**UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460**  **OFFICE OF PREVENTION, PESTICIDES AND TOXIC SUBSTANCES** 

**MEMORANDUM Date:**  November 25 , 2009 **SUBJECT:** Fluopy

ram : Report of the Cancer Assessment Review Committee **PC Code:** 080302 **DP Barcode:** N/A **Decision No.:** N/A **Registration No. :**  N/A **Petition No.:** N/A **Regulatory Action:** N/A **Risk Assessment Type:** Cancer Assessment **Case No.:**  N/A **T XR No**  **.:** 0055261 **CAS No.:** 658066 - 35 - 4 **MRID No.:** N/A **40 CFR:** N/A **FROM:** Jessica Kidwell , Executive Secretary

Cancer Assessment Review Committee Health Effects Division (7509P) **THROUGH:** Mary Manibusan, Co - chair

Cancer Assessment Review Committee Health Effects Division (7509P) **TO:** My

ron Ottley , Toxicologist Paula Deschamp, Branch Chief Registration Action Branch II I Health Effects Division (7509P) Cy nthia Giles - Parker Fungicide Branch Registration Division (7505P) The Cancer Assessment Review Committee met on Ju ly 8, 2009 to evaluate the cancer classification of Fluopy

ram i n accordance with the EPA’s Final Guidelines for Ca rcinogen Risk Assessment (March, 2005). Attached please find the final Cancer Assessment Document.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

2 CANCER ASSESSMENT DOCUMENT

EVALUATION OF THE CARCI NOGENI C POTENTIAL OF **FLUOPYRAM** PC CODE 080302

November 25 , 2009 

### C ANCER 

### A SSESSMENT 

### REVIEW 

### C OMMI TTEE

HEALTH EFFECTS DIVI SI ON OFFICE OF PESTI CIDE PROGRAMSFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

3 DATA PRESENTATI ON:

____________________________________ My ron Ottley , Toxicologist DOCUMENT PREPARATI ON:

____________________________________ Jessica Kidwell, E xecutive Secretary COMMI TTEE MEMBERS I N ATTENDANCE: (Signature indicates concurrence with the assessment unless otherwise noted.) Gregory Akerman

____________________________________ Lo ri Brunsman, Statistician

____________________________________ Marion Copley

____________________________________ Kit Farwell

____________________________________ Ray Kent

____________________________________ Mary Manibusan , Co - Chair

____________________________________ Karly n Middleton

____________________________________ Rob Mitkus

____________________________________ Esther Rinde

____________________________________ NON - COMMI TTEE MEMBERS I N ATTENDANCE: (Signature indicates concurrence with the pathology report) John Pletc her, Consulting Pathologist

___ _______________

__________________ OTHER ATTENDEES: L eung Cheng, Barry O’Keefe, Joel Patterson (PMRA observer) , Meheret Negussie, Steve Funk, Whang Phang, Doug Wolf, Sheila Healy , Paula Deschamp, Michael Khan; On conference phone with Global Review Partners - Germany (Philip Marx St ö lting, David Schumacher) and Canada (Catherine Adcock, Mi chael Honey man , Martin Gerrits, Carmen Cheung )FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

4 **TABLE OF CONTENTS**  EXECUTIVE SUMMARY...................................................................................5 I. Introduction ................................ ................................ ................................ ................................ .. 8 II. Background Information ................................ ................................ ................................ ............. 8 III. Evaluation of Carcinogenicity ................................ ................................ ................................ ... 9 1. Combined Chronic Toxicity / Carcinogenicity

Study in Rats ................................ ............ 9 2. Carcinogenicity

Study in Mice ................................ ................................ ...................... 13 IV . Toxicology ................................ ................................ ................................ .............................. 1 6 1. Metabolism ................................ ................................ ................................ .................... 1 6 2. Mutagenicity ................................ ................................ ................................ .................. 1 7 3. Structure Activity

Relationship ................................ ................................ ...................... 1 8 4. Subchronic and C hronic Toxicity ................................ ................................ ................. 1 9 5. Mode of Action .... ...........................................................................2 7 V. Committee’s Assessment of the Weight - of - the Evidence ................................ ....................... 31 VI . Classification of Carcinogenic Potential ................................ ................................ ................. 3 3 VI I. Quantific at i on of Carcinogenicity ................................ ................................ ......................... 3 3 VI II. Bibliography ................................ ................................ ................................ .......................... 3 3FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 5 of 35 **EXECUTIVE SUMMARY** On July 8, 2009, the Cancer Assessment Review Committee (CARC) of the Health Effects Division of the Office of Pesticide Programs met to evaluate the carcinogenic potential of f luopy ram . This is the first assessment of fluopy ram by the CARC. Fluopy

ram is a global review chemical in partnership with Canada and Germany . Germany has the work share lead for review of toxicology

data. My ron Ottley

of Risk Assessment Branch I II presented the chronic toxicity /carcinogenicity

study in Wistar rats and the carcin ogenicity study in C57BL/C6 mice. Ina combined chronic toxicity and carcinogenicity

study , groups of 60 male and female Wistar rats were fed a diet containing 0, 30, 150 and 750 ppm (males) and 0, 30, 150 and 1500 ppm (females) fluopy ram for 24 months. In males, the top dose level of 750 ppm had to be reduced to 375 ppm from week 85 onwards because of the high mortality in this group. Over the whole study period, these dietary concentrations corresponded to a mean daily intake of 0, 1.20, 6.0 and 29 mg/kg bw in male rats or 1.68, 8.6 and 89 mg/kg bw in females. In addition, satellite groups of 10 animals per sex and dose were subject to interim sacrifice after 1 year. Groups of 60 male and female C57BL/6J mice were fed diets containing 0, 30, 150 or 750 ppm of fluopy ram (corresponding to a mean compound intake of 0, 4.2, 20.9 and 105 mg/kg bw/day in males and 0, 5.3, 26.8 and 129 mg/kg bw/day in females, respectively

) for up to 78 weeks. After 52 weeks, 10 males and 10 females from each group which had b een allocated to the chronic phase of the study were killed and necropsied. The remaining 50 animals per sex and group (allocated to the carcinogenicity phase) continued treatment until scheduled sacrifice after at least 78 weeks of treatment. In formation on mutagenicity , structure activity relationship and mode of action data for thy roid and liver tumors was also presented. The CARC concluded the following: **Carcinogenicity** Rat

**·** Administration of fluopy ram resulted in the induction of liver tumors in female Wistar rats. There were statistically

significant trends for l iver adenomas (p <0.01), carcinomas (p<0.05) and combined l iver adenomas and carcinomas (p< 0.01). There were sign ificant pair - wise comparisons of the 1500 ppm dose group with the controls for liver adenomas at p< 0.05 and for combined liver adenomas and carcinomas at p< 0.01. When compared to historical control data (uncensored data) from the testing laboratory , the i ncidence of hepatocellular adenomas in the female high dose group (9/5 5 , 16 %) was outside the range of the historical control group (range, 0 - 5%; average, 1.9%) . Similarly , the incidence of hepatocellular carcinomas in the female mid (2/56, 4%) and high dose group s (3/5 5 , 5%) , while not statistically significant by

pair - wise comparison, exceeded the range of the historical control group (no carcinomas observed in 10FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 6 of 35 studies from 2000 - 2006) and was considered to be biologically relevant . There were no statistically

significant trends or significant pair - wise comparisons of the dosed groups with the controls for the male rats. **The CARC considered the liver tumors in female Wistar rats to be treatment**  **- related.**  · Adequacy of Dosing : The doses tested were considered to be adequate and not excessive to assess carcinogenic potential in both sexes. **In female rats** , the highest dose tested of 1500 ppm was considered to be adequate. This was based on a 20% decrease in mean body weights an d increased liver weights (mean: +39%, absolute: +56%), which were dose - related and correlated well with findings such as prominent lobulation and hy pertrophy . Other effects that occurred at the high dose included thy roid f ollicular cell hy

pertrophy and c olloid alteration , c hronic progressive nephropathy

and kidney tubular dilatation , and r etinal atro phy and lenticular degeneration of the eye. **In male rats** , the top dose level of 750 ppm, an excessive dose, had to be reduced to 375 ppm from week 85 onwards because of the high mortality in this group. At the termination of the study , mortality among high - dose males was 50% of control values. The dose of 375 ppm, which is roughly one - half the excessive dose of 750 ppm, however, was considered to be adequate. This was based on decreased mean body weight, increased liver weight, liver hy pertrophy , f

ollicular cell hy

pe rtrophy and colloid alteration, c hronic progressive nephropathy and tubular dilatation and tubular hy pertrophy (males) all observed at the high do se. Mouse

· Administration of fluopy ram resulted in the induction of thy roid follicular cell tumors in male C57BL/6J mice. Male mice had a statistically significant trend at p<0.01 and a significant pair wise comparison of the 750 ppm dose group with the controls at p<0.05 for thy roid follicular cell adenomas. There were no statistically significant trends or significant pair - wise comparisons of the dosed groups with the controls for the female mice. When compared to historical control data (uncensor ed data) from the testing laboratory , the incidence of thy roid follicular cell adenomas in the male high dose group ( 7/48 , 15 %) was outside the range of the historical control group (range, 0 - 2 %; average, 0.4 %) . **The CARC considered the thyroid follicular cell**  **adenomas in male C57BL/6J**  **mice to be treatment**  **- related.** 

· Adequacy of Dosing : The high dose of 750 ppm was considered to be adequate, and not excessive, to assess carcinogenic potential in both male and female mice, based on t he following: 1) Increased absolute and relative liver weights in males and females at the high - and mid - dose levels. These changes were dose - related, and correlated well with macroscopic findings (dark/enlarged livers) and non - neoplastic histopathologica l lesions (eosinophilic foci [females], hepatocellular hy

pertrophy [both sexes], cholestasis [males], single cell degeneration/necrosis [males]); 2) Increased incidence of thy roid follicular cell hy

perplasia in both sexes at the high dose; and 3) Decreased absolute and relative kidney weights in both sexes at the high dose level, along with increased incidence and/or severity of kidney

lesions (bilateral cortical basophilic tubules, hyaline casts and interstitial mononuclear cell infiltrates, glomerular con gestion/ hemorrhage, and amyloid deposition) in females.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 7 of 35 **Mutagenicity** There is no concern for mutagenicity . **Structure -** 

**Activity Relationship (SAR)**  The SAR data do not inform the mode of action. **Mode of Action** The CARC concluded that insufficient data were provided to support definitive modes of action for the induction of liver tumors in female rats or thy roid follicular cell tumors in male mice. The main deficiency included a lack of dose - response concordance with key events and tumors. **Classification and Quantification of Carcinogenic Potential** Inaccordance with the EPA’s Final Guidelines for Carcinogen Risk Assessment (March, 2005), the CARC classifi ed f

luopy ram as **“Likely to be Carcinogenic to Human s**  **”** based on tumors in two species and two sexes: a treatment - related increase in thy roid follicular cell adenomas in high dose male mice and liver tumors in female rats at the high dose, with incidences exceeding that of the laboratory ’s historical controls. There is no mutagenic concern for fluopy ram. The available data do not support the proposed modes of action for the thy roid or liver tumors. The CARC recommended the use of a linear low dose extrapolation model applied to the animal data (Q1*) for quant itative estimation of human risk. The unit risk, Q 1 * (mg/kg/day ) - 1 , of Fluopy ram based upon female rat liver combined adenoma and carcinoma tumor rates is 1.55 x 10 - 2 in human equivalents.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 8 of 35 **I. INTRODUCTION** On July 8, 2009, the Cancer Assessment Review Committee (CARC) of the Health Effects Division of the Office of Pesticide Programs met to evaluate the carcinogenic potential of Fluopy ram . **II. BACKGROUND INFORMATION** Fluopy ram is a global review chemical in partnership with Canada and Germany . Germany has the work share lead for review of toxicology data. Fluopy ram is a new broad - spectrum sy stemic fungicide of the carboxamide group (FRAC Group 7). Itacts on cell respiration in the fungus by inhibiting succinate dehydrogenase (mitochondrial respiration Complex I I), thus blocking electron transport . The main use of fluopy

ram is the selective control of a variety of fungal diseases like powdery mildew species, Botrytis cinerea, and Alternaria solani on grape vines and tomatoes. It is also to be used on ornamentals and non - residential turf. Its structure and other pertinent information are depicted in the following table. Chemical structure: 13 14 15 16

11 12 10 N

H 9 O 8 7 2 N 1 6 5 4 3 Cl C F 3 C F 3 Empirical formula: C 16 H 11 Cl F 6 N 2 O Common name: Fluopy ram Company code: AE C656948 IU PAC name: N - {2 - [3 - chloro - 5 - (trifluoromethyl)py ridin - 2 - yl ]ethyl} - 2 - (trifluoromethyl)benzamide CAS name: Benzamide, N - [2 - [3 - chloro - 5 - (trifluoromethyl) - 2 - py ridinyl]ethyl] - 2 - (trifluoromethyl) - (9CI) CAS no.: 658066 - 35 - 4 PC Code 080302FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 9 of 35 **III. EVALUATION OF CARCINOGENICITY STUDIES** 1. Combined Chronic Toxicity /Carcinogenicity

Study with Fluopy

ram in Wistar Rats **Reference** : Chronic Toxicity and Carcinogenicity

Study of AE C656948 in the Wistar Rat by Dietary Administration for 24 months . Bayer Crop Science Project Number: SA/04312, M/298339/01/2. Report No. 04312 February 29, 2008. MRID 47372501 A. Experimental Design Ina combined chronic toxicity and carcinogenicity

study , groups of 60 male and female Wistar rats were fed a diet containing 0, 30, 150 and 750 ppm (males) and 0, 30, 150 and 1500 ppm (females) fluopy

ram for 24 months. In males, the top dose level of 750 ppm had to be reduced to 375 ppm from week 85 onwards because of the high mortality in this group. Over the whole study period, these dietary

concentrations corresponded to a mean daily intake of 0, 1.20, 6.0 and 29 mg/kg bw in male rats or 1.68, 8.6 and 8 9 mg/kg bw in females. In addition, satellite groups of 10 animals per sex and dose were subject to interim sacrifice after 1 year. B. Discussion of Mortality and Tumor Data Mortality Male rats showed a statistically significant trend (p <0.01) for mortality with increasing doses of Fluopy ram, as well as a sign ificant pair - wise comparison (p< 0.05) of the 750 ppm dose group with the controls. There was no evidence of significant differences in mortality in female rats (Memo, L . Brunsman, Jun e 10 , 200 9, TXR #0055218). Tumors Female rats had statistically significant trends for l iver adenomas (p <0.01), carcinomas (p<0.05) and combined l iver adenomas and carcinomas (p< 0.01). There were significant pair - wise comparisons of the 1500 ppm dose group with the controls for liver adenomas at p< 0.05 and for combined liver adenomas and carcinomas at p< 0.01 (Table 1) . There were no statistically significant trends or significant pair - wise comparisons of the dosed groups with the controls for the male rats. The statistical analyses of the tumors in the female rats were based upon Fisher’s Exact Test for pair - wise comparisons and the Exact Test for (Memo, L . Brunsman, Jun e 10 , 2009, TXR #0055218).FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 10 of 35 Table 1. Li ver Tumor Rates + and Fisher’s Exact Test and Exact Test for Trend Results in Female Rats Dose (ppm) 0 30 150 1500 Adenomas (%) p = 2/59

(3) 0.00049** 2/57

(4) 0.67765 0/56

(0) 1.00000 9 a

/55 (16) 0.01978* Carcinomas (%) p = 0/59

(0) 0.02134* 0/57

(0) 1.00000 2/56

(4) 0.23494 3 b /55

(5) 0.10910 Combined (%) p = 2/59

(3) 0.00015** 2/57

(4) 0.67765 2/56

(4) 0.67083 11 c/55

(20) 0.00536** +Number of tumor bearing animals/Number of animals examined, excluding those that died or were sacrificed before we ek 54. aFirst adenoma observed at week 75, dose 1500 ppm. b First carcinoma observed at week 97, dose 1500 ppm. c One animal in the 1500 ppm dose group had both an adenoma and a carcinoma. Note: Significance of trend denoted at control .

Significance of pair - wise comparison with control denoted at dose level. If * , then p < 0.05. If ** , then p < 0.01. When compared to historical control data (uncensored data) from the testing laboratory , the incidence of hepatocellular adenomas in th e fe male high dose group at week 104 ( 9 / 5 5 , 16.4 %) was outside the range of the historical control group (range, 0 - 5 %; average, 1.9% ) . Similarly , the incidence of hepatocellular carcinomas in the female mid and high dose group at week 104 ( 2/56 (4%) mid and 3/5 5 , 5% , high ) exceeded the range of the historical control group (no carcinomas observed in 10 studies from 2000 - 2006). C. Non - Neoplastic L esions in the Li ver Non - neoplastic lesions of the liver are presented in Tables 2a and 2b .FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 11 of 35 **Table 2a**  **: Incidence and severity of microscopic changes i**  **n the liver, all animals of the**

**carcinogenicity phase (p**  **art 1 of 2 )**  **Sex Males**  **Fem ales**  **Dose level fluopyram (ppm** **) 0**  **30 150**  **750/375 0**  **30 150**  **1500 Dose level fluopyram** **(m g/kg)**  **0 1.20**  **6.0 29.0/15.0**  **0 1.7**  **8.6 89**  **Num ber of anim**  **als exam ined**  **60 60**  **60 58**  **60 60**  **60 59**  Centrilobular to panlobular hepatocellular hypertrophy: diffuse Minimal 1 1 14 15 0 0 0 6 Slight 0 0 0 15 0 0 0 21 Moderate 0 0 0 0 0 0 0 21 Total 1 1 14** 30** 0 0 0 48** Focus(i) of hepatocellular alteration: clear: focal/multifocal Minimal 8 6 6 11 0 3 3 8 Slight 2 2 1 5 1 1 1 3 Total 10 8 7 16 1 4 4 11** Focus(i) of hepatocellular alteration: eosinophilic: focal/multifocal Minimal 12 20 18 15 23 17 14 25 Slight 4 3 10 12 6 7 15 13 Moderate 0 1 2 1 0 2 1 7 Marked 0 0 1 0 0 0 0 3 Total 16 24 31* 28** 29 26 30 48** Hepatocellular vacuolation: focal/multifocal Minimal 10 4 13 7 5 10 7 13 Slight 0 2 3 0 1 4 2 9 Moderate 0 1 0 0 0 0 0 0 Total 10 7 16 7 6 14** 9 22** *: p ≤0.05, **: p≤0.01 **Table 2b**  **: Incidence and severity of microscopic changes i**  **n the liver, all animals of the**

**carcinogenicity phase (part 2 of**  **2) Sex**  **Males Fem**  **ales Dose level of fluopyram** **(ppm )**  **0 30**  **150 750/375**  **0 30**  **150 1500**  **Dose level fluopyram** **(m g/kg)**  **0 1.20**  **6.0 29.0/15.0**  **0 1.7**  **8.6 89**  **Num ber of anim**  **als exam ined**  **60 60**  **60 58**  **60 60**  **60 59**  Increased number of mitoses Present 0 0 0 0 6 1 5 33 Total 0 0 0 0 6 1 5 33** Multinucleated hepatocytes with anisocaryosis Present 1 0 1 0 4 2 6 38 Total 1 0 1 0 4 2 6 38** Hepatocellular single cell necrosis: focal/multifocal Minimal 2 0 1 1 0 3 1 25 Slight 0 0 1 0 0 1 0 12 Total 2 0 2 1 0 4 1 37** Hepatocellular brow

n pigment(s): focal/multifocalFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 12 of 35 **Sex Males**  **Fem ales**  **Dose level of fluopyram (ppm** **) 0**  **30 150**  **750/375 0**  **30 150**  **1500 Dose level**  **fluopyram (m**  **g/kg) 0**  **1.20 6.0**  **29.0/15.0 0**  **1.7 8.6**  **89 Num** **ber of anim als exam** **ined 60**  **60 60**  **58 60**  **60 60**  **59** Minimal 0 0 0 0 1 1 2 22 Slight 0 0 0 0 0 0 0 2 Total 0 0 0 0 1 1 2 24** Accumulation of brown pigments in Kupffer cells: focal/multifocal Minimal 6 2 9 5 8 7 9 27 Slight 1 0 1 2 4 3 2 4 Moderate 0 0 0 0 0 0 0 1 Total 7 2 10 7 12 10 11 32** Hepatocellular macrovacuolation: centrilobular to midzonal: diffuse Minimal 0 0 0 9 0 0 0 5 Slight 0 0 2 1 0 0 0 4 Moderate 0 0 0 0 0 0 0 2 Total 0 0 2 10** 0 0 0 11** Extramedullary hematopoiesis: multifocal Minimal 16 7 10 15 17 24 21 30 Slight 1 3 1 1 2 1 3 3 Moderate 1 0 0 0 0 0 0 0 Total 18 10 11 16 19 25 24 33 : p ≤0.01 D. Adequacy

of the Dosing for Assessment of Carcinogenicity The doses tested were considered to be adequate and not excessive to assess carcinogenic potential in both sexes . Infemale rats , the highest dose tested of 1500 ppm was considered to be adequate . This was based on a 20 % decrease in mean body weights and increased liver weights (mean: +39%, absolute: +56%), which were dose - related and correlated well with findings such as prominent lobulation and hy pertrophy. Other effects that occurred at the high dose included thy roid f ollicular cell hy

pertrophy and colloid alteration , c hronic progressive nephropathy and kidney tubular dilatation , and r etinal atro phy and lenticular degeneration of the eye. Inmales , the top dose level of 750 ppm, an excessive dose, had to be reduced to 375 ppm from week 85 onwards because of the high mortality in this group. At the termination of the study , mortality

among high - dose males was 50% of control values. The dose of 375 ppm, which is roughly

one - half the excessive dose of 750 ppm, however, was considered to be adequate. This was based on decreased mean body weight, increased liver weight, l iver hy pertrophy , f

ollicular cell hy

pe rtrophy and colloid alteration, c hronic progressive nephropathy

and tubular dilatation and tubular hy pertrophy (males) all observed at the high dose.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 13 of 35 2. Carcinogenicity

Study in C57BL/6J Mice Reference : Carcinogenicity Study of AE C656948 [fluopy

ram] in the C57BL/6J Mouse By Dietary Administration for 18 months. Bayer CropS cience.Project Number: SA/05094 . Report No.05094 . December 14, 2007 MRID 47372450 A. Experimental Design Groups of 60 male and female C57BL/6J mice were fed diets containing 0, 30, 150 or 750 ppm of fluopy ram (corresponding to a mean compound intake of 0, 4.2, 20.9 and 105 mg/kg bw/day in males and 0, 5.3, 26.8 and 129 mg/kg bw/day in females, respectively

) for up to 78 weeks. After 52 weeks, 10 males and 10 females from each group which had been allocated to the chronic phase of the study were killed and necropsied. The remaining 50 animals per sex and group (allocated to the carcinogenicity phase) continued treatment until scheduled sacrifice after at least 78 weeks of treatment. B. Discussion of Mortality and Tumor Data Mortality There was no evidence of significant differences in mortality w ith increasing doses of Fluopy ram in male or female mice (Memo, L . Brunsman, Jun e 10 , 2009, TXR # 0055218). Tumors Male mice had a statistically significant trend at p<0.01 and a significant pair - wise comparison of the 750 ppm dose group with the controls at p<0.05 for thy roid follicular cell adenomas (Table 3) . There were no statistically significant trends or significant pair - wise comparisons of the dosed groups with the control s for the female mice. The statistical analyses of the tumors in the male and female mice were based upon Fisher’s Exact Test for pair - wise comparisons and the Exact Test for trend (Memo, L . Brunsman, Jun e 10 , 2009, TXR # 0055218).FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 14 of 35 Table 3. Thy

roid Foll icular Cell Tumor Rates + and Fisher’s Exact Test and Exact Test for Trend Results in Male Mice Dose (ppm) 0 30 150 750 Adenomas # (%) p = 1/49

(2) 0.00357**  1/47

(2) 0.74211 3 **a**/48

(6) 0.30076 7/48

(15) 0.02758* +Number of tumor bearing animals/Number of animals examined, excluding those that died or were sacrificed before we ek 54. #No carcinomas were observed. aFirst adenoma observed at week 79, dose 150 ppm. Note: Significance of trend denoted at control .

Significance of pair - wise comparison with control denoted at dose level. If * , then p < 0.05. If ** , then p < 0.01. When compared to historical control data (uncensored data) from the testing laboratory , the incidence of thy

roid follicular cell adenoma s in the male high dose group at week 78 ( 7/48 , 14.6 %) was outside the range of the historical control group (range, 0 - 2 %; average, 0.4 %) . C. Non - Neoplastic L esions The non - neoplastic lesions observed in liver and thy roid of both sexes of mice are presented in Table s 4 and 5 . **Table 4**  **: Incidence**  **and severity**  **of microscopic**  **changes in**  **the liver,**  **all animals,** 

**carcinogenicity phase Sex**  **Males Fem**  **ales Dose level of fluopyram** **(ppm )**  **0 30**  **150 750**  **0 30**  **150 750**  **Dose level of fluopyram** **(m g/kg)**  **0 4.20**  **20.9 105**  **0 5.30**  **26.8 129**  **Num ber of anim**  **als 49**  **49 49**  **50 48**  **50 50**  **50** Eosinophilic focus(i) of altered hepatocytes: focal/multifocal Minimal 1 0 0 0 0 1 2 0 Slight 0 0 0 2 0 0 0 2 Moderate 0 0 0 0 0 0 0 2 Marked 0 0 0 0 0 0 0 1 Total 1 0 0 2 0 1 2 5* Centrilobular to panlobular hepatocellular hypertrophy: diffuse Minimal 0 0 16 3 0 0 18 24 Slight 0 0 22 11 0 0 0 2 Moderate 0 0 0 36 0 0 0 0 Total 0 0 38** 50** 0 0 18** 26** Number of animals 49 49 49 50 48 50 50 50 Hepatocellular cholestasis: focal/multifocalFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 15 of 35 **Sex Males**  **Fem ales**  **Dose level of fluopyram (ppm** **) 0**  **30 150**  **750 0**  **30 150**  **750 Dose level of**  **fluopyram (m**  **g/kg) 0**  **4.20 20.9**  **105 0**  **5.30 26.8**  **129 Num** **ber of anim als**  **49 49**  **49 50**  **48 50**  **50 50**  Minimal 0 0 2 29 0 0 0 0 Slight 0 0 0 2 0 0 0 0 Total 0 0 2 31** 0 0 0 0 Hepatocellular single cell degeneration/necrosis: focal/multifocal Minimal 1 2 7 28 1 1 0 1 Slight 0 0 0 12 0 0 0 0 Moderate 0 0 0 0 0 0 0 1 Total 1 2 7* 40** 1 1 0 2 Interstitial mixed cell infiltrate: focal/multifocal Minimal 18 15 19 39 8 8 10 8 Slight 0 1 0 1 2 3 2 0 Total 18 16 19 40** 10 11 12 8 Eosinophilic inclusion bodies: focal/multifocal Minimal 2 3 5 18 0 0 0 0 Slight 0 0 0 1 0 0 0 0 Total 2 3 5 19** 0 0 0 0 Multinucleated hepatocytes: focal/multifocal Minimal 3 1 3 25 1 0 0 0 Slight 0 0 1 2 0 0 0 0 Total 3 1 4 27** 1 0 0 0 Hepatocellular vacuolation: mainly centrilobular: diffuse Minimal 21 15 13 0 3 4 1 0 Slight 6 19 11 2 2 5 0 1 Moderate 0 4 3 1 0 1 0 0 Total 27 38* 27 3** 5 10 1 1 *: p ≤0.05, **: p≤0.01 **Table 5**  **: Incidence**  **and severity**  **of microscopic**  **changes in**  **the thyroid**  **gland, all**  **animals,**

**carcinogenicity phase Sex**  **Males Fem**  **ales Dose level**  **offluopyram (ppm** **) 0**  **30 150**  **750 0**  **30 150**  **750 Dose level of fluopyram** **(m g/kg)**  **0 4.20**  **20.9 105**  **0 5.30**  **26.8 129**  Number of animals 50 50 50 50 48 50 50 50 Follicular cell hyperplasia: focal/multifocal Minimal 0 2 10 18 11 4 10 10 Slight 3 2 5 6 5 2 5 12 Moderate 1 2 2 5 1 2 3 6 Marked 0 0 3 3 0 0 1 4 Severe 0 0 1 0 0 0 0 1 Total 4 6 21** 32** 17 8* 19 33** **: p ≤0.01FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 16 of 35 D. Adequacy

of Dosing for Assessment of Carcinogenicity The high dose of 750 ppm was considered to be adequate , and not excessive , to assess the carcinogenic potential in both male and female mice , based on the following:

 In creased absolute and relative liver weights in males and females at the high - and mid dose levels. These changes were dose - related, and correlated well with macroscopic findings ( dark/enlarged livers ) and non - neopl astic histopathological lesions ( eosinophilic foci [ females ] , h epatocellular hy pertro phy [ both sexes ] , cholestasis [males], single cell degeneration/necrosis [males]) ;  In creased incidence of thy roid follicular cell hy

perplasia in both sexes at the high dose;

 Decreased absolute and relative kidney weights in both sexes at the high dose le vel, along with increased incidence and /or severity

of kidney

lesions (bilateral cortical basophilic tubules, hyaline casts and interstitial mononuclear cell infiltrates, glomerular congestion/hemorrhage, and amyl oid deposi t ion) in females. **IV. TOXICOLOGY**  **1. Metabolism**  Ina two ADME studies, two quantitative whole body autoradiography

studies with the two different labels and an organ depletion study with [py ridyl - 2,6 - 14 C]fluopy

ram ( MRIDs 47372509, 47372510, 47372511, 47372512, 47372513 ) , it was shown that fluopy ram was quickly

absorbed (with an absorp tion half

- life of 0.1

– 0.5 h) and excreted ( 90 - 98% within 72 h) . There was evidence of signif

icant enterohepatic circulation . In general, to xicokinetics w as not different betwe en a single dose study and a study

with repeated administration , suggesting a low, if any , risk for accumulation. In particular, elimination kinetics was not significantly altered. Only minor sex specific differences in toxicokinetics have been observed , although sy stemic exposure was higher and enterohepatic circulation more pronounced in females. While significant differences in the toxicokinetic data existed, depending on the part of the molecule that had been radiolabeled, routes of excretion were slightly differ ent depending on the label. When labeled in the phenyl ring, excretion via the feces accounted for 47 % - 64% , whereas renal excretion ranged from 35% - 45% in both sexes. With the py ridyl - label, excretion was 53 % fecal and 45% urinary in m ale rats but 39% fecal and 60 % urinary in females. Exhalation was negligible in all tests. Following absorption from the GI tract, fluopy

ram was widely

distributed. Organs with highest residues were the liver, the kidneys, Harderian gland and, in some studies and to a l esser extent, ery throcy tes, adrenals, thy

roid, ovaries and carcass. The analysis of metabolism revealed a number of metabolites including fluopy ram - 7 - hydroxyFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 17 of 35 and - 8 - hydroxy , fluopy

ram - py ridyl - acetic acid and fluopy ram - benzamid being the most abundant. **2 .**  **Mutagenicity** Fluopy ram was tested in a minimum battery of standard genotoxicity

and mutagenicity

tests in vitro and in vivo . The available studies demonstrate that fluopy ram has no genotoxic potential. There was no indication of gene mutation either in the presence or absence of metabolic activation in both the bacterial reverse mutation and mammalian gene mutation tests. The in vitro chromosome aberration test and the in vivo mouse micronucleus test were both negative and, thus, a clastogenic potential may be excluded. **Study type Metabolic**  **a ctivation**  **Concentrations**

**that could be assessed**  **Results C**  **lassification A.**  **In vitro tests**  Ames Test with strains TA 1535, TA 100, TA 1537, TA 98 and TA 102 (Wirnitzer, 2006 , ASB2008 - 5552 ) MRID 47372502 +/ - 16, 50, 158, 500, 1581 and 5000  g/plate Negative Acceptable/Guideline Ames Test with strains TA 1535, TA 100, TA 1537, TA 98 and TA 102 (Herbold, 2008 , ASB2008 - 5554 ) MRID 47372503 +/ - 16, 50, 158, 500, and 1581  g/plate Negative Acceptable/Guideline HPRT Test (V79 cells) ( Herbold, 2006, ASB2008 - 5556 ) MRID 47372504 +/ - 1.95 to 256  g/mL Negative Acceptable/Guideline Chromosome aberrations (V79 cells) (Nern, 2005, ASB2008 - 5555 ) MRID 47372505 +/ - 30, 60, 120, 180 and 240  g/mL Negative Acceptable/Guideline **B. In vivo**  **tests** Bone marrow

micronucleus assay in mice

– intraperitoneal administration ( Herbold, 2005, ASB2008 - 5557 ) MRID 47372506 Not relevant; in vivo test Dose levels =

250, 500 & 1000 mg/kg bw Negative Acceptable/GuidelineFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 18 of 35 **3. Structure**  **- Activity Relationship**  Fluopicolide , f

luazinam and b oscalid are considered possible analogs for Fluopy ram because they have a benzamide sub - structure and/or a py ridine sub - structure , and share a common use as fungicides . Among the three analogs, f luopicolide is the closest based on chemical structural similarity . The SAR , however, does not inform the mode of action for fluopy ram. **Chemical Name Structure**  **Fluopyram** Benzamide, N - [2 - [3 - chloro - 5 - (trifluoromethyl) - 2 - py

ridinyl]ethyl] - 2 - (trifluoromethyl) - (9CI) CAS Registry

No.: 658066 - 35 - 4 13 14 15 16

11 12 10 N

H 9 O 8 7 2 N 1 6 5 4 3 Cl C F 3 C F 3

**Fluopicolide** ( AE C638206 or 2,6 - dichloro -N - [[3 - chloro 5 - (trifluoromethyl) - 2 - py ridinyl]methyl]benzamide) CAS Registry

No.: 239110 - 15 - 7 Cl

Cl N H N O Cl F 3 C

**Fluazinam; IKF -**  **1216** 3 - Chloro -N

- [3 - chloro - 2,6 - dinitro - 4 - (trifluoromethyl)phenyl] - 5 - (trifluoromethyl) - 2 - py ridinamine Caswell#: 959 CAS Registry

No.: 79622 - 59 - 6 N N H C F 3 Cl

Cl C F 3 N O 2 O 2 N **Boscalid** BAS 510; 3 - py

ridinecarboxamide, 2 - chloro N - (4’chloro[1,1’biphenyl] - 2 - yl ) CAS No. 188425 - 85 - 6 Inaccordance with the EPA’s Final Guidelines for Carcino gen Risk Assessment (March 2005), the CARC classified **Fluopicolide** as **“Not Likely to be Carcinogenic to Humans”** based onFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 19 of 35 convincing evidence that a non - genotoxic, mitogenic mode of action for liver tumors was established in the mouse and that the carcinogenic effects were not likely at doses that do not cause perturbations of the liver . Inaccordance with the EPA’s Final Guidelines for Carcinogen Risk Assessment (March 2005), **Fluazinam** wa s classified as a **“suggestive evidence of carcinogenicity to humans”**  based on 1) statistically

significant increases in hepatocellular adenomas, carcinomas and combined adenomas/carcinomas in male CD1 mice (MRID 42208405); 2) statistically increased incidence of follicular cell adenocarcinomas and combined follicular cell adenomas/ adenocarcinomas in Sprague - Dawley

rats (MRID 42248620); equivocal evidence of increased incidence of hepatocellular adenomas, carcinomas or combined adenomas/carcinomas for the male mice (MRID 44807222) Fluaz in am is negative in mutageni city tests. Inaccordance with the EPA’s Draft Guidelines for Carcinogen Risk Assessment ( 199 6 ), **Boscalid** wa s classified as **“suggestive evidence of carcinogenicity, but not sufficient to assess human carcinogenic potential”**  based on increased incidence of follicular cell adenomas in male (MRID 45404827) and female (MRID 45414828 ) rats . No evidence of carcinogenicity was found in mice (MRID 45404901) . Boscalid is negative in mutagenicity tests . **4. Subchronic and Chronic**  **Toxicity a) Subchronic Toxicity**  Rat (MRID 47372516) Ina 28 - day

oral toxicity

dose - range finding study in rats (MRID 47372516), groups of Wistar rats (5/sex/dose) were administered fluopy ram (98.6% purity

) in the diet at concentrations of 0, 50, 400 and 3200 ppm (0, 4.0, 31.0 and 254 mg/kg/day in males and 4.6, 36.1 and 263 mg/kg/day in females ) for 28 day s. The results showed that at the fluopy ram did not produce mortality or clinical signs of toxicity . There were no changes in terminal body weight s at any

dose in either sex. Increases in platelet counts and prothrombin times were reported in 3200 ppm males. Total cholesterol and triglyceride levels were in increased in 3200 ppm males and females. L iver weights were increased in the 400 and 3200 ppm dose groups in both sexes compared with controls, the effect being slightly

more pronounced in females. At 3200 ppm, the increase was more than 50 % compared to controls. This increased weight was associated with enlarged and dark livers at macroscopic ex amination and with minimal to moderate centrilobular hepatocellular hy pertrophy in most animals in both sexes. Similar effects were observed at 400 ppm, however with lower magnitude, incidence and severity . In addition to the liver effects, the thy roid gland weights (absolute and relative) were higher in males at 3200 ppm and 400 ppm. In association, hy pertrophy of the follicular cells was observed in 3/5 males. Thy roid gland weight was not affected in females although 2/5 animals showed a minimal follicula r cell hy pertrophy .FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 20 of 35 Inaddition, a dose - related slight increase in total cy tochrome P - 450, BROD and PROD activities was observed in both sexes at 3200 and 400 ppm. No significant effects were observed at 50 ppm. For establishing the NOAEL and LOAEL for this study , the results of the 90 - day oral toxicity study should be considered along with those of this study . In this study minimal effects were seen at 400 ppm. However, in the 90 - day oral toxicity

study , no effects were seen at 200 ppm. At 1000 ppm, si milar toxic effects were found as those seen in the 3200 ppm of the 28 day dose - range finding study . Therefore the NOAEL for the 28 - day dose - range finding study lies between 400 ppm and 200 ppm (equivalent to 31 to 12.5 mg/kg/day ), while the L OAEL is 400 ppm (31 mg/kg/day

). Rat (MRID 47372441) Ina subchronic oral toxicity study (MRID 4737 2441 ) f

luopy ram was administered in the diet for at least 90 day s to 10 animals/sex/dose group at 0, 50, 200, 1000, and 3200 ppm (equating approximately

to 0, 3.06, 12.5, 60.5 and 204 mg/kg/day in males and 0, 3.63, 14.6, 70.1, and 230 mg/kg/day

in females ). An additional 10 males and 10 females fed either 0 or 3200 ppm of test diet for at least 90 days were maintained for a minimum of 28 days to examine the reversibi lity of any effects seen. Following exposure at the prescribed dose level, animals were sacrificed and subjected to toxicological examination . There were no treatment - related clinical signs in either sex throughout the study . Food consumption rates were essentially similar to controls in all dose groups. At 3200 ppm mean overall bo dy

weight gain was reduced by 9 % in males (p ≤0.05) and 17% in females (p≤0.01). The effect on mean body

weight was still observed after 4 weeks of recovery in both sexes, as the magnitude of the decrease was similar to that observed at the end of the treatment phase. At 1000 ppm, mean body

weight parameters were unaffected by the treatment in either sex, with the exception of a slight decrease of 15% (not statistically signific ant) in mean body weight gain per day in females during the first week of treatment, compared to controls. Higher mean prothrombin time was noted at 3200 ppm in males only (+74 %, p ≤0.01), when compared to the control values. Slightly lower mean hemoglobin concentrations were noted at 3200 ppm in both sexes and at 1000 ppm in males only . These variations were a ssociated with lower mean hematocrit in males and lower mean corpuscular volume and mean corpuscular hemoglobin in females at 3200 ppm. Additionally , higher mean platelet (+24 %, p ≤0.01) and reticulocy te (+50 % for absolute count and +42 % for percentage, p ≤0.01) counts were noted at 3200 ppm in females. There was no evidence of treatment - related changes at 1000 ppm in females or at 200 and 50 ppm in either sex. Hematological changes were reversed at the end of the recovery

period, with the exception of me an hemoglobin concentrations, which remained slightly lower than controls.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 21 of 35 Several clinical chemistry variations were statistically significant at 1000 ppm or 3200 ppm, and included increased total cholesterol ( > 45%), decreased total bilirubin ( > - 30%), in creased glutamyltransferase (+625%) and triglycerides (+102%). After one month of recovery in the high dose group, there was a general tendency towards reversibil i ty . Nevertheless, statistically significant differences were still noted in mean total chole sterol, globulin concentrations and albumin/globulin ratio in females (+19 %, +14 % and - 14 % respectively , p ≤0.05), compared to the control values. The incidence and severity of cellular casts in urine were increased in all groups of fluopy

ram treated ma les in a dose - related manner, and correlated with hyaline droplet nephropathy

observed at histopathology examinations. However, these effects were largely reversible. At Week 3, TSH levels were elevated in both sexes at 3200 ppm (+63 %, p ≤0.05 and +71 %, p ≤0.01, respectively

), together with an increase in mean T3 and T4 levels in females (+24 %, p ≤0.05 and +54 %, p ≤0.01, respectively ), whereas at Week 13, only an increase in mean TSH and T3 levels was noted in males (+88 and +40 %, respectively , p ≤0.01). At 1000 ppm, mean TSH level was increased by 54 % (not statistically

significant) and T4 level was significantly increased by

43 % in males on Week 13. A dose dependent increase of T3, T4 and TSH was observed in both sexes at most time points and for most parameters at all dose levels when compared to controls, however, without being significant or of relevant magnitude at the lower two dose levels of 200 and 50 ppm. These changes were reversed in the recovery phase. Mean body

weights were not statistical ly changed by

fluopy ram administration. Mean absolute and relative liver weights were statistically increased in males and females at 1000 ppm and 3200 ppm (+ 20 to +25%, and +53% to +74%, respectively ). Mean absolute and relative kidney weights were statistically increased predominantly in males (+25% - +34%) at 1000 ppm and 3200 ppm. Thy

roid weights were also statistically higher (+10% to +22%) in males and females at 3200 ppm. These various increases were only partially reversed after the one - month r ecovery period. Microscopic effects of treatment with fluopy ram were seen in the liver, kidney , thy roid gland and lung. In the liver, minimal to moderate centrilobular hepatocellular hy pertrophy was observed with a dose - related increase in incidence and s everity at 3200 and 1000 ppm in both sexes and minimal centrilobular hepatocellular hy pertrophy was observed at 200 ppm in males. In addition, minimal to moderate periportal to midzonal hepatocellular macrovacuolation was observed in females at 3200 and 10 00 ppm. In the kidney , hyaline droplet nephropathy (characterized by basophilic tubules, hyaline droplets in proximal tubules and granular casts in the medulla) and hyaline casts were higher at 3200 and 1000 ppm in males, in comparison with controls. Hyal ine droplet nephropathy

was also slightly

higher at 200 ppm in males. In the thy roid gland, a higher incidence of minimal to slight diffuse hy pertrophy of follicular cells was seen at 3200 and 1000 ppm in both sexes compared to controls and internal histo rical control data . After the recovery period, liver and thy

roid gland were comparable between the high dose and control groups, indicating that the changes noted after 90 day s of treatment were reversible in these organs. In the kidney

in the high dose groups, basophilic tubules, hyaline droplets in proximalFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 22 of 35 tubules, granular casts in the medulla and hyaline casts were persistent in the kidney of some males. Inthis study

in rats the lowest dose level of 50 ppm represented the No Observed Effect Level (NOEL) in males and females (equating to 3.06 / 3.63 mg/kg body weight/day

). At the next higher dose level of 200 ppm (12.5 / 14.6 mg/kg bw/d), however, treatment - related effects were confined to a rather adaptive liver weight increase due to hepatocellular hy pertrophy that was associated with non - significant changes in clinical chemistry parameters. Because these findings were not considered adverse, the NOAEL for both sexes was set at 200 ppm (12.5 / 14.6 mg/kg bw/day ), and the L OAEL is 1000 ppm (60.5 / 70.1 mg/kg bw/day ) based on organ weight changes , and clinical chemistry and histopathological findings in liver, thy roid and kidneys. Mouse (MRID 47372517) Ina 28 - day

oral toxicity

dose - ran ge finding study in mice (MRID 47372517), groups of C57BL/6J mice (5/sex/dose) were administered fluopy ram (98.4% purity

) in the diet at concentrations of 0, 150, 1000 and 5000 ppm (0, 24.7 , 162, and 747mgkg/day in males and 0, 31.1, 197, 954 mg/kg/day i n females ) for 28 day s. The mg/kg dose equivalent for the high dose males (747 mg/kg/day

) was calculated for weeks 1 - 3 only due to mortality

or early sacrifice in this group. At 5000 ppm, all males and 3/5 females were sacrificed prior to scheduled sacri fice. All unscheduled sacrificed animals presented the following clinical signs: reduced body weights, decreased motor activity , hunched posture, piloerection, wasted appearance and/or coldness to touch in both sexes together with labored respiration, and distended abdomen. The two surviving females at 5000 ppm, had elevated total cholesterol (+118 %) and total protein (+16 %) concentrations, and alanine aminotransferase activities (+384 %). Mean absolute and relative liver weights were higher at 5000 ppm i n females and at 1000 and 150 ppm in both sexes. These changes were found to be dose - related. Gross pathology showed enlarged and dark livers males and females, the Thy

mus size was reduced in 4/5 males and 1/3 females, and distended abdomen was noted in 3/5 males. Red liquid was observed in the thoracic cavity in all males. Histopathology

indicated. Histopathology demonstrated hy

pertrophy of the hepatocy

tes in males and females and hepatocellular focal necrosis was al so noted in surviving females. At 100 0 ppm, there was a slight decrease in the daily mean body

weight gain during study Week 2 in males. No treatment - related effect on mean body weight and mean body

weight change parameters was noted at 1000 ppm in females or 150 ppm in either sex. No effect on mean food consumption was noted in animals surviving to terminal sacrifice. Enlarged liver was seen in all males and in 4/5 females. Hepatocellular hy pertrophy was seen in all 1000 ppm males and 2/5 females, and focal necrosis was found in 2/5 females. At 150 ppm, there was a slight increase in liver weights, and liver hy pertrophy was seen in all males and 3/6 females.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 23 of 35 Inconsideration of NOAEL and L OAEL, the results show that hepatocellular hy pertrophy at 150 ppm (24.7 mg/kg/day

) (LDT), but currentl y liver hy pertrophy is considered to be an adaptive effect. However, more pronounced liver effects (hy pertrophy and focal necrosis) are seen at 1000 ppm and above. NOAEL for this study can be established at 150 ppm (24.7 mg/kg/day ). LOAEL is 1000 ppm (162 mg/kg/day ) based on increased incidence of liver hy pertrophy and focal necrosis in the liver. Mouse (MRID 47372442

) Ina sub - chronic oral toxicity study (MRID 47372442 ), f

luopy ram was administered in the diet for at least 90 days to C57BL/6J mice at the following doses

– 0, 30, 150 and 1000 ppm (equating approximately

to 0, 5.4, 26.6 and 188 mg/kg/day in males and 0, 6.8, 32.0 and 216 mg/kg/day

in females). A negative control group received plain diet. Animals were then sacrifice and subjected to toxicol ogical examination. There were no treatment - related clinical signs of toxicity observed in animals at any dose level. No treatment - related mortality occurred in the test animals at any dose level. Body

weight and food consumption were not affected by treatment. Blood analy sis: At 1000 ppm, mean alanine aminotransferase activity was higher in males and females (+205 and +109 % after removal of an outlier in the female control values, respectively, p ≤0.01), compared to the control groups. In addition in ma les, mean alkaline phosphatase activity was higher (+21 %, p ≤0.01), mean albumin and mean total cholesterol concentrations were lower ( - 12 and - 40 %, respectively , p ≤0.01) and a tendency towards higher values was noted in aspartate aminotransferase activity (+46 %, p ≤0.05). In females, a tendency towards lower mean albumin concentration was also noted ( - 9 %, p ≤0.01). At 150 treatment - related changes consisted in a lower mean total cholesterol concentration in both sexes ( - 41 and - 30 %, r espectively , p ≤0.01) and lower total protein and albumin concentrations in males ( - 6 % and

– 7 %, p ≤0.01), associated with lower total bilirubin in males and females ( - 43 %, significant only in males) compared to the control group. At 30 ppm the only observed finding was a s ignificant decrease in total cholesterol in males ( - 30 %, p ≤0.01). There was no relevant change in terminal body weight in either sex. Mean absolute and/or relative liver weights were statistically significantly

higher at 1000 ppm (+36% to +45%) and 150 ppm (+14% to +28%) in both sexes, with a dose - related effect. At 30 ppm, mean absolute liver weight and relative liver to body weight ratio were statistically significantly

higher (+11% to +13%) in females only , but this change was considered not to be to xicologically relevant since it was not associated with any histological change. Mean absolute and relative adrenal gland weights were increased by between 87 % and 92 % at 1000 ppm in males compared to controls, the difference being statistically signifi cant for mean adrenal gland to body weight ratio only (p ≤0.05). At 1000 ppm, enlarged livers were observed in 8/10 males and 9/10 females, and dark livers were observed in 5/10 males and 10/10 females.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 24 of 35 Effects of treatment with fluopy ram were seen in the liver and adrenal gland in both sexes. In the liver, there was a minimal to moderate hy pertrophy of centrilobular hepatocy tes in both sexes at 1000 and 150 ppm. This finding correlated with the enlarged livers observed at necropsy in animals given 1000 pp m and the statistically significant increase in liver weight seen in animals given 1000 or 150 ppm. Also observed was a greater incidence of minimal or slight focal necrosis in both sexes at 1000 ppm, when compared to controls. Minimal or slight focal nec rosis was present in 3/10 males and 6/10 females given 1000 ppm, compared to one female in the control group. In the adrenal glands at 1000 ppm, there was a lower incidence of cortical st eroid pigment in males and a greater incidence of minimal to slight cortical vacuolation in females, compared to controls. The change noted in males at 1000 ppm was considered to be slight compared to the magnitude of increase in adrenal gland weights seen in this sex. No treatment related changes were observed in the adre nal glands at 150 or 30 ppm in either sex. In the fore stomach of males given 1000 ppm, there was a slightly greater incidence of focal epithelial hy perplasia than in controls. However, as these changes in males were only focal and isolated, they were con sidered not to be treatment - related. In females, the incidence and severity of this finding were comparable between controls and treated animals. The NOAEL for this study is 150 ppm (equating to 26.6 and 32.0 mg/kg bw/d ay in males and females respectively

). The LOAEL is 1000 ppm (approximately 118 or 216 mg/kg bw/d ay for males and females, respectively ) based on liver effects (hy pertrophy , necrosis, changes in related clinical chemistry

parameters). **b) Chronic Toxicity** R at (MRID 47372501) Ina combined chronic toxicity and carcinogenicity

study (MRID 47372501), groups of 60 male and female Wistar rats were fed a diet containing 0, 30, 150 and 750 ppm (male animals) and 0, 30, 150 and 1500 ppm (females) fluopy ram for 24 m onths. In males, the top dose level of 750 ppm had to be reduced to 375 ppm from week 85 onwards because of the high mortality in this group. Over the whole study period, these dietary

concentrations corresponded to a mean daily intake of 0, 1.20, 6.0 and 29 mg/kg bw in male rats or 1.68, 8.6 and 89 mg/kg bw in females. Inaddition, satellite groups of 10 animals per sex and dose were subject to interim sacrifice after 1 year. Overall, there was a statistically significant increase in mortality in males at 750/375 ppm during the study and after 24 months although no clear cause for these premature deaths could be established. At the highest dose levels of 750/375 in males or 150 0 ppm in females , respectively , mean body

weights were significantly reduced in both sexes at various times throughout the study . The m ain target organs were the liver, kidney s and the thy roid gland, and also the eyes. Li ver toxicity

became apparent by an increase in organ weight at the two upper dose levels inFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 25 of 35 male rats and in the highest dose group in females . This was sometimes accompanied by gross pathological findings such as nodules/masses which correlated histologically with neoplastic changes . Clinical chemistry

findings suggesting hepatotoxicity were minor in nature and confined to the respective top dose level in both sexes. They comprised occasionally

higher mean triglyceride concentrations and slightly lower mean glucose concentrations in th e females. Activity of alkaline phosphatase was reduced in both sexes throughout the study but achieved statistical significance only

occasionally . Histological changes included a higher incidence of altered hepatocy tes (eosinophilic foci) and hepatocellul ar brown pigments, focal or multifocal hepatocellular vacuolation, increased number of mitoses, centrilobular to panlobular hy pertrophy and hepatocellular single cell necrosis with females being much more affected. Again, this might be due to the higher do se that the female rats were fed. At the mid dose level of 150 ppm, however, histopathological lesions (hy pertrophy ) were confined to male rats and, thus, were in line with the increase in organ weight that was noted in the same sex only . Inthe kidney s, marked degenerative changes such as chronic progressive nephropathy or focal/multifocal (medullar or cortical) tubular dilatation, together with an increased incidence of tubular golden/brown pigments (mainly in females) and collecting ducts hy perplasia, were observed at the high dose level. In addition, a higher incidence of hyaline droplets and of renal cy sts was noted in male rats. At the mid dose level of 150 ppm, male rats still displayed a higher frequency of tubular hy

pertrophy or dilatation. During the first year of treatment, but not thereafter, urinalysis revealed higher incidences of abnormal color of urine (orange to red) in females and a higher incidence and severity of cellular casts in males. This latter finding was also confirmed in male rat s receiving the intermediate dose. Effects of fluopy

ram on the thy

roid gland were demonstrated by an increased organ weight at the highest dose level in both sexes. This finding was associated with histopathological changes (follicular cell hy

perplasia and/or hy

pertrophy , colloid alteration/depletion) that were of less severity or frequency

but yet to be noted at the mid dose level, too. Inaddition, the eyes were affected by long - term treatment. Ophthalmologic examination revealed abnormal color of the retinal fundus in females after 12 months. At the 24 - month examination, this condition was observed in females and males, together with small retinal vessels. In addition, hy perreflectivity

in the retina was noted in females and corneal opacity, edema of the cornea and nuclear opacity

in males. These effects were more severe at the top dose level and less pronounced but still present after 2 years in the male and female groups receiving the intermediate dose. Histologically , bilateral retinal atrophy was n oted at the highest dose level , together with a higher incidence of lens degeneration. A tendency

towards lower ery

throcy te parameters (hemoglobin concentration, mean corpuscular volume, hematocrit and/or mean corpuscular hemoglobin) was observed in the female high dose group throughout the study confirming evidence from short - term studies that the red blood cellsFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 26 of 35 might be an additional target. The same tendency was observed in high dose males at most time points, however, statistical significance was not achieved. The assumption of an effect on the blood was further substantiated by a more frequent occurrence of extramedullary hematopoiesis in the livers of high dose females. In contrast to the subchronic study , prothrombin time was this time shorter in the female high dose group with the difference being significant at months 6 and 12. Therefore, the NOAEL was 30 ppm (1.2 and 1.7 mg/kg bw/day in males and females, respectively

). The LOAEL of 150 ppm (6.0 and 8.6 mg/kg/day , males and females), is based on nephropathy

in the kidney , follicular cell hy pertrophy in the thy

roid, and increased liver weight with gross pathological and histopathological findings. Mouse (MRID 47372450) Ina mouse carcinogenicity study (MRID 47372450), groups of 60 male and fe male C57BL/6J mice were fed a diet containing 0, 30, 150 or 750 ppm of fluopy ram (corresponding to a mean compound intake of 0, 4.2, 20.9 and 105 mg/kg bw/day in males and 0, 5.3, 26.8 and 129 mg/kg bw/day

in females, respectively

) for up to 78 weeks. Afte r 52 weeks, 10 males and 10 females from each group which had been allocated to the chronic phase of the study were killed and necropsied. The remaining 50 animals per sex and group (allocated to the carcinogenicity phase) continued treatment until schedul ed sacrifice after at least 78 weeks of treatment. There were no unscheduled deaths or clinical signs occurring during the study that could be attributed to treatment. The survival rate was not different among the control and dose groups. Body weight gai n was decreased only in high and mid dose males and only during the second trimester of the study

(weeks 26 to 54). Afterwards, some compensatory growth was observed resulting in a mean final body weight that was similar to the control group value. A slightly higher mean platelet count was determined in top dose males but no other changes in hematological parameters were seen. Pathological examination revealed that the liver, the thy roid gland and the kidneys were the main target organs in mice. Mean absolute and relative liver weights were markedly increased in high and mid dose males and females at interim sacrifice as well as at study termination. The increment exhibited a clear dose response. At these two upper dose levels, gross necropsy find ings such as dark and enlarged livers were corroborated by an increase in non - neoplastic histopathological lesions such as centrilobular to panlobular hy pertrophy , hepatocellular cholestasis, single cell degeneration/necrosis or eosinophilic foci. A few of these non - neoplastic effects on the liver were observed only

in males pointing to a higher vulnerability of this sex with regard to hepatotoxicity .FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 27 of 35 Toxic effects on the thy

roid were noted at the top dose level in both males and females and in the mid dos e male group. The main non - neoplastic finding, i.e. , follicular cell hy perplasia became apparent in male mice at interim sacrifice already . In line with that, the only neoplastic change in this study consisted of a higher incidence of follicular cell adenoma in high dose males (7/50) as compared to the control group (1/50). This difference was statistically significant (p ≤0.05). Mean absolute and relative kidney weights were decreased at the 750 ppm dose level in both sexes. In addition, a higher inci dence and/or severity of bilateral cortical basophilic tubules, hyaline casts(s) and interstitial mononuclear cell infiltrates, glomerular congestion/ hemorrhage(s), and more pronounced amyloid deposition (mainly in the glomerular interstitium) was noted at this dose but only in females. Thus, the No Observed Adverse Effect Level (NOAEL) in this study was 30 ppm in males (equivalent to 4.2 mg/kg/day ) and females (equivalent to 5.3 mg/kg/day ). The LOAEL was 150 ppm (equivalent to 20.9 mg/kg bw/day in males and 26.8 mg/kg bw/day in females) based on hepatocellular hy

pertrophy in the liver in both sexes and follicular cell hy perplasia in the thy

roid gland in males. **5. Mode of Action** 1. Mode of Action for Thyroid Follicular Cell Tumors in Male Mice The regi strant, Bayer Cro pScience, submitted a proposed mode of a ction for the thy roid follicular cell tumors seen in male mice using the International Programme on Chemical Safety (IPCS ) framework ( MRID 47753406 ) . The ir postulated MOA for fluopy ram - induced thy

roid follicular cell tumors in mice involved the perturbation of homeostas is of the pituitary

–thy roid axis by

an extra thy roidal mechanism that is not the result of genotoxicity . The key events proposed included: 1) increases in liver enzy me activity , s imilar in pattern to the p henobarbital - induced enzy me activity

pattern , 2 ) alterations in thy roid and pituitary

hormone levels, 3 ) i ncrease in thy roid growth, and 4 ) lesion progression in the thy roid from hy

perplasia to neoplasia. Thy roid follicular cell a denomas in male mice were associated with chronic exposure to 105 mg/kg/day fluopy ram. Inaddition to the subchronic and chronic studies, the registrant submitted MOA studies to further elucidate the MOA for fluopy ram exposing mice for short durations (3 , 4 and 14 day s) to doses of 300 mg/kg/day (MRID No. 47567128, 47372519 , 47690101 ). A summary of the results of th is MOA analy sis is provided in the table below. **Mode of Action --**  **Thyroid Tumors in Mice Key Events**  **Anim al Evidence**  **Dose Concordance Tem** **poral Association** FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 28 of 35 **Mode of Action --**  **Thyroid Tumors in Mice Key Events**  **Anim al Evidence**  **Dose Concordance Tem** **poral Association**  Induction of Liver Enzymes 3 - day day mechanistic study at 2000 ppm, 308 mg/kg/day [MRID 47567128] 14 - day mechanistic study at 2000 ppm or 314 mg/kg/day [MRID 47372519] Increased Total CYT P450, PROD , EROD and BROD at 2000 ppm **No** . Studies we re performed at dose levels h igher than the tumorigenic dose of 105 mg/kg/day. Yes

Hormone Changes (Decreased T3, T4, Increased TSH) 3 - day day mechanistic study at 2000 ppm, 308 mg/kg/day [MRID 47567128] 4 - day mechanistic study at 200 ppm [MRID 47690101] 14 - day mechanistic study at 2000 ppm or 314 mg/kg/day [MRID 47372519] No decreases in T3 observed . Some decreases in T4 observed . Increase in T4 clearance over 24 hrs. Increase in TSH . **No.** Studies were performed at dose levels higher than the tumorigenic dose. Yes

Increases in cell growth (Increased thyroid we ight, Increased thyroid hypertrophy and hyperplasia) Mouse carcinogenicity Study 0, 30, 150, 750 ppm (0, 4.2/5.3, 20.9/26.8, 105/129 mg/kg/day, Males/Females) Thyroid hyperplasia observed at 20.9 and 105 mg/kg/day at 1 year and 18 months. Not seen at 90 days [MRID: 47372450] No increase in thyroid weight and no thyroid hypertrophy observed in subchronic or chronic mo use studies. **Yes** , some evidence of thyroid hyperplasia observed at and below

the tumorigenic dose of 750 ppm

(105 mg/kg/day) Yes

Thyroid follicular cell tumors Mouse carcinogenicity Study [MRID: 47372450] Tumors observed at 750 ppm (105 mg/kg/day) -- **The CARC evaluated the submitted MOA and concluded that there is insufficient data to**  **support the proposed MOA for fluopyram induced thyroid follicular cell tumors**  **. The**  **major deficiency in**  **the analysis was**  **the lack of d ose**  **- response concordance**  **with key events**  **and tumors**  **. The MOA studies were performed at doses**  **that were above the tumorigenic**  **dose. There is no data**  **for several key events at or below the tumorigenic dose**  **, which is required as experimental support**  **for the MOA**  **. While temporal association was**  **estab lished, the missing dose concordance data precludes an acceptable MOA.**  **In addition, while a comparison to Phenobarbital MOA is informative, the MOA for fluopyram must**  **stand on its own.**FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 29 of 35 2. Mode of Action for Liver Tumors in Female Rats The registrant, Bayer CropScience, submitted a proposed mode of action for the liver tumors in female rats using the IPCS framework (MRID 47753406 ) . The y postulated a mitogenic MOA for fluopy ram - induced liver tumors . It involved the induction of liver enzy mes, including cy tochro me P450s (CYP450) iso zy mes, consequent liver cell proliferation and hy pertrophy , and altered hepatic foci, ultimately progressing to neoplasia. The specific MOA for fluopy ram was considered by

the registrant to be similar to the phen obarbital MOA for liver tumors in rats. Li ver tumors in female rats were associated with chronic exposure to 89 mg/kg/day (1500 ppm) fluopy ram. Inaddition to the subchronic and chronic studies, th e registrant submitted a MOA study to further elucidate the MOA for fluopy ram exposing rats for short durations (7 day s) to a dose of 190 mg/kg/day (3000 ppm) (MRID 47372520 ). A summary of the results of the MOA analy sis is provided in the table below. **Mode of Action —** **Mitogenesis Liver Tumors in Rats**  **Key Events Anim**  **al Evidence Dose Concordance**  **Tem poral**  **Association** Nuclear Receptor Activation No -- -- Increased Induction of CYP Phase I Enzymes 7 - day mechanistic study at 3000 ppm, 193 mg/kg/day [MRID 47372520] 28 - day subchronic study with liver enzyme induction at 50, 400 and 3200 ppm , 4.6, 36.1 and 263 mg/kg/day [MRID 47372516] No. Study wa s performed at dose levels higher than the tumorigenic dose of 1500 ppm , 89 mg/kg/day. Yes. BROD and PROD elevated at 400 ppm, below tumorigenic dose Yes Measured at 7 days and 28 days Increased Liver Weight 7 - day mechanistic study at 3200 ppm, 193 mg/kg/day [MRID 47372520] 28 - day subchronic study at 50, 400 and 3200 ppm , 4.6, 36.1 and 263 mg/kg/day [MRID 47372516] 90 - day subchronic. at 50, 200,1000, 3200 ppm , 3.63,14.6, 70.1, 230 mg/kg/day [MRID 47372441] No. Studies were performed at dose levels higher than the tumorigenic dose of 1500 ppm , 89 mg/kg/day . No. Liver weight increase seen only at 3200 ppm, higher than tumorigenic dose. Yes. Liver weight changes seen at 1000 and 3200 ppm Yes Measured at 7 days, 28 days, 90 days and 2 yearsFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 30 of 35 **Mode of Action —** **Mitogenesis Liver Tumors in Rats**  **Key Events Anim**  **al Evidence Dose Concordance**  **Tem poral**  **Association** Increased Liver Hypertrophy and Hyperplasia 28 day subchronic study at 3200 ppm [MRID 47372516] 90 subchronic study at 3200 ppm [MRID 47372441] Chronic/Oncogenicity study at 1500 ppm [MRID 47372501] No. Hypertrophy seen only at 3200 ppm, higher than tumorigenic dose. Yes. Hypertrophy seen at 1000 and 3200 ppm Yes. Hypertrophy seen at 1500 ppm , the tumorigenic dose. Yes Hypertrophy observed at 7 days, 28 days, 90 days and 2 years Increased Cell Proliferation 7 - day mechanistic study [MRID 47372520] No. Studies were performed at dose levels higher than the tumorigenic dose. No time - course data on cell proliferation . Tested only at 7 days Inhibition of Apoptosis No -- -- Liver Tumors Chronic/Oncogenicity study [MRID 47372501] Tumors observed at 1500 ppm (89 mg/kg/day) **The CARC evaluated the submitte d MOA and concluded that there was**  **insufficient data to support the proposed MOA**  **for fluopyram induced liver tumors .**  **The major deficiency in the analysis was the lack of dose**  **- response concordance with key events and tumors.**  **The MOA studies were performed at doses above the tumorigenic dose. There is no MOA data**  **at or below the tumori genic dose, which is required as experimental support for the MOA.**  **In addition, there is no time course data on cell proliferation. Ideally, a study of several**  **weeks duration i s needed with early time points**  **(minimum of 2 time points of 90 days or less) to**  **demonstrate that cell proliferation is not sustained. The data submitted was not sufficient and tested only 1 dose and 1 time point.**  **While temporal association was established**  **for several key events , the missing dose concordance data**  **and adequate cell pr**  **oliferation data preclude an acceptable MOA.**  **In addition, while a comparison to Phenobarbital MOA is informative, the MOA for fluopyram must stand on its own.** FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 31 of 35 **V. COMMITTEE’S ASSESSMENT OF THE WEIGHT -**  **OF -**  **THE -**  **EVIDENCE** The Committee considered the following for a weight - of - evidence determination of the carcino genic potential of fluopy ram: 1. Carcinogenicity Rat ● Administration of fluopy ram resulted in the induction of liver tumors in female Wistar rats. There were statistically

significant trends for l iver adenomas (p <0.01), carcinomas (p<0.05) and combined l iver adenomas and carcinomas (p< 0.01). There were significant pair - wise comparisons of the 1500 ppm dose group with the controls for liver adenomas at p< 0.05 and for combined liver adenomas and carcinomas at p< 0.01. When compared to historical control data (uncensored data) from the testing laboratory , the incidence of hepatocellular adenomas in the female high dose group (9/5 5 , 16 %) was outside the range of the historical control group (range, 0 - 5%; average, 1.9%) . Similarly , the incidence of hepatocellular carcinomas in the female mid (2/56, 4%) and high dose group s (3/5 5 , 5%) , while not statistically significant by

pair - wise comparison, exceeded the range of the historical control group (no carcinomas observed in 10 studies from 2000 - 2006 ) and was considered to be biologically relevant . There were no statistically

significant trends or significant pair - wise comparisons of the dosed groups with the controls for the male rats. **The CARC considered the liver tumors**  **in female Wistar rats to be treatment**  **- related.**  ● Adequacy of Dosing : The doses tested were considered to be adequate and not excessive to assess carcinogenic potential in both sexes. **In female rats** , the highest dose test ed of 1500 ppm was considered to be adequate. This was based on a 20% decrease in mean body weights and increased liver weights (mean: +39%, absolute: +56%), which were dose - related and correlated well with findings such as prominent lobulation and hy pertr ophy . Other effects that occurred at the high dose included thy roid f ollicular cell hy

pertrophy and colloid alteration , c hronic progressive nephropathy

and kidney tubular dilatation , and r etinal atro phy and lenticular degeneration of the eye. **In male rats**  , the top dose level of 750 ppm, an excessive dose, had to be reduced to 375 ppm from week 85 onwards because of the high mortality in this group. At the termination of the study , mortality among high - dose males was 50% of control values. The dose of 375 ppm, which is roughly one - half the excessive dose of 750 ppm, however, was considered to be adequate. This was based on decreased mean body weight, increased liver weight, liver hy pertrophy , f

ollicular cell hy

pe rtrophy and colloid alteration, c hronic p rogressive nephropathy and tubular dilatation and tubular hy pertrophy (males) all observed at the high dose.

MouseFLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 32 of 35 ● Administration of fluopy ram resulted in the induction of thy roid follicular cell tumors in male C57BL/6J mice. Male mice had a statis tically significant trend at p<0.01 and a significant pair wise comparison of the 750 ppm dose group with the controls at p<0.05 for thy roid follicular cell adenomas. There were no statistically significant trends or significant pair - wise comparisons of the dosed groups with the controls for the female mice. When compared to historical control data (uncensored data) from the testing laboratory, the incidence of thy roid follicular cell adenomas in the male high dose group ( 7/48 , 15 %) was outside the range of the historical control group (range, 0 - 2 %; average, 0.4 %) . **The CARC considered the thyroid follicular cell**  **adenomas in male C57BL/6J**  **mice to be treatment**  **- related.**  ● Adequacy of Dosing : The high dose of 750 ppm was consider ed to be adequate, and not excessive, to assess carcinogenic potential in both male and female mice, based on the following: 1) Increased absolute and relative liver weights in males and females at the high - and mid - dose levels. These changes were dose - re lated, and correlated well with macroscopic findings (dark/enlarged livers) and non - neoplastic histopathological lesions (eosinophilic foci [females], hepatocellular hy

pertrophy [both sexes], cholestasis [males], single cell degeneration/necrosis [males]); 2) Increased incidence of thy roid follicular cell hy

perplasia in both sexes at the high dose; and 3) Decreased absolute and relative kidney weights in both sexes at the high dose level, along with increased incidence and/or severity of kidney

lesions (bil ateral cortical basophilic tubules, hyaline casts and interstitial mononuclear cell infiltrates, glomerular congestion/ hemorrhage, and amyl oid deposi t ion) in females. 2. Mutagenicity There is no concern for mutagenicity . 3. Structure - Activity Relationship The SAR data do not inform the mode of action. 4. Mode of Action The CARC concluded that insufficient data were provided to support definitive modes of action for the induction of liver tumors in female rats or thy roid follicular cell tumor s in male mice. Deficiencies included a lack of dose - response concordance with key events and tumors.

**VI. CLASSIFICATION OF CARCINOGENIC POTENTIAL**FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 33 of 35 Inaccordance with the EPA’s Final Guidelines for Carcinogen Risk Assessment (March, 2005), the CARC classified fluopy ram as **“Likely to be Carcinogenic to Human s**  **”** based on tumors in two species and two sexes : a treatment - related increase in thy roid follicular cell adenomas in high dose male mice and liver tumors in female rats at the high dose , with incidences exceeding that of the laboratory ’s historical controls. There is no mutagenic concern for fluopy ram . The available data do not support the proposed modes of action for the thy roid or liver tumors. **VII. QUANTIFICATION OF CARCINOGENIC POTENTIAL**  The CARC recommended the use of a linear low dose extrapolation mo del applied to the animal data (Q 1 *) for quantitative estimation of human risk . 1 The unit risk, Q 1 * (mg/kg/day ) - 1 , of Fluopy ram based upon female rat liver combined adenoma and carcinoma tumor rates is 1.55 x 10 - 2 in human equivalents (Memo. L . Brunsman, 11/17/09, TXR No. 0055280). **VIII BIBLIOGRAPHY**  47372441 Kennel, P.; AE C656948

– 90 - day toxicity study in the rat by

dietary administration; Bayer CropScience S.A., Sophia Antipolis, France; Report No.: SA 04048; Document No.: M - 250946 - 01 - 2; December 07, 2007; Pages: 709 47372442 Kennel, P.; AE C656948

– 90 - day toxicity study in the mouse by

dietary administration; Bayer CropScience S.A., Sophia Antipolis, France; Report No.: SA 04052; Document No.: M - 251136 - 01 - 2; February 20, 2008; Pages: 324 47372450 Wason, S. M.; Carcinogenicity study of AE C656948 in the C57BL/6J mouse by dietary administration; Bayer Cr opScience S.A., Sophia Antipolis, France; Report No.: SA 05094; Document No.: M - 295688 - 01 - 2; January 15, 2008; Pages: 2477 47372501 Kennel, P.; Chronic toxicity and carcinogenicity

study of AE C656948 in the Wistar rat by dietary administration; Bayer C ropScience S.A., Sophia Antipolis, France; Report No.: SA 04312; Document No.: M - 298339 - 01 - 2; March 14, 2008; Pages: 4088 47372502 Wirnitzer, U.; AE C656948 - Salmonel l a/microsome test plate incorporation and

preincubation method; Bayer HealthCare AG, Wuppertal, Germany ; Report No.: AT02911; Document No.: M - 269978 - 01 - 2; December 03, 2007; Pages: 56 47372503 Herbold, B.; AE C656948 (project: AE C656948) - Salmonella/microsome test -

Plate incorporation and preincubation method; Bayer HealthCare AG, Wu ppertal, 1 At the July 8, 2009 meeting , the CARC recommended a classification of “Likely” with no quantific ation. Following this meeting, as per the recommendation of the CARC chair, a revote was requested to be consistent with the cancer guidelines . Subsequently, the CARC recommended a “ Likely” classification with quantification.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 34 of 35 Germany ; Report No.: AT04419; Document No.: M - 298529 - 01 - 2; April 14, 2008; Pages: 65 47372504 Herbold, B.; AE C656948 - V79/HPRT - test in vitro the detection of induced forward

mutations; Bayer HealthCare AG, Wuppertal, Germany ; Report No.: AT02 875; Document No.: M - 268775 - 01 - 2; December 03, 2007; Pages: 40 47372505 Nern, M.; AE C656948 (project: AE C656948) - Invitro chromosome aberration test

with chinese hamster V79 cells; Bayer HealthCare AG, Wuppertal, Germany ; Report No.: AT02798; Document No.: M - 266066 - 01 - 2; December 03, 2007; Pages: 51 47372506 Herbold, B.; AE C656948 - Micronucleus - test on the male mouse; Bayer HealthCare

AG, Wuppertal, Germany ; Report No.: AT02753; Document No.: M - 263710 - 01 - 2; December 03, 2007; Pages: 51 47372509 Klempner, A. (2008) [Phenyl - UL - (Carbon 14)]AE C656948: Absorption,

Di stribution, Excretion and Metabolism in the Rat. Project Number: MEF/07/508, M11819165, M/298614/01/2. Unpublished study prepared by

Bayer CropScience AG. 366 p. 47372510 Koester, J.; Klempner, A. (2008) [Py ridyl - 2,6 - (Carbon 14)]AE C656948 - Metabolism in Organs and Tissues of Male and Female Rats (3 Timepoints). Project Number: MEF/08/115, M1824540/4, M/298834/01/2. Unpublished study prepared by Bayer CropScience AG. 268 p. 47372511 Klempner, A. (2008) [Py ridyl - 2, 6 - (Carbon 14)]AE C656948: Absorption, Distribution, Excretion and Metabolism in the Rat. Project Number: MEF/07/486, M31819167, M/298924/01/2. Unpublished study prepared by

Bayer CropScience AG. 266 p. 47372512 Koester, J.; Kelmpner, A. (2008) [Py ridyl - 2,6 - (Carbon 14)]AE C656948: Distribution of the Total Radioactivity in Male and Female Rats Determined by Quantitative Whole Body Autoradiography

(QWBA), Determination of the Exhaled [(Carbon 14) Dioxide]. Project Number: MEF/07/457, M/1819149/3, M/296485/ 01/2. Unpublished study prepared by

Bayer CropScience. 125 p. 47372513 Koester, J.; Klempner, A. (2008) [Phenyl - UL (Carbon 14)]AE C656948:

Distribution of the Total Radioactivity in Male and Female Rats Determined by Quantitative Whole Body Autoradiogr aphy

(QWBA), Determination of the Exhaled [(Carbon 14) Dioxide] and Metabolic Profiling in E. Project Number: MEF/07/456, M/1811509/5, M/296623/01/2. Unpublished study prepared by

Bayer CropScience. 201 p.FLUOPYRAM CANCER ASSESSMENT DOCUMENT FINAL

Page 35 of 35 47372516 Kennel, P.; AE C656948 - Exploratory 28 - day toxicity study in the rat by

dietary administration; Bayer CropScience S.A., Sophia Antipolis, France; Report No.: SA 03332; Document No.: M - 085510 - 01 - 3; January 14, 2008; Pages: 224 47372517 Kennel, P.; AE C 656948 - Preliminary 28 - day toxicity study in the mouse by

dietary administration; Bayer CropScience S.A., Sophia Antipolis, France; Report No.: SA 04013; Document No.: M - 088486 - 01 - 3; January 14, 2008; Pages: 171 47372519 Rouquie, D.; AE C656948 - Mech anistic 14 - day toxicity study in the mouse by dietary administration (hepatotoxicity and thy roid hormone investigations); Bayer CropScience S.A., Sophia Antipolis, France; Report No.: SA 07215; Document No.: M - 299522 - 01 - 2; April 14, 2008; Pages: 262 47372 520 Blanck, M.; AE C656948 (AE C656948) - 7 - day mechanistic study

in the female Wistar rat by

dietary administration; Bayer CropScience S.A., Sophia Antipolis, France; Report No.: SA 07323; Document No.: M - 299274 - 01 - 2; April 03, 2008; Pages: 160 47567128 Rouquie, D.; AE C656948 - Mechanistic 3 - day toxicity study in the male mouse (pharmacokinetic investigations of the clearance of intravenously administered 125I thy roxine); Bayer CropScience, Sophia Antipolis, France; Report No.: SA 08159; Docum ent No.: M - 308369 - 01 - 2; October 07, 2008; Pages: 78 47690101 Rouquie, D.; AE C656948 - Definitive mechanistic 4 - day toxicity study in the male mouse (pharmacokinetic investigations of the clearance of intravenously administered 125I - thy

roxine); Bayer Cr opScience, Sophia Antipolis, France; Report No.: SA 08288; Document No.: M - 328662 - 01 - 2; February 16, 2006; Pages: 82 47753406 Van Goethem, D.; Wason, S.; Mileson, B. (2009) Fluopy ram: Weight of Evidence Evaluation of Thy

roid Carcinogenesis in Mice and L i ver Carcinogenesis in Rats Using the IPCS Mode of Action Framework. Project Number: 051809, M/347600/01/1. Unpublished study prepared by

Bayer CropScience LP. 49 p. Memo L. Brunsman. 2009. Fluopy ram Qualitative Risk Assessment Based on W istar Rj :WI (IOPS HAN) Rat and C57BL/6J Mouse Dietary Studies. June 10, 2009. TXR # 0055218 . Memo L. Brunsman. 2009. Fluopy ram Quantitative Risk Assessment Based on W istar Rj:WI(I OPS HAN) Rat Dietary Study . November 17, 2009. TXR # 00552 80 .