UNITED STATES ENV IRONMENTAL PRO TECTION AGENC Y WASHINGTON, D.C
. 20460 O FFICE OF C H EMICAL SA FE TY AND POL L UTION PREVENTION

MEMORANDUM DATE: February 8, 2011 SU BJECT: I sopyrazam : Repo rt o f th e C ancer Assess ment Review C ommittee P C Code : 1 29222 Decis ion No. : N / A P etition No. : N/ A R i sk Ass essment T ype : Ca ncer A ssessment T X RN o .: 0055 6 19 MRID No .: N/A D P Barcode: N/ A R egist rat ion N o . : N /A R e gulatory Action : N / A Case No.: N/ A C AS No .: N/A 40 CFR: N /A FROM :

T HROUGH:

T O : J es si ca K idwell , E xecutive Sec re tary Ca ncer Ass essment Re view Co mmittee () He alth Effects D ivision (75 09P) J ess R owland, Co -Chair

## ~~~ C a nc er Assess ment Re view Co mmittee He alth Effects D ivision ( 7509P) Mar quea Kin g,

## Xi**OI gi** **st X-** **;i** RAB IV, He alth Eff ects D ivi si on (75 09P) 

# U S haja J oyner , RM 2 0 , and Shaun ta H ill
. RM 21 Fungicide Branch, Reg i s tration Di vision (75 05P) T he C ancer Assess ment Review C ommittee m et o n Janu ary 5 , 2 011 to eva luate th e canc er cl assification of i s op y ra za m i n accor dance with th e E PA 's Final Guidelinesfor Ca rci noge n Risk A s s essment (Ma rch , 2 005)
. A ttached pl ease find the final Ca ncer Ass essment D ocument.I SOPYRA ZAM C ANCER ASSESSMENT DOCU MENT



### CANCER ASSESSMENT DOCUMENT

EVALUATION OF TilE CARCINOGENIC POTENTIAL OF 

### ISOPYRAZAM Pceod,129222



### FINAL February 8, 2011 CANCER ASSESSMENT REVIEW COMMITTEE ilEAL Til EFFECTS DIVISION OFFICE OF PESTI CIDE PROGRAMS Page 2 of 41 F INAL

### DOCUMENT PREPARATION : FINAL CAN CE R AS SESS ME NT DOCUMEN

.. 

### DATA PRES ENTATION: I S OPYRAZAM



### Ie ic a 

### Kidwell , Executive Secretary

Marion Copley Ki t 

### Farw e ll Ray Kent K arl 

### yo Middlet on 

### Es ther Rinde Je ss Row land , Co - Cha i r P .V
. S hah
NON-COMM ITTEE ME MBER S IN A TTE NDANCE : (S i g nature ind icate s co ncurrence w ith the p atho logy rep ort) Jo hn 

### P letcher, Co nsulti n g P athologist OTHER ATTE ND EES : R ebecca D e iss
. 

### A bda llah Kh a s awinah, 

### J onathan Ch e n, Je ss ica Ry man

Page 3 of 

### 41FINAL ISOPYRA ZAM CANCE R ASS ESSMENT DOCUMENT 

### Ta ble of Co nten ts Exec utive Sum mary ... ...............................................................................................•.. •• •..••••.........5 I. I nt roductio n 7 II. Backg rou nd Information ••.•••• ••................................................................... . ........................•.. 7 III. Eva l uation of Ca rcinoge nicity St udies
. .......................................................................••....... 8 I. 

### Comb ined C hron ic Toxicity /Carci noge nicity Study in Rats 8 2. Ca rclncgenlctty Study i n Mi ce 18 IV. Toxicology
.. .................................. . ..... .....••. ........•..•. .•...•..••. ........... ...... .................................. 21 

### I. Metabo lism 21 2. Mutagenicity 24 3 . St r ucture Activit), Re lationshi p 26 4
. S ubchr o nic a nd Ch r onic Toxic ity 27 5. Mode of Action •....• .•.•....... .. .. .. ....... ... .. .. ... ..... ..•. • .•. .... ..... ..... ••• •.. ••.. .. ...3 5 V. Co mm ittee's Assessment o C th e W e ight -oC-th e Evid ence 3 6 VI
. Classifica tion of Carcin ogenic Pote ntial 3 7 VII. Qua ntificat io n of Car cin ogenic P oten tial 37 VJJJ
. Bibliog raphy
. •.. . ....•.•. . ........... . .. . . .. .. . .. .... .. . . . .. . . ... .. . . .. . .. .. . ... . .. . .. . .. ........ . .. ....•..••..........•..... . . .. .. . 38

P age 4 of 41ISOPYRAZAM C ANCER ASS ESSME NT DOC UMENT
E XECU TI VE SU M M ARY FINAL

O n January 5 , 20 11 , th e Can cer Asses sme nt Re view Co mm ittee (CARC) of th e Health Ef fects D ivisio n ( HED) of th e Office o f P e st icide Programs (O PP) m et to eval uate the c arc inogenic po tential of I scpyrazam . Marquea King of Risk As ses smen t B ranch IV pr esented the chronic tox icity /carcinog enicity study in W i star rats and th e c arcinoge n icity s tudy in C57 B UI**OJ~D** m ice
. In a combi ned c hronic tox icity /carcinogenicity s tudy , 52 Wis tar rat s/s e x/do se we re exposed to Iso pyrazam (96.4 % a.i
. ; Lot No
. : SMU6A P00 1) fo r up to 2 years i n th e di et a t c oncentrations of 0 , 1 00, 500 , o r 30 00 ppm (e quiv alent to 0/ 0, 5 .5/6. 9 , 27.6/3 4.9, and 1 73.5 /2 32 .8 m g/kg b w/d ay in mal es/ females , r e s pectively)
. A dditiona lly, 1 2 rats / s e x/d ose w e r e treated s imilarly for u p to I year an d t hen s acrificed. In a carci nogenici ty s tu dy, i sopy razam (96.4 % a . i .; Batch No
. SM U6A POO I) wa s adminis tered in the d iet to C5 7 B UIOJ ,cD - I mice ( 50/s ex/ dose) for up to 80 wee ks at do ses o f 0 , 70 , 500 , or 3500 ppm ( equivalent 10 0/0 , 7 .8/9 .9, 56
. 2174.9, and 4 32 .6/553 .6 m g/kg bw /day in male s/females ). Muta genic ity data and s tructure activity rela tion sh ip we re al so di scussed
. The CA RC cons ide red the follo wing fo r a we igh t-of -evidence d etermin ation of the ca rcinoge nic p otential o f i sopyrazam . Carcinogenicity Rat

• Th yroid Tum ors : Male Wi star rats had a s tatistically s ignificant tre nd for thyroid fo llicular cell ade nomas , and s i g ni ficant pair -w ise co mparisons of the 3000 p pm d o s e grou p with the co ntr o ls for th yroid follic ular ce ll ade noma s a nd combined a denoma s and/o r ca rcinomas , all at p < 0.05 . T here was a lso a s ig nificant p air-wi s e co mparison o f the mid -d o s e gro up with th e co ntro ls f or th yroid fo ll icu l ar c e ll carcin omas an d co mbined a deno mas and /or c arcino mas , b oth at p 

### < 0 .05. The CA RC co nside red t he t hyroid t u m ors t o be treatm en t-re lated in male rat s.

• Liver Tumor s : Female Wi star rats had s tati st i ca ll y sig n i ficant trend s, and significant pai r-wi se co mparisons o f the 30 00 ppm do se group with the controls, fo r liver aden omas and co mbined adenomas an d/or carcinom as, both a t p < 0 .01
. The tumors were s upported by non -neoplastic lesion s ( eos i no p hilic foc i)
. T he CA RC c onside red the liver tumors t o be t reatm ent-rel at ed in fe ma le rat s.

• U terine Endo m e trial E pi thelial T umors : Fe male Wis tar rats had s tatis tica lly s ig n i fi c ant trends, a nd s i g nifi c ant p air

- wise co mpari so ns o f th e 3000 ppm d o s e gro up wi th th e con trols, ut erine e ndometrial e pithelial ade nocarci no mas and co mbined ade no mas and/or a de nocarcinomas , bot h at p < 0 .0 1. T he CA RC con sid ered the ut e rin e tumo rs t o b e trea tment -relate d i n f emale r at s.
Page 5 of 4 1ISOPYRAZAM CANCER ASSESSMENT DOCUMENT F INAL



### • A dequacy of Dosi ng : In b oth m ales and fe male rats, the hi gh d ose o f 3 000 ppm was co nsidered to b e ad equ ate, b ut no t excess ive. In m ales, b ody w eight d ecreases ran ged f rom 4-14 % thr oughout th e s tudy. Ove rall bod y w eight ga in in m ales w as d ecreased 1 6% (wee ks 1 -105). In fe males, b ody w eight an d b ody w eight g ain e ffects wer e m ore m arked
. A t 3 000 ppm in femal es, bod y w eight d ecreases ranged from 4- 30% thr oughout th e s tudy
. Ov erall bod y we ight g ain w as decreased 37% ( weeks 1-10 5). A lthough th e o bserve d b ody w eightlbody w eight gai n s c h anges e xceeded t he co nventional 10 % crit eria e mployed for assess ment o f b ody w eight c hanges in carc inogenicity s tudies, th e CARC d id not c onsider th e hig h do se (3 000 ppm ) to be excess ive in e ither se x s ince th ere w as no ot her c o rro borative ev idence of excess ive toxicity. A t thi s do se, th ere we re no ad verse cli nical s i g n s o f toxic it y o r s ignificant c hanges in th e clinic al c hemis try param eter s , s urvival was not affec ted, an d ther e w ere n o a dverse non-ne oplastic lesion s ( e.g., n o necrosis). Mo use • T here w ere n o tr eatment-r elated i ncrea s e s in tum ors in ei ther m ale o r fe male mi ce. 

### • A dequacy of D osing : Th e do ses t ested w ere c onsidered to b e a dequate and not excess ive i n b oth m ale and f emale m ice. Th is w as b ased on d ecreased b ody we ight, b ody w eight gai n, f ood utili zation, and m inimal clin ical s i g n s, cl inical ch emistry c h anges, liver toxicity and non-ne oplastic le sions in the na sal cavi ty and g all bladder. 

### M utagenicity: The re i s n o co ncern f or mut agenicity. 

### St ructure Activity Relations hip : I sopyrazam i s a member o f th e p yrazol e-carbo x amide cl ass of s uccinate de hydrogenase inhib itor fung icides. Th ere are no SA R dat a av ailable f or thi s c hemical at th i s tim e . 

### Classification and Q uantification o f Ca rcinogenic P ote ntial In accorda nce wi th th e E PA' s 

### Final Guidelinesf or Ca rcinogen R isk Assess ment (M arch, 2 005), th e CA RC cl assified i sopyrazam as " Likely t o b e Ca rci nogenic t o Hu ma ns"
. T his cla ssification i s b ased on the pr esence of th yroid fo llicular ce ll tumor s in mal e rat s, and liver and ut erine tum ors in fe male rat s at d oses that w ere a dequate to ev aluate the car cinogenic po tential o f i sopyrazam . No tre atment-r elated tum or s w ere see n in mi ce . T here is n o muta genic c oncern fo r i sopyr azam .

A lin ear low-dose e xtrapolation m odel 

### ( Q , *) will be u sed f or quantifi cation of c ancer ri sk to hum ans.

P age 6 o f 41ISOPYRAZAM 

### I. INT RODUCTION C ANCE R ASS ESSM ENT DOCU MENT FI NAL

On Jan uary 5 , 2 01 1, the Cance r Asse ss ment Review Committee of the Health Eff ects D ivision o f t he O ffice of Pes tic ide Pr ograms m et to eva luate th e c arc i nogenic p otential o f I sopyrazam . 

### II
. B ACK GR O UND INFORMATION Isopyrazam i s a new che mical with a pet ition for the e stab l ishment of an import to lerance on b ananas . 

### Th e PC Code i s 129222 and the 

### CAS number is 

### 881685 -58 -1.

S tr uct ure o f IPyrazole-5-J)'n 1'C) -lsopy razam 

### I s opyrazam ( SYN5204 53) is a pyrazole carboxamide w hich is u sed as a broa d s pectrum f un gicide b elong in g to th e cl as s of orth o - s ub s tituted p henyl am ides (DPA s). This cl a ss of fun gicide s i s und erstood to be inhibit or s o f s uccinate dehy droge na se (S DH) , w hich is a funct ional part of th e tri carboxy l ic ac id cyc le 

### ( Te A) an d th e mitoc hond rial e lectron tran sport c hain
. I sopyr azam h a s tw o di a st eri o i s omers (e pimers): syn an d anti i s omer s. Bot h are bi ologicall y ac tive a nd t he spec ification f or tec hnical Iso py razam covers the r ange of sy n:an t i- isome r conte nt o f 70 % sy n: 30% ant i t o 

### t 00 % s y n. The core package o f mamma lian toxic ity and A DM E studie s wa s con ducted wi th a te st materia l con tai ning 9 2 .8 % (w/w ) of the s yn i some r and 7
. 2% (w /

w) o f the anti- i somer. Fo r both i some rs excret ion w as f airly rapid ( almost co mplete by 7 2 hours po st d osing) with a s ignificant proportion of the adm ini stered rad io activity being excreted by 24 hours po st d osing
. I s opyrazam w as widely di stributed t hrougho ut the bod y withi n 2 hours and no s ex diffe rence was o bserved
. Is op yrazam i s o f low a cute to xicity by the o ra l, de rmal, and i nhalation rou te s and i s not i rri tating to the e ye s and s ki n . How ever , b ased on a s tudy no t s ub m itt ed to EPA , b ut reviewed b y th e U K i t h a s a s ki n se n sitizat ion po tential. Negative re sults were found i n bo th in v ivo an d in vitro g eno tox i c ity, mu tagenicity , and cl astogenicity s tudies. The prim ary t arget o rgan is the l iver and a ll s tu di es p erf orm ed co nsistently demo nstrate th at, irr e s pective 

### Page 7 of 41I SOPYRA ZAM CANCE R A SSESSMENT DO CUMENT FINAL
o f th e s pecie s and dur ation of expo sur e, th e principl e effect i s in creased l iv e r weight a nd c entril obular h epatocyte h ypertroph y . Li ver toxicity i s g enerally acc ompani ed by bod y w eight and fo od c on sumption reductions
. 

### 1II. E VALUAnON O F CARC INO GEN ICIT Y STUDI ES 1 . C ombined C hro n ic To xicity/C arcin og enicity S tudy i n Ra ts R eferen ce : Mi lburn , G
. (2008) S YN52045 3

- 2 Year Dietary T oxicity and C arcino ge nicit y S tudy in Rats : Final R eport . Proje ct N wn ber : T011 963 /0 5, PR135 3 /R EG, PR1 3 5 3. Unpubli s hed s tudy prepar ed b y C entral T oxico lo gy Lab . ( Syngenta), CE M A na l ytica l S ervice s, Ltd. a nd ProPath ( UK), L td
. 4098 p
. M RID 4774 6851. A
. E xperime n ta l D e s ign In a co mbined chron ic toxicit y/carcinogenicity s tud y , 5 2 W i star r at s/ se x/do se we re expo sed to Is opyrazam ( 9 6 .4% a.i .; Lot N o.: SM U 6AP OO I) for up to 2 y ear s in the d iet at conc entrat ion s of 0 , 10 0 , 5 00 , o r 3 000 ppm ( equiva le ut to 010, 5
. 5 /6 .9, 27
. 6 /

34
. 9 , aud 1 73
. 5 /

232. 8 m g/kg b w/day i n ma l e s/f ema l e s , r e s pectiv ely). A ddit io na lly , 12 r at s/ s ex /do se w ere tr eated s imilar ly f or up t o 1 y e ar a nd then sac rificed
. B
. Dis cussion o f Tum or Da ta 

### Su rvival Ana lyses T here w ere n o s tatisti c all y s ignificant s urviv al di sp ariti es am ong th e d os e g roup s with i ncrea sin g d ose s of l s opyrazam in m al e ( Table I ) or fe ma l e ( Tab le 2 ) rat s (Mem o, 

### L. Brun sman , TXR N o . 00 55586)
.

Page 8 o f 4 1IS O PYRA ZAM C A NC ER A SSESSMENT IXX: U MENT FI NAL T ab le 1. l so pyr azam - W i star Ra t Stu dy ( MRlD 4 77 4 6 8 51 ) M a le M orta lity R ate s " an d Cox or G e ne raliz ed KfW T e st R esults Weeks

"N u m be r o f ani mal s th at d i ed dunng int e rval /N umb er o f a ni ma ls a hve at t he b eg inning of t he i nterv al . Do se (ppm ) 1- 2 6 27

- 52 5 3 i 53

- 78 79

- 105 ' To tal 0 0/64 0/64 12 / 64 4/ 52 10 /48 14 / 52 (2 7) 100 0/64 0/64 1 2/64 1 /52 13/5 1 14 / 5 2 ( 27) 50 0 0/64 0/64 1 2/ 64 3 / 52 8 /4 9 I I / 52 (2 1 ) 3000 0/64 0/ 64 1 2/ 64 4/ 5 2 8/48 12 / 52 (23)



### . ilnt erim sac rific e at w eek 5 3 . ( Fi na l sac rific e at week 10 5 . ( ) Percent.

No te : T ime interval s were se lected f o r di s play purpo ses o nly . S i gnificance o f tre nd d en o ted at c o ntrol.**fic ance**  o f pai r-wi s e co mpariso n with co n trol de n oted at d os e level. If , then P < 0
. 0 5. I f" , tb en 

### p < 0 .01.

P ag e 9 of 41I SOPYRA ZAM C ANCER A SSESSMENT D OCUMENT FI NAL Tab le 2
. I s opyra zam - Wi star Rat S tudy (MR ID 4774 6851) F ema le Mortality Rat es " and Cox or Ge neralized K/W Te st Re sults Weeks

"N umber ofamm a l s tha t di ed dunng mtervallNumb er o f ammals ah ve at the be gmnmg o f the mterval. D ose (ppm) 1-26 2 7 -52 5 3 ' 5 3-78 79 -105 ' Total 0 0/64 0/64 1 2/ 64 2/52 12 / 50 14 /52 ( 27) 100 0/64 2/64 1 2/ 6 2 1/50 14 /49 1 7/ 5 2 ( 33) 5 00 1/ 6 4 1/63 12 /62 4 /50 16 /46 22/ 52 (42) 3 000 0/64 1/ 64 12 / 63 2/ 51 16 /49 19/ 52 ( 37)



### . i Interim sac rifice at wee k 5 3 . [F

ina l s acrifice at w eek 105. ( ) Percent.
N ote: Time i ntervals w ere s elected for d isp l ay purp oses o n ly . S ignificance o f trend denoted at con trol. S ii nificance o f**air-~i** c omparison w ith contr ol denoted at do se le v el. If , th en p < 0 .05. If , then p < 0 .0 1. T umor A nalyses M ale r ats had a s tatistically s ignificant tr end for t h yroid f ollicu lar ce ll adenoma s, and s ignificant pair-wi se co mparisons o f th e 3 000 p pm dose g roup with the c ontrols f or thyroid f ollicu lar cell adenomas and c ombined adenoma s and/or carci noma s, a ll at p < 0 .05. There was a l so a s ignificant p air-wise compari son o f th e mid -do se g roup w ith the c ontro ls for thyroid f ollicu lar ce ll carc inomas and combined adenomas and/or car cinomas , both at p 

### < 0 .05 . The s tatistical anal yses 

### of the tumor s in the m ale rat s w ere based upon Fisher's Exact Tes t for pair-wi se compari sons and the Exact Test for trend ( Table 3 ) ( Memo , L. Brun sman , TXR N o . 005 5586) . Female r at

s had s tatistically s ignificant tre nds, and sig nificant pair-wi s e com parisons 

### of the 3 000 P age 10 o f 41ISOPYRAZA M C ANCER ASSESSME NT DOC UMENT FINAL ppm dose group w ith th e co ntrols, for liver ade nomas and combi ned a denom as and/o r c arcinomas, an d ute rine en dom etrial e pithe lial ade nocarc inomas an d co mbined a denomas and/or a de nocarc inomas, all at p < 0
. 01
. Th e sta tistical analyses of th e tum ors i n the fema le rat s we re b ased u pon F is he r's Exact T est f or pair

- w i se co mparisons and th e Ex act Te st f or trend (Ta bles 4 an d 5) (Me mo, 

### L. Brun sman, TXR No
. 0055586)
. 

### Historical Co ntrol Data H istorical con trol dat a l fo r li ver . th yroid an d uteri ne tum ors in Han Wi star rat s fro m four 10 4 w eek d ietary s tud i es conduc ted at the Synge nta CTL fac ility fro m 2002 to 2006 were s u b m itte d after the CARC meeti ng . For live rtumors in female rat s , th e hi storica l control range was 0 -1.9 % range fo r liver a denomas ; no liver carcino mas were s een . Fo r thyroid follicular cell twnors in male rats , the hi storical co ntrol range w as 0 -9 .6 % for th yroid follic ular cell a de nomas and 0 -1 .6 % fo r carcino mas, and 1
. 6-9
. 6 % fo r co mbi ned
. For uterine e ndo metrial tu mors in femal e rats, the h istorical con trol ran ge w as 1 .9 -7 .8 % f or carci nomas and co mbined ; n o a denomas we re s ee n . Th e inc idences of thyro i d c arc inomas i n mal e rats, l iv er ade nomas and carc inomas in fema le rats an d uterine carci nomas in female ra ts all e xceeded the laboratory hi sto rical c ontr ols. Th e c on current co ntrol s wer e all w ithin th e range of th e lab oratory hi storica l co ntro ls.

1 Peff er , R and Pars ons, P
. 201 1. l s opyra zam - Histortc Co ntrol D ata f or US E PA on M icropathology in 2 Year Die tary Toxicity an d C a rc inoge nicity Study in Rats ( Adde n dum to MRID 4774685 1 ). Asse ssmen t.Janua ry 1 3 , 20 11 . Submitter: Syng enta C rop Protect ion , LL C. P age 11 o f 4 1ISOPYRAZAM CA NCER A SSE SSME NT DO C

UMENT FI NAL Tabl e 3
. I sop yrazam - Wi star Rat S tudy (MRID 477468 51) 

### Male Thyroid Follicular Ce ll Tumor Rate s " 

### and Fi sher' s Exact Test and Exact Trend Test Results D ( )



### + Numb e r of tum or 

### beanng animal s/Number 

### of ammals examined , 

### excludmg those that died before week 5 2. 

### a First a denoma o bserved at week 53, d ose 3000 ppm . bF irst carcinoma o bserved at w eek 98 , do se 500 ppm . 

### o se {ppm 0 100 500 3 000 Adenomas 1/64 4/63 2/64 7' /64 ( %) ( 2) (6) ( 3) (11) 0 .0214' 0.1776 0.50 00 0.0 309' 

### Carcinomas 0 /64 0 /63 5 ' /64 0 164 ( %) ( 0) (0 ) (8) (0) 0 .2326 1.0000 0 .0288' 1.0000 

### C

ombined 1/64 4 /6 3 7 /64 7 /64 (%) ( 2) (6) ( 11) (11 ) 0 .0726 0 .1776 0 .0309' 0 .0309'



### .

N ote: S ignificance 

### of trend denoted at control. 

### Sii nificance o f**air-\\:~** 

### compari son with co ntrol denoted at do se level. If , th en p < 0 .05
. If • th en p < 0 .01.

Pag e 12 

### o f 41ISOPYRAZAM C ANCER AS SES SMENT DOC UM ENT FINAL Ta b le 4
. I s o p yr azam - Wis tar R at Study (M RID 4 7746851) 

### F

ema le Li verT umor R a

t es+ 

### and F isher's Exa ct T est and E xac t T ren d Test R esult s

+ Number of t umor beanng arnmal s/Number of 

### ani mal s examined, excl uding those that died o r 

### were sac rificed befo re week 54 . D ose ( porn) 0 10 0 500 3000 Adeno mas 0/52 11 50 1/ 50 I 1 ' /51 ( %) ( 0) ( 2) ( 2) ( 2 2) p = 0 .0000" 0 .4902 0.4902 0 .0002" 

### Carc inomas 0/52 0/50 0 /50 I h / 5 I ( %) (0) ( 0) (0) ( 2) 0 .2 5 12 1.00 00 1.0000 0
. 4 952 

### Co mbined 0/52 115 0 1/ 5 0 12/ 5 1 (%) (0) (2) (2) (2 4) p = 0 .0000" 0 .4902 0.490 2 

### 0 .000 1"



# . . .

' F irs t adeno ma o bserved at wee k 9 2, d ose 3000 ppm. 

### bF irst carcinoma o bse rved a t week 10 3 , dos e 3000 ppm .

N ote : S ignificance o f trend den o t ed at c on trol. Sic nificance o f p air-wi s e c o mpariso n 

### with c o ntr o l de noted at d ose le vel. ¥
.. If , th en p < 0
. 05
. If , th en p < 0.0 1. Page 13 of 41ISOPYRAZAM CANCER A SSESSME NT OO CUMENT
T ab le 5. I sop yrazam - Wi star Rat Stud y (M RID 4 77468 51) 

### Fe male U terine Endometrial Epithelial T umor Rat es + 

### a

nd Fi sher's E xact T est and E xact T rend Tes t R esult s FI NAL

Dos e ( ppm) 0 100 50 0 3 000 Ad enomas 1 ' / 63 0/62 1 ' /62 0/ 63 ( %) ( 2) (0 ) (2 ) (0 ) 0 .3745 1.0 000 0. 7480 1. 0000 A denocarcinomas 1/63 2 ' / 62 3/ 62 1 5/63 ( %) (2) (3 ) ( 5) ( 24) O

.OOOO*' 0.4940 0
. 3034 0 .000 1*' 

### Combined 2 / 63 2/ 62 4/ 62 15/6 3 ( %) (3 ) (3 ) (6) (24 ) O

.O OO O*' 0 .6845 0
. 3323 0
. 00 06*' +N umber of tum or b eanng a nimals/N umber o f animal s examined , e xcludmg t hos e t hat died befor e week 51.
a First adenoma o bserved a t w eek 105, s imultaneous ly at do ses 0 and 5 00 ppm. bF irst c arcinoma o bserve d at w eek 5 1, d o s e 100 ppm.

No te: Si gnificance o f trend denoted at control. 

### S i¥nificance o f**pair ~i** ***se** 

### co mparison with co ntro l d enoted a t d o s e level. If , th en p 

### < 0 .05. If , th en p 

### < 0 .01. Pag e 14 

### of 41I SOPY RAZAM C ANC ER ASSESS MEN T DOC U MENT F IN AL 

### C. No n-Ne oplas tic L esions
a. Non.neoplastic: A t the interim s acrifi c e , in crea sed (p :5: 0 .0 1) i nc i dence s wer e n oted in th e fo llowi ng 

### findin g s in the liver ( # 

### affected/12 in trea ted v s controls ; Tab le 9 ) : (i ) 

### h epatocyte vacuolation in the 5 00 and 3 000 ppm m al e s ( 10- 1 1 v s I ; mi nimal to s light) and fema le s ( 8 eac h v s 0 ; min imal) ; ( ii) minimal to s light h epatoc yte h ypertrophy in the 500 and 3 000 pp m male s (4-1 2 v s 0 ; N S at 500) and f emale s ( 9

- 1 2 vs 0 ) ; and ( p oS O .0 5) ( iii) inc reased hepat ocyte/Kupffer ce ll p i g mentation i n th e 3 0 0 0 ppm m ale s (6 v s 0 ; min ima l t o s l i g ht ) an d fem a les (10 v s 3 ; m inima l ) . I nc idenc e s o f othe r fin dings in th e tr e ated gr oups we re s im ilar t o co ntrol s . 

### At the 

### t erm in a

l sacri fice incl uding deced ents, incre ased ( pg) .O l) in cidences were n oted in th e follo wing findi ngs in the li ver (# 

### affect ed/52 in treated v s 

### co n tro ls; Ta ble 1 0) : (i ) 

### m in

i mal t o m oderate c en trilob ular h e pat oce llular h ypertr o phy i n a ll m ale s ( 12 -49 vs 0) an d a ll f emales ( 22 -50 v s 0 ) ; ( ii) mi nimal to m odera te c en trilob ular hepa tocellu lar v acuo lat ion i n the 5 00 an d 3 000 ppm m ale s ( 32 ·3 9 vs 3) ; ( ii i ) bile d uct hyperplasia in all m ale s (1 1 -19 v s 3 ; min imal to s lig ht) and th e 3000 p pm fem ales ( 2 3 

### v s 1 5 ; N S; m inimal to m oderate ); (iv) mi nimal to s light b ile d uct fibro si s i n the 5 00 an d 3 000 ppm m ales ( 10-12 v s I ) and 50 0 and 3 00 0 ppm fe ma le s ( 1 1

- 12 v s 6; N S ); and ( v) foca l e o sin op hilic a lt ered hep at ocyte s in the 500 and 30 00 ppm m ale s (23 -3 2 v s 7; min imal to m oderate) a nd all fem ales (15 - 29 v s 9 ; N S at 1 00 ppm ; min imal to s light) . Th e in cid en ce of m in imal to s eve re f oca l f ollic u lar h yp erp la sia wa s i ncr eased (NS ) in th e thyroid g l and ofth e 30 00 p pm m a le s ( 1 8 / 52 trea ted v s 1 1/5 2 c ontrol s) . Th e in c iden ce o f min imal to s light brown pi gm en t i n ren al tubule s w as incre as ed (p oS O .05 at 5 00 p pm , poSO
. OI at 3000 ppm , e xcept at 100 p pm ) in a ll fe male s ( 24-4 2/ 52 trea ted 

### vs 18/52 c ontrols) . Inciden ces of o ther find ings i n th e treated gro ups w ere s imilar to co ntrols .

Page 15 of 41I S OPYRA ZA M C ANC ER ASS ESSMENT D OCUMENT FINAL

P en: mt dLge l,1!oce fro m C OIl1l 1ll:. ullldu ded III Da

ta Wl!re o b:.a. aed n om T ab.e _4**OIlp.l~** _-4+ _56 m

_ lRl!) 4 7 ,4 685l. par

eutaese s (calc ul ated by r e riew ers ). Si guifu: .mtl '!- '**diiferen (p~** **il 05)**  n o m the e ean e l p oup:;

.. Sipli6.c .mtl'!- ' di.fferen r tJI ;,; C .O l ) 

### nol:C ":he ccerr c l !I O"olpS 

### • 

## • TABLE 9
. Se lected**mic ro** **cap :'::**  Ii ,w**(llldint** iD l'J ':!i ( 10**aff'~ ct.d** **12** treat ed wi th**kop;. T~**  in:J:.e diet for up ttl I )' U I,-
. • (Pp",) P u :noeter 

# • I 

## 10. != OO 3000 

# ''''''' H epal0l: ) 1e \"i.l: UObt:Oll (total) 1 1 i o« 1 1 "" niuima.l 1 1 

# • II**ig bt**  C 

### o 

### , o • Hep aeccyte hypenrophy ( 10':3 :> C 

### o 

# , 12'" miui..ma l C 

### o 

# , 

# ,**bI** C 

### o 0 7**luct.~** htpa IOl:)u K'.lpt'Sn c en piJme n:ariou (lo ta : ) C 

### o 

# • " miaiIw.1 C 

# • 0 

### s**ig hl**  C 

# • • 1 F e mn..l e s H ep.llOl: )1e \" a..:UOIa t:Oll (total : Dll.lli..m.al) C 

### o 

# ", " , H epa l0l:) 1e bYP!! lUophy ( to -:,a: ) C 

### o 

# ", n '" ",;";,,w 

### c o 

# , o**:ig:bI** C 

### o 0 11**fucre~** hepatOl: )1e.X:.1pt!er**c~llpipne l1utiou**  (lOla!) 3 

### s 

# , 10· ",;";,,w 3 

# , , ic

!J. ighl C 1 

### o o , 

## , , , 

# , , -

Pag e 1 6 of 4 1I SOPYRAZAM C AN CER ASSESSMENT DOC U MENT F INAL TA BLE 1 0 .**Sd«ta1~** In"tf 6ndmI!:s mnR ( If rm \\;l m**I~iD** Ihr: dM:t fur up D:l 2**)'~ 2B**  1 Dow (pp m ) 

# -_

... 0 I 100 I 

# ,.. JOOI) lWM U\'ft H )pe[trOphy . btpatoed1u1ar , cmtrilobular ( f oul) 0 11 " 4 S " 

# .. . , 

# """""" 0 12 13 4



# .,

"'" 0 0 2 0 

# " """""" 0 0 2 13 uver Va cuola tion,**htpat oc~nu1aI**  r enmlobular ( 100.1 ) J 7 32" 39 " minimal J 7 Jl J4 slighr 0 0 0 4 moderate 0 0 0 1 Ln"**}' ta** **si a.**  bik durn (t Ol a!) J II" 19" 1 2' minimal J 10 18 12 

# ""'" 0 1 1 0 ill" F ibr osis, bile dum ( total) 1 5 1 0 " 12-- 

# """""" 1 5 

# • 1 2 

# ""'" 0 0 1 0 ill"**Al~ 0C}** **ta.** toStDOphilJc . foca.I (t otal) 7 10 23" " 32 '" """"",1 

# • 10 1 8 20 

# ""'" 1 0 

# , • """"" " 0 0 0 J Th) TOid gland.**lasia.** follicu1ar , focaJ (tota l) 11 11 

# • 18 

# """""" 8 

# • 2 7



# .""" 2 l 1 J

""""" te 1 0 2 5

awt<d 0 1 1 J severe 0 1 0 0 F

fo DJ. ,I" li= H ypenroph y. bepa tocellular , c cnlrilo bubr (t otal) 0 2 2" " 49"" 5 0" 

# """""" 0 22 IJ 1**i.ghI** 0 0 JS IJ 

# """"" " 0 0 1 50 ill" Hyprrp l n ia. bile durn ( tofal) I S 12 17 lJ 

# """""" 15 11 

# " 20



# ""'" 0 1 1 1 

# """"" " 0 0 0 2 ill" Flb1 O!1is. bik dum ( total) 

# • • 12 11 

# """""" 

# • • 11 11 

# ""'" 0 0 1 0 ill"**Al~** bepa1oc}1e-... toSinopbllK, focal (total) 

# • 1 5 26 " 29" 

# """""" 

# • 14 21 

### zz 

# .""" 0 1 5 7 

### Page 1 7 0f41I S OPYRA ZAM CANCE R A SSESSMENT DOC UMENT FI NAL
TABLE 1 0. Select ed mi croscopi c liver findln l!'> in ra ts ( # rats afi'«ted.' S2) trea ted v .. i lh I soo vrazam in tbe diet for up 1 0 2 y e ars. t Dow (ppm) P a rameotn' 0 100 5 00 3000 Kidne y Pi gmmt, br own , mbul es 18 

# ,. 30' 4 2"'- minimal 18 

# ,. 3 0 J9

,light 0 0 0 3 

## ,. Data were obtained from Tab le 00 pages 37 4 -_,6 In**~tRID** 4 774 68 51
. Percent**di ffet** **en c~**  from c ontrols IS mcl uded III p

arenth eses (calculated by revi ewers)
. Sigwficmtly di fferent (pSO ,05) from**th** cootrol gr oup ..

... Significantly di fferen t (pg) ,Ol) from contro l gr oups 

### • a

D . A dequacy o f th e Dos ing f or Ass essment o f Ca rcinogenicity In both male s and female rat s, the high do se of 3 000 ppm was c onsidered to be a dequate , but not e xcessive. In mal es, bod y wei ght decrea ses rang ed from 4- 14% throughout the s tudy. Ov erall bod y weight g ain in mal es was decr eased 16% ( week s 1 -105 ). In f emales, bod y w eight and b ody weig ht g ain eff ects w ere mor e marked . A t 3 000 ppm in fe ma les, bod y weight de creases ranged f rom 4- 30% throughout the s tudy. Ov erall body w eight gain wa s decreased 37 % (week s 1 -105) . A lthough th e o bserved bod y we i ght/body w eight g ains chan ges ex ceeded th e c onventional 1 0% criteri a e mp loy ed f or ass essment o f bod y w eig ht c h anges in car cinogenicity s tudies, the C ARC d id n ot co nsider the high do se (3 000 ppm ) to b e e xcessive in e ither se x s ince th ere wa s no oth er c orroborative eviden ce o f exc essive tox icit y. A t thi s dose , there were no adv erse clinical s igns o f toxicity or s ignificant c hange s in th e clinical ch emistry p aram eters, s urvival w as not aff ected, a nd th ere wer e no adv erse n on -neop la stic le s io n s (e .g., n o necrosis). 2. Carcin ogeni city S tudy in M ice 

### R eference : Pi nto , 

### P.l
. SYN5204 53-80 w eek dietar y c arcinogenicity s tudy in the mou se. C entra l Tox ico l ogy Laboratory, C hes hi re, U K and Prop ath UK Limited , Hereford , U K. Laboratory Repo rt/ Stud y N o: P MI352-R EG /P M1352, Augu s t 15 , 2008. M RlD 4 7746849
. U npublished. 

### A
. Expe rimental Des ign In a c arcinogenicity s tudy (MRID 47746849) , isopyrazam (9 6.4% a. i .; Bat ch No. SMU 6APOOl) wa s admini s tered in th e di et to C57 BLl IOJtCD- 1 mi ce (50 / sex/do se) f or up to 80 week s at dos es o f 0 , 70 , 50 0, or 3 500 ppm (e quivalent to 010 , 7. 8 /

9
. 9, 56 .2/74.9, and 432.6 /

553.6 mglkg bw ld ay in m ales /females)
. 

### B. D iscussion o f T umor Dat a A t th e doses t ested, there was not a treatment-re lat ed in crea se in tumor in cidence w hen c ompared to Page 18 o f 41ISOPYRA ZAM con trol s . 

### C No n-N eoplas tic Lesion s C ANCER A SSES S\tE NT DOC UMENT FI NAL

N on - neo pla stic: A t th e terminal sa crifi ce includin g de cedents, incre ased**~O. I)**  inc idences wer e noted in the f ollowin g findin gs (# aff ecte d/50 in tr eated v s co ntrols, unl ess n oted o therwise; Ta ble 7 ): ( i) min imal to s light m id

- zonal hepatocellul ar h ypertrophy in the 3 500 ppm m ale s ( 40 vs 0); (ii) mi nima l to m oderate periportal h epatocellular h ypert roph y i n the 500 an d 35 00 ppm fema les (13- 47 vs 0) ; (iii ) m inimal to moderate inflammation/exudates f rom th e n asolacrimal du ct i n th e 3500 pp m male s ( 38 v s 6 ); ( iv) m inimal to sligh t eo sinoph ilic droplets in the ep ithelium of th e n asal cavity/pharynx in the 3 500 ppm female s ( 36 v s 21 ); and ( v) minimal t o moderate eo sinophi lic drople t s i n the epithelium in the gall bladder i n the 35 00 ppm female s ( 25/ 50 vs 14 /48 at p g)
. 05)
. In cidence s o f o ther fi nding s in t he t reated grou p s were similar to controls
.

Page 1 9 of 4 1I SO PYRA ZAM CANC E R A SSESS ME N T DO C U M E N T F I NAL TABLE 7, Sele cted mic roscopic find ing s in mice treat ed wirh hopyrazam ill the diet for up to 80 weeks
. ' Dow (ppm) Pa rameter 0 

### I 70 

### I 500 

### I 35 00**l\Jal** Liver H ype rtr op hy, hep ato cellular , mi d -zona l ( tott l) 0 /50 0· ' 50 5 /50 4 0-'50 " minimal 0 0 

### s 

# " s light 0 0 0 I '**~a al**  c avity. pharynx I uflam mari orr ex udates , nas olac rimal du ct (tou l) 6 /50 9 ·'5 0 13. '50 3 8 :' 50" minima l 3 9 11 

# " sbghr 2 0 I 1 3 mo derate I 0 1 4 F emales liver H ypertroph y , hepatocellu lar , peri portal (tottI) 0 / 50 0-'5 0 J3, ' S O " 4 7/50 '" mi

nima l 0 0 IJ 

### s slight 0 0 0 35 mod er

ate 0 0 0 4 Nasal c avi ty.p harynx E o sinophilic drople ts. epithelium ( to t :l. l) 2 1
. '50 30
. :50 15 /50 36 :' 50 '" ruimma l 19 

# " 2J 

# "

s bght 2 

# , , 4 • G

all bladder E o sin ophilic drople ts , epithelium (tO t al ) 14.:48 1 1 :'4 9 1 5 "' 50 25.'50 · minimal 9 1 0 

# , 18

s li ght I 0 

### s 3 moderate 4 0 1 

# ,

mark ed 0 1 1 0

, 

### . 

## , D ata w ere obtaine d from Ta ble 15 on pag es 2b-
... 8 8 1lI MRID 47 7-1 6849
. S igni ficant ly different**(p (>-** **05)** fromthe control group s

... S igni ficantly diff eren t (pgl .O l) fr om the control group'io • a

D , Ade quacy o f Dos ing f or As sess ment o f C arcinogenicity Th e d o se s te sted we re c onsidere d to b e ad equate a nd n ot exce ssive i n b oth s exes . Ther e w ere no ad ver s e , tr eatment-related eff ect s noted o n m orta l it y. f ood co nsumption, o r l eukocyte or le uko cyte di fferentia l co unt s. At th e d ose s tested , there wa s n ot a tre atm ent-rel ated in cre a se i n tum or in cidence w h en co mpared to co ntrol s, D o sin g ( 350 0 p pm) w as c onsidered a dequate b ased o n th e f ollowing : e ye di s charg e in m ales; de creased ( p ::'; 0
. 05) bod y w eights w e re ob serv e d throughout tr eatment i n m al e s ( 1 2

- 2 3%) an d from Week 4 unti l termination in fe ma le s (! 1-11 % ); b o dy we ight g ain s durin g th e fir st 1 3 w eek s we re d ecreased i n m ale s b y 1 8 % a nd in f ema l es b y 11 % ; ov erall ( Wee ks 1- 8 1) bod y we ight ga in s w e re d ecreased in th e m ale s b y 4 2 % and in th e fe male s b y 2 2%; f ood utili zation wa s d ecrea sed (p ::'; 0 .0 5) b y 16- 26 % in mal es duri ng We ek s 1-4 and 5 -8, and w as de creased ( p::'; 0. 05) b y 14- 22% in th e 5 00 and 35 00 ppm fe male s du ring Wee ks 1 -4 ; f ood utili zation for Wee ks 1 -12 w as Page 2 0 of 41I SOPYRAZAM CANCER AS SESSMENT DOCUMENT F INAL decreased (p 50 .01 ) by 21 % 

### in the 3500 ppm males and by 9 -17 % in the 500 and 3500 ppm females ; there was al so eo sinophilic drop lets in the nasal cavity and gall bladde r in female s. 

### IV
. TOXICOLOGY 1. M e tabo lism In a se ries of rel ated me tabo lism studies (M RID s 4 7746867-47746874), H an Wistar rats were d osed ora lly b y g avage u s ing [P yrazolyl-5- 14 C]-S YN520453 in co rn oil , radio labeled in ei ther the Sy n i so mer**~98.6%** radioch emical puri ty, B atc h 

### # BPM -X XXIII -31 o r RDR-II-74 ) o r th e A nti i somer**~9 .4%**  radio chemical p urity , Batch # R DR -II-7 )
. In th e m ain ma ss balan ce /excretion! m etabolism s tud y, tw o gro ups ofrats ( 4 s ex/ d ose) we re giv en a si ngle o ral d ose o fe 4C] -SYN5 20453 (Sy n-isomer) a t I a nd 7 5 m glkg, and radi oactivity in th e exc reta wa s mo nito red fo r up to 7 days p o st- dose
. In a bil iary excre tion s tudy, two group s of bil e du c t -cannu lated ra t s ( 4/ s e xldo se) w ere g iven a s ingle o ral d ose Of[ 14 C ]_ SYN5 20453 (Syn-i so mer) at I an d 75 mglkg
. I n a s econd bili ary e xcretion s tudy, f our grou ps of bil e d uct-cannulated rat s (4 /s ex/ dose) wer e g iven a s i ng l e oral d o s e of 

### [ 14 C ]

- SYN5 2 0453 ( S yn- o r A nti

i somer) at 2 and 75 m glk g . In a t issue d i stributi on s tudy, two group s ofrats (IS / se x/do se gro up ) we re given a sing le oral d ose o f ["C] -SYN520453 ( Syn

- iso m e r) at 1 and 75 m glkg , and 3 rat s/ se x/interval were s acrificed fro m 6 to 96 hours po st-do se to examine the eliminati on o f radio activity from ti ssues . In an othe r ti ssue d i stributi on s tudy , 30 ma le rats ( 3 / sampling in terval ) were d osed o ra ll y with [ 1 4C ]. SYN520453 ( Syn -iso me r) a t I mg/kg fo r 14 co n secut ive day s, and radi oactivity in ti ssues w as de te rmined a t i nterval s from 3 to 57 day s ( 43 days po s t-dose)
. I n a b lood k inetic s s tudy, tw o grou p s o f rats ( 9/ sex/ dose) w ere given a s i ngle o ral do se of [ 14 C]-SYN520453 (Syn -iso mer } a t 1 an d 75 mglkg, an d radioactivity in w hole b lood an d pl asma we re m on itored fro m 0 .2 5 to 4 8 hour s po s t-dose
. Fo llowing a s i ngle o ral do se of [ 14C]- SYN520453 at 1 o r 75 m g/kg , r adioactivity wa s rapidl y a b sorbe d and e xcreted b y m ale and fe male rat s. The m ajority of th e a dministered radioactivity ( 92.3-104 .6% of th e admini stered d ose, A D) was e xcre ted within 48 hours of do sin g, and 95 .6· 106 .3% o f th e A D wa s rec overed in the exc reta b y 7 d ay s p ost-do s e . R adioactive re sidu es in th e m ajority o ft issue s w ere b elow th e lim it of detection « LOD) b y 7 da ys p o st -d o s e, with th e e xceptions o f th e G .I. tr act , liv er and ki dney
. Th e ro utes an d r ate s of e xcretion w ere gene rally s i m ilar fo r ma les a nd fe male s, w ith f ece s (7 7.3 -83 .0 % A D) bei ng th e m ajor rou te of el iminati on at both d o s e l evels
. Uri nary e xcret ion wa s s l ig htl y hi gh er fo r f ema les (17 .5-27 .1 % A D ) th an for m ale s ( 13.3- 1 9
. 6%) and w as al so s lig htly hi gher i n th e low - do s e gro up ( 19 .6 -27
. 1 % AD) co m p ared t o th e high d os e g roup (13 .3 -17
. 5 % A D )
. Data fro m a preliminary s tudy a lso ind ica ted that rad ioactiv i ty was not recovered in exhale d ai r as e ither C0 2 o r vo latile o rganics
. At bo th dose l evel s , the maximum concentrati on of radi oactiv ity i n the pl as m a (C max) and wh ole bl ood of male and fem ale rat s w as attai ned within 3 -6 h ours po s t-do se . B ased o n th e area un der th e c o ncentr a tion cu rve ( AUC ( Q-t ) for pl asma, th e s ys temic exposu re o f fem a le s to radioac tivity was 1
. 26- 2 .54x higher than for m ales a t bo th d o s e le vel s, and th e s ys t e mi c expo s ure of bo th s exes in creased Pa ge 21 o f 41I SOPYRAZAM C ANCE R ASSESS MEN T DO C UMENT FINAL 

### prop ortionally from I to 7 5 mg/kg , Radi oactivity in pla sma and whole blood d eclined with 

### a me an termin al e limination hal f-life (T Yzel) o f 4.6-8 .7 hours. T reatment o f bil e du ct ca nnulated rat s with a s ingle o ral do se of [ 1 4C]-SYN520453 at I o r 7 5 mglk g s howed a s imilar p attern of excretion to non- cannulated rat s and ind icated that enterohepatic c irculation pla yed a maj or rol e in th e e xcretion o f 

# C 4 Cj - S YN 52045 3
. Within 48 hours o f do sing , 91.0-10 2.5% o f the A D w as re covered in the ex creta o f bile du ct c ann ulated rats , w ith the majorit y o f th e d ose bein g recovered in th e bile ( 47.8-57 .9% A D). Fe ce s acc ounted f or 21 .2-35 .7 % o f th e AD , and th e l evels o f radi oacti vity in urine (7.3 -15.9% A D) were s imilar for c ann ulated and non-cannulated rat s. Ba sed o n th e amount of radi oacti vity r ecovered in the urine, bile and carca ss, rat s absorbed 63. 1-7 2.9% of the A D w ithin 48 h ours of d o sin g, with mo st o f the do se being ab sorbed wit hin th e fir st 24 h ours (58 .8-71. 6% A D). The absorpt io n and elim ination o f radi oact ivity w ere s imilar b etween s exes and th e two d o s e level s. Simil ar re sult s w ere obtain ed from the bil e duct cannulated rat s d osed o nce orall y with either th e 1 4

C

_ Syn or 14 C _Anti i s omer s o f S YN52045 3 at 2 a nd 75 mglkg . For eac h i somer at both do se level s , 4 9- 77 % of the AD w as ab sorbed , and e limination o f th e do sed radioactivit y was rapid and almost complete within 2 4 hour s of d osing. For both is omers, the re sidual c arc asses and G J. tract ( with co ntents) ac counted for S O.3 and S O. 1 % o f th e AD , resp ectively , b y 72 hour s p o st-do s e . Irre specti ve o f do se lev el or s e x, th e ab sorbed d o se wa s eliminated primaril y v ia the b ile. For rat s d osed w ith the 1 4

C

_ A nt i i somer , 36 .5-61.1 % o f the A D w as recovered in th e bil e , 2 0.4-38 .3% o f the A D wa s rec overed in the fece s, and 12 .2-22.1% of th e AD was recovered in th e urin e. For r at s d o sed w ith th e 1 4 C

_ Syn is omer , 41 .6-58
. 0 % o f the A D w as recovered in the bile , 19 .8-48 .3% of the AD w a s re c o vered i n the f eces, and 7
. 0-26.0% o f the AD wa s reco vered in the urine
. Th e p att ern o f exc retion w as also s imilar foll owing repe ated do sing
. For male r ats d osed orall y onc e a da y for 14 con secutive da ys , t he p att ern o f e xcretion f ollowing the 1 s t do se was s imilar to the pattern of e xcretion foll owin g th e 14th do se . In th e 24 hour s f ollowing do sing o n Day 1, the uri ne and f eces a ccounted for 19 .7 and 4 7.9% o f the dail y A D , re spectively. In th e 2 4 hour s f ollowing d o sin g on D ay 14 , th e urin e and fec es ac counted f or 21.6 and 88 .6 % of the dail y AD , re specti vely. In th e ti ssue di stribution tim e-cour se s tudy, radi oactivity was w idely di stributed in th e ti s sue s foll owing a s ing le o ral d o s e at 1 mg /k g . Peak c oncentrations o f radi oactivity in tis sues w ere ob served at th e fir st s ampling time (6 ho urs po st-do se), w ith c oncentrations b eing th e hi gh e st in li ver , kidn ey and ad renal s o f male s and in li ver, kidne ys , fat and adr enals o f female s. Higher l evels o f radioactivit y w ere dete cted in th e ti ssue s offemal e rat s at 6 hours p o st-d ose than in tis sue s o f mal e rat s at the s am e tim e point , indi catin g fas ter ab so rption and di stribution in f emale rat s. A fter 6 hours p o st-d ose , co ncentration s in all ti ssues declined , w ith s imilar o r lower levels o f radio activit y o bserved in th e ti s sue s o f fe male s compared w ith the mal e s at th e r esp ectiv e time point s. The T Y2el in the majori t y o f ti s su es ran ged from 3 to 87 hour s, and low le vels of radio a ctivit y w ere d et ectabl e in m ost ti ssues tested by 9 6 hours p ostdo se . Re sidual radi oactivity in ti s sues and car cass o f male and f emal e rats at 96 hours po st-dose ac counted f or a mean o f 0 .76% and 0.6 2% o fthe A D, re specti vely. P age 22 of 41ISOPYRAZAM C ANCER A SSESSMENT IX>C UM ENT FlN AL 

### Following a s ingle oral d ose at 75 mg/kg, the di stribution o f radi oactiv ity in ti ssues and organ s was ge nerally s im ilar to the r elative di stribution o bserved for th e low-do se rats . P eak tis s ue co ncentrat ions we re atta i ned at the fir st sa mpl i n g t ime ( 10 ho urs po st-dose), and co ncentrati o n s in all t issue s dec lined th ereafter. At th e 10-hour s ampling i nterv al , concentrations were highe st i n the li ver, kidney and ad renal s of ma le s and in t he li ver, kidn ey s , fa t , ad rcnals, panc rea s, ova rie s and ute rus of f emale s. The hi gher co ncentrations of radioactivity i n t he t i ss ues an d o rgan s of th e hi gh do s e grou p we re ge nerally proportional t o the in crease in the dose rate . The el imination ha lf lives in the majority o f ti ss ue s we re in the r ange of 3 to 4 9 hours . B y 96 h our s po st-dose , re si dual radioact ivity pre se nt in t iss ues and carca ss of ma les and fema les avera ged 0 .6 1 and 0.2 4 % of the AD , re spectively
. A s at the low dose, re sid ues in t i ssues o f the fema le s were initially higher th an in the male s, but the level s fo r fem ale s were generally s imilar o r lower than f or the m ales by 48 hours pos t-dose
. Follow in g repeated oral d osing of ma le rats with [' 4C]-SYN520453 at I mglk glday f or 14 day s, the h ighe st conce ntration s o f radi oac tivity were o bserved i n the li ver , kidney s an d b lood ( 0
. 361 , 0 .1 21 and 0 .033 11g equiv .lg , re spect ively) o n Day s 10-14
. S teady s tate concen tration s o f radioactivity i n mo st ti ss ue s were reac hed by D ay 10 , wit h the exceptions of li ver and renal fat (Da y 14)
. Mean c oncentration s o f radioactiv ity in t issue s dec lined ra pidly f o llowing cessa tion of d osing o n Day 14
. Co ncentration s o f rad ioactivity we re quan tifiable in o nly li ver (0 .049 Ilg equ iv .lg) an d ki dney ( 0
. 020 p g eq uiv .lg ) by 14 d ays po s t-dose ( Day 28) ; and radioactivit y was d etectable onl y in ki dneys (0 .011 u g equiv .l g) and s p l een (0 .00 7 pg equiv .lg) by 4 3 day s po st-dose ( Day 57 )
. Rad ioactive re s idue s in excreta we re adequ ately identifie d/ characterized
. Following a s ingle oral d ose of[ 14 C]SYN520453 at I o r 7 5 m glkg, 7 9 .1- 95.4 % of the AD w as identified in excreta from n ormal rats and bile duct c annulated rat s; and f ollow ing repeated do sing o f m ale rat s at I m glkg, 74 .8 -7 9.4 % of the rad ioactivity w as identified i n e xcreta co llected from 0

- 24 hour s after the l S I 

### and 14 th 

### d oses . In all d ose gro up s, [ 1 4

### C ]SYN52045 3 was ex ten s ively metabolized b y rat s via ox idation o f the i so propyl and bicycle m oieties giving ri se to a r an g e o f hydroxy , d ihydroxy , ac id and hydroxy acid m etabolite s , along wi th th eir g lucuronide an d s u l fate co njugate s. Oxidatio n of th e deme thylated m etab olite of SYN52045 3 pr oduced an e quivalent range of oxi dized de methylated m etabolites. No cleavage o f the p arent mol ecule was o bserved. Th e major ro ute s of b iotransformation we re ge nerall y i ndependent of do se level and s ex, and were si milar for the Syn and Anti i somers . Metabolism o f the Sy n i somer al so appeared to he s im ilar fo llowin g ei ther a s ingle dose or repe ated d os ing. l sopyrazam ( either the 9 3 % s yn:7 % ant i material or the 70 % s yn :3 0 % anti s pecifi c atio n ) w as exten s ively metabo lised gi ving rise to up to 25 metabolite types (including c onjugate s) w ith potenti al for multiple i somers in most gro up s. The bi otransformation of I sopyrazam was po s tulated to proceed by hydrox ylation in the bicy cle

- i s opropy l re gion, followed b y f urthe r o x i dat io n to f orm the car boxylic acid an d/or to give ri se to mu ltiple hydroxyl moie ties wi th s ub s eq ue n t format io n o f g lucuronic acid o r su lphate co n jugates. This oxida tive pathway also occurre d foll owi ng N -demethy lation of I s opyrazam g iving ri se to a co rre spo nding s et of oxidative N -desmethyl metaboli tes and their co njugates
. No cl eavage of th e molecule wa s o bserved
. In ge neral, un conjugated metabolite s we re e liminated in urine an d co njugated metabo lite s in h ile
. (MRlD s 4 7746867-47746874) Page 2 3 of 4 \IS OPYRAZ AM 

### 2
. Muta gen icity CANC ER ASSESSMEN T DOCUM ENT FINAL



### As series o f 1 3 G enetic T oxicology s tudies ( 7 on the par ent, i sopyrazam a s SYN5 20453 and 3 e ach o n the metab olites, CSC D465008 and CSC D459488) were s ubmitted by the Regi strant . Th e te sts inclu ded bact erial reverse g ene mutation ass ays in S almo nella typ himurium 

### and E scherichia coli 

### te ster s trains; in v itro 

### m ammalian ce ll f orward g ene mu tation ass ays in L51 78Y mouse l ymphoma ce lls a nd in v itro 

### cyt ogenetic assays in peripheral blo od human lymphocytes. The onl y i n vi vo 

### a s say ( mouse b one marrow m icronucleus assa y) wa s conduct ed w ith SYN 520453
. Ne ither th e parent co mpound nor th e t wo metabolites w ere muta genic in b acteria o r mamm alian ce lls or cl astogenic i n mammalian cell s . S imilarly , i sopyrazam was n ot cl astogenic or aneu genic in whole animal bone marr ow cell s. A ll o f th ese s tudies w ere cl as sifi ed a s a ccept ab le/guideline , and fu lly sa tisfy th e requirement for the 1991 FlF RA Tes t G uideline 84-2 f or muta genicity data
. B ased o n th ese co nsi derations, there i s no mut agenic conce rn for th e par ent mol ecule or th e tw o metabolites. S ummaries o ft he 13 Genetic T oxicology s tudies a re pre sented below : S YN520453 GENE MU TA TIONS (i) 

### In tw o indep endent tri al s of th e b acterial rev er se g ene mu tation assay (M RID 4 7746854) , is opyrazam, as SYN 520453 (90 .8%), was in soluble at ?:2 500 

### ug /plate +1

- S9 (plat e inc orporation test) o r ?: 1 000 uglplate 

### +1

- S9 (prei ncubation test) but f ailed to induc e a mut agenic response ove r a co ncentration ran ge 

### 0 0- 5000 ugl plate +/

- S 9 i n 

### S. typ himurium 

### T A1535 , T A1537, T A98 or T AIOO or i n E. coli 

### WP 2 (pKMlOl) or WP 2 u vrA 

### (pKMlOl ). ( ii) 

### Similar re sults w ere o btained in th e i ndependent tri als of the b ac terial re verse mutati on a ssay, u sing the pl ate in corporation t est and th e preincubation m odification t o th e s tandard method , which w ere perf ormed by a diff erent lab oratory ( MRID 4 774685 5). In ag reement with th e ear lier findin g s , is opyrazam, as SYN52 0453 (9 6.4%) , was g enerally i nsoluble a t**00** 

### ugl plate +/ -S 9 (both pro cedures) but failed to indu ce a mutageni c r e sp onse at co ncentrations ran ging f rom 100 to 50 00 uglplate in 

### S. typhimuriu m 

### T A1535 , T A1537, T A98 o r T AIOOo r in E. coli 

### WP 2 (pKM lO l) o r W P2 u vrA 

### (PK MlOl ). (iii) 

### In an in v itro 

### mammalian ce ll f orward ge ne mut ation a ssay «MR ID 4774 6858), i sopyrazam , as S YN520453 (9 6.4%), w as not mut agenic in L5187 Y (TK + 1 ° ) 

### ce lls i n three ind ependentl y perf ormed tests up t o c ytotox i c c oncentration s ( 25- 3 0 ugl mL

- S9 , 40- 50 ugl mL + S 9 ). ( iv) Dat a from an other mou se lymphoma f orward g ene mutation assay «MRlD 4 7746859), perf ormed in a diff erent laboratory, wer e in go od agreement with the earli er re sult s. T hese fi ndings indic ated that i sopyrazam, as S YN520453 ( 90
. 8%), pr ecipitated at ?: 1 7 5 u glmL 

### +1 -S 9 but w as n ot mutag enic up to cyto toxic c oncentration s ( 33- 44 ugl mL - S9, 44-88 uglmL + S9 )
. Page 2 4 o f 4 1ISOPYRAZAM C AN CER ASS ESSMEN T IXX UMENT FI NAL CHROMOSOME ABERRATIONS (v) In an in vitro chrom o some as say ( MRID 47746862 ) , human lymphocytes, derived from the peripheral blood of two male and two female don ors were exposed to i scpyrezam , as SYN52045 3 ( 96.4 %), fo r 3 hours at 20, 3 0, or 40 u glmL

- S 9 or 20 , 30 o r 50 u glmL + S9 ( Trial I ) or co ntinuou sly fo r 20 h ours to 1 0, 1 5 o r 20 u glmL

- S9 or f or 3 hours to 20 , 30 or 50 u glm L + S9 ( T ri al 2)
. Higher co ncen tratio n s reduce the mitotic indices by > 50 % , and were not s cored
. Sl ight b ut s ignificant (p <0.0 5) inc rease i n the p er centage of abe rrant ce ll s was see n a t 20 and 30 u g/mL

- S 9 after 3 h ours of expo su re. The response w as , h owever , not s ee n at the h igh er leve l o r in the 

### zu- hour ex p os ure trial. The re was no evi dence o f a cl astogenic respo nse i n th e pre s ence of S9 ac tivation
. The refore, isopyrazam i s not co nsidered to b e cl astogenic in th is in v itro h uman l ymphocytes test s ys te m. (iv) Th e find ing s from the e arlier assay, d i sc u s sed above we re confi rmed in the a dditional in vitro chro mos ome a ssay (M RlD 477 46863) i n h uman lymphocyte s performed by an i ndependent laboratory o n i so pyrazam . a s SYN520453 ( 90
. 8%)
. I n this s tudy, hu man l y mphocytes , derived from the periphe ral bl ood of o ne m ale and one female d onor s, were expo se d fo r 4 hours to test co ncentrati o n s of 16 .9, 29 .6 or 51 .7 u glmL

- S9 or to 29 .6, 51 .7 , o r 90.5 u gl mL + S9 ( Trial I ) or c ontinuously for 22 h o urs to 3 .0 , 5
. 2, 9
. 1 o r 1 6 .0 u glmL

- S9 or for 4 hours and to 25 .0 , 5 0 .0 , or 7 5 .0 u gimL + S9 (Trial 2)
. Co mpound precipitati on an d/or 

### S 50 % reduced mitotic indices ( MI s) were apparent at 2: 90.5 u g/m L

- 5 9 o r 2:51.7 u g/mL + S 9
. Slight but s i gnificant ( p <O. 0 5) increas e in the percent age o f abe rrant cell s w as seen at 51
. 7 and 90 .5 u g/m L + S 9 after 4 hours of expo sure . The re sponse w as , h oweve r , within the hi storical contro l range fo r the perf orming la boratory and not replicated i n the repeat trial. There w as a l so no evidence of a cla sto genic respon s e in the ab sence ofS9 a ctivation
. Acco rdi ngly , i so p yraza m i s conside red not to be cl as togen ic i n this in vitro hu man lym phocyte s te st s yste m . (v) In a r at bo ne m arrow micro nucleus as s ay ( MRlD 4 776864) , five m ale Wi s tar rat were o rally d oses wi th isopyrazam , as SYN5 2 0 453 (96.4 %), p repared i n 0 .5 % ca rboxy methyl cellulo se up to t he li mit do se . Th e te st arti cle wa s no t ove rtly toxic to the te st an imals or cytotoxic to the targe t ce ll and d id not i nduce a cla stogenic or ane ugenic re spon s e i n bo ne marro w cell s h arve ste d 2 4 an d 4 8 h ours after tr e atment. CSCD46 5008, METABOLITE o f SYN 520453 GENE MUTATIONS (i) In two independent trial s o fthe bacterial re ver se g ene mutati on assa y ( MRID 4774685 2) , C SCD465008, (9 4 %), w as neither cyt otoxic nor muta genic in either the pl ate inc orporation o r the prein cubation te st wh en as say ed up t o the lim it co ncentration ( 5000 u g/plate + /

- S9 ) in 

### S. typh imurium T A I535, TAI 537, T A98 o r T A IOO or i n 

### E. co li WP2 (PKMI01 ) o r WP2 uVfA (pKM IOI ). (ii) In an in vitro mammalian c ell forward g ene mutati on assal, 

### «MRID 4774 6856 ) , CSCD465 008 , (9 4 %) was n either cyt ot oxic nor m utagenic i n L5 I 8 7Y ( T K + ") cells i n two indepe ndently perf ormed te st up to t he l imit con centratio n ( 1 OmM, equiv a lent to 1760 u gl mL +/- S 9). P a ge 2 5 of 41I SOPYRAZAM C ANC ER AS SES SMENT DOCUM ENT F INAL 

### CHROMO SOM E AB ERRATIONS (iii) 

### In an in vitro 

### c hromo so me as say (MR ID 4 7746860) , human lymphocytes , d eri ved fro m the p eripheral b lood o f two f emale donors w ere e xpo s ed to CSCD 465008 (9 4%) f or 4 h ours at 57 4 .7, 1 005
. 7 and 1 760 .0 ug/ mL

- S 9 o r 32 8 .4 , 57 4 .7 o r 1005
. 7 u g/mL + S9 (Trial I ) or co ntinuou s ly for 22 h ours to 57 4.7 , 1 005.7 and 1 760.0 u g/mL

- S9 o r f or 4 hour s to 57 4.7, 1 005.7 and 1 760
. 0 u g/mL + S9 (Trial 2 )
. T he hi ghe s t d o se te sted ( 1 760.0 u g/mL) repr e s ent s t he li mit co ncentration (l Om M) f o r th i s te st s ys te m. S light but s i g ni fican t (p<O .05) in crea se s in th e per centage o f aberrant ce lls we re see n at 57 4 .7 uglm L

- S 9 an d at 1 760 .0 uglm L +S9 after 4 h ours o f ex posure . Howev e r , the response s w ere w ithin th e hi st orical range o f th e performin g laboratory or not replicated
. Acco rdingly, C SC D46 5 008 i s not co n si dered to be c1as togenic in th is in vit ro 

### hu man ly mphocytes te st s ys tem . CS CD4 59488, METABOLIT E of S Y N 520453 GE NE MU TA TIO NS (i) 

### In t wo indep endent tr ial s of th e b acterial re verse ge ne mu tation assay (M RID 477 4685 3), C SC D4 59 48 8 , (99 % ), w as neither c ytotoxic no r mut agenic i n e ither the pl ate in corporation or the pre incubation te s t w hen a ssayed u p to th e l imit co ncentration (5 000 ug/plate +/

- S9 ) i n S
. ty phimurium 

### TA 15 35 , TA 15 37, TA98 or TAI OO o r i n 

### E . coli 

### WP 2 (p KM IOI) or WP2 

### uvrA (pKM IO I). C o mp o und precipitation wa s s e en at ::: 1000 u g/p late +/

- S9 u si ng b oth te s t methods. (ii) 

### In an in v itro 

### mammalian ce ll f orward ge ne mutati on a ssay «( MRID 4 77468 5 7) , CSC D459488 , (99%) wa s neither cytotoxic n or muta g eni c in L5 187Y (T K + 1 - ) 

### ce ll s in two i ndepende ntl y p erform ed t e sts up t o th e i n so luble c o n ce ntratio n of 4 00 u g/mL +/

- S9. C H ROMOSOME ABERRA TIO NS ( iii) 

### In an in vitr o 

### c hromo so me as say (MRID 4 7746861), human lymphocyte s , d erived fro m th e peri pheral bl ood o f two f emale donors were e xposed to C SC D4 5 94 8 8 ( 99%) f or 4 hour s at 1 70.6 , 298
. 5 and 52 2.4 u g/mL +/

- S9 (T ri a l 1 ) o r co ntinuou s ly f or 22 hours t o 3 1. 8, 55 .7 , 2 98.5 and 5 33. 4 u g/m L

- S9 and 1 70.6, 298 .5 and 522 .4 u g/mL + S9 for 4 h our s (Tri al 2 )
. Co mpound precipitation wa s ap par e nt at 522 .4 u glmL +/

- S9
. The test material w as not cl astogenic at an y co ncentration w ith o r wi thout S 9 acti vation or und er an y expo s ure con dition s. Acco rdingly, CS CD 459488 i s n ot cl a stogenic in thi s in vit ro 

### hum an lymphocyt e s te st s ys te m . 

### 3
. St ru cture
- Act ivity Relationsh ip I s op y razam i s a m emb er o f the pyrazole-c arboxam ide clas s o f s u cc in ate deh ydrogenase i nhibitor fungicide s . The r e ar e n o SA R d ata ava ilable f or t his c he mical at th is t ime
.

P age 2 6 o f 4 1ISOPYRAZAM C ANCER AS SESSMENT DOC UMENT F INAL 

### 4. Subchronic a nd C hronic T oxicity a) S ubchronic T oxicity (1 ) RA T In a s ubchronic o ral t oxicity s tudy (M RID 4 7746833), 10 W istar ra t s/sex/dose w ere administered e ither S YN520453 (89 .5 % S yn: 6 .9 % Anti ; 96 .4 % a.i ., B atch No
. SM U6APOO I) o r SYN52 0453 ( 63
. 3% 

### Syn : 2 7.5 % An ti; 90 .8% a .i ., Batc h No
. SM U7D POI7) v ia th e d iet a t co ncentrations of 0, 1 00 , 250, o r 2000 pp m (equivalent to approximately 

### 8.3 /8.2, 20.3 /20
. 8, and 

### 158
. 71163 .3 m glk gi day [MIF] ofSYN 520453 [8 9.5 % Syn : 6 .9% An ti] and 

### 9 .9 /9.5, 24.1/24 .1, and 192 .9/197 .0 mglk giday [MIF] ofSYN5 20453 [63.3 % Syn : 27 .5 % Ant i]) fo r at least 90 days
. Th e p urpose of th i s s tudy w as to compare the oral toxicity effec t s of tw o te st materials con taining diff erent co ncentrations of the Sy n- a nd Anti e p imers of SYN5 20453
. No tre atment-related effe cts we re no ted o n m ortality, clin ical sig ns o f tox icity , o phthalmoscopic e xaminat ions , F OB , motor ac tivity, hem atology , urinal ysis , o r gross path ology at any do se i n e ither s ex . Th e target o rgan w as th e l iver. A t 2 000 p pm , abs olute li ver weig hts w ere increase d by 1 5-18 % in bo th s exe s i n bo th t e s t material grou p s . Additionally at thi s do s e, relative (to body) li ver wei ght s we re increased by 15 -19% in both s exes in bot h te st mat erial group s . The i ncide nce (# a ffec ted/ l 0 vs
. 0 110 controls ) o f m inimal to moderate cen trilobular hype rtrophy was increased i n the ma les ( 8 i n eac h te s t m aterial gro up ) and female s ( 10 in e ach te st m aterial group )
. Additionally at this dos e, the incidence of h epatocyte mi dz on al vac uolation w as inc reased in the ma le s (3-4 per t est ma terial group ; min imal to m ild sev erity) an d fema le s (4 i n e ach te st m aterial grou p ; m inimal severi ty)
. A t 20 00 ppm , c umulative bo dy w eight ga ins we re d ecreased by 11-1 9% and 11-1 6% (SYN5204 53 [89 .5% S yn : 6. 9% Anti] an d S YN5204 53 [ 63.3% Syn : 27 .5% A nti], re spectively) i n the fem ales beginning the sec ond w eek o f treatment and c ontinuing throu ghout the s tudy
. A dditionally at thi s dose, b ody we ights we re d ecreased (p< 0.05) b y 6
- 10% on D ays 14 -91 in the SYN 520453 (89 .5% Syn : 6 .9% Anti ) tr eated f emales
. B ody we ights wer e a lso d ecreased thr oughout th e s tudy in th e f emales treated wi th SYN520453 (63 .3% Syn : 2 7.5 % A nti) at thi s dose , b ut o n l y achieved s tati s tical sig nificance ( p<0.05) o n Day s 77 and 91 ( deer 6 -7 %)
. In t he m ales treated with SYN52045 3 (63 .3 % Sy n : 27 .5 % An ti ) a t 2000 ppm, body weight s were decreased by 6 -7 % o n Days 7 , 35
. and 56-77, and cwnulative body weight gains were decreased by 10-14 % du ring Days 0 -7 and 0

- 70 . In the female s. overall ( Weeks 1- 13 ) food ut ilization was al so dec reased by 1 1-15 % i n bot h te s t ma terial grou p s . N o treatment-related fin ding s we re o bserved at 1 00 o r 250 ppm in e ither sex in e ither t est m aterial group . T here w ere no d ifference s in th e toxicological pr ofile o f th e tw o te st m aterials i n th is c omparative s tudy.
Page 2 7 of 41I SOPYRAZAM CA NCER A SSESSMENT DO CUMENT FI NAL T he LOA EL is 20 00 ppm (e quivalent to 1 58.7/163.3 and 192
. 9/197.0 mg /kg/day [ M/ F] f or SYN5 2045 3 [89 .5 % Syn : 6 .9% A nti ] an d SYN 520453 [6 3.3% Sy n: 2 7.5% A nti]
. re spective ly) ba sed o n liv er to xicity ( increased ab solut e an d relat i ve l i ve r wei gh t s, and incre ased incid ence o f ce ntrilobu lar h ypertroph y and hepatoc yte mid zona l v acuolation) and d ecrea ses in b ody wei ght g ain and foo d uti li zat ion in femal es . T he N O AE L i s 250 ppm ( equivalent to 20
. 3/20 .8 and 24 .1/2 4
. 1 mg/k g/day [M/F] f or SYN5 20453 [ 8 9
. 5 % Sy n: 6 .9% A nti] and S YN52045 3 [6 3 .3% Syn: 2 7.5% Anti]
. re spective l y)
. 

### (2) RAT In a 90-da y diet ary s tudy (MRlD 477468 34) in H sdRc cHan:WIST s tra in rat s , 4 - 5 we ek s o ld , were randoml y a s sig ned to the te s t gr oups a s s hown in the table below
. The test and c ontro l diet s wer e prepared u sing RM I di et. Th e homo geneity of I so py raza m in th e di et wa s e stabli shed for the l ow and high do se l e vel s. T he sta bi l ity o f th e te st s ubstance f or the period of u se wa s e stablished at room tempe rature for th e di ets u sed for the 80 00 ppm do se group i n a 28 da y s tudy. T he ac hieved conc e ntr ations of Isopyra zam i n th e anal yzed s ample s of test di e t u s e d o n the s tudy were t yp icall y w ithin 8 % o f th e nominal conc en tration
. Group Dietary leve l (ppm) o f C o lour c ode I dentities of ra ts SYN520453 ma les females 1 (co n tro l) 0 Blue 4 t -52 89-tO O 2 3 00 Green 5 3

- 64 10 1-112 3 1 5 00 Yellow 6 5

- 76 1 13- 1 24 4 6000 Red 77

- 88 125-136 D etailed clinic al o bserv a tion s w ere re corded at th e s ame tim e that the bod y we i ghts w ere record ed
. C age-side o bs ervat io n s whi ch included re cordin g an y c hanges in clinical c ondit io n or behav ior were mad e twice da il y. Detail ed observat i o n s (during w h ich eac h rat was remo ved from it s cage and phy sically e xamined f or chan ge s in g eneral h ealth s tatus), quanti tati ve a s se s sments o f landing f oot s p lay mu scl e we akne ss (f ore - a nd h indlimb grip s tre ngth and s e nsor y perception (tail -fli ck te st) and a me a surement ofl ocomotor acti vity u sin g a utomated re cordi n g equipm ent , w ere made in week 1 2 o r 13
. T he b ody w eigh t o f e ach rat wa s recorded weekly . Food con sumpti on wa s m ea sured we ek l y throu ghout the s tud y an d f ood utili zation , in term s o f g b odyweig ht ga in/ IOO g food co n s umed , was c alculated
. Th e e ye s o f a ll animal s we re e xamined prior to t reatment and a gain f or the c ontrol and hi gh do s e an im a l s during the la st w eek o f tre atme nt. Indi vidual urine s amples were c ollec te d f rom all rat s dur ing the w e ek prior to terminati on and a s tandard ran ge of param e ter s wer e m easure d. C ardiac b lood s ampl es w ere tak en from a ll rats s u rv iv in g to s ch e dule d termination and a s tand ard ran ge o f ha ematolog y and cl ini ca l c h emistry parameter s we re me asured
. A n ecrop s y wa s c o nducted o n all anima l s the en d o f th e tr eatmen t p erio d
. T he we ight s o f m ajor organ s Page 28 o f 4 1ISOPYRAZAM CANCER ASS ES S'-1 ENT DOC UMENT FINAL were recorded
. Macroscopic changes were recorded
. A standard range of organs and t is sues were removed and fixed
. All tis sues fro m the co ntrol and 6000 ppm gro up toge ther with the li ver only fo r the 300 and 1500 ppm group s were exam ined 

### by light microscopy
. 

### Received dose s (based on nominal d ietary levels o f I sopyrazam) were calcu lated in t erms of mg I s op yraz am/k g bo dy we ight. A ll anim als s urvived to th e en d of th e s tudy
. Th ere were n o tre atmentrel ated cl i nica l c han ges . B ody we ight ga in was clearly reduced in fema les at 1 500 and 6000 p pm , w ith m axi mum diffe rence s from con trol of 7 % and 19 %, re spect ively
. Males at 6000 ppm gro up had s ignificantly lower body weights during weeks 2 and 3 (maximum effect 8 % in week 2 ) and bod y we i ghts remai ned approx imately 5 % lower than contro l s thereafter
. There were n o adverse effects o n bodyweight in male s at 1500 ppm o r in e ither s ex in the 3 00 ppm g roup . Food consumption was reduced i n fema les at 1 500 and 1600 pp m, wi th max imum e ff ects of 1 2 % and 23 %, respectively
. Foo d co nsumption wa s reduced i n males at 6000 ppm du ring th e fi rst week of the s tudy bu t wa s ge nera lly s lightl y hi gher than cont rol for the rem a inder o f th e s t u dy
. The re were no effe cts o n f ood con sumpti on in mal es at 1 500 ppm group o r in e ither sex receiving 3 00 ppm . Foo d util isation ove r the 1 3 w eek s tudy p eriod , in term s o f g bo dy weig ht g ai n/l 00 g foo d c ons umed, w as s ignificantly reduced at 6000 p pm in bo th s exes. Th ere were no treatment related op htha lmo sco py change s . There we re no u rine chemistry d ifferences considered to 

### be treatment related
. The re were n o differen ces in haemat ological parameter s which were con sidered to be related t o treatment . A marginally lower value f or e osin ophil s for male s in the 6000 ppm group i s cons idered n ot to 

### be re lated to treatment as all va lue s obtained for the high d ose group we re within the con current c on trol group range and the re w as no s im ilar finding in females
. The re we re a n umber of change s in cli nical che mistry parameter s at 1 500 and 6000 pp m . A t the h ighest d ose l evel t here we re i ncrea ses in th e activ ities of a num ber o f p lasma e nzyme s ; th e se w ere a lan ine am inotran sfer ase and ga mma-glutamy l tr ansferase i n b oth s e xes, a nd as partate a minotrans fera se and c reatine ki nase in m ales
. In the case of a lanine am inotran sf erase the d ifference i n th e mal es i s d ue to one ex tremely h igh value
. Sligh tly hi gher g am m a-g l utamyl transfe rase ac tivity was a lso see n in female s a t 1 500 ppm, w hich was exagge rated by a low va lue for a s ing le co ntrol fe male
. The level s of the p lasma ions, ch loride , sodium and p h o sphoru s we re slightly increase d in ma les in the 6000 pp m group, but as s im il ar effects were n o t p resen t i n o ther repeat dose toxic ity s tudies the se d ifference s wer e con sidered unlikely to 

### be treatment -related
. There w as a tr e a tment-related increase i n li ver we ight i n both s exe s in the 15 00 and 6000 ppm g roup s and in fe male s i n t he 300 pp m gro up . The inc reases i n li ver weight (a d ju s ted for b odyweight) we re 24 an d 1 9 % i n ma le s an d fe ma les at 6000 p pm, 14 % in both sexes at 1 500 p pm and 14 % in females at 300 pp m . Differe nces we re al so s ee n in the weights of heart, kidney and th ymus in tr eated gro ups, b ut in th e absence of any s upporting h i st op athol ogical fi nding s th e s e we re co n s idered to b e incidental to tre atment. A s ma ll numbe r o f macro sco pic lesions we re observed at n ecropsy b ut n one wa s conside red to be Page 2 9 of 4 1ISOPYRAZAM CANCE R ASS ESSMENT DOCUM ENT F INAL related to treatment. T reatment-related mi croscopic findin gs were co nfined to th e liv er and co nsi sted of hypertrophy of ce ntrilobular he patocytes, gra ded e ither mi nimal o r s light, affecting all anim al s at 1 500 an d 6000 p pm . No signi ficant c han ges we re ob served i n a nimal s in t he 3 0 0 ppm gro up . Die tary a dmini s tration of I sopyrazam (93% sy n :7% an ti) for 90 days at a conce ntrations of 1 500 an d 6000 ppm caused a red uction i n bo dyweight gain and foo d co n s umption and li ver c hanges in th e f orm of ce ntrilobular h epatocyte hypertrophy an d increased li ver we ight. T h ese c hange s we re acco mpanied b y s lig ht d iffere nc es s o me cl inical c hemi stry param e te rs , whic h may be re lated to the effects o n t he l ive and th erefore , leaving a L OAEL of 1 500 pp m. A t th e lowe s t ex po sure l evel of 300 pp m th e l i ver weigh t in f emale s wa s in creased ( by ]4 % i n co mparison w ith co ntrols), an effec t th at is c onsidered to 

### be a dverse. A lthough the e ffec t wa s se e n in iso latio n of othe r ef fect s t he li ver i s consid ered t he p rimary target o rgan . Therefore a s tudy NO AEL i s cannot be identified. The L OAEL is 300 ppm ( int ake of abou t 29 ru g/k g/d ay i n f e males). (3) MI CE In a 90 -day p reliminary c arcinogenicity stu dy ( M RID 4 774632) i n C57BL/ lOJfCD- I s t rai n mice, 4

- 5 weeks o ld, were r andomly a ss igned to th e t e st g roups a s s ho wn in th e table bel ow . T he te s t an d con trol diet s were prepared u s in g CTI di et. Th e ho mogeneity of I s opyrazam i n t he die t was es tabli s hed for the l ow an d hi gh do se gro up co nce ntrations. Th e s tab ility of th e te st s ubstance in th e die t for the pe riod o f use was es tablis hed pr ior to th e st udy. T he ac hieve d co ncentrations of I sopyr azam in th e an alyzed s ampl es of test die t u s ed o n th e s tudy we re all w ithin 4 % o f th e no minal co ncentration. M ean Dos e Rec eived (mg/kg/day)

- weeks 1 -13**=5~0453** (pp m) 500 250 0 7 000 

### Ma les 7 6.5 390 .8**13 8~**  

### Fe males 87.2 448.9 1 759
. 6 D etailed cl inica l observation s we re reco rded at th e s ame tim e that th e b ody we ight s we re reco rded. C a g e -side o b serva tions w hich inclu ded record ing an y c h ange s i n cl inical co ndition o r be havior w e re m ade
. Th e bod y we i ght o f eac h m ou s e was re corded wee kly
. F ood co n s umption was mea sured wee k ly thr oughout th e s t udy and food u tilis ation , in terms o f g b odyweight ga in/ l 00 g foo d co n s umed, was ca lculated
. C ardiac b lood s ampl es were take n fro m a ll m ice s urviv ing to sc heduled termination f or cl inical p athol o g y . A s tand ard r ange o f hae matology ( five m ice f rom eac h gro up ) and cl inical c h e mi s try ( five mi c e p er gro up) p arameters w e re measured
. A nec rop sy wa s con ducted o n a ll ani mal s the end o f th e treatment perio d
. The we ights of m ajor o rgan s w ere r ecorded
. M a crosco pic c hange s were re cord ed . A s tand ard range o f organ s and ti ssues were Page 30 of 41I SOP YRAZAM C AN CER A SSESS MENT IX>C U MENT F INA L rem oved and fix e d
. A ll ti ss ue s fr om th e con trol gr oup , th e 7000 p pm gro up and deceden ts t o geth er wi th the l iver o nly for the 500 and 250 0 pp m g rou ps w ere exami ned b y li ght m icrosc op y . Recei ved d ose s ( b ase d o n nom inal d ietary leve l s of Is opyrazam ) were c a lculated i n term s of m g Is opyrazamlkg bo dywei g ht. There were no tr eatm ent rela ted dea th s, a lthough tw o mice ( o ne fem ale a t 5 00 ppm and one co ntrol m ale) were k illed prem aturel y due to po or clinical c onditi on . There were n o tr e a tment related cli ni cal s ign s. B od ywei ght s f or bot h s exe s at 7 000 ppm were lo wer than c ontro l s through out the s tu dy, wi th a m ax i mum differen ce of

- 10% toward s th e en d o f th e s tudy
. B odywei g ht s f or b oth sexe s at 2 500 ppm we re l ower than c ontrol s , with a maximum di fference of -6% towa rd s the end o f the stud y . Food co nsumption for b o th s exes at 70 00 pp m w a s h igh er th an co ntrols thro ughout t he s tudy
. Fo od c o ns umptio n for ma le s at 250 0 pp m w as a l so hi gh er than co ntrol, with s tati s tical s i gnificance b e in g a c hieved fr om we ek 6 o n l y . Food ut ilizati on o ve rall ( week s 

### I -B ) w as l e ss e fficient compared wi th c ontro l f o r male s in a ll t reated g ro ups and f em ales a t 250 0 and 7 000 ppm , with a do se re s po n s e ev ident
. a s s h own in the fo ll o wi ng t able
. Po s sib l y the differen ce i n th e f ood co n s umpt io n parameter s w as d ue t o i ncrease s p illage o f food i n th e Is opyr azam g ro up s . For the ha ematol ogy p arameters th ere w ere n o diffe rences c o n s idered to be tre atm ent rel ated
. Any o bse rved d i fference s w ere s m all, v alue s w ere ge n erally w ithi n t he ran ge of h i s to ric al m eans and a ffecting only one s ex, o r s ho we d n o c o n sis tent p att ern . To consi der th e cli nica l c he mi s try c hange s , in f em ales a t 7 000 ppm pl as ma a lbum i n/g lobulin rati o w as lower than c ontro l
. tri g ly cerid e s were high er th an con tr ol an d p l as m a alani ne ami not ransfe rase ac tivity w as h i g h e r than c ontr o l. The se s tati s tica lly s ign i fi cant differenc e s m ay be rel a ted to tr e at ment. Pl as ma c holes te ro l was s tat is t ic ally s ig n i fi cant l o w er t han c o ntro l s fo r mal e s a t all d o s e le ve ls, bu t there w a s n o d o se re s pon s e re l ation sh ip and i n the absence o f a ny o the r cl i nica l c hem i st ry di ffe rence s a mo ng m ales thi s d ifference i s c o nsidered t o be in ciden ta l to tre atme nt. Liver wei ght a dj us ted f o r fi nal bodyweight w as increased co mpared w ith co ntrol in bo th s exes at a ll d o s e level s , w ith a clear do se re s pon s e ev ident. Brain weight for m a le s a t a ll do se level s was s tat i s tica lly s i g nifi can tl y diffe rent f rom co ntrol s . H ow ev er, t here w as no d o se re sponse . Furtherm o re, th e c ontrol mean w as low co mp ared to h i st orical co n trol d ata, where as mean v a lues fo r a ll trea ted g roup s were within the ran ge o f hi storical c ontro l me an we ig hts
. In a ddition, th ere w ere n o p atho lo g i ca l fi n d ing s . Th erefore , th e s e d iffere nce s are co nsidered to be i ncide ntal to tre a tm ent. At necrops y a num ber o f c o mm o n s po ntaneous m acr os cop ic le sio n s w as o bserved , n one o f w h ich w as re lated t o treat ment with I sop yr azam . Mi c r osco pic ally , al l mi ce at 7000 p pm we re foun d w ith m od erate hepatoc yte h y pertro phy
. Am o ng ma le s t he h ypertro phy h a d a c entril obular di stributio n , w hil s t in th e females th i s change h ad d iffus e di stributi o n throu ghout the lobe s . A t 2500 p pm, hepatocy t e h ypert rophy w as a lso pre sent in al m os t a ll animal s o f bo th sex e s, b ut w as l e ss p ron oun ced ( m a inly minim al o r s l i ght )
. I n the m ale s th e pa ttern w as again c entril o bular and in th e f ema les it w as pred om inantl y c entri lo bu lar . A t the l ow do se ( 500 p pm) hepato cyte h ypertroph y ( min i m al/s li g ht) w as pre sent o nly in Page 31 of 41ISOPYRA ZAM
m ale s. C ANC ER A SSESSM ENT DO CUMENT FI NAL

Dietary admini stration of Isopyrazam (93% sy n : 7% anti) for 90 days at a concentration s of 500 , 250 0 and 7000 ppm caus ed a reduction in bodyweight gain and liver change s in the fonn of centrilobular hepatocyte hypertroph y and increased l iver weight. The se change s were accom panied by s l ight difference s some clinical chemi stry parameters, whic h may be re lated to the e ffects on the l iver. A s ad verse treatment-related effect s were s een at the low est expo sure leve l of 500 ppm a s tudy NOAEL cannot be identified. The LOAEL i s 500 ppm (intake of about 76 and 87 mglkg/day in ma le s and female s, re spectivel y). The s tudy provided th e ba sis for the s election of th e dose level s for a s ubsequent 80 w eek carcinogenicity s tudy. T he selection of a high dose le ve l of between 2 500 and 7000 ppm can be e xpected to elicit cl ear treatment-re lated effec t s on body weight and liver weigh t but without impact on s urviv a l over an 80 week p eriod
. b) C hronic T oxicity (4) RA T In a c ombined chronic toxicit y/carcinogenicity s tudy ( MRlD 477468 51), 52 Wi star rat s/ se x/do se were e xpo s ed to I sop yrazam (96.4 % a. i .; Lot N o.: SMU6APOOI) for up to 2 year s in t he diet a t concentrati ons of 0, 100 ,500, or 3000 ppm (equivalent to 0/0, 5. 5/ 6 . 9, 27 .6/34 .9 , and 17 3.5 /2 32 .8 mgikg bw /day in ma les /fema les ). Additiona lly, 12 rats/ se x/ dose were treated s imilarly for up to I y ear and then s acrificed
. N o adv erse, treatment-related effect s were ob served on s urviva l, hematolog y , or urin aly si s . The incidence o f the clinical observatio n " thinne ss " wa s increas ed in the 500 and 3000 ppm fema les (22-27 ob servations in 10-13 anima l s) compar ed to contro l s (10 ob s ervations in 5 animal s). Bod y weight s were decrea sed genera ll y from Week 5 through 1 05 in the 500 ppm females and throughout treatment in the 30 00 ppm m ales and females
. Body weig ht gains during the fir st 1 3 week s were decreased by 6 -20% in the 500 and 3000 ppm female s and b y 1 1% in the 3000 ppm male s . O verall (W eeks 1 -105) body weight g ains w ere decreased by 15

- 37% in the 5 00 and 3 000 ppm female s and by 1 6% in the 3000 ppm male s . A t 3 000 ppm , f ood c on sumption wa s s poradically decreas ed by 3-1 1 % in th e male s, genera lly during the first 1 1 week s and by 4 -8% in th e femal es, ge nera ll y during the firs t 14 week s. F ood uti l ization wa s decreas ed by 4-13 % in ma le s and 11 -16% in females during Week s 1-4,5-8 , and 9 -12
. Food utili zation for W eek s 1 -12 wa s decreased by 7 % in males and 1 5% in female s. In the 30 00 ppm fema le s, hind

- limb grip strength wa s decrea sed b y 29 % . Motor acti vity was incr eas ed by 95 % during Minute s 3 6
-40 , contributing to an incr ease of24 % in the o verall (Minute s I -50) activit y. Page 32 of 41ISOPYRAZAM C ANCER A S SESSMENT DOCU MENT FI NAL In fe males , i ncreased (s tatistical an aly s i s no t pe rfo rmed) incidence s of s mall li near opac ity in the co rnea was n oted at 3000 ppm (14 % treated v s 2% c ontro l s), a nd promin ent s uture lin e s in th e ey e we re not ed at 5 00 an d 3000 ppm (11-1 6% treated v s 2 % co ntrol s). The fo llowing findings may be indic ative of h epatotoxic ity . Incre ased y

- g l utamy l transferase was noted in the 5 00 ppm ma le s d uring Week s 53 , 79 , and 105, in th e 30 00 ppm ma le s during the interim kill and during Week s 5 3, 79. and 105, and in the 3 000 p pm f emales during Week 1 05
. Increased alanine amin otransfe rase was noted in the 3000 ppm ma le s du ring Weeks 53 and 105
. Adju sted li ver weight s w ere i ncreased i n the 3000 pp m grou p at the in terim an d te rmina l kill s. In the 3000 ppm fe males, increased i ncidence s of l iver m as s (9 /64 treated v s 0 /64 co ntrol s) and pa le s po tls or are a/s on li ver ( 10 /64 tre ated vs 1/ 64 co ntro l s) we re o b se rved. A t th e int erim sac rifice , i n c rease d i nc idence s wer e noted in the follow ing fin dings in th e l iver (# a ff ected/ 12 in treated v s co ntrol s) : ( i) h epatocyte vac uolation in the 500 and 3000 pp m ma le s ( 10

- 11 v s I ; m inimal to s light ) and females ( 8 eac h v s 0; mi nimal ); ( ii) mi nimal to s light hepa tocyte hype rtrophy in th e 500 and 3000 ppm males ( 4 -12 v s 0 ) and females ( 9 -12 v s 0); and ( iii) incre ased hepatocytelKupff e r cell p i gmentation in the 3 000 ppm male s (6 v s 0 ; minimal to s light) and f emales (1 0 vs 3 ; minimal )
. A t the terminal sac rifice including d ecedent s, increased incidence s were noted i n t he f ollo wing finding s in th e l iver (# affecte d/52 in treated v s co ntrol s): (i) m inimal to moderate centrilob ular hepato cell u l ar hype rtrophy i n the 500 and 3 000 pp m ma le s (45

- 49 v s 0) and fe male s (49 -50 v s 0 ); ( ii) m inimal to m oderate ce n tri lobu lar h epatocellu lar vac uo latio n i n th e 5 00 an d 300 0 ppm m a les ( 3 2-39 v s 3) ; ( i ii) b ile du ct h yperp la si a in 5 00 and 30 00 pp m m a les ( 12

- 19 v s 3; min imal to s li ght) and the 3000 pp m f e male s (2 3 v s I S; NS ; m inimal to mo derate); (iv ) m inimal to s li g b t b i le d uct fi bro sis i n th e 500 and 3000 ppm ma le s ( 10-12 v s I ) an d fe male s ( 1 1- 1 2 v s 6 ); and ( v) f oeal eo sinophili c alte red hepat ocyte s i n th e 500 and 30 00 ppm male s ( 23-32 v s 7 ; minim al to m oder ate) and female s ( 26-29 vs 9 ; m inimal to s light)
. Increased plasm a urea w as noted in the 3 000 ppm female s during Week s 53 and 1 05
. At th e termi nal sac ri fice in cluding dece dent s ( n= 52), th e incidence o f minimal to s li ght brown pigment in renal tub ule s wa s i ncrea s ed in th e 500 and 3000 p pm fema les 

### (30-42 /5

2 trea ted vs 

### 18 /

52 c o ntrol s). Dec reased pla s ma tri glyceride level s we re noted in 3000 ppm ma les througho ut th e s tudy and in 3000 pp m female s during Wee k s 53 , 7 9, and 105
. In t he 3000 ppm fe ma les , i ncre as ed inc idence s of pale s pot/s or are a/s o n l ung ( 21/ 64 treated v s 1 1/64 c ontrol s) w ere o bserved
. At the terminal sac rifice including deceden ts. the i ncidence of min im al to s eve r e foc al f ollicular hyperpl asia was increased in the th yroid gland of the 3 000 ppm male s 

### ( 18 /

52 treated vs 

### 11 152 c ontr ols)
. Th ese finding s were with out c o rrobo rating evidence of an a dve rs e effec t. Regar ding the non -n eopl ast ic effec t s, conce ntration s of 1 00 , 500 and 3 00 0 p pm Is opyr azam caused l iver c h ange s i n th e fo rm o f hepatocell ular h ypert rophy , vac uo lat i o n and p i gme nt ation
. A t 5 00 a nd 30 00 pp m th ese c h an ges were accompanied by reduced bodyweigh t gai n , i ncreased l iver weight, and al so by s light differe nce s in s o me cl inical chemi s try pa ramete rs which may be related to the effec ts on the liver
. Addit ional ly, at 3 000 ppm there was an i nc reased incide n ce o f s inus erythrocyt osis in the me senteri c lymph node s in male s and haem atological chan ges (reduced haem ogl obin, haemat ocrit and red blood cell co unt, incre ased p latelet co un t) in fem ales
. A s a dverse treatment-related effects were se en at the Page 33 of 4 1I SOPYRAZAM C ANC E R A SSESSMENT DO CUME N T FIN AL 

### lowe st e xposure level a s tudy NO AEL for non-ne oplastic ch anges c annot be identifi ed
. Th e L OAEL i s 100 ppm ( intake o f about 5 and 7 m g/kg/day in male s and f emales , re spectively). 

### (5) M ICE In a c arcinogenicity s tudy ( MRID 477 46849) , i s opyrazam ( 96.4% a .i
. ; Batch No
. S MU6AP001) wa s admini stered in th e di et to C 57BU IOJ rC D

- I mice (50 / sex/dose) for up to 80 week s at d ose s o f 0 , 7 0, 5 00 , or 35 00 ppm (equivalent to 0/0 , 7. 8 /9.9, 56 .2/74 .9, and 4 32 .6/55 3.6 m g/kg bw/da y in males/f emales)
. N o adverse , treatment -r elated e ff ec t w as noted on mortality , food con sumption , o r leukoc yte o r leukocyte diff erential c ounts . In the 3 500 ppm ma le s , the incid ence o f di scharge from ey e w as incre ased ( 326 ob servations in 3 6 anima l s) c ompared to contro l s ( 36 o bservations in 5 an imal s). Thi s finding wa s firs t noted in the co ntrol gr oup during We ek 24 and in the tr eated group s during Week 48
. At necrop s y , th e inciden c e o f ey e di schar ge was increa sed ( 26/ 50 treated vs 2/5 0 c ontro l s)
. A t the terminal s acrifice includin g de cedent s, i nc reased (p < =0.05 ) i ncidences w ere n oted in minima l to moderate i nflammation/exudate s from the na s olacrima l duct (3 8/50 treat ed v s 6/5 0 c ontro l s)
. At 3 500 ppm , decr ea sed (p <=0.05) bo dy weig ht s were ob served thr oughout treatm ent i n male s (deer 2

- 23%) and from Week 4 until termination in female s (deer 1 -11%)
. B ody we ight gain s during the fir st 13 week s w ere d ecrea sed i n ma le s by 1 8 % and in f emales by 1 1 % . O vera ll (Week s 1 -81 ) bod y w eight g ain s we re decr ea s ed b y 42 % in th e ma les and b y 22 % in the fema le s. A t 3 500 ppm , food uti lization was de creased (p <= 0.05) by 1 6-2 6% in ma les durin g W eek s 1-4 and 5 -8 , and w as d ecrea s ed**(p <~0** **.0 5)**  

### b y 1 4-22 % in the 5 00 and 35 00 ppm f emale s du ring Weeks 1-4
. Foo d utili zation f or We eks 1 -12 w a s decreas ed (p <= O .O I ) by 1 7-21% in b oth s exes. At the termin a l s acrifice including decedent s, in creased (p <= 0 .05 ) i ncidences w ere noted in the fo llowin g finding s (# 

### a ffectedl # e xam i ned in treated v s c ontrols) in th e 3500 ppm fe ma les : minimal to s light eo sinophi lic dropl ets in the epithelium of the nasal c avity/ ph ary nx (36/ 50 v s 2 1/50) ; and minim al to mod erate e osinophil ic dr op let s in th e e pith elium in the ga ll bl adder (25 / 50 vs 14/48 )
. A fte r 80 week s of tr eatment, terminal b ody w eigh t s w ere s tati s tically s imilar to co ntrols. Increa sed (p <=0. 05 ) adjusted (for b od y weigh t) l iv er wei ght s were noted in the 3 500 ppm ma les (incr 24%) and 5 00 and 35 00 ppm fe male s (1 3

- 38%)
. Incr eased**(p ~0** **.0 5)**  

### ab so lute li ver we ight s w ere not ed in the 50 0 and 3 50 0 ppm f ema le s ( incr 1 1- 3 0 % )
. At the t cnn ina l s acrifice i ncluding decedent s , increased (p <= 0
. 05) in cidence s w ere noted in the fo llowin g finding s in the liver (# affected/50 in treated v s co ntrol s) : minim a l to s light m id

- zona l hepatoc ellu l ar h ypertrophy in the 3 500 ppm ma le s (40 vs 0 ) ; and minima l to m oderate p eriportal hepatoce llular h ypertrophy in the 500 and 3 500 ppm female s (13-4 7 vs 0)
. Increa sed l i ver w eight and hypertrop h y in the ab sence o f an y oth er eff ect on th e liver i s not co n s id e red adv erse . Pag e 3 4 of 4 1I SOPYRA ZAM CANCE R ASSES SM ENT DOCU MENT F I NAL 

### The LOAEL is 3 500 ppm ( equivalent to 43 3/554 m g/kglday in m ale s/ females) bas ed o n ey e d ischarge i n 

### male s; decreased bod y weight s, bod y weight ga ins, and f o od utilizat ion in bo th 

### s exes; and eosinophilic d

roplet s in the n asal cavity an d ga ll bl adder in fem ales . The 'KO AEL is 500 ppm ( equivalent t o 56/75 m g/k glda y in ma le s/fema les)
. 5. Mode of Action 

### A formal mode o f action framework for the se tumo r s wa s not provided by the reg istrant f or consideratio n by the C A Re.

Pa ge 35 o f 41ISOPYRAZAM CANCER A SSESSMENT DOCUMENT FINA L 

### V
. COMMIT TEE'S ASS ESSMENT OF THE WEIGH T -OF- THE-EVIDENCE The Committee considered th e fo llowing for a weight -of-evidence determination of the carcinogenic potentia l of isopyrazam
. I
. Carcinogenicity Rat



### • Thyroid Tumors : 

### Ma le W i star rat s had a statistically significant trend for thyroid follicu lar cell ad enomas, and s ign ificant pair-wi se compari sons of the 3 000 ppm dos e group with the contro ls for thyroid follicu lar cell adenomas and combined adenomas and/or carcinoma s, all at p 

### < 0 .05. There wa s a lso a significant pair -wi se compari son ofthe mid-do se group (500 ppm) with the controls for thyroid fo llicular cell carcinomas and combined adenomas and/or carcinom as, both at p < 

### 0 .05. The CARC considered the thyroid tumors to be t reatment-related in male rats.

• Liv er Tumors: 

### Female Wi star rats had s tatistically s ignificant trend s, and s ignificant pair

- wise comparisons o f the 3000 ppm do se group wi th the contro l s, for l iver adenomas and comb ined ade noma s and/or carcinomas, bo th at p < 

### 0.01. The tumor s were s upported b y non-neop lastic le si ons ( eos i nophil ic foci) . T he C ARC considered the liver tumors to be treatment-related in fema le rat s. • U terine Endometria l Epithelial Tumo rs: 

### Fema le Wi star rat s had stati stically significant trends , and s ignificant pair-wise compari sons of th e 3000 ppm do se group wi th the co ntrol s, uterine endometrial epithe lial adenocarcinom as and combi ned adenoma s and/or ade nocarcinoma s, both at p < 

### 0 .01 . T he CAR C con sidered the uterine tumors to be treatme nt -related in female rats .

• Ad equacy o f Dosing : 

### In both ma le s and female rat s, the high dose o f 3000 ppm wa s con sidered to be adequate , but not e xcessive
. In mal es , bod y weight decrease s ranged from 4 -14 % throu ghout th e s tudy. Overa ll body weig ht gain in ma le s wa s decreased 16% (week s 1 -10 5 )
. In female s, body we i ght and body weight gain e ffects were more marked. At 30 00 ppm in fema les , body we ight decrea ses ranged from 4- 30% throug hout the s tudy. Ove ra ll bod y w eight gain was de creased 3 7% (we eks 1-1 05)
. A lthough the ob served bod y we ight/body weig ht gain s chan ges exceeded the convent ional 1 0% criteria emp loyed for as sessmen t o f body weig ht change s in carc inogenicity s tud ies, the C ARC did not con s ider th e h igh dose ( 3000 ppm ) to be exce ssive in ei ther s ex s ince there wa s no other corrobora tive evidence of e xcessive toxicity . At thi s do se, ther e were no adver se clinical s igns of toxicit y or s ignificant changes in the cli nical c hemi stry parameters , s urvival was not affected , and there w ere no adverse non -neoplastic le s ion s (e.g ., no n ecro s i s). Mo u se

• There were no treatment-re lated increase s in tumors in either male or fema le mice
.

• Ad equacy of Do sing: 

### The do ses t ested we re con sidered to be adeq uate and not e xcessive in both ma le Page 36 of 41ISO PYRAZAM CANC ER ASSESSMENT DOC UM ENT FINAL 

### an d f emale mi ce. Th is was based o n dec reased body weight, bo dy weight gai n, food uti lization, and mini mal cli nical sign s, clinical che mistry c hanges, liver toxicity and non -neo plastic lesion s in the nasal cav ity and ga ll bladder
. 2
. M utagenicity: The re i s n o co ncern fo r m utagenic ity . 3
. S tructure Ac tivity Rel ationship : Is opyrazam is a me mber o f the p yrazole-carboxam ide class of s uccinate de hydrogenase inhi bitor fun gicides
. Th ere ar e no S AR d ata avai lable fo r thi s c hemical at thi s t ime. V I. C LASS IFI CATIO N O F C AR CI NOGENIC PO T ENTIAL In ac cordance wi th the EPA's Fin al G uidelines/or Ca rcinogen Ri sk As sess ment (March, 2 005) , th e C ARC cl assified i sopyrazam as "Likely to be Ca rcinogenic to Humans"
. Thi s cl assification i s ba sed o n the pre sence o f t hyroid fo llicular c ell t umors in mal e rats, and liver and uterine tum ors i n fe male rats at d oses that we re adequate to eva luate the carci nogenic po tential o f i sopyrazam . No tr eatment-related t umors we re see n in m ice
. The re is no mut agenic concern f or i sopyrazam . V II. Q UANTI FICATI ON OF CAR CI NOG ENIC PO TE NTI AL A l inear low-dose extrapo lation mode l ( Q, * ) 

### will be used for quantification of cance r ri sk to human s .

P age3 7 of 4 1I SOPYRA ZAM 

### VIII B IBLIOGRAPHY CANCE R ASSESS MEN T D OCUM E

NT FI NAL

M RID No. 4774 6832

4 7746833

4 7746834

4 7746835

4 7746836

4 7746848

4 774684 9

4 7746851

4 7746852

4 7746 853 C ITA TION 

### Pint o , 

### P .I

. (2008) 9 0 Day Pr elimin ary Ca rcinogenicity St udy i n th e Mo u se. P roject Nu mber : P M I 3 48. T0 11961-05
. Un published study 

### pr epared by Cen tra l Toxicology L aboratory . 3 19 p . S hearer, J
. ; Wood , M
. (2 008) SYN520 453 (89.5% Syn:6.9% A nti), S YN5204 53 (63.3% Sy n :27.5% A nti)

- 13 W eek Co mparative D ietary Tox icity S tudy i n Rats. Project N umber: 4 58440 , T00 9147-06. Un published s tudy pr ep ared b y C harles R iver. 570 p . M ilburn, a. ( 2007) 90 D ay Diet ary To xicity S tudy in Rats . P ro ject N umber : P R-1349, T 0037 12-05. Unpublished st udy prepared b y Ce ntral 

### T

oxicology Labor atory . 920 p. J ackson , A .M. ( 2008) SY N52 0453 - 13-Week O ral (Ca p s ule) T oxicity Stu dy in th e Dog
. Project N umber : B4 3852 , T00088 2-07
. U npublished s tudy pr epared b y RC C , Ltd. 359 p. Br ammer, A
. ( 2007) 9 0 D ay O ral T oxicity S tudy in Dog s. P roj ec t N umb e r: PD135 1, TOI1 964-05. Unpublished s tudy pr epared b y Ce ntral 

### Tox icology L ab orato ry . 41 9 p . J ackson , A .M. (2 008) S YN5204 53

- 52 -Week O ral (Ca p s ule) T oxicity S tudy in th e Beag le Dog. Proj ect N umber: B 09336, TOI1 966-05
. U npublished s tudy pr epared b y RCC , Ltd
. 4 36 p . P into , P J
. (20 08) S Y N52 04 53

- 80 Wee k Die tary C arci no geni c i ty S tudy in t he M ouse . Pr oject N umber : PM 1352 , T OI 19 62-05. U npublished s tudy pr epared by CEMAS
. 2 , 235 p
. Mi lburn, a.M
. (200 8) SYN52 0453

- 2 Ye ar D ietary T oxicity and Carci nogenicity S t udy in Rats . P ro jec t N umbe r : PR 1 35 3, T0 11963-05
. U npublished s tudy p repared b y CE MAS
. 4 ,08 8 p
. Sok olowski, A
. (200 8) CSC D465 008

- 

### Sal monell a typh imurium and E

scherichia coli Reve rse Mutation Assay. P roject N umber : 11 29601 , T 012725-05. U n pu b l is he d s tudy pr epared b y RCC, Ltd
. 44 p . So kolowski, A. (2008) CS CD 459488

- 

### Salmonella ty phimurium an d E

scherichia c oli 

### R everse Mutation Assay. P rojec t Num ber 11 64800, Pag e 3 8 

### of 41I SOPYRAZAM 

### M RIDN o . C ANCER A SSESSMENT DOCU MENT 

### CITATION FI NAL



### 4 7746 854

4 7746855

4774685 6

4 774 6 85 7

477 46858

47746859

4 77468 6 0

477 468 6 1

4 774686 2

4 774 6 8 63 TO I2726 -05 . U npu b lis h ed s t ud y pr epared b y RCC, L td
. 44 p . 

### Soko lo wsk i , A. (2 008) Sal monella typ h i murium 

### a nd E sch eric hia coli 

### R

everse Mutation A ssay with S YN520453 . Pr oject Numbe r: 110 4001 , T00 0877 -07
. Unpu b l is hed s tud y p rep a red b y RCC , Ltd
. 4 5 p . 

### Calland e r

, R.D. (2006 ) Bacte ri al M utat i o n A ssay in 

### S. typhimuriu m 

### and E. co li
. 

### Pr o

ject Nu mber : YV 7254, T0037 1 5 -05 . U n pu b l ish ed st udy p repared b y Ce ntra l T oxicology Labora t o ry . 3 1 p Wo llny. H
. ( 2008) C SCD465008

- C e ll M utatio n A ssay at the Th ymidine Ki nase Loc u s (T K +1 -) 

### i n Mouse Lymphoma L5 178Y Cell s . Project Number: 1 129 60 3, T 01272 2

- 05
. U np ublish ed st ud y prepared by R CC , Lt d
. 56 P Woll ne y , H
. (20 08) C SCD4 59 4 88

- Ce ll M utation Assay a t th e Thy midine K in ase Loc u s ( T K +1

- ) 

### i n Mou s e L ympho m a L5 17 8Y C e lls
. 

### P

roject Num ber: C lay, P
. (2006 ) L5 178Y T K + 1

- 

### M o u se Lymph oma Mu tation A ssay . Proj ect Numbe r : VV0342 , T003 7 17-05
. U npu blis h ed st udy prepared by C entra l T oxico l ogy Laboratory
. 3 3 p
. Wo llney, H
. ( 2008) SYN520453

- C e ll Mu tation A ssay at th e Thym id ine Kinase Loc u s (T K +1

- ) 

### in M o use L ymphoma L5 178Y Cells
. Pro j ect Nu mber : 1104003 , T00 0879-07
. Unp u b lis h e d st ud y p repared b y R CC, Lt d
. 56 p . B oh nenb erger , S . (2 008) C SC D4 65008 - Chro moso me Aberrat io n Te st i n H uman Ly mph ocyte s I n Vitro . 

### Project N umber : 1 12 9602, TO I27 13

- 05
. U n p ub l i sh e d s tud y p re pared by RCC, L td
. 4 3 p . B ohnenberger , S . ( 2008) CSC D459488

- C hrom oso me Abe rratio n Te st in Human Lymp hocyte s In V itro . 

### Project N umber : 1 19200 1 , T0 12714

- 05
. Unpublished s tudy prepared by RCC, Ltd
. 41 p . 

### Fox, V . (2006) I n V itro 

### C ytog en et

i c A ssay in Human Ly mphocytes . P

roje c t N umber: S V1348, T00 3720

- 05
. Unp ubli s he d st udy prepared by 

### Centr al Tox i cology L ab oratory. 3 1 p. 

### Bo hn enb erger, S . (20 08) Chr omo so me Aberr a tion Tes t in H uman L

ymphocytes in Vitro 

### with SYN5 20453
. P roject N umber: 11 04 0 02 , Page 39 o f 4 1ISOPYRAZAM M RID No. C ANC ER A SS E SS ME N T D OCUMEN T C ITATI ON F IN AL

4 7746864

4 7746867

4 7746868

4 7746869

4 7746870

4 7746871

4 7746872

4 774687 3 T 000878-07 . U npubl is hed s tudy pr epared b y RC C , Ltd
. 4 2 p . 

### Fo x , V . (2 006) Rat Bone Marr ow Mi cronucleu s Test. Proj ect N umber; S R 1330, TOI9 063-04 . U npublished s tudy pr epared b y C e ntral 

### To xicology Labor ator y . 2 6 p. A llan, 

### L. ( 2009) S YN 52 045 3

- Pharmacokin etic s in th e R at Fo llowing S ingle O ral A dministration of[ 14C]- SYN5204 53 (I and 75 mg/k g)
. Pro ject N umber : 18 6713, TOI1 960-05 . U npubl is he d s tudy pr epared b y 

### Ch ar les R iver La b

ora tori e s. 6 5 p. Gr een , M .A. ( 2009) SYN520 453 -In vesti gation ofthe N ature and 

### Id entit y o f Radi olab elled 

### M et

abolite s Present in Pla sma, U r

ine, F aeces and B ile Co llect ed fr om Rat s Fo llowing Ora l Admini s tration o f [1 4C]SYN5204 53
. Proje ct N umber : 21 2072 , T00 9151-06. U np ublish e d 

### study p repared by C ha rl es Ri ver Laboratorie s . 23 8 p. A llan , 

### L. ( 2008) SYN52 045 3

- The Ti ssue Depletion o f [ " C]- 

### S YN52 0453 in th e Rat Fo llowing a Si ngle O ra l Admini stration (I 

### and 75 m g/kg)
. Pro je ct N umber : 186954, T002 I8 0-07. U npubli s hed s tudy 

### prepared b y Ch ar les Riv er L aboratori es. 133 p . All an , 

### L. (20 08) SYN5 2045 3

- T he Tis s ue D istribution an d E li minat ion o f 

# e 4

C]

- S YN52 04 5 3 in th e Rat f ollo wing Multi ple O ral A dmini s tration ( I m g/k g/d ay)
. Pr o j e ct N umber : 186603, TOI1 955-05
. U npubli s hed 

### stud y pr epared by C har le s Ri ver Lab or atori es. 112 p . S ilcock , R
. C. (20 07) Wh ole B ody A uto radiography a nd E xpire d Air S tudy i n th e Rat
. P ro ject N umber: U R0911 , T OI 1950-05 . U npubli s hed 

### stud y pr epared by Centr a l T oxicology L aborator y . 36 p. S haw , J
. (2 008) S YN 52045 3

- T he Biliary E limination o f To tal 

### Rad ioacti vity in t he Rat Fo llowing S ingle Oral Administration o f [ 14C 

### ]_ SYN 5204 53 (I m g/k g and 75 mg/k g)
. Proje ct N umber : 186467 , T OI 1951-05 . U np ublis hed s tudy pr epared by C harle s Riv e r 

### L

ab oratorie s. 50 p. S h a w, 

### J
. (2 008) SYN5 20453

- The Biliary E limination o f To tal 

### R

adi oactivity in t he R at F o llow ing S ing le Oral Admini stra tion o f Syn o r A nti [ 14C]-S YN5204 53 (2 m g/kg a nd 75 m g/kg)
. Pro jec t N umber : 18 7 586 , T00088 6 -0 7
. U npublished Pag e 40 of 41ISOPYRAZAM MRiD No. C ANC ER A SSESSMENT DOC UMENT C ITATION FIN AL

47746874



### Memo Shaw , J
. (2008) SY N52045 3

- The E xcretion and Ti ssue Di stribution of [ 1

4C]-SYN520453 in the Rat Followi ng Single Oral Admini stration (I mg/kg and 75 mg/kg ). Project Number : 186692 , TOI 1957-05
. 

### Unpublished study prepared b y Charles River Laboratori es. 56 p Brun sman, L. 

### 2010. I sopyrazam Qualitative Risk Asses sment Based On Wi star Rat Dietary Stud y. 12 /16 /2010
. TXR N o. 00 55586
.

Page 41 

### of 41