 43522301 . Fogleman , R.W . (1971 ) Evaluatio n o f th e Subchroni c (90-day ) Toxicity Effect s o f SAN 978 9 i n Rats : Contrac t

No . 120-546-9-70 . Fina l rept . (Unpublishe dstud y receive dNo v 14 , 197 2 unde r

3G1310 ; prepare db y Affiliate dMedica l Enterprises , Inc. , submitte db y Sandoz , Inc. , Homestead , Fla. ; CDL:092233-F) . EP A MRI D 00091055 . Harris , D.L . (1972 ) 9 MonthFeedin g Stud y i n Rats : F O Generation . Fina l rept . (Unpublishe d stud y receive dNo v 14,197 2 unde r 3G1310 ; prepare db y WAR FInstitute , Inc. , submitte db y Sandoz , Inc. , Homestead , Fla. ; CDL : 092233-G) . EP A MRI D 00091056 . Harris , D. ; Nees , P . (1973 ) Lon g TermFeedin g Study-Rat : 1 Year Report : Compoun d9789 . (Unpublishe dstud y receivedMay 13 , 197 4 unde r 4F1506 ; prepare db y WAR FInstitute , Inc. , submitte db y Sandoz-Wander , Inc. , Homestead , FL ; CDL:093994-G ; 093995) . EP A MRI D 00111617 . Hartman , H.A. ; Hrab , R . (1972) : Compoun d9789 : Investigatio n o f Teratogeni c Potentia l i n th e Rat : Report

No . T-1-8/7/72 . (Unpublishe dstud y submitte db y Sandoz , Inc.) . EP A MRI D 00063621 . Houck , K . (2009).What

Can ToxCast TM Tel l U s abou t Endocrin e Disruption ? Presente dat th e Internationa l Society o f Regulatory Toxicolog y an dPharmacolog y 200 9 Endocrin e Workshop , Septembe r

9-10 , 2009 . Nationa l Institute s o f Health , 860 0 Rockvill e Pike , Bethesda , MD . Hrab , R. ; D e Raismes , M. ; Yao , C . (1983a) . Dos e Range-findin g Stud y o f Norflurazo n i n Pregnan t Rabbits : Sando z Projec t

No . T-1793 . (Unpublishedstud y submitte db y Sandoz , Inc. , Cro p Protection) . EP A MRI D 00131151 . Hrab , R. ; Halbert , F. ; Quagliotti , M . e t al . (1983b) : Investigatio n o f Teratogeni c Potentia l o f Norflurazo n i n th e Rabbit—Segmen t II : Sando z Projec t

T-1794 . (Unpublishe dstud y submitte db y Sandoz , Inc. , Cro p Protection) . EP A MRI D 00131152 . Institut e fo r

Environmen t

an dHeath(IEH) . (2005) . Chemical s purporte dt o b e endocrin e disrupters : A compilatio n o f publishe dlists . IE H We b Report 20 . Marc h2005 . Report Number : TK002342 5 Pag e 2 6 o f 4 3

### http://www.cranfield.ac.uk/health/researchareas/environmenthealth/ieh/ieh%20publications /

w20.pd f Kavlock , R. ; Dix , D. ; Houck , K. ; Judson , R. ; Knudsen , T. ; Reif , D. ; an dMartin , M . (2009) . Biological Profilin g o f

Endocrin e Relate dEffect s o f Chemical s i n ToxCastTM . Presente dat ToxCas t Dat aAnalysi s Summit , RTP , NC , Ma y 14-15,2009 . http://www.epa.gov/NCCT/toxcast/summit.html . Klotzsche , C. ; Carpy , S. ; D e Rijk ; e t al . (1973) . San 978 9 (Zorial) , 6 Month s Feedin g Study i n Dogs : Report

Nr . 56/73 . (Unpublishe dstud y receive dMay 13 , 197 4 unde r 4F1506 ; prepare db y Sandoz-Wander , Inc. , Homestead , FL ; CDL : 093995-A) . EP A MRI D 00111618 . USEPA . 2002 . Reregistratio n Eligibilit y Decisio n (RED ) Documen t fo r Norfiurazon . LIS T A . CAS E 0229 . May 31 , 2002 . http://www.epa.gov/oppsrrdl/REDs/0229.pd f WAR FInstitute , Incorporate d(1975a ) Compoun d9789 : Reproductio n Studie s i n Rat s an d Mice : Volum e I : Reproductio n Studie s o n Rats . Fina l rept . (Unpublishe dstudy , includin g submitte r summary , receive dMay 8 , 197 5 unde r 5F1602 ; submitte db y Sandoz , Inc. , Homestead , Fla. ; CDL:094369-A) . EP A MRI D 00080750 . WAR FInstitute , Incorporate d(1975b ) Compoun d9789 : Reproductio n Studie s i n Rat s an d Mice : Volum e 2 : Reproductio n Studie s i n Mice . Fina l rept . (Unpublishe dstudy , includin g submitte r summary , receive dMay 8 , 197 5 unde r 5F1602 ; submitte db y Sandoz , Inc. , Homestead , Fla. ; CDL:094368-A ) EP A MRI D 00080751 . WAR FInstitute , Incorporate d(1975c ) Compoun d9789 : Tw o Year Feedin g Stud y i n Rats . Fina l rept . (Unpublishe dstudy receive dMay 8 , 197 5 unde r 5F1602 ; submitte db y Sandoz , Inc. , Homestead , Fla. ; CDL : 094370-A ; 094371 ; 094372 ; 094373 ; 094374) . EP A MRI D 00082019 . WAR FInstitute , Inc . (1975d) : Compoun d9789 : Tw o Year Feedin g Study i n Mice . Fina l Report . (Unpublishe dstud y submitte db y Sandoz-Wander , Inc.) . EP A MRI D 00111649 .

TK0023425-EDSPWaiver.do c sbh : 2/4/1 0 verl. O



### Report Number : TK002342 5 Pag e 2 7 o f 4 3

### APPENDICE S SECTIO N

Report Number : TK002342 5 Pag e 2 8 o f 4 3

### APPENDI X 1 Titl e Page s o f Cite dStudie s

Report Number : TK002342 5 Pag e 2 9 o f 4 3:GbNF.£DEMTIAI.-T.aSD E SECRE T 

### OOM804 7 

### o )' . 

### ? Guideline : requiremen t : 

### i s <- f iiljfeilled . CRED »

SUBMITTED ; T O "EAST ,

REPOR T *3W-82-5:-ll : 6: 6 Aquati c 7bxicoi-a : gy , Labo'ratoir?/ - 

### 790 : 

### Mai n Strs^- t 'fa-reh-aa ; Massapaus e tc s

'



### Report Number : TK002342 5 Pag e 3 0 o f 4 3SECRET

SANDOZ , INC . EAS T HANOVER , NE

WJERSE Y |fil V THE'SAINBQJfSROO T Oti NORFUJRAZO N

......... . t : SftNDOZ: : J?;ROafEGT--T-i"t;J'3i ; ); i MID : 0011804 8 Classified : Guidelin e requiremen t i s fiilftile d(KED , 1996 )

REPOR T 1BW-82-5-116 5 •EG&G , Bionomic s Aquati c Toxicology : Laborator y 79 0 Mai n Stree t 'Vfar.eham- , Ma$sachus,etl: s Ma y 2Q'. t 193 2

Repor t Number : TK002342 5 Pag e 3 1 o f 4 3S A N D O 2 i A G R0 LT D DEVELOPMEN T REGIO N BASL E 

# .OESiMt Hiss ) ? F& l Glassi f led : Acceptibl^/.Guide l ine H 0 R P" L 0 R AZ 0 H . REPORT : -W12 5 %DSX :: • $5255 6



### TV O 

### RKPROMJCTIO

H STUD T

FIHALRKPOB T

Dat aRequirenents t OEO ) Guideline s 41 6 I?g3 ) Sponso r : SANDO Z CRO P PROTECTIO NCORPORATIO

N De s Plaines , U.S.A. . Authors: : B . Eschbac h R . Aern i P . Bert ! P . Performin

g Laborator

y SANDO Z AGR O Ltd. , Developmen t Region

Basle , Pepar^fflcnt o f toxicology , Basle/Switzerlan

d Sando z Projec t Numbe r Stud yinitiate d : Repor t complete d o n : 44 0 R Octobe r 198 9 Decembe r 199 1

Page 1 o f 137 6 Report Number : TK002342 5 Pag e 3 2 o f 4 3MR10 : 0009105 5

Clas«ified{ : Gor e supplementary/ ; registran t wa s -requested , t o -verif y dos e level s -ftopor t 

# o O P Tti£ . SUHCimOKIC i O F

Affilinto dMiiilicis l Entori>iris<i S f Inc . Priiicofeon , Jfe w Jarso y

Jul y 20 . 197 1

Report Number : TK002342 5 Pag e 3 3 o f 4 3MR1D : 6009105 6

Classified : Cot

e : Supplementar

y

•Fi'ha l R:cpor t

h!i'clwt>»', . Tnc . Midvson . 'V/ii5c"oni»s!h , U.S . A .



### Report Number : TK002342 5 Pag e 3 4 o f 4 3. Ssfo a biofe .

^ 0011161 7 Classified : G6r e minimu man dsatisfie s guideliii e r^quii:eine|nts r

Sandoz-Wander , tec . .Compoun d

Lon g "Ter

m;Fee<iin g Stod^

r 1 Year Report

WAR FInstitute , lac . Madison , "Wisconsin ; U.S.A .

Repor t Number : TK002342 5 Pag e 3 5 o f 4 3, INC . EXE.- ; #6.42 4 DEVELOPMEN

T pSEARC H DEPARTMEN T EAST;HANOVER , NE W .JERSE Y

MRID : 0006362 1 Classified : Cor e minimu mand satisfie s guidelin e requirement s



### 978 9 Ihyestigatip

n o f Teratogeni c pqtehtiai : i n tli e Ra t

TOXICOLOGY-PATHOLOG Y SECTIO

N DATE : Augus t ?. , 197 2



### Report Number : TK002342 5 Pag e 3 6 o f 4 3OON|! 1 OtN f I AL-TRAD E SECRE T SANDOZ , INC . PHARMACEUTICA L RESEARC H AN D DEVELOPMEN T DEPARTMEN T O FPRECLINICA L SAFET Y ASSESSMEN T (PSA ) EAS T HANOVER , NE W JERSE Y 0793 6

: 0013115 1 Classified ? Cor e minimu mand : satisfies ^ guidelin e requirement s 

## DOSE ; RANGE-FIWP'ING ; STUDY;O PNPRFLURSZON ; ;xt f SREISNANT^RABBIT S (SANDQ Z PROJEC T T* 1

: 

### C o r i tgtbu t <> r s '•' ; -R; ; 

### -Hr.ab; ; Resgonsibi e Technician s M . <3 e Ralsme' s PSASystem s Leader : C . Yao , M.S .

Repor t Number : TK002342 5 Pag e 3 7 o f 4 3CONFIDENTIAL-TRAD E SECRE T PHARMACEUTICAI , RESEARC H AN D DEVELOPMEN T DEPARTMENT O P PRBCLINICA L SAFET Y ASSESSMEN T (PSA ) EAS T laSKWER , KfEWJESSE * 07?3i > ......... . ' .... . MRID : 0013115

2 Cor e minimu m^n dsatis * ie s guidelin e INVESTIGATIO N O FTERATOGENI C POTENTIA L O FNORFLDRA20 N IN TH E ^Blt-SEGMEH T I I { SANDO Z PROJEC T T-17S4 )



### Cdntrtbutors t Stud y Directoir : Responsibl e Technicians : PS A System s Leader : R . Hrab , H.S . P . Halber t M . Quagliott i C . 

### Yao , M.S .

. Augus t 31 > 1§8 3

Repor t Number : TK002342 5 Pag e 3 8 o f 4 3

### 00ll618 ,

GlassiMed i Cor e mittlmu mand ;

CRO P OR ; laARKET ; Toxicolog y 

## EXPTNO,; .

TYPEREPORT L CONTACT : (Name , Position . Organizatio n an dLocation )

L' ySiiic : 

### iaoivt

h JPigeaiq g Stud y'in Doc s reporte d PREPARE D RYit * 

### . , _....San<ic>g - JLti3: . • . :-BasIe> . 'Switzerland: :

" " " " ~ ~ " ^^^^ ^ 10/9/7 3 APPROVED; !

"DATE><>g»ygf

g SUMMAR Y

\ S 

### SA

N9789 , th e activ

e ingredien t o f ZORIAL , wa s fe d t o Beagl e dog s a t level s o f 50 , 150 , an d 45 0 ppm , respectively . Th e actua l averag e dail yintak e i n mgAg/dail ywa s highe r than calculated fro

mth e ppm-value s (1.25 , 3.75 , an d 11.2 5 rag/kg/dail^) 

### :

A ^ B C femal e 1.5 4 

### 5i.02 i 14.2 7 Mal e 1.5 9 4.77 " 17.7 5 

### I n al l thre e doses , weight , behavior , hematolog yan d iiririalysi s wer e within , norma l ranges- . Gholesterb l showe d s'pjn e highe r values , bu t als o i n th e controls . I n ith

e middl e and : hig h dose, , 'the - SAP: ' activit y-did ' .not decreas e i n -the -^sam e manne r a s the i control s an d the ; lowdose , Bu t ther e wa s n o increas e i n : 

### -activit

y\ An : 

### inductio n -o f live r enzyme s wa s no t observedi . 'No : 'increas e o f wate r or protei n i n thi s tissue coul d b $ detected . I n the necropsy , no gros s change s wer e seen . The live r weights , both absolut e an d relative wer e increasin g signifi cantl yi n the hig

h dos e o f bot h sexes . Histopatholog yshowe d an iyticrease

d quantit yo f colloi d vacuole s i n the thyroi d o f animal s on the high dos e level . .—....... . ,Thii¥a'ft/:fJ.-in'iHngH ' rtasulte a in a no-e f fec,t-J.e.VeJ-..O!f - ISO ppm.. . VFB:s a 10/9/7 3

313 H 

### Report

Number : TK002342 5 Pag e 3 9 o f 4 3MR1D : 0008075X )



### Classified : UnaC'eep.tfllble ; / Guidelin e fcedaus e ri 6 parenta l tbxtblt y wa s foun d

SANDdZ-WANDER , ;

GOMPOITKI ) 'IN ' SRM! S .JtN D MIJE E VOLUM E1 REPRCiDUCTiO N STUDIE S O N RAT S

WAliFINSTIT0TE > INC . MADISON , WISCONSIN ;



### Report Number : TK002342 5 Pag e 4 0 o f 4 3

### MRifi : 0008075 1 Classified : Supplementary

SANpOZ-WANDER,; : INC. : ^GOMEQUN

D 9^8 9 ; STSJDIEi aI N 'RATS , -AN D JHCCE - 

### '



### : 2 STUDIES ? I N 'MICE :

WARFINSTITUTE D -INC .

, WISGONSI N



### Report Number : TK002342 5 Pag e 4 1 o f 4 3

## RE£(5R D NO T REMOVE INDKW C STSIBi s. . HRID : 0008201 9 iGlassi f i6d : Got e niinlmu miti dsstti^fie s guidelin e requi y exist s .

SANDOZ~WANDER > ING, , Thf e Recor d ha s

COMPOUND 

### TWO ; 

### YEA RFEEDING ; INRAT S

SUMMAR Y DAT A

WARFINSTiTtJTE , INC . MADISON , WISCONSI N

CONFIDENTIA L Thf e i s proprietar

y informatio n o f

Sando z Crop protection CorpOraUon . Th e us « o f thi s Informatio n b y other* , withou t th e permissio n o f

Sando z C| P o r

without : jus t compenaatlbi i pai dt o Sondo z Cj P t o obtai n govemm«n t approva l for

to * sat e of . competHiv e product s 1 $ i »lof» - Do n o f this , pro'prtetar

y right . Thi s Inlormallo n sheuk) no t B e disclose dto anyon e withou t Ih ap«rmlssior

i o f SawtorCrop : Protectio n Carporstion . Report Number : TK002342 5 Pag e 4 2 o f 4 3

### pfiNFiOENTlft t UUINr-V"-:., . • . _ • (Ojettof t 

# MID , mu mW9tei 

# S^^S S Classified : Gore minimum , and , satisfie s 'Slf ^ permissio n^^^^SS* 1 

### ^ guidelin e Tequirement s 't&&*$^^$^*^'6«^k?^ ,pp,ova:ior:tr»;^w^^ . ^ ^^tswvW

, INC .

<TVK > ''& & ' J |EEiP>IN G:SSOl>Y : 'IN . MICE ;

•FIHAE 'STUiJ V WRITE-U P SUMMARY DATA



### WAR F INSTITUTE , INC . MA0ISi6N

f 

### WISCONSIN '

Repor t Number : TK002342 5 Pag e 4 3 o f 4 3