Document 2qyy4rjD8KMVm8X4Q5r16o10L

TSCA SANITIZED p. 1 ,9cu6-3ss3 CHARLES RIVER LABORATORIES fted'rnicaJ Services 29 May 2007 Steven C. Gordon, PhD, DABT ----3i~1-f.cnpozate-Toxrcobo~gY-~RegnlsodrYSecviees.._. .. 3M Medical Departnent 3M Canter; Bldg. 2'IO-6Fr03 .St Paul, MN 55144-1000 Icl: 651 :736..548S Fax: 651,.733 .1773 &aaait: acgordo~mmm_oom RE: Protocol ZJCA000'7'7 - Oral (Gavage) Developmental Toxicity Screening Study of' MID>D 6675 in Rats Sponsoi's Reference No . 06-.376 Dear Dt. Goidon: Baclosed are unaaadited draft summary tables fionn the end of the in~-life phase of-the screening study., The followlng ir*rnnation summeriae:Jhe results of* this atndy. Mab:!= .!- m .. t Ga! .r:.t'. tio! Rob Sixty pmumed pregnant Cs1:CD(SD) no. 10 per group, were randomly assigaed to six dossEge groups. Fumulations of-the too article, MTD1D 06-75.C 1, and/or the vehicle, revage osmosis metnlnmo processed deionized waft (ILO. deionizad wata), weze administered once daily via oral gavage at dosage leyels 0 (Vabicle),10, 30,90, 270 and 500 megraay. rlo dour volume was S mr.nm, daily on the basis of the individual body waiglrts recorded immediately before Female rats were administered the test article uWor the vdhicle once daily on day 0 of'premuned gestation (DG 0) thcough the day of delivery or DG 24 (*s that do not deliver a Hum). Checks for- viability were made twice daily. Clinical obs4vations were recorded wealdy during the acclimationperiod, once ptior to randomiz~tio~ d~ the predoaage period and daily bsfo~~e a+~ministiaticm during the dosage period. Po~tdc, observations were reeorded at appzox-- ly hoiniy intervals for the four hours and at the end of the normal worldng day fox the fast nine days of the dosage period, Beginning on,the tenth day of'the dosage period, postdosage observations were reoorded within 1 to 2 baotu* of dosage and at the end of the normal wotlturg day.. Maternal behavior was evaluated daily and recorded on days 1, 4 and 6 oflactation (Us 1, 4 and 6). Abnormal behaviox~ was recorded dailyupon observation. 905 Sheehy Drive, Bldg.. A, Horsham, PA 19044 " 215 443-8710 9 FAX : 215..443 8587 0 www .criver com TSCA SANITIZED p. 2 Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 2 of 16 Body weights were recorded weekly during the acclimation period and on DG 0, daily during the dosage period, daily during the postdosage period and on the day of euthanasia. Feed consumption values were recorded on DG 0, daily during the dosage and postdosage periods, and the remaining feed value was recorded on the day of euthanasia. Beginning on DG 21, FO generation female rats were continually evaluated (12 hours per day; 0800 to 2000, and at 2300, 0200 and 0500) for the following parameters : clinical observations during parturition (time each pup is observed was recorded); duration of gestation (DG 0 to the time the first pup is observed); length of parturition (time of observation of last pup minus the time of observation of the first pup divided by N-1 pups in each litter); litter size (defined as all pups delivered); and pup viability at birth. Clinical signs and the time of death for each pup were recorded. These observations continued to be recorded through the first 24 hours after parturition on the previously defined schedule. On DLs 2 through 5, clinical signs and the times of any deaths were recorded at least three times each day. Whenever parturition was complete after 1800 hours, the following day was considered DL 1 . Any pups found partially cannibalized or that were in the process of being cannibalized were euthanized, as needed, and the carcass was retained for possible future evaluation in the appropriate fixative for the age of the pup. Rats that were found dead or were euthanized before scheduled termination were examined for gross lesions on the day the observation was made . The lungs, trachea and esophagus were perfused and saved in neutral buffered 10% formalin for possible future evaluation . When not precluded by autolysis, the heart, liver, kidneys, stomach and spleen were retained in neutral buffered 10% formalin for possible histological evaluation . Pregnancy status and uterine contents offemale rats were recorded . Conceptuses in utero and/or delivered pups were examined to the extent possible. Uteri of apparently nonpregnant rats were stained with 10% ammonium sulfide to confirm the absence of implantation sites. Rats that did not deliver a litter were euthanized on DG 25 and examined for gross lesions. Uteri of apparently nonpregnant rats were stained with 10% ammonium sulfide to confirm the absence of implantation sites. After completion of a 5-day postpartum period, all surviving female rats were euthanized via carbon dioxide asphyxiation on DL 6, and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The number and distribution of implantation sites were recorded. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443 .8710 9 FAX 215.443 .8587 p. 3 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 3 of 16 Methodology - F 1 Generation Pup s F 1 generation pups were not directly given the test artiole or the vehicle but may have been exposed to the test article or the vehicle during maternajl gestation (in utero exposure) or via maternal milk during the lactation period . Litters were observed for dead pups at least three times, daily. The approximate times of death were recorded . The pups in each litter were cou#ted three times daily, except on DL 1, in cases where parturition was complete after 1800 houJrs and on DL 6, when pups in each litter were counted once prior to euthanasia. Clinical observations were recorded at least three times daily during the preweaning period . Body Weights were recorded on postnatal days l, 4 and 6 (PNDs 1, 4 and 6). Pups that died before examination of the litter for pup viability were evaluated for vital status at birth. The lungs were removed and immersed in water; pups with lungs that did not float were identified as stillborn and pups with lungs that did float were identified as liveborn, and to have died shortly after birth. Pups were preserved in Bouin's solution for possible future evaluation. Pups that died or were euthanized prior to scheduled teihnination were examined for cause of death or condition and gross lesions on the day the observation was made . Pups found on PNDs 2 to 4 were preserved in Bouin's solution for possible future evaluation. Surviving pups were euthanized via intraperitoneal injection of sodium pentobarbital on PND 6 and examined for gross lesions. Necropsy inelulded a single cross-section ofthe head at the level of the frontal-parietal suture, and examination of the cross-sectioned brain for apparent hydrocephaly . Results Mortalil:ies Two FO generation female rats assigned to the 500 mg/kg/day dosage group were found dead on DG 2 (day 2 of dosing). Dosage administration was subsequently terminated for the 500 mg/kg/day dosage group due to these deaths and adverse clinical observations that were occurring . The remaining eight FO generation female rats in this dosage group were euthanized . Four FO generation female rats assigned to the 270 mg/!Kg/day dosage group were found dead on either DG 3 or DG 5, and one rat in this dosage group was euthanized due to adverse clinical condition on DG 21 . 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215 .443.8710 " FAX 215.443 .8587 p. 4 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 4 of 16 All of these mortalities were attributed to MTDID 6675 because the incidences were dosage dependent. One FO generation female rat in the 10 mg/kg/day dosage group was euthanized on DG 23 due to adverse clinical condition. The dam apparently had problems in delivering its litter, resulting in the fetuses dying in utero, leading to its adverse clinical condition. This single event was not considered related to the test material because it did not occur in a dosage-dependent manner and problems in delivery are known to occur in this strain of rat. Information for these female rats is included in the table on subsequent pages. All other FO generation rats survived to scheduled euthanasia . 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 9 215 .443.8710 9 FAX 215 .443 .8587 p. 5 Dosage Group 500 MKD TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 5 of 16 Rat Number 23960 23959 23958 Day of Gestation Mode of Death Number of Doses Admini Scheduled DG 1 EuthEaanralsyia - 1 Termination Scheduled DG 1 EuthEaarnlaysia- 1 Termination Scheduled DG 1 EuthEaanrlaysia- 1 Termination Clinical Observations, Body Weights, Feed Consumption, Necropsy Observations and Uterine Contents Clinical Observations: Localized alopecia (limbs) and cold to touch (DGs 0 and 1); dehydration, hunched posture and decreased motor activity (DG 1). Body Weigits : Body weight loss of 33 g from DG 0 to DG 1. Feed Consumption: Feed consumption reduced from DG 0 to DG 1 . Neeropsy Observations: Spleen appeared small; all other tissues appeared normal. Uterine Contents: Pregnancy status could not be confirmed due to early 'onal e. ClinkalObservations : Localized alopecia [limbs (DGs 0 and 1)]; urine-stained abdominal fur and cold to touch (DG 1). Body Weights: Body weight loss of 16 g from DG 0 to DG 1 . Feed Consumption: Feed consumption reduced from DG 0 to DG 1 . Necropsy Observations: All tissues appeared normal . Uterine Contents: Pregnancy status could not be confirmed due to early esmtional e. Clinks[ Observations: Dehydration (DG 1). Body W: eights Body weight loss of 19gfrom DGOtoDG 1 . Feed Consumption: Feed consumption reduced from DG 0 to DG 1. Necropsy Observations: All tissues appeared normal. Uterine Contents: Pregnancy status could not be confirmed due to early estational age. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215 .443.8710 " FAX 215.443 .8587 p. 6 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No . 06-376 29 May 2007 Page 6 of 16 Dosage Group 500 MKD Rat Number 23957 23956 23955 Day of Gestation Mode of Death DG 1 Scheduled Euthanasia - Early Termination DG 1 Scheduled Euthanasia - Early Termination DG 2 Scheduled EuthEaarnlaysia- Termination Number of Doses Administered 1 1 2 Clinical Observations, Body Weights, Feed Consumption, Necropsy Observations and Uterine Contents Clinical Observations: Urinestained abdominal fur and cold to touch (DGs 0 and 1) ; and rales (DG 1). Body Weights : Body weight loss of 28 g from DG 0 to DG 1 . Feed Consumption: Feed consumption reduced from DG 0 to DG 1 . Necropsy Observations: All tissues appeared normal. Uterine Contents: Pregnancy status could not be confirmed due to early es~tional age. Clinical Observations : Dehydration and cold to touch (DG 1). Body Weights: Body weight loss of 23 g from DG 0 to DG 1. Feed Consumption : Feed consumption reduced from DG 0 to DG 1. Necropsy Observations : All tissues appeared normal. Uterine Contents : Pregnancy status could not be confirmed due to early estati.onal e. Clinical Observations : Rales (DG 1); and dehydration (DGs 1 and 2). Body Weights: Body weight loss of 6 g from DG 0 to DG 1 . Feed Consumption: Feed consumption reduced from DG 0 to DG 2. Necropsy Observations: All tissues appeared normal. Uterine Contents: Pregnancy status could not be confirmed due to early estational Mg . 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443 .8710 " FAX 215 .443 .8587 p. 7 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 7 of 16 Dosage Group 500 MKD Rat Number 23954 23953 23952 Day of Gresation DG 2 DG 2 DG 2 Mode of Death Scheduled Euthanasia- Early Termination Found Dead Scheduled Euthanasia - Early Termination Number of Doses Administe* 2 2 2 Clinical Observations, Body Weights, Feed Consumption, Necropsy Observations and Uterine Contents Clinical Observations: Urinestained abdominal fur and dehydration (DGs 0 and 1); ungroomed coat, cold to touch, and decreased motor activity (DG 1). Body Weights: Body weight loss of 34 g from DG 0 to DG 2. Feed Consumption: Feed consumption reduced from DG 0 to DG 2. Neeropsy Observations: Stomach distended with gas; all other tissues appeared normal. Uterine Contents: Pregnancy status could .not be confirmed due to early Clinical Observations : Urinestained abdominal fnr (DGs 0 to 1); rales, dehydration, cold to touch, pale extremities, ptosis and decreased motor activity (DG 1). Body Weights : Body weight loss of 33gfrom DG0toDG 1. Feed Consumption: Feed consumption reduced from DG 0 to DG 1 . Necropsy Observations: Stomach distended with gas; all other tissues appeared normal . Uterine Contents: Pregnancy status could not be confirrned due to early astational e. Ciinkal0bservations: Dehydration and cold to touch (DGs 1 and 2); ungroomed coat and decreased motor activity (DG 2). Body Weights: Body weight loss of 39 g from DG 0 to DG 2. Feed Consumption: Feed consumption reduced from DG 0 to DG 2' Necropsy Observations : Stomach distended with gas; all other tissues appeared normal . Uterine Contents: Pregnancy status could not be confirmed due to early estational e. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443 .8710 " FAX 215.443.8587 Dosage Group 500 MKD 270D p. 8 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 8 of 16 Rat Number 23951 23950 Day of Gestation Mode of Death DG 2 Found Dead DG 3 Found Dead Number of Doses Administered 2 3 Clinical Observations, Body Weights, Feed Consumption, Necropsy Observations and Uterine Contents Clinical Observations : Urinestained abdominal fur (DGs 0 and 1) and dehydration (DG 1) . Body Weights: Body weight loss of 30 g from DG 0 to DG 1. Feed Consumption: Feed consumption reduced from DG 0 to DG 1 . Necropsy Observations: Thymus contained red areas too numerous to count, ranging in size from pinpoint to 0.2 cm x 0.1 cm; all other tissues appeared normal . Uterine Contents : Pregnancy status could not be confirmed due to early estational e. Clinical Observations : Rales (DG 0); dehydration and cold to touch (DGs 1 and 2); urine-stained abdominal fur and ungroomed coat (DG 2). Body Weights: Body weight loss of 36 g from DG 0 to DG 2. Feed Consumption : Feed consumption reduced from DG 0 to DG 1 . Necropsy Observations : All tissues appeared normal. Uterine Contents : Pregnancy status could not be confirmed due to early estational age. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443.8710 9 FAX 215 .443.8587 p. 9 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No . 06-376 29 May 2007 Page 9 of 16 Dosage Group 270 MKD Rat Number Day of Gestation Mode of Death 23947 DG S Found Dead 23946 DG 3 Found Dead Number of Doses Admi ' red S 4 Clinical Observations, Body Weights, Feed Consumption, Necropsy Observations and Uterine Contents CliniealObservations: Urinestained abdominal fur (DGs 1 through 3); cold to touch (DGs 1 through 4); dehydration (DGs 1 through S); ungroomed coat (DGs 2 through 4); pale extremities, and decreased motor activity (DG 3). Body Weights: Body weight loss of 30gfromDGOtoDG4. Feed Consumption: Feed consumption reduced from DG 0 to DG 1 . Necropsy Observations: Stomach distended with gas; all lobes ofthe lungs mottled red and dark red; lungs appeared to be inflated prior to perfusion; trachea and esophagus contained a pink foamy substance; all other tissues appeared normal. Uterine Contents: Pregnancy status could not be confirmed due to early Clinical Observations: Urinestained abdominal fur (DG 0, 2 and 3); dehydration and cold to touch (DGs 1 through 3); rales (DG 2); ungroomed coat, bradypnea, perioral substance, pale extremities, discoloration, ptosis, perinasal substance, decreased motor activity, and impaired righting reflex (DG 3). Body Weights : Body weight loss of 60gfrom DG0toDG3. Feed Consumption: Feed consumption reduced from DG 0 to DG 1 . Neeropsy Observations: Stomach distended with gas; spleen appeared small ; mucosal surface ofthe fundic region of the stomach contained red areas too numerous to count ranging in size from pinpoint to 0.4 to 0.5 cm; renal pelvis of both kidneys red; both adrenals red; abdominal adipose tissue pink; all other tissues appeared normal. Uterine Contents : Pregnancy status could not be confirmed due to early esCational age. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443.8710 " FAX 215.443 .8587 TSCA SANITIZED p. 10 Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 10 of 16 Dosage Group 270 D Rat Number 23945 23941 Day of Gestation Mode of Death DG 21 Unscheduled Euthanasia DG 3 Found Dead Number of Doses Administered 22 3 Clinical Observations, Body Weights, Feed Consumption, Necropsy Observations and Uterine Contents Clinical Observations : Ungroomed coat (DGs 0, 1 and 2); dehydration (DGs 1 through 4 and 21); urine-stained abdominal fur (DGs 2, 4 and 6) ; cold to touch (l)Gs 3 through 6 and 21); rales (DG 4); red perioral substance, pale extremities, low carriage and decreased motor activity (DG 21). Body Weights: Body weight loss of 36 g from DG 19 to DG 21 . Feed Consumption: Feed consumption reduced from DG 19 to DG 21 . Necropsy Observations : All tissues appeared normal. Uterine Contents : There were 12 dead fetuses and one early resorption in utero. All fetuses appeared normal for develo pmental e. Clinical Observations : Dried feces, urine-stained abdominal fur, dehydration, cold to touch, pale extremities, lacrimation and decreased motor activity (DG 2). Body Weights : Body weight loss of 55gfrom DGOtoDG2 . Feed Consumption: Feed consumption reduced from DG 0 to DG 2. Necropsy Observations : All tissues appeared normal . Uterine Contents : Pregnancy status could not be confirmed due to early estational age . 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443 .8710 " FAX 215.443 .8587 P. 11 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 11 of 16 Dosage Group Rat Number 14 MKD 23919 Day of Gestation Mode of Death DG 23 Unscheduled Euthanasia Number of Doses Administered 23 Clinical Observations, Body Weights, Feed Consumption, Necropsy Observations and Uterine Contents Clinical Observations: Localised alopecia on limbs (DGs 14 through 16); alopecia no longer apparent (DG 17); cold to touch, pale extremities, perivaginal substance (DG 23). Body Weiahts : Body weight gain of 152 g from DG 0 to DG 23. Feed Consumption: Feed consumption reduced from DG 22 to DG 23 . Necropsy Observations: Red gelatinous material present in the right horn ofthe uterus at the cervix; all other tissues appeared normal. Uterine Contents: There were 2 live and 14 dead fetuses in utero. One fetus had the left hindlimb rotated medially ; all other fetuses appeared normal for developmental Clinical observations Clinical observations in rats in the 500 mg/kg/day dosage group were described previously in the mortality table. Clinical signs observed in rats in the 270 mg/kg/day dosage group included decreased motor activity, impaired righting reflex, chromorhinorrhea, ptosis, lacrimation, perinasal substance, low carriage, dehydration, cold to touch, pale extremities, discoloration, perioral substance, rales, bradypnea, urin6-stained abdominal fur, ungroomed coat and dried feces; all of these signs were attributed th treatment with the test article because the signs occurred in the highest surviving dosage group. Urine-stained abdominal fur was also observed in the 90 mg/kg/day dosage group and was attributed to MTDID 6675. Cold to touch and pale extremities were observed in the 10 mg/kg/day dosage group but were not attributed to MTDID 6675 because: 1) the incidences ofthese clinical signs in this dosage group did not demonstrate a dosage-dependent pattern of effect; and 2) both clinical signs occurred in only one rat in this group. Chromodacryorrhea, perivaginal substance, sparse hair coat (limbs, back and/or underside) and soft or liquid feces were observed in the 0 (Vehicle), 10, 30 and 90 mg/kg/day dosage groups and were not attributed to MTDID 6675 because: 1) the incidences for these findings did not demonstrate a dosage-dependent pattern of effet; and/or 2) the clinical sign occurred in a limited number of rats in any dosage group. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443.8710 " FAX 215 .443.8587 p. 12 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No . 06-376 29 May 2007 Page 12 of 16 During the lactation period, sparse hair coat (limbs, back and/or underside) and localized alopecia (limbs) were observed in the 0 (Vehicle), 10, 30 and/or 90 mg/kg/day dosage groups . These clinical observations were not attributed to treatment with MTDID 6675 because: 1) the signs were observed after completion ofthe dosage period; and 2) the incidences of these findings did not demonstrate a dosage-dependent pattern ofeffect . Necropsy Observations With the exception of gross lesions described previously in the mortality table, necropsy examinations revealed no additional gross lesions. Body Weights/Body Weight Changes Following the initiation of the dosage period, an overall body weight loss occurred on DGs 0 through 3, and body weight gains were reduced on DGs 3 through 6 in the 270 mg/kg/day dosage group, in comparison with the respective control group values . However, body weight gain began to rebound in this dosage group on DGs 6 through 9. Reductions in body weight gains also occurred in the 90 mg/kg/day dosage group on DGs 0 through 3 and DGs 6 through 9, in comparison with the respective control group values . Body weight gains were reduced in the 90 and 270 mg/kg/day dosage groups during the overall gestation dosage period (calculated as DGs 0 through 20). On DG 20, mean body weights were 97.8%, 98.7%, 96.4% and 85 .7% of the control group value in the 10, 30, 90 and 270 mg/kg/day dosage groups, respectively . During the overall lactation period (calculated as DLs 1 through 6), body weight gains were increased over the control group value in the 90 and 270 mg/kg/day dosage groups . On DL 6, mean body weights were 98.6%, 101 .1%, 98.4% and 95.7% of the control group value, in the 10, 30, 90 and 270 mg/kg/day dosage groups, respectively. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443.8710 " FAX 215.443 .8587 p. 13 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 13 of 16 Absolute Cs/day) and Relative (gIg/dav) Feed Consunption Values Absolute (g/day) and relative (g/kg/day) feed consumption values could not be recorded accurately for the 270 mg/kg/day dosage group on DG9 0 through 3 and 3 through 6 because the rats in this dosage group were provided with moiste6ned feed during these periods at the recommendation of the veterinary staff. Absolute (g/day) and relative (g/kg/day) feed consumption values were reduced in the 90 and 270 mg/kg/day dosage groups during the gestation dosage period (calculated as DGs 0 through 20), in comparison with the respective control group values, as well as most of the intervals within the gestation dosage period. During the overall gestation dosage period (calculated 4s DGs 0 through 20), absolute feed consumption values were 99.1%, 100.0%, 94.1 % and gi.9% and relative feed consumption values were 100.5%,100.6%, 96.1% and 93.0% ofthe control group means in the 10, 30, 90 and 270 mg/kg/day dosage groups, respectively. Absolute and relative feed consumption values were reduced during the overall lactation period (calculated as DLs 1 through 6) in the 10, 30, 90~ and 270 mg/kg/day dosage groups, in comparison with the respective control group values . On DLs 1 through 6, absolute feed consumption values were 92.0%, 92.6%, 92.0%, and 76.1%and relative feed consumption values were 91 .6%, 90.7%, 93.7% and 79.8% of the control group value, in the 10, 30, 90 and 270 mg/lcg/day dosage groups, respectively. Natural Delivery and Litter Parameters Nine, eight, ten, ten and five dams delivered litters with one or more liveborn pups in the 0 (Vehicle), 10, 30, 90 and 270 mg/kg/day dosage groups, respectively . There were 2, 3, 2, 1 and 1 dams with stillborn pups in each of the 0 (Vehicle), 10, 30, 90 and 270 mg/kg/day dosage groups, respectively. One dam in the control group and two dams in the 270 mg/kg/day dosage group had all pups die between Ms 1 through 4. The numbers of stillborn pups, pups found dead or presumed cannibalized on DL 1 and on DLs 2 through 4 were increased, and the number of livobom pups, viability index, the number of surviving pups on DLs 1, 4 and 6, live litter bizes and pup weights per litter on DLs 1, 4 and 6 were reduced in the 270 mg/kg/day do*e group, in comparison with the respective control group values . Mean pup weights per' litter were also reduced in the 90 mg/kg/day dosage group on DLs 1, 4 and 6. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443.8710 " FAX 215 .443 .8587 p. 14 TSCA SANITIZED Protocol ZXA00077 Sponsor's Reference No . 06-376 29 May 2007 Page 14 of 16 Mean length of parturition was reduced in the 270 mg/kg/day dosage group, in comparison with the control group mean. Information on the length of parturition is included below in the table. Dosage Level (mg/kg/day) Number of Rats Per Group Number Mated Number Pregnant Parturition Parameters - Mean f S.D . 0 Vehicle 10 30 10 10 _ 10 10 10 10 9 9 10 minutes 90 10 10 10 Length of Parturition (N) 7.57t2.57 2 Time Between Deliveries - 9.90t4.06 Partial 6 Time Between Deliveries - 7.57f2.57 Com lete 2 N - Number ofdams evaluated 13 .OOt6.55 5 19 .9910 .90 3 _ 13 .OOt6.55 5 9.98f1 .24 4 9.63t3.71 6 9.981 .24 4 9.27t2.86 5 8.96t3.04 5 9.27f2.86 (5 -270 10 10 6 5.04f1 .83 2 6.18t0.0 1 5.04t1 .83 2) 500 10 10 Not applicable Not applicable Not licable Not applicable Pup weights per litter were considered reduced in the 90 and 270 mg/kg/day dosage groups . On PND 1, average pup weights were 96.8%, 93 .7%, 87.3% and 73 .0% ofthe control group value, and on PND 6, average pup weights were 100.9%, 99.1%, 90.1% and 74 .8% for the 10, 30, 90 and 270 mg/kg/day dosage groups, respectively . The values in the 90 and 270 mg/kg/day dosage groups on DL 1 (5 .5 and 4.6, respectively) were below the historical control range (6 .Og to 6.8 g) for this Testing Facility based on data collected from June 2004 to January 2006. The duration of gestation was generally comparable among the dosage groups . 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215 .443.8710 " FAX 215 .443 .8587 p. 15 TSCA SANITIZED Clinical and Necropsv Observations - F 1 Generation Puus Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 15 of 16 Two pups from two litters in the 270 mg/kg/day dosage group had body regions that were discolored purple; the lower midline, both hindlimbs a#id tail were discolored purple in one pup, and the right hindlimb and hindpaw were discolored purple in the other pup. The nose was discolored purple in three pups from one litter in 3b mg/kg/day dosage group. One pup from the 10 mg/kg/day dosage group and two littermates in the 90 mg/kg/day dosage group were cold to touch. One of the two pups from the 90 mg/kg/day dosage group was also observed on DL 4 as pale, not nesting and not nursing. One pup from the 30 mg/kg/day dosage group was missing the tip ofthe tail. Purple discoloration of body regions in the pups in the 270 mg/kg/day dosage group was attributed to MTDID 6675 because: 1) the clinical obsfrrvation occurred in the highest surviving dosage group; and 2) the discoloration was more extensive in these pups . All other clinical observations were not attributed to treatment w~th the test article because the incidences did not demonstrate a dosage-dependent pattern of effect. All other pups appeared normal . Among pups that were found dead, 0, 0, 1, 1 and 10 pups in the 0 (Vehicle), 10, 30, 90 and 270 mg/kg/day dosage groups, respectively, had no milk present in the stomach. The increased incidence in the 270 mg/kg/day dosage group! was attributed to 1VITDID 6675. 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215.443 .8710 " FAX 215 .443.8587 TSCA SANITIZED p. 16 Protocol ZXA00077 Sponsor's Reference No. 06-376 29 May 2007 Page 16 of 16 Conclusion Mortalities, adverse clinical signs and decreases in maternal body weight and feed consumption occurred during gestation at 90 and/or 270 mg/kg/day; body weight rebounded during the lactation period in these dosage groups, but feed consumption remained reduced. A reduction in pup weight occurred in the 90 mg/kg/day dosage group, and doses of 270 mg/kg/day also produced reproductive and developmental toxicity as evidenced by shortened parturition time, pup death and decreased pup weights. Therefore the no-observable-adverse-effect-level (NOAEL) for both maternal toxicity and developmental toxicity was 30 mg/kg/day of MTDID 6675 . If you have any questions, please do not hesitate to contact me . Sincerely, AMH:mbc Attachments J Alan M. Hoberman, Ph.D., DABT, Fellow ATS Director of Research Study Director 905 Sheehy Drive, Bldg. A, Horsham, PA 19044 " 215 .443.8710 " FAX 215.443.8587 . H_ IPTG V IZ-1 T. 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