Document B236vMERVaD99jVKyDZz6D9w

418-011: PAGE F-1 M W rO Z rt FINAL PILOT REPORT Study Title Oral (Gavage) Dosage-Range Developmental Toxicity Study of N-EtFOSE in Rats Sponsor's Study Number: T-6316.7 Author Raymond G. York, Ph.D., DABT (Study Director) Study Completed On 10 December 1998 (Final Pilot Report) Performing Laboratory Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Laboratory Project ID Argus Research Laboratories, Inc., Protocol Number: 418-011P 003750 418-011: PAGE F-2 418-011 pipage 2 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS TABLE OF CONTENTS SUBJECT ABSTRACT I. Purpose II. Methods III. Results IV. Conclusion F ig u re ! Maternal Body Weights Table ! Clinical and Necropsy Observations - Summary Table 2. Maternal Body Weights - Summary Table 3. Maternal Body Weight Changes - Summary Table 4. Maternal Absolute Feed Consumption Values (g/day) Summary Table 5. Maternal Relative Feed Consumption Values (g/kg/day) - Summary Table 6. Caesarean-Sectioning Observations - Summary PAGE 4 6 6 7 9 10 11 13 15 17 19 21 003751 418-011 PAGE F-3 418-011P:PAGE 3 SUBJECT Table 7. Litter Observations (Caesarean-Delivered Fetuses) Summary Table 8 Fetal Gross External Alterations - Summary Table 9. Clinical Observations - Individual Data Table 10. Necropsy Observations - Individual Data Table 11. Maternal Body Weights - Individual Data Table 12. Maternal Feed Consumption Values - individual Data Table 13. Caesarean-Sectioning Observations - Individual Data Table 14. Litter Observations (Caesarean-Delivered Fetuses) Individual Data Table 15. Fetal Sex, Vital Status and Body Weight Individual Data ATTACHMENT 1 - PROTOCOL AND AMENDMENT PAGE 23 25 27 31 35 42 46 50 54 58 003752 418-011 PAGE F-4 418-011P:PAGE 4 TITLE: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS ARGUS RESEARCH LABORATORIES, INC. PROTOCOL NUMBER: 418-011P SPONSOR'S STUDY NUMBER: T-6316.7 ABSTRACT Fifty-six presumed pregnant Crl:CDBR VAF/Plus (Sprague-Dawley) rats were randomly assigned to seven dosage groups [eight per group (Groups I through VII)]. Suspensions of the N-EtFOSE were administered orally via gavage once daily to these naturally-bred female rats on days 6 through 17 of presumed gestation (DGs 6 to 17) at dosages of 0 (Vehicle), 1, 5, 10, 20, 25 and 35 mg/kg/day. The dosage volume was 5 ml_/kg, adjusted daily on the basis of the individual body weights recorded immediately before administration of the test article. Checks for viability were made twice daily. Clinical observations were recorded daily before dosage, approximately one hour after administration and then four to six hours later. These observations were also recorded once daily during the postdosage period. Body weights were recorded daily during the dosage and postdosage periods and feed consumption values were recorded on DGs 0, 4, 6, 8, 10, 12, 14, 16, 18 and 20. All rats were sacrificed on DG 20 and examined for the number and distribution of corpora lutea, implantation sites and uterine contents. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Fetuses were weighed and examined for gross external alterations and sex. All rats survived until scheduled sacrifice. Clinical observations during gestation considered treatment-related were limited to single rats with chromorhinorrhea, chromodacryorrhea or emaciation at the 20 and 35 mg/kg/day dosages. Clinical observations of localized alopecia and emaciation were confirmed at necropsy. No additional necropsy observations occurred. 003*753 418-011 PAGE F-5 418-011P:PAGE 5 Body weight gains and absolute and relative feed consumption values for the entire dosage period (calculated as DGs 6 to 18), the entire interval after initiation of treatment (DGs 6 to 20) and the entire gestation period (DGs 0 to 20) were reduced in the 10 mg/kg/day and higher dosage groups, compared to the control group. Reduced fetal body weights occurred at the 25 and 35 mg/kg/day dosages, relative to the control group (-8.5% and -13.8%, respectively). No other Caesarean-sectioning or litter parameters were affected by dosages of the test article as high as 35 mg/kg/day. One litter in the 35 mg/kg/day dosage group consisted of two early resorptions and 13 fetuses with cleft palate (two also had whole body edema). These fetal findings were considered genetic in origin and not test article-related. Based on the results of this dosage-range finding study, dosages of 0, 1, 5, 10 and 20 mg/kg/day were recommended for the full developmental toxicity study of N-EtFOSE in rats. 003754 418-011 PAG E F-6 418-011P:PAGE 6 I. Purpose: The purpose of this study was to provide information for the selection of dosages to be used in the developmental toxicity (embryo-fetal toxicity and teratogenic potential) study of N-EtFOSE administered orally via gavage to Crl:CDBR VAF/Plus presumed pregnant female rats. II. Methods1: The test article, N-EtFOSE [lot/batch number FM-3929 (30035, 30037, 30039)], was received on 20 May 1998, and stored at room temperature. The vehicle was 2% Tween 80 in reverse osmosis membrane processed deionized water (R.O. deionized water). The Tween 80 was received from J.T. Baker, Philipsburg, New Jersey, on 22 May 1998, and was stored at room temperature. The R.O. deionized water is available from a continuous source at the Testing Facility and is maintained at room temperature. The prepared vehicle was stored at room temperature. Test article formulations were prepared daily. Fifty-six presumed pregnant Crl:CDBR VAF/Plus (Sprague-Dawley) rats were randomly assigned to seven dosage groups [eight per group (Groups I through VII)]. Suspensions of the test article were administered orally via gavage once daily to these naturally-bred female rats on days 6 through 17 of presumed gestation (DGs 6 to 17) at dosages of 0 (Vehicle), 1, 5, 10, 20, 25 and 35 mg/kg/day. The dosage volume was 5 mL/kg, adjusted daily on the basis of the individual body weights recorded immediately before administration of the test article. Checks for viability were made twice daily. Clinical observations were recorded daily before dosage, approximately one hour after administration and then four to six hours later. These observations were also recorded once daily during the postdosage period. Body weights were recorded daily during the dosage and postdosage periods and feed consumption values were recorded on DGs 0, 4, 6, 8, 10, 12, 14, 16, 18 and 20. a. Detailed descriptions of all procedures used in the conduct of this study are provid^l in the appropriate sections of this report and in the attached protocol an& amendments. Deviations from the Protocol and Standard Operating Procedures Of the Testing Facility are available in the raw data. 003755 418-011 :PAGE F-7 418-011pipage i All rats were sacrificed on DG 20 and examined for the number and distribution of corpora ltea, implantation sites live and dead fetuses and early and late resorptions. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Fetuses were weighed and examined for gross external alterations and sex. III. Results: A. Mortality. Clinical and Necropsy Observations (Summary - Table 1: Individual Data - Tables 9 and 101 A.1. Mortality All rats survived until scheduled sacrifice. A.2. Clinical Observations Clinical observations during gestation considered treatment-related were limited to single rats with chromorhinorrhea, chromodacryorrhea or emaciation at the 20 and 35 mg/kg/day dosages. All other clinical observations were considered unrelated to the test article because: 1) the incidences were not dosage-dependent; and/or 2) they occurred in only one rat. These observations included localized alopecia on the limbs, underside, back and/or neck. A. 3. Necropsy Observations Clinical observations of localized alopecia and emaciation were confirmed at necropsy. No additional necropsy observations occurred. B. Maternal Body Weights and Body Weight Changes (Figure 1: Summaries - Tables 2 and 3: Individual Data - Table 111 Maternal body weight gains in the 10 mg/kg/day and higher dosage groups were decreased, compared to the control group on gestation days (DGs) 6 to 8 and 8 to 10. Body weights were decreased for the 25 mg/kg/day and higher dosages on DGs 10 to 12 ,12 to 14 and 14 to 16. Maternal body weight gains in the 1, 5, 10, 20, 25 and 35 mg/kg/day dosage groups were generally comparable to control values postdosage (DGs 18 to 20). 003756 418-011: PAGE F-8 418-011P:PAGE 8 Reflecting these effects of the test article, weight gains for the entire dosage period (calculated as DGs 6 to 18), the entire interval after initiation of treatment (DGs 6 to 20) and the entire gestation period (DGs 0 to 20) were reduced in the 10 mg/kg/day and higher dosage groups, compared to the control group. Maternal body weights and body weight gains for the 1 and 5 mg/kg/day dosage groups were generally comparable to control values over each interval tabulated. C. Maternal Absolute fo/davl and Relative (o/ko/dav) Feed Consumption Values (Summaries - Tables 4 and 5: Individual Data - Table 121 Absolute and relative feed consumption values for the 10 mg/kg/day and higher dosage groups were decreased, compared to the control group at all tabulated intervals during the dosage period (DGs 6 to 8, 8 to 10, 10 to 12, 12 to 1 4 ,1 4 to 16 and 16 to 18). Maternal feed consumption values for the 1, 5, 10, 20, 25 and 35 mg/kg/day dosage groups were generally comparable to control values postdosage (DGs 18 to 20). Reflecting these effects of the test article, absolute and relative feed consumption values for the entire dosage period (calculated as DGs 6 to 18), the entire interval after initiation of treatment (DGs 6 to 20) and the entire gestation period (DGs 0 to 20) were reduced in the 10 mg/kg/day and higher dosage groups, compared to the control group. Absolute and relative feed consumption values for the 1 and 5 mg/kg/day dosage groups were generally comparable to control values over each interval tabulated. D. Caesarean-Sectioning and Utter Observations (Summaries - Tables 6 through 8: Individual Data - Tables 13 through 15) Caesarean-sectioning observations were based on 8 (100%), 7 (87.5%), 8 (100%), 8 (100%), 8 (100%), 8 (100%) and 8 (100%) pregnant rats with live litters in the seven respective dosage groups. Reduced fetal body weights occurred at the 25 and 35 mg/kg/day dosages, relative to the control group (-8.5% and -13.8%, respectively). This observation was considered an effect of the test article because it was dosage-dependent 003757 418-011 .PAGE F-9 418-011P:PAGE 9 and occurred at the dosages in which there was maternal toxicity observed (decreased body weight and feed consumption values). No other Caesarean-sectioning or litter parameters were affected by dosages of the test article as high as 35 mg/kg/day. The litter averages for corpora ltea, implantations, litter size, live fetuses, resorptions (early and late), dams with any resorptions and percent live male fetuses were comparable among the 0 Vehicle), 1, 5, 10, 20, 25 and 35 mg/kg/day dosage groups. No dams had all resorbed conceptuses, there were no dead fetuses and all placentae appeared normal. Totals of 109, 93, 116, 110,118, 116 and 114 live fetuses were evaluated for external gross alterations in the seven respective dosage groups. No fetal gross external alterations were observed at dosages up to 25 mg/kg/day. One litter (10650) in the 35 mg/kg/day dosage group consisted of two early resorptions and 13 fetuses with cleft palate (two also had whole body edema). These fetal findings were considered genetic in origin and not test article-related. IV. Conclusion: Based on the results of this study, dosages of 0 (Vehicle), 1, 5, 10 and 20 mg/kg/day of N-EtFOSE are recommended for the developmental toxicity study in rats (418-011). The 1 mg/kg/day dosage is expected to be a no observable-effect-level (NOEL) for both maternal and embryo-fetal toxicity, and the 20 mg/kg/day dosage is expected to produce maternal toxicity (decreased maternal body weight and feed consumption values) and minimal developmental toxicity (decreased fetal body weights and possibly delayed ossification). Study Director 003758 418-011:PAGE F-10 (O) 1H0I3M 003759 MATERNAL BODY WEIGHTS Figure 1 400 380 y V 0 (VEHICLE) MG/KG/DAY 360 1 MG/KG/DAY 340 O 320 5 MG/KG/DAY A 300 10 MG/KG/DAY 280 X 20 MG/KG/DAY 260 25 MG/KG/DAY 240 220 i i i i i i ; i ! 35 MG/KG/DAY 10 11 12 13 14 15 16 17 18 19 20 DAYS OF GESTATION 00 TJ 5O m PROTOCOL 416-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 1 (PAGE 1): CLINICAL AND NECROPSY OBSERVATIONS - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a I 0 (VEHICLE) II 1 III s IV 10 MAXIMUM POSSIBLE INCIDENCE 120/ 8 120/ 8 120/ 8 120/ 8 MORTALITY 0000 LOCALIZED ALOPECIA: * ft TOTAL LIMBS UNDERSIDE BACK NECK 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 01 0 0/ 0 0/ 0 15/ 2 8/ 1 7/ 1 0/ 0 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 EMACIATION 0/ 0 0/ 0 0/ 0 0/ 0 CHROMORHINORRHEA 0/ 0 0/ 0 0/ 0 0/ 0 CHROMODACRYQRRHEA 0/ 0 0/ 0 0/ 0 0/ 0 PERSISTENT ADVERSE CLINICAL OBSERVATIONS HERE CONFIRMED AT NECROPSY, NO ADDITIONAL GROSS LESIONS HERE IDENTIFIED MAXIMUM POSSIBLE INCIDENCE (DAYS X RATS) /NUMBER OF RATS EXAMINED PER GROUP ON DAYS 6 THROUGH 20 OF PRESUMED GESTATION. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS HITH OBSERVATION, a. Dosage occurred on days 6 through 17 of presumed gestation. 418-011 P A G E F-1 003760 0oI0 TJ 5 mO PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 1 (PAGE 2) : CLINICAL AND NECROPSY OBSERVATIONS - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a V VI VII 20 25 35 MAXIMUM POSSIBLE INCIDENCE 120/ 8 120/ 8 120/ 8 MORTALITY 000 LOCALIZED ALOPECIA: , TOTAL LIMBS UNDERSIDE BACK NECK 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 7/ 1 0/ 0 7/ 1 6/ 1 0/ 0 20/ 2 12/ 1 8/ 1 8/ I 0/ 0 EMACIATION 0/ 0 0/ 0 4/ 1 CHROMORHINORRHEA 0/ 0 0/ 0 1/ 1 CHROMODACRYORRHEA 3/ 1 0/ 0 0/ 0 PERSISTENT ADVERSE CLINICAL OBSERVATIONS HERE CONFIRMED AT NECROPSY, NO ADDITIONAL GROSS LESIONS HERE IDENTIFIED MAXIMUM POSSIBLE INCIDENCE - (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP ON DAYS 6 THROUGH 20 OF PRESUMED GESTATION. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION, a. Dosage occurred on days 6 through 17 of presumed gestation. T9& C 00 00 6 1O "D T> > 0 DG> m m T1| to ro 418-011 pipag e 13 003762 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OP N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 2 (PAGE 1) : MATERNAL BODY HEIGHTS - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a I 0 (VEHICLE) RATS TESTED N 8 PREGNANT N8 BODY HEIGHT (G) DAY 0 MEAN+S.D. 235.9 + 10.3 DAY 4 MEAN+S.D. 262.4 + 15.2 DAY 6 MEAN+S.D. 266.4 + 13.9 DAY 7 MEAN+S.D. 269.4 + 16.6 DAY 8 MEAN+S.D . 274.6 + 17.0 DAY 9 MEAN+S.D. 278.9 17.2 DAY. 10 MEAN+S.D. 285.8 + 19.0 DAY 11 MEAN+S.D. 290.4 + 19.4 DAY 12 MEAN+S.D. 295.4 + 18.2 DAY 13 MEAN+S.D . 302.6 + 20.5 DAY 14 MEAN+S.D. 306.8 + 21.9 DAY 15 MEAN+S.D . 313.1 + 21.9 DAY 16 MEAN+S.D. 323.0 + 24.0 DAY 17 MEAN+S.D. 334.4 + 24.0 DAY 18 MEAN+S.D . 350.1 + 29.0 DAY 19 MEAN+S.D . 367.2 + 29.0 DAY 20 MEAN+S.D. 385.6 + 31.8 DAY = DAY OF GESTATION a. Dosage occurred on days 6 through 17 of gestation. II 1 8 7 236.4 + 12.6 254.7 10.3 264.4 + 16.3 266.3 + 14.9 270.4 + 16.6 276.6 + 16.2 282.7 + 15.5 284.4 + 17.9 291.7 + 18.6 296.6 + 16.5 302.4 + 16.5 307.8 + 17.8 320.0 + 21.6 330.1 + 22.8 348.0 + 23.6 360.6 + 26.6 378.7 + 32.3 III 5 8 a 236.4 + 8.4 258.0 _+ 9.0 263.9 + 10.2 266.5 + 9.4 271.8 + 8.7 276.1 + 9.8 281.4 + 9.8 286.9 + 11.0 288.4 +_ 8.5 293.4 9.9 297.5 + 8.9 304.4 + 9.6 314.5 + 9.5 326.1 + 10.0 342.5 + 8.5 357.8 + 11.2 375.0 + 9.9 IV 10 8 8 236.0 + 8.1 255.0 + 11.8 257.8 + 12.9 259.0 _+ 14.3 260.9 18.6 264.4 16.5 268.9 + 17.2 274.8 + 18.7 280.4 + 18.0 2B3.4 + 20.4 288.2 + 19.6 296.1 + 21.8 303.8 + 20.7 313.4 22.6 326.2 + 22.1 341.2 + 25.5 359.1 + 27.6 -fe. 00 PROTOCOL 416-OUP: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 2 (PAGE 2): MATERNAL BODY WEIGHTS - SUMMARY 418-011P:PAG E 14 003763 DOSAGE GROUP DOSAGE (MG/KG/DAY)a V 20 RATS TESTED N 8 PREGNANT N8 BODY HEIGHT (G) DAY 0 MEAN+S.D. 236.2 + 8.0 DAY 4 MEAN+S.D. 258.5 + 8.2 DAY 6 MEAN+S.D . 262.1 + 8.2 DAY 7 MEAN+S.D . 260.8 + 8.1 DAY 8 MEAN+S.D. 263.1 + 9.0 DAY 9 MEAN+S.D. 268.5 12.5 DAY 10 MEAN+S.D . 270.1 +_ 12.6 DAY 11 ' MEAN+S.D. 276.5 + 13.7 DAY 12 MEAN+S.D . 280.9 + 12.5 DAY 13 MEAN+S.D. 285.6 + 15.3 DAY 14 MEAN+S.D . 288.1 + 12.9 DAY 15 MEAN+S.D . 295.1 + 17.1 DAY 16 MEAN+S.D . 305.6 + 18.2 DAY 17 MEAN+S.D. 318.2 + 18.6 DAY 18 MEAN+S.D . 330.9 + 21.3 DAY 19 MEAN+S.D. 344.2 + 22.5 DAY 20 MEAN+S.D . 363.5 22.1 DAY * DAY OF GESTATION a. Dosage occurred on days 6 through 17 of gestation. VI 25 8 8 236.0 + 8 .5 257.9 + 12 .6 264.0 + 14 .2 262.0 + IS .8 264.4 + 16 .5 269.2 + 16 .3 272.1 + 15 .2 274.1 + 15 .0 276.0 + 17 .8 277.0 + 16 .7 276.8 + IB .7 281.0 + 19 .2 289.9 + 21 .6 301.6 + 23 .5 318.0 + 22 .3 332.0 + 22 .4 345.8 + 20 .4 VII 35 8 236.5 9.2 255.2 + 11.3 261.1 + 10.5 261.9 + 9.4 262.6 + 7.2 265.0 + 9.3 265.6 11.4 266.9 + 10.9 267.2 12.2 267.8 + 16.9 269.4 + 20.7 274.0 + 25.8 283.8 + 30.1 289.0 + 34.6 302.0 39.7 318.1 +_ 41.8 336.1 41.6 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR1S STUDY NUMBER: T-6316.7) TABLE 3 (PAGE 1) : MATERNAL BODY WEIGHT CHANGES - SUMMARY o <o p O 4* DOSAGE GROUP DOSAGE (MG/KG/DAY) a I 0 (VEHICLE) RATS TESTED N 8 PREGNANT N8 BODY HEIGHT CHANGE (G) DAYS 0 - 6 8: DAYS 6 - MEAN+S.D . MEAN+S.D . +30.5 + 10.3 +8.2 + 4.3 DAYS 8 10 MEAN+S.D. li.i + 2.9 DAYS 10 - 12 MEAN+S. D. +9.6 + 4.0 DAYS 12 - 14 MEAN+S.D. +11.4 + 5.0 DAYS 14 - 16 MEAN+S.D. +16.2 + 6.7 DAYS IS - 18 MEAN+S.D. +27.1 + 7.4 DAYS . 6 - 18 MEAN+S.D . +83.8 + 20.5 DAYS 18 - 20 MEAN+S.D . +35.5 + 7.0 DAYS 6 - 20 MEAN+S.D . +119.2 + 24.4 DAYS 0 - 20 MEAN+S.D. +149.8 + 33.1 DAYS = DAYS OF GESTATION a. Dosage occurred on days E through 17 of gestation. II 1 8 7 +28.0 + 7.2 +6.0 + 3.4 +12.3 + 1.5 +9.0 + 5.4 +10.7 _+ 5.8 +17.6 8.0 +28.0 + 4.7 +83.6 + 10.4 +30.7 + 11.4 +114.3 + 21.2 +142.3 + 26.4 III 5 8 8 +27.5 5.8 +7.9 2.3 +9.6 + 3.1 +7.0 + 2.9 +9.1 + 3.1 +17.0 + 2.2 +28.0 + 4.5 +78.6 + 8.8 +32.5 + 5.1 +111.1 11.1 +138.6 + 12.2 IV 10 8 8 +21.8 + 6.4 +3.1 + 11.4 +8.0 + 5.0 +11.5 + 6.7 +7.9 3.2 +15.5 + 3.2 +22.5 4.0 +68.5 + 13.1 +32.9 + 6.8 +101.4 + 17.7 +123.1 + 20.5 00 *- 9o " TJ T>3 > Q m o m cn tn PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 3 (PAGE 2) : MATERNAL BODY WEIGHT CHANGES - SUMMARY 003765 DOSAGE GROUP DOSAGE (MG/KG/DAY)a RATS TESTED PREGNANT N N V 20 e 8 BODY WEIGHT CHANGE (G) DAYS 0 - 6 MEANS.D. +25.9 + 7.6 DAYS 6 - 8 MEAN+S.D. +1.0 + 4.8 DAYS 8 - 10 MEAN+S.D. +7.0 + S.6 DAYS 10 - 12 MEAN+S.D. +10.8 3.6 DAYS 12 - K DAYS 14 - 16 MEAN+S.D. MEAN+S.D. +7.2 2.4 +17.5 + 7.9 DAYS 16 - 18 MEAN+S.D. +25.2 + 4.8 DAYS 6 - 18 MEAN+S.D. +68.8 + 15.4 DAYS 18 - 20 MEAN+S.D. +32.6 _+ 3.4 DAYS 6 - 20 MEAN+S.D. +101.4 16.1 DAYS 0 - 20 MEAN+S.D. +127.2 + 21.0 DAYS = DAYS OF GESTATION a. Dosage occurred on days 6 through 17 of gestation. VI 25 s 8 +28.0 + 9.4 +0.4 + 4.5 +7.8 + 4.5 +3.9 + 7.6 +0.8 + 6.2 +13.1 + 11.3 +28.1 + 9.3 +54.0 + 13.9 +27.8 + 7.6 +81.8 + 11.9 +109.8 + 14.0 VII 35 8 e +24.6 + 6.6 +1.5 + 6.7 +3.0 + 7.2 +1.6 J+ 10.5 +2.1 + 13.4 +14.4 + 11.1 +18.2 + 11.7 +40.9 + 42.3 +34.1 + 4.5 +75.0 + 44.6 +99.6 + 44.4 .b. oo -tt ?O "D > O 5Om m o> CD 418-011 PAGE F-17 003766 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 4 (PAGE 1) : MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a I 0 (VEHICLE) RATS TESTED N 8 PREGNANT N8 FEED CONSUMPTION (G/DAY) DAYS 0 - 6 -DAYS 6 8 MEAN+S. D. MEAN+S. D. 23.6 + 3.6 25.9 5.4 DAYS 8 - 10 MEAN+S.D . 25.4 3.2 DAYS 10 - 12 DAYS 12 - 14 DAYS 14 - 16 MEAN+S. D. MEAN+S. D. MEAN+S.D . 26.2 + 3.7 C 7]b 28.1 + 8.2 t 71b 24.2 _+ 2.3 DAYS lfi 18 MEAN+S.D. 25.9 + 2.6 DAYS 6 - 18 MEAN+S.D. 25.9 + 3.4 DAYS 18 - 20 MEAN+S.D, 26.2 3.0 DAYS 6 - 20 MEAN+S.D. 25.9 3.2 DAYS 0 - 20 MEAN+S.D. 25.2 3.1 DAYS = DAYS OF GESTATION [ ] - NUMBER OF VALUES AVERAGED a. Dosage occurred on days 6 through 17 of gestation. b. Excludes values that were associated with spillage. II 1 8 7 21.9 + 2.9 23.5 + 2.4 24.1 + 2.5 23.1 2.0 27.9 + 5.2 25.1 + 3.0 25.5 + 4.4 24 .B + 1.6 26.2 +_ 2.0 25.0 1.6 24.1 + 1.8 hi 5 8 8 23.5 + 1.0 22.0 4.6 24.2 l.i 24.4 2.2 23.5 2.5 23.4 +_ 2.5 25.4 + 1.7 23.8 + 1.4 24.8 2.2 24.0 1.3 23.8 0.9 IV 10 8 8 21.3 + 2.4 19.1 5.6 20.8 2.7 22.8 2.0 22.7 3.3 21.1 + 2.9 +22.2 2.6 21.4 + 2.6 24.0 + 3.1 21.8 f 2.6 21.7 2.3 00 5O m PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER; T-6316.7) TABLE 4 (PAGE 2) ; MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - SUMMARY n!9 C 0 0 DOSAGE GROUP DOSAGE (MG/KG/DAY)a V 20 RATS TESTED N 8 PREGNANT N8 FEED CONSUMPTION (G/DAY) DAYS 0 - 6 MEAN+S.D. 24.8 + 3.3 DAYS 6 - 8 MEAN+S.D 21.2 + 3.3 DAYS 6 - 10 MEAN+S.D. 22.2 + 4.4 DAYS 10 -- 12 MEAN+S.D. 23.1 + 3.0 DAYS 12 - 14 MEAN+S.D. 23.1 +_ 3.4 DAYS 14 - 16 MEAN+S.D. 22.6 + 5.4 DAYS 16- - 18 MEAN+S.D. 23.4 4 3.7 DAYS 6 - 18 MEAN+S.D. 22.6 4 3.3 DAYS 18 - 20 MEAN+S.D . 23.8 4 2.1 DAYS 6 - 20 MEAN+S.D. 22.8 4 3.1 DAYS 0 - 20 MEAN+S.D. 23.4 + 2.8 DAYS = DAYS OF GESTATION a. Dosage occurred on days 6 through 17 of gestation. VI 25 8 B 22.0 + 2.7 19.8 + 3.7 20.9 + 2.4 20.1 + 2.4 16.6 +_ 3.3 17.8 _+ 5.8 21.0 + 3.3 19.4 +_ 2.6 21.5 1.6 19.7 4 2.3 20.4 + 2.1 VII 35 8 8 22.6 1.5 21.1 2.7 20.1 3.5 18.9 + 5.3 20.1 + 8.7 18.1 + 7.2 18.6 + 5.6 19.5 + 4.2 24.2 + 4.3 20.2 4 4.1 20.9 4 3.0 CO 6 a 9o u > o 5 mG) m T! 00 CD 418-011-.PAGE F-19 003768 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE S (PAGE 1) : MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a I 0 (VEHICLE) RATS TESTED H 8 PREGNANT N8 FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 6 DAYS 6 - a MEAN+S.D. MEAN+S.D. 92.2 11.2 95.5 +_ 16.5 DAYS 8 - 10 MEAN+S.D. 90.7 + 7.5 DAYS 10 - 12 DAYS 12 14 DAYS 14 - 16 MEAN+S.D. MEAN+S.D. MEAN+S.D. 90.0 + 8.8 [ 7]b 93.4 + 26.7 I 7]b 77.1 + 7.2 DAYS 16 - 18 MEAN+S.D. 77.3 + 5.8 DAYS 6 - 18 DAYS 18 - 20 MEAN+S.D. MEAN+S.D. 86.4 + 9.6 71.4 + 7.8 DAYS 6 - 20 MEAN+S.D. 83.8 + 8.9 DAYS 0 - 20 MEAN+S.D. 83.2 + 7.6 DAYS = DAYS OF GESTATION [ ] NUMBER OF VALUES AVERAGED a. Dosage occurred on days 6 through 17 of gestation. b. Excludes values that were associated with spillage. ii 1 8 7 86.7 + 9.8 8B.1 + 8.3 87.2 7.6 80.5 + 4.2 95.1 + 23.5 80.8 + 7.6 76.3 + 9.6 84.2 + 5.0 72.4 + 3.3 82.1 + 4.5 80.8 4.5 III 5 8 8 93.1 + 2.7 82.5 + 17.8 87.6 + 4.5 85.6 + 7.8 80.3 + 9.2 76.5 8.1 77.7 + 5.6 81.4 + 5.7 69.2 + 5.5 79.1 + 5.1 80.4 3.6 IV 10 8 8 85.1 + 7.1 72.9 + 19.5 78.4 + 6.2 82.8 + 3.6 79.7 8.5 71.3 + 7.6 70.4 + 4.1 75.5 + 5.3 69.9 + 5.9 74.5 + 4.7 75.5 + 3.9 A ?O v mo CO 418-011: PAGE F-21 003770 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 6 (PAGE 1) : CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a I 0 (VEHICLE) RATS TESTED N 8 PREGNANT N (%) 8(100.0) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 20 OP GESTATION N 8 CORPORA LUTEA MEAN+S.D. 16.0 2.6 IMPLANTATIONS MEAN+S.D . 14.0 4.5 LITTER SIZES MEAN+S.D. 13.6 + 4.6 LIVE FETUSES N MEAN+S.D. 109 13.6 + 4.6 DEAD FETUSES N 0 RESORPTIONS MEAN+S.D. 0.4 + 0.7 EARLY RESORPTIONS N MEAN+S.D . 3 0.4 + 0.7 LATE RESORPTIONS N MEAN+S.D. 0 0.0 + 0.0 DAMS WITH ANY RESORPTIONS N(t) 2 ( 25.0) DAMS WITH ALL CONCEPTUSES RESORBED N 0 DAMS WITH VIABLE FETUSES N(%) 8(100.0) PLACENTAE APPEARED NORMAL Nil) 8 (100.0) a. Dosage occurred on days 6 through 17 of gestation. II 1 8 7( 87.5) 7 15.7 + 2.6 13.6 + 3.2 13.3 + 3.0 93 13.3 + 3-0 0 0.3 + 0.5 2 0.3 + 0.5 0 0.0 + 0.0 2( 28.6) 0 7 (100.0) 7(100.0) III 5 8 8(100.0) 8 16.0 + 1.8 15.2 + 1.8 14.5 + 1.7 116 14.5 + 1.7 0 0.8 + 1.2 6 0.8 + 1.2 0 0.0 + 0.0 3( 37.5) 0 8(100.0) 8 (100.0) IV 10 a 8(100.0) 8 16.0 + 2.7 14.4 + 1.7 13.8 + 1.8 110 13.8 + 1.8 0 0.6 + 0.7 4 0.5 + 0.8 1 0-1 + 0.4 4( 50.0) 0 8(100.0) 8(100.0) o00 m>O-0 ro PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE e (PAGE 2) : CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a V 20 RATS TESTED N 8 PREGNANT N(%) 8(100.0) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 20 OF GESTATION N 8 CORPORA LUTEA MEAN+S.D. 15.5 2.1 IMPLANTATIONS MEAN+S.D , 15.1 + 1.7 LITTER SIZES MEAN+S.D. 14.8 + 1.8 LIVE FETUSES N MEAN+S.D . 118 14.8 + 1.8 DEAD FETUSES N 0 RESORPTIONS MEAN+S.D . 0.4 + 0.5 EARLY RESORPTIONS N MEAN+S.D . 3 0.4 + 0.5 LATE RESORPTIONS N MEAN+S.D . 0 0.0 + 0.0 DAMS WITH ANY RESORPTIONS N(%) 3( 37.5) DAMS WITH ALL CONCEPTUSES RESORBED N 0 DAMS WITH VIABLE FETUSES N(t) 8(100.0) PLACENTAE APPEARED NORMAL N(%) 8(100.0) a. Dosage occurred on days 6 through 17 of gestation. VI 25 a 8(100.0) 8 15.9 + 2.8 14.9 + 2.8 14.5 + 2.6 116 14.5 + 2.6 0 0.4 + 0.7 3 0.4 + 0.7 0 0 . 0 + 0.0 2( 25.0) 0 8(100.0) 8(100.0) VII 35 8 8(100.0) 8 16.2 + 3.4 15.1 + 2.2 14.2 + 2.4 114 14.2 + 2.4 0 0.9 + 1.1 7 0.9 + 1.1 0 0.0 + 0.0 4( 50.0) 0 8(100.0) 8(100.0) 0oI0 p m5o K> M 418-011: PAGE F-22 003772 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 7 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a I 0 (VEHICLE) LITTERS WITH ONE OR MORE LIVE FETUSES N 8 IMPLANTATIONS MEAN+S.D. 14.0 + 4.5 LIVE FETUSES N MEAN+S.D. 109 13.6 + 4.6 LIVE MALE FETUSES N 49 t LIVE MALE FETUSES/LITTER MEAN+S.D. 43.1 + 14.3 LIVE FETAL BODY HEIGHTS (GRAMS) /LITTER MEAN+S.D. 3.40 + 0.47 MALE FETUSES MEAN+S.D. 3. S3 + 0.47 FEMALE FETUSES MEAN+S.D. 3.30 + 0.44 % RESORBED CONCEPTUSES/LITTER MEAN+S.D. 2.7 + 5.6 a. Dosage occurred on days S through 17 of gestation. II 1 7 13.6 + 3.2 93 13.3 + 3.0 42 42.8 + 22.2 3.43 + 0.22 3.56 + 0.23 3.36 + 0.21 1.8 + 3.0 III 5 8 15.2 + 116 14.5 + 59 1.8 1.7 50.5 + 13.0 3.28 + 0.17 3.37 + 0.21 3.18 + 0.15 4.6 + 6.9 IV 10 8 14.4 + 110 13.8 + 50 1.7 1.8 46.7 + 13.8 3.34 + 0.16 3.42 + 0.19 3.25 + 0.16 4.4 + 5.0 A ooo 00 o T> o m>oi) > O m ro co PROTOCOL 41B-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 7 (PAGE 2) : LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a V 20 LITTERS WITH ONE OR MORE LIVE FETUSES N B IMPLANTATIONS MEAN+S.D. 15.1 + 1.7 LIVE FETUSES N MEAN+S.D. 118 14.8 + 1.8 LIVE MALE FETUSES N 66 % LIVE MALE FETUSES/LITTER MEAN+S.D. 55.3 + 9.1 LIVE FETAL BODY HEIGHTS (GRAMS) /LITTER MEAN+S.D. 3.24 + 0.15 MALE FETUSES MEAN+S.D. 3.34 + 0.17 FEMALE FETUSES MEAN+S.D. 3.13 + 0.14 % RESORBED CONCEPTUSES/LITTER MEAN+S.D. 2.6 + 3.6 a. Dosage occurred on days 6 through 17 of gestation. VI 25 8 14.9 + 116 14.5 + 59 2.8 2.6 49.2 + 13.8 3.11 + 0.18 3.21 + 0.21 3.03 + 0.20 2.4 + 4.4 VII 35 8 15.1 + 2.2 114 14.2 + 2.4 58 50.4 + 15.3 2.93 + 0.46 3.03 + 0.45 2.83 + 0.44 5.9 + 7.5 003773 0o10 -o > T3 O 5 G) m m *in ro M -fk PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 8 (PAGE 1) : FETAL GROSS EXTERNAL ALTERATIONS - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a I 0 (VEHICLE) LITTERS EVALUATED FETUSES EVALUATED LIVE N8 N 109 N 109 BODY: EDEMA LITTER INCIDENCE FETAL INCIDENCE N (%) Nil) 0( 0.0) 0( 0.0) PAIATE: CLEFT LITTER INCIDENCE PETAL INCIDENCE N (%) N (%) 0( 0.0) 0( 0.0) a. Dosage occurred on days 6 through 17 of gestation. II 1 7 93 93 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) III 5 8 116 116 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) IV 10 8 110 110 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 003774 -N 0oI0 TJ > I0 m n m NJ NJ 01 U1 PROTOCOL 418-OUP: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 8 (PAGE 2) : FETAL GROSS EXTERNAL ALTERATIONS - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY)a V 20 LITTERS EVALUATED FETUSES EVALUATED LIVE N8 N 118 N 118 BODY: EDEMA LITTER INCIDENCE FETAL INCIDENCE N(%> N (%) 0( 0.0) 0( 0.0) PALATE: CLEFT LITTER INCIDENCE FETAL INCIDENCE N (%) Nil) 0( 0.0) 0( 0.0) a. Dosage occurred on days 6 through 17 of gestation. b. Fetus 106S0-1 had other gross external alterations. c. Fetus 10650-10 had other gross external alterations. VI 25 8 116 116 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) VII 35 8 114 114 1( 12.5) 2 ( 1.8)b,c It 12.5) 13 ( 11.4 )b, c 003775 0oI0 1* O T3 > 13 O 5O m m jn 6o> PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 9 (PAGE 1) ; CLINICAL OBSERVATIONS - INDIVIDUAL DATA RAT U DOSAGE GROUP I 10601 10602 10603 10604 10605 10606 10607 10608 DOSAGE GROUP II 10609 10610 10611 10612 10613 10614 10615 10616 DG DAY OF PRESUMED GESTATION DESCRIPTION 0 (VEHICLE) MG/KG/DAY NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS 1 MG/KG/DAY NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS 418-011: PAGE F-27 003776 00 o "0 '>ij mo M PROTOCOL 41B-011P: ORAL (GAVAGB) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OP N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 9 (PAGE 2) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA RAT DESCRIPTION DOSAGE GROUP III S MG/KG/DAY 10617 10618 10619 10620 10621 10622 10623 10624 DG ( 13- 20) DG ( 14- 20) DG( 20 ) LOCALIZED ALOPECIA: NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LIMBS a UNDERSIDE a NECK a DOSAGE GROUP IV 10 MG/KG/DAY 1062S 10626 10627 10628 10629 10630 10631 10632 NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS DG DAY OP PRESUMED GESTATION a. observation confirmed at necropsy. 003777 00 6 9o TJ > o 5 ma m p to W oo 00 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 9 (PAGE 3): CLINICAL OBSERVATIONS - INDIVIDUAL DATA RAT # DESCRIPTION DOSAGE GROUP V 20 MG/KG/DAY 10633 10634 10635 10636 10637 10638 10639 10640 DG( 15- 17) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMODACRYORRHEA NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS DOSAGE GROUP VI 25 MG/KG/DAY 10641 10642 10643 10644 10645 10646 10647 10648 DG ( 14- 20) DG ( 15- 20) NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS UNDERSIDE a BACK a DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 418-011:PAGE F-29 003778 * O ;p o m rcoo PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 9 (PAGE 4): CLINICAL OBSERVATIONS - INDIVIDUAL DATA RAT DESCRIPTION DOSAGE GROUP VII 35 MG/KG/DAY 10649 10650 10651 10652 10653 10654 10655 0656 DG ( 13- 20) DG ( 13- 20) DG ( 17- 20) DG ( 8- 19) DG ( 20 ) DG ( 12 ) NO ADVERSE FINDINGS LOCALIZED ALOPECIA: UNDERSIDE a LOCALIZED ALOPECIA: BACK a EMACIATION a NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS ALOPECIA NO LONGER APPARENT NO ADVERSE FINDINGS NO ADVERSE PINDINGS CHROMORHINORRHEA DG - DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 003779 fe. 00 6 o U > 0 >m mCD T| 1 too PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 10 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP (MG/KG/DAY) I 0 (VEHICLE) II 1 RAT DAY OF PREGNANCY DOSAGES f NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a 10601 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10602 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10603 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10604 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10605 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10606 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10607 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10608 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10609 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10610 DG 20 NP 12 ALL TISSUES APPEARED NORMAL. 10611 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10612 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10613 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10614 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10615 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10616 DG 20 P 12 ALL TISSUES APPEARED NORMAL. P = PREGNANT NP * NOT PREGNANT DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 9} for external observations confirmed at necropsy. 003780 6oo ? O "0 > CD m 5O -n m 0 CO * PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 10 (PAGE 2) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a III 10617 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10618 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10619 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10620 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10621 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10622 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10623 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10624 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10625 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10626 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10627 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10628 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10629 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10630 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10631 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10632 DG 20 P 12 ALL TISSUES APPEARED NORMAL. P = PREGNANT NP * NOT PREGNANT DG - DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 9) for external observations confirmed at necropsy. 003781 *> 00 6 9o O > Tl o 5m om i co wM N) 003782 PROTOCOL 418-011P; ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 10 (PAGE 3) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a 10633 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10634 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10635 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10636 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10637 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10638 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10639 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10640 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10641 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10642 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10643 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10644 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10645 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10646 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10647 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10648 DG 20 P 12 ALL TISSUES APPEARED NORMAL. P = PREGNANT NP * NOT PREGNANT DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 9) for external observations confirmed at necropsy. 0O0 9 O *0 > TJ o ?Om m CO 10 CO to PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 10 (PAGE 4) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a VII 35 10649 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10650 DG 20 p 12 ALL TISSUES APPEARED NORMAL. 10651 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10652 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10653 DG 20 p 12 ALL TISSUES APPEARED NORMAL. 10654 DG 20 P 12 ALL TISSUES APPEARED NORMAL. 10655 DG 20 p 12 ALL TISSUES APPEARED NORMAL. 10656 DG 20 P 12 ALL TISSUES APPEARED NORMAL. P - PREGNANT NP - NOT PREGNANT DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 9) for external observations confirmed at necropsy. 003783 th. 00 6 s "D > TJ o TJ m >Q T 1 m CO -N 003784 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 11 (PAGE 1): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA RAT * DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAY 0 4 6 7 8 9 10601 P 10602 P 10603 P 10604 P 10605 P 10606 P 10607 P 10608 P 218. 227. 242. 245. 238. 232. 250. 235. 236. 253. 276. 278. 261. 273. 273. 249. 244. 263. 286. 280. 260. 275. 269. 254. 242. 270. 292. 290. 261. 271. 272. 257. 247. 273. 300. 293. 263. 280. 277. 264. 252. 281. 303. 299. 267. 284. 280. 265. DAY 17 16 19 20 10601 P 10602 P 10603 P 10604 P' 10605 P 10606 P 10607 P 10608 P 310. 338. 377. 353. 315. 348. 310. 324. 319. 361. 400. 371. 329. 363. 315. 343. 335. 371. 412. 390. 347. 392. 332. 359. 361. 399. 432. 414. 362. 405. 338. 374. P - PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 10 257. 287. 314. 306. 270. 294. 285. 273. 11 265. 291. 318. 312. 274 . 301. 291. 271. 12 270. 295. 321. 315. 282. 310. 288. 282. 13 274. 308. 335. 322. 285. 313. 295. 289. 14 274. 311. 341. 325. 289. 321. 301. 292. 15 282. 318. 348. 334. 297. 325. 301. 300. 16 295. 331. 364. 344. 301. 333. 304. 312. -tk. o4--Vkk. Gc>OmTTnO> CGDO OCO>mTrTi?OJ|i 418-011:PAGE F-36 003785 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 11 (PAGE 2) : MATERNAL BODY WEIGHTS - INDIVIDUAL DATA RAT DOSAGE GROUP II 1 MG/KG/DAY PREGNANCY STATUS DAY 0 4 6 7 8 9 10609 P 10610 NP 10611 P 10612 P 10613 P 10614 P 10615 P 10616 P 241. 235. 256. 219. 245. 230. 225. 239. 267. 247. 250. 246. 270. 256. 243. 251. 276. 250. 285. 251. 280. 258. 241. 260. 278. 251. 282. 255. 282. 262. 243. 262. 286. 255. 289. 260. 285. 267. 245. 261. 289. 248. 295. 265. 291. 274. 250. 272. DAY 17 18 19 20 10609 P 10610 NP 10611 P 10612 P 10613 P 10614 P 10615 P 10616 P 351. 266. 353. 325. 354. 311. 297. 320. 373. 270. 374. 342. 369. 328. 318. 332. 386. 265. 389. 357. 384. 331. 325. 352. 412. 265. 409. 383. 408. 337. 344. 358. P - PREGNANT NP > NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY - DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 10 296. 256. 301. 272. 296. 281. 259. 274. 11 302. 260. 306. 276. 297. 276. 256. 278. 12 309. 261. 307. 283. 309. 288. 258. 288. 13 313. 268. 313. 290. 310. 291. 268. 291. 14 321. 259. 314. 292. 321. 290. 279. 300. 15 330. 263. 321. 298. 324. 300. 280. 302. 16 342. 271. 340. 315. 342. 309. 288. 304. -N 00 oi fm5t Ol o> PROTOCOL 418 -011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 11 (PAGE 3) : MATERNAL BODY HEIGHTS - INDIVIDUAL DATA RAT DOSAGE GROUP III 5 MG/KG/DAY PREGNANCY STATUS DAY 0 4 6 7 8 9 10617 P 10618 P 10619 P 10620 P 10621 P 10622 P 10623 P 10624 P 242. 244. 223. 229. 230. 236. 240. 247. 265. 264 . 243. 258. 256. 260. 248. 270. 270. 273. 249. 265. 251. 268. 258. 277. 272. 276. 253. 266. 256. 270. 260. 279. 277. 260. 260. 272. 263. 274 . 264. 284. 281. 285. 264. 277. 267. 276. 267. 292. DAY 17 18 19 20 10617 P 10618 P 10619 P 10620 P 10621 P 10622 P 10623 P 10624 P 316. 339. 327. 324. 321. 328. 313. 341. 334. 351. 341. 342. 339. 340. 334. 359. 344. 366. 355. 352. 358. 365. 345. 377. 371. 384. 380. 377. 373. 369. 357. 389. P - PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY DAY OF PRESUMED GESTATION ALL WEIGHTS HERE RECORDED IN GRAMS (G) . 10 286. 291. 274. 280. 270. 280. 272. 298. 11 287. 299. 278. 286. 279. 287. 273. 306. 12 291. 300. 283. 290. 276. 288. 280. 299. 13 292. 306. 293. 302. 280. 288. 282. 304. 14 295. 310. 294. 300. 289. 294. 287. 311. 15 302. 316. 304. 307. 293. 297. 296. 320. 16 312. 327. 316. 317. 305. 309. 302. 328. A o010 o9 --* " > TJ O T m > O m n1 CO G~->4 418-011: PAGE F-38 003787 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-631S.7) TABLE 11 (PAGE 4) : MATERNAL BODY WEIGHTS - INDIVIDUAL DATA RAT DOSAGE GROUP IV 10 MG/KG/DAY PREGNANCY STATUS DAY 0 4 6 7 8 9 10625 P 10626 P 10627 P 10628 P 10629 P 10630 P 10631 P 10632 P 242. 223. 237. 235. 244. 234. 227. 246. 268. 234. 258. 254. 270. 254. 244. 258. 275. 237. 264. 253. 269. 257. 243. 264. 275. 239. 266. 256. 276. 244. 248. 268. 281. 240. 270. 262. 282. 233. 248. 271. 285. 243. 273. 265. 286. 246. 252. 265. DAY 17 18 19 20 10625 p 10626 P 10627 p 10628 P 10629 P 10630 P 10631 P 10632 P 341. 278. 311. 319. 340. 297. 294. 327. 345. 289. 327. 330. 356. 306. 315. 342. 365. 299. 332. 349. 377. 321. 330. 357. 384. 312. 353. 364. 398. 342. 342. 378. P = PREGNANT NP NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL HEIGHTS WERE RECORDED IN GRAMS (G) . 10 295. 250. 276. 270. 286. 249. 252. 273. 11 298. 252. 279. 278. 299. 256. 255. 281. 12 301. 256. 278. 285. 307. 268. 262. 286. 13 309. 259. 285. 289. 308. 260. 264. 293. 14 311. 262. 287. 292. 316. 269. 272. 297. 15 317. 264. 292. 302. 322. 276. 279. 317. 16 325. 272. 299. 311. 333. 285. 291. 314. 4k 4 0i9 O O) 00 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-631S.7) TABLE 11 (PAGE 5) : MATERNAL BODY WEIGHTS - INDIVIDUAL DATA RAT ft DOSAGE GROUP V 20 MG/KG/DAY PREGNANCY STATUS DAY 0 10633 P 10634 P 1063S P 10636 P 10637 P 10638 P 10639 P 10640 P 239. 231. 224. 228. 244. 237. 240. 247. 4 259. 260. 244. 260. 259. 250. 266. 270. 6 267. 260. 247. 269. 262. 254. 268. 270. 7 268. 256. 248. 268. 260. 254. 272. 260. 8 273. 264. 251. 273. 260. 251. 271. 262. 9 277. 269. 251. 283. 266. 255. 285. 262. DAY 17 18 19 20 10633 P 10634 P 10635 P 10636 P 10637 P 10638 P 10639 P 10640 P 326. 327. 277. 338. 318. 316. 331. 313. 334. 342. 285. 356. 326. 326. 347. 331. 351. 352. 293. 366. 340. 342. 362. 348. 372. 376. 314. 387. 363. 358. 376. 362. P PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 10 278. 272. 246. 284. 266. 262. 284. 269. il 282. 280. 249. 290. 275. 270. 293. 273. 12 289. 282. 255. 288. 278. 278. 298. 279. 13 295. 290. 255. 299. 281. 281. 304. 280. 14 296. 290. 261. 292. 290. 283. 306. 287. IS 304. 303. 257. 308. 294. 295. 311. 289. 16 312. 318. 266. 321. 302. 302. 322. 302. 003788 4--k. 00 A 6-A ?o ~ TO > CD TO > Q m T| fn w<o ID3 418-011P:PAGE 40 003789 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 11 (PAGE 6) : MATERNAL BODY HEIGHTS - INDIVIDUAL DATA RAT ft DOSAGE GROUP VZ 25 MG/KG/DAY PREGNANCY STATUS DAY 0 4 6 7 8 9 10641 P 10642 P 10643 P 10644 P 10645 P 10646 P 10647 P 10648 P 240. 221. 247. 236. 244. 233. 228. 239. 256. 237. 265. 262. 270. 249. 249. 275. 264 . 240. 270. 270. 275. 249. 260. 284. 256. 236. 268. 267. 275. 247. 261. 286. 262. 239. 269. 264. 278. 246. 266. 291. 269. 242. 276. 270. 282. 250. 273. 292. DAY 17 18 19 20 10641 P 10642 P 10643 P 10644 P 10645 P 10646 P 10647 P 10648 P 304. 281. 333. 273. 325. 276. 299. 322. 314. 297. 345. 303. 337. 285. 320. 343. 329. 304. 360. 324. 356. 302. 329. 352. 349. 323. 377. 340. 359. 318. 336. 364. P - PREGNANT NP NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL HEIGHTS HERE RECORDED IN GRAMS (G) . 10 265. 247. 281. 278. 283. 255. 276. 292. ii 274. 247. 288. 277. 285. 258. 274. 290. 12 273 . 244. 288. 281. 301. 261. 272. 288. 13 275. 245. 292. 282. 294. 262. 276. 290. 14 274. 248. 291. 270. 301. 256. 279. 295. 15 286. 261. 298. 263. 303. 254. 284. 299. 16 299. 272. 311. 260. 313. 267. 286. 311. 00 <b 418-011 PAGE F-41 003790 PROTOCOL 418-OUP: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-63I6.7) TABLE 11 (PAGE 7) : MATERNAL BODY HEIGHTS - INDIVIDUAL DATA RAT It DOSAGE GROUP VII 35 MG/KG/DAY PREGNANCY STATUS DAY 0 4 6 7 8 9 10649 P 10650 P 10651 P 10652 P 10653 P 10654 P 10655 P 10656 P 244. 237. 222. 227. 246. 231. 238. 247. 252. 253. 245. 243. 269. 256. 249. 275. 258. 266. 251. 248. 276. 261. 254. 275. 260. 264. 254. 250. 275. 268. 252. 272. 264 . 269. 262. 249. 269. 267. 255. 266. 264 . 261. 267. 251. 276. 273. 254. 274. DAY 17 18 19 20 10649 P 10650 P 10651 P 10652 P 10653 P 10654 P 10655 P 10656 P 291. 207. 316. 293. 300. 313. 289. 303. 305. 208. 334. 304. 316. 327. 302. 320. 316. 220. 350. 316. 331. 347. 324. 341. 332. 239. 374. 343. 350. 364. 337. 350. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL HEIGHTS HERE RECORDED IN GRAMS (G) . 10 267. 268. 268. 256. 274. 274. 242. 276. 11 268. 260. 275. 258. 281. 276. 248. 269. 12 274 . 252. 278. 260. 279. 281. 252. 262. 13 278. 235. 279. 266. 280. 259. 258. 287. 14 281. 223. 285. 269. 277. 289. 266. 265. 15 286. 216. 292. 275. 289. 297. 271. 266. 16 295. 213. 303. 289. 297. 306. 275. 292. O TJ 3 om 418-011: PAGE F-42 003791 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER! T-6316.7) TABLE 12 (PAGE 1) : MATERNAL FEED CONSUMPTION VALUES - INDIVIDUAL DATA PREGNANCY STATUS DAYS 0 - 4 4-6 6-8 8 - 10 10 - 12 12 - 14 14 - 16 16 - 18 18 - 20 RAT DOSAGE GROUP I 0 1(VEHICLE) MG/KG/DAY 10601 p 10602 P 10603 p 10604 P 10605 P 10606 p 10607 P 10608 P 76. 45. 40. 44, 45. 44. 47. 54. 49. 93. 56. 49. 49 a a 48. 53. 54. 97. 56. 56. 54 56. 61. 54 . 61. 54. 115. 62. 64. 61 60. 55. 45. 53. 61. 87. 44. 40. 43, 44. 46. 44 . 50. 52. 103. 51. 56. 53 57. 55. 51. 54. 56. 89. 47. 68. 57 60. 90. 55. 46. 53. 72. 37. 42. 46 45. 43. 43. 44. 40. RAT DOSAGE GROUP II 1 MG/KG/DAY 10609 p 10610 NP 10611 P 10612 P 10613 P 10614 P 10615 P 10616 P 97. 50. 49. 50 88. 44. 44. 39 77. 55. 51. 52 94. 46. 51. 54 93. 53. 46. 49 94. 43. 51. 50 61. 38. 41. 40 76. 42. 40. 43 50. 51. 50. 53. 53. 43. 42. 37. 49. 36. 49. 47. 50. 59. 57. 47. 52. 52. 57. 54 . 46. 54. 60. 62. 57. 48. 58. 53. 44 . 47. 38. 78. 41. 42. 51. 45. 51. 45. 40. 48. P > PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS * DAYS OF GESTATION ALL HEIGHTS HERE RECORDED IN GRAMS (G) . a. Spilled feed precluded the calculation of this value. 9 o 5 Om 4k. NJ 418-011: PAGE F-43 003792 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 12 (PAGE 2) : MATERNAL FEED CONSUMPTION VALUES - INDIVIDUAL DATA PREGNANCY STATUS DAYS 0 - 4 4 - 6 6 - 8 8 - 1 0 1 0 - 1 2 12 - 14 14 - 16 16 - 18 18 - 20 RAT * DOSAGE GROUP III 5 MG/KG/DAY 10617 P 1061B P 10619 P 10620 P 10621 P 10622 P 10623 P 10624 P 90. 48. 45. 45 94. 56. 22. 48 86. 49. 49. 51 95. 50. SO. 49 90. 46. 44. 47 95. 47. 47. 48 90. 44. 45. 47 103. 46. 50. 52 45. 40. 45. 49. 49. 50. 52. 51. 55. S3. 56. 54 . 53. 56. 56. 54. 48. 46. 48. 44. 46. 51. 48. 51. 52. 46. 42. 36. 47. 50. 44. 46. 48. 53. 43. 50. 43. 47. 48. 50. RAT # DOSAGE GROUP IV 10 MG/KG/DAY 10625 P 10626 P 10627 P 10628 P 10629 P 10630 P 10631 P 10632 P 91. 50. 45. 48 66. 33. 31. 38 90. 56. 47. 43 85. 42. 44. 45 85. 45. 47. 49 85. 46. 14. 35 77. 40. 36. 38 88. 43. 41. 37 48. 46 . 44 . 48. 50. 40. 35. 35. 37. 36. 45. 40. 34. 46. 50. 48. 50. 47. 46. 52. 50. 55. 46. 51. 55. 44. 40. 38. 39. 49. 40. 47. 45. 39. 41. 49. 50. 49. 49. 51. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF GESTATION ALL HEIGHTS HERE RECORDED IN GRAMS (G). 0oI0 U 5 o m CO 418-011 P:PAGE 44 003793 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 12 (PAGE 3) : MATERNAL FEED CONSUMPTION VALUES - INDIVIDUAL DATA PREGNANCY STATUS DAYS 0 - 4 4 - 6 6 - 8 8 - 1 0 10 - 12 12 - 14 14 - 16 16 - 18 18 - 20 RAT DOSAGE GROUP V 20 MG/KG/DAY 10633 P 10634 P 10635 P 10636 P 10637 P 10638 P 10639 P 10640 P 88. 50. 47. 46. 47. 48. 46. 51. 46. 89. 47. 41. 44. 46. 43. 47. 49. 44 . 78. 48. 31. 28. 33. 31. 21. 35. 41. 105. 62. 45. 57. 44. 48. 59. 59. 53. 87. 48. 41. 42. 50. 48. 48. 52. 51. 140. 47. 40. 44. 51. 48. 46. 43. 44. 95. 54. 54. 54. 52. 50. 46. 41. 51. 103. 47. 41. 40. 47. 53. 49. 44. 49. RAT DOSAGE GROUP VI 25 MG/KG/DAY 10641 P 10642 P 10643 P 10644 P 10645 P 10646 P 10647 P 10648 P 86. 48. 36. 40. 41. 33. 38. 37. 40. 79. 42. 32. 34. 35. 25. 41. 41. 38. 83. 47. 40. 45. 46. 38. 44. 53. 48. 94. 51. 38. 45. 43. 25. 11. 40. 46. 91. 54. 51. 45. 47. 40. 46. 39. 45. 73. 38. 31. 35. 37. 27. 26. 32. 43. 75. 38. 38. 45. 38. 39. 40. 48. 42. 102. 54. 50. 46. 34. 39. 38. 46. 42. P = PREGNANT NP NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS DAYS OF PRESUMED GESTATION ALL HEIGHTS HERE RECORDED IN GRAMS (G). A--X 00 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 12 (PAGE 4): MATERNAL FEED CONSUMPTION VALUES - INDIVIDUAL DATA PREGNANCY STATUS DAYS 0 - 4 4 - 6 6 - 8 0 - 1 0 10 - 12 12 - 14 14 - 16 16 - 18 18 - 20 RAT * DOSAGE GROUP VII 35 MG/KG/DAY 1064 P 10650 P 10651 P 10652 P 10653 P 10654 P 10655 P 10656 P 86. 48. 41. 41. 43. 41. 39. 38. 43. 87. 49. 45. 39. 17. 2. 1. 13. 34 . 94. 53. 49. 48. 45. 42. 45. 51. 52. 80. 43. 40. 38. 41. 39. 42. 44. 46. 95. 55. 48. 43. 47. 46. 41. 35. 62. 89. 46. 44. 46. 45. 46. 40. 36. 52. 82. 47. 37. 25. 39. 64. 43. 43. 55. 90. 42. 33. 42. 26. 42. 39. 38. 44 . P = PREGNANT NP NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS - DAYS OF PRESUMED GESTATION ALL HEIGHTS HERE RECORDED IN GRAMS (G) . 003794 cn--bk. <b ? O U > U 0 m n T| mL Ui 01 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 13 (PAGE 1): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT # M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 10601 6 7 6 7 13 0 0 0 0 0 0 0 0 0 6 7 13 6 7 13 10602 5 10 11 4 15 0 0 0 1 0 1 0 0 0 12 4 16 12 5 17 10603 12 5 9 8 17 0 0 0 0 0 0 0 0 0 9 8 17 11 8 19 10604 5 10 3 12 15 0 0 0 0 0 0 0 0 0 3 12 15 4 12 16 10605 6 5 8 3 11 0 0 0 0 2 2 0 0 0 8 5 13 8 6 14 10606 6 9 5 10 15 0 0 0 0 0 0 0 0 0 5 10 15 5 11 16 10607 1 3 i 3 4 0 0 0 0 0 0 0 0 0 1 3 4 5 8 13 10608 8 11 10 9 19 0 0 0 0 0 0 0 0 0 10 9 19 10 10 20 DOSAGE GROUP II 1 MG/KG/DAY 10609 11 5 7 9 16 0 0 0 0 0 0 0 0 0 7 9 16 7 11 18 10610 NOT PREGNANT 10611 2 13 8 7 15 0 0 0 0 1 1 0 0 0 8 8 16 9 9 18 10612 5 9 7 7 14 0 0 0 0 0 0 0 0 0 7 7 14 9 9 18 10613 9 6 9 6 15 0 0 0 0 1 1 0 0 0 9 7 16 10 7 17 10614 1 6 1 6 7 0 0 0 0 0 0 0 0 0 1 6 7 5 7 12 10615 7 6 4 9 13 0 0 0 0 0 0 0 0 0 4 9 13 5 9 14 10616 7 6 6 7 13 0 0 0 0 0 0 0 0 0 6 7 13 6 7 13 M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-011: PAGE F-46 0oI0 TJ T) > G) m j05. 96C 00 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OP N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 13 (PAGE 2): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT ft M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP III 5 MG/KG/DAY 10617 7 6 7 6 13 0 0 0 0 1 1 0 0 0 7 7 14 7 7 14 10618 3 10 4 9 13 0 0 0 1 1 2 0 0 0 5 10 15 7 10 17 10619 7 9 10 6 16 0 0 0 0 0 0 0 0 0 10 6 16 10 7 17 10620 8 7 6 9 15 0 0 0 3 0 3 0 0 0 9 9 18 9 9 18 10621 9 6 9 6 15 0 0 0 0 0 0 0 0 0 9 6 15 10 6 16 10622 7 8 7 8 15 0 0 0 0 0 0 0 0 0 7 8 15 7 8 15 10623 7 5 4 8 12 0 0 0 0 0 0 0 0 0 4 8 12 4 9 13 10624 11 6 6 11 17 0 0 0 0 0 0 0 0 0 6 11 17 6 12 18 DOSAGE GROUP IV 10 MG/KG/DAY 10625 5 10 11 4 15 0 0 0 1 0 1 0 0 0 12 4 16 13 8 21 10626 7 3 3 7 10 0 0 0 0 0 0 0 1 1 3 8 11 3 9 12 10627 6 7 9 4 13 0 0 0 0 1 1 0 0 0 9 5 14 9 5 14 10628 8 7 5 10 15 0 0 0 0 0 0 0 0 0 5 10 15 6 10 16 10629 6 9 8 7 15 0 0 0 0 0 0 0 0 0 8 7 15 8 7 IS 10630 8 5 10 3 13 0 0 0 0 0 0 0 0 0 10 3 13 10 5 15 10631 5 10 11 4 15 0 0 0 0 0 0 0 0 0 11 4 15 12 6 18 10632 5 9 6 8 14 0 0 0 2 0 2 0 0 0 8 8 16 8 9 17 M = MALE F FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-011P:PAGE 47 b. 00 I PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 13 (PAGE 3): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT * M P HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP V 20 MG/KG/DAY 10633 9 5 4 10 14 0 0 0 0 0 0 0 0 0 4 10 14 4 11 15 10634 9 7 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 8 8 16 10635 5 6 7 4 11 0 0 0 0 1 1 0 0 0 7 5 12 7 5 12 10636 8 7 7 8 15 0 0 0 1 0 1 0 0 0 8 6 16 a 8 16 10637 8 7 9 6 15 0 0 0 0 0 0 0 0 0 9 6 15 9 6 15 10638 6 8 6 8 14 00 0 0 0 0 0 0 0 6 8 14 6 8 14 10639 9 7 8 8 16 0 0 0 0 1 1 0 0 0 8 9 17 8 9 17 10640 12 5 7 10 17 0 0 0 0 0 0 0 0 0 7 10 17 8 11 19 DOSAGE GROUP VI 25 MG/KG/DAY 10641 4 9 7 6 13 0 0 0 0 0 0 0 0 07 6 13 7 6 13 10642 9 7 7 9 16 0 0 0 0 0 0 0 0 0 7 9 16 7 10 17 10643 8 7 9 6 15 0 0 0 0 0 0 0 0 0 9 6 15 9 8 17 10644 13 5 8 10 18 000 1 12 000 9 11 20 9 11 20 >10645 7 7 6 8 14 00 0 0 0 0 0 0 0 6 8 14 8 8 16 10646 7 6 5 8 13 0 0 0 0 0 0 0 0 0 5 8 13 7 8 15 10647 3 7 2 8 10 0 0 0 1 0 1 0 0 0 3 8 11 3 8 11 10648 8 9 10 7 17 0 0 0 0 0 0 0 0 0 10 7 17 11 7 18 M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 003797 -b00 t- <b 9o" ? TJ O m am T1 A oo 00 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 13 (PAGE 4): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT # M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP VII 35 MG/KG/DAY 10G49 3 8 6 5 11 0 0 0 1 1 2 0 0 0 7 6 13 7 6 13 10650 5 B 5 8 13 0 00 2 1 30 0 07 9 16 8 9 17 10651 9 8 10 7 17 0 0 0 0 0 0 0 0 0 10 7 17 10 8 18 10652 8 6 7 7 14 00 0 0 0 00 0 0 7 7 14 7 8 15 10653 6 7 8 5 13 00 0 1 0 10 0 0 9 5 14 9 5 14 10654 7 10 9 8 17 0 0 0 0 1 1 0 0 0 9 9 18 9 13 22 10655 9 3 5 7 12 00 0 0 0 0 0 0 0 5 7 12 5 7 12 10656 11 6 9 8 17 0 0 0 0 0 0 0 0 0 9 a 17 9 10 19 M * MALE F * FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERHISE. 418-011 P:PAGE 49 003798 00 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 14 (PAGE 1) : LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA NUMBER OF LIVE FETUSES AVERAGE FETAL BODY HEIGHT (G) RAT # MALE FEMALE TOTAL MALE FEMALE TOTAL a D0SAGE GROUP I 0 (VEHICLE) MG/KG/DAY 10601 10602 10603 10604 10605 10606 10607 10608 6 7 13 3.54 3.43 3.48 5 10 15 3.71 3.50 3.57 12 s 17 3.91 3.71 3.85 5 10 15 3.68 3.44 3.52 6 5 11 3.94 3.57 3.78 6 9 15 3.62 3.42 3.50 1 3 4 2.45 2.30 2.34 8 11 19 3.38 3.06 3.20 DOSAGE GROUP II 1 MG/KG/DAY 10609 10610 10611 10612 10613 10614 10615 10616 11 5 NOT PREGNANT 2 13 59 96 16 76 76 16 3.37 3.29 3.35 15 4.01 3.64 3.69 14 3.53 3.48 3.50 15 3.50 3.39 3.46 7 3.31 2.96 3.01 13 3.69 3.44 3.58 13 3.50 3.33 3.42 a. TOTAL = SUM OF FETAL HEIGHTS/NUMBER OF LIVE FETUSES. -- CONCEPTUSES ..... RESORBED N N% 13 0 0.0 16 1 6.2 17 0 0.0 15 0 0.0 13 2 15.4 15 0 0.0 4 0 0.0 19 0 0.0 16 0 0.0 16 1 6.2 14 0 0.0 16 1 6.2 7 0 0.0 13 0 0.0 13 0 0.0 003799 00 <b O 0 > o 5m o ti m CD Ool o 008C00 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 14 (PAGE 2) : LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA NUMBER OF LIVE FETUSES AVERAGE FETAL BODY WEIGHT (G) RAT * MALE FEMALE TOTAL MALE FEMALE TOTAL a DOSAGE GROUP III 5 MG/KG/DAY 10617 10618 10619 10620 10621 10622 10623 10624 7 6 13 3.63 3.32 3.49 3 10 13 3.40 3.19 3.24 7 9 16 3.18 2.91 3.03 8 7 15 3.38 3.36 3.37 9 6 15 3.62 3.29 3.48 7 8 15 3.02 3.07 3.05 7 5 12 3.44 3.08 3.29 ii 6 17 3.31 3.21 3.28 DOSAGE GROUP IV 10 MG/KG/DAY 10625 10626 10627 10628 10629 10630 10631 10632 5 10 15 3.08 3.30 3.22 7 3 10 3.73 3.19 3.57 6 7 13 3.42 3.33 3.37 8 7 15 3.47 3.38 3.43 6 9 15 3.51 3.26 3.36 8 5 13 3.42 3.14 3.32 5 10 15 3.43 3.45 3.44 5 9 14 3.27 2.92 3.05 a. TOTAL SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES. -- CONCEPTUSES ..... RESORBED N N% 14 1 7.1 15 2 13.3 16 0 0.0 18 3 16.7 IS 0 0.0 15 0 0.0 12 0 0.0 17 0 0.0 16 1 6.2 ii 1 9.1 14 1 7.1 15 0 0.0 15 0 0.0 13 0 0.0 15 0 0.0 16 2 12.5 418-011:PAGE F-51 4k 00 o "0 T3 S mat 003S01 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 14 (PAGE 3): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA NUMBER OF LIVE FETUSES AVERAGE FETAL BODY HEIGHT (G) RAT MALE FEMALE TOTAL MALE FEMALE TOTAL a DOSAGE GROUP V 20 MG/KG/DAY 10633 9 5 14 3.54 3.24 3.43 10634 9 7 16 3.26 3.04 3.16 1063S 10636 5 6 11 3.25 3.11 3.18 a 7 15 3.54 3.37 3.46 10637 8 7 15 3.17 2.92 3.05 10638 6 8 14 3.50 3.19 3.32 10639 9 7 16 3.33 3.16 3.25 10640 12 5 17 3.12 3.01 3.09 DOSAGE GROUP VI 25 MG/KG/DAY 10641 4 9 13 3.46 3.31 3.35 10642 9 7 16 3.12 2.89 3.02 10643 8 7 15 3.45 3.35 3.40 10644 13 s 18 2.90 2.80 2.87 10645 7 7 14 3.16 3.02 3.09 10646 7 6 13 3.05 3.02 3.03 10647 3 7 10 3.43 3.02 3.14 10648 8 9 17 3.10 2.85 2.96 a. TOTAL - SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES. -- CONCEPTUSES -.... RESORBED N N% 14 0 0.0 16 0 0.0 12 i 8.3 16 1 6.2 15 0 0.0 14 0 0.0 17 1 5.9 17 0 0.0 13 0 0.0 16 0 0.0 15 0 0.0 20 2 10.0 14 0 0.0 13 0 0.0 11 1 9.1 17 0 0.0 -fe. oo o ? TJ CD TJ m m> o Tl i Ol NO PROTOCOL 41B-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 14 (PAGE 4): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA NUMBER OF LIVE FETUSES AVERAGE FETAL BODY WEIGHT (G) RAT MALE FEMALE TOTAL MALE FEMALE TOTAL a DOSAGE GROUP VII 10649 10650 10651 10652 10653 10654 10655 10656 3 5 9 8 6 7 9 ii 35 MG/KG/DAY e 11 2.80 8 13 1.98 8 17 3.24 6 14 3.23 7 13 3.25 10 17 3.21 3 12 3.30 6 17 3.25 2.54 1.85 3.16 2.94 3.04 2.95 3.07 3.07 2.61 1.90 3.20 3.11 3.14 3.06 3.24 3.19 a. TOTAL - SUM OF FETAL HEIGHTS/NUMBER OF LIVE FETUSES. -- CONCEPTUSES.... RESORBED N N% 13 2 15.4 16 3 18.8 17 0 0.0 14 0 0.0 14 1 7.1 18 1 5.6 12 0 0.0 17 0 0.0 418-011: PAGE F-53 003802 0i3 O TJ TJ m>O PROTOCOL 418-OUP: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 15 (PAGE 1): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA 418-011 P:PAGE 54 003803 FETUS ft 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY RAT CLs 10601 6/ 7 10602 12/ 5 10603 11/ a 10604 4/12 10605 8/ 6 10606 5/11 10607 5/ 8 10608 10/10 FA MA FA MA FA FA /' FA MA FA FA MA MA MA 3.34 3.64 3.28 3.61 3.40 3.68 3.34 3.24 3.65 3.30 3.49 3.64 3.65 MA FA FA MA MA FA E FA FA FA FA FA / FA FA MA MA 3.37 3.38 3.55 3-31 3.69 3.19 3.57 3.58 3.29 3.59 3.71 3.52 3.63 4.00 4.18 FA FA MA FA MA MA MA MA MA / MA FA MA MA MA MA FA MA 3.59 3.84 4.14 3.79 3.70 3.92 4.03 3.93 3.69 3.82 3.74 3.80 3.85 3.90 3.90 3.58 4.26 MA FA FA / FA MA FA FA MA FA FA MA MA FA FA FA 3.81 3.54 3.86 2.62 3.74 3.35 3.38 3.73 3.39 3.13 3.53 3.60 3.72 3.58 3.81 FA FA FA MA MA FA FA MA / MA E E MA MA 3.41 3.67 3.69 4.08 3.82 3.69 3.41 3.90 3.95 3.85 4.06 FA FA FA FA MA /' MA MA FA MA FA FA FA FA MA MA 3.80 3.56 3.48 3.61 3.68 3.71 3.68 3.47 3.57 3.07 3.40 3.07 3.31 3.42 3.64 MA / FA FA FA 2.45 2.35 2.41 2.14 JMA FA FA MA FA FA PA FA FA MA ' MA FA MA MA MA FA MA FA FA 3.13 3.19 3.22 3.22 2.76 3.29 3.29 2.93 2.95 3.66 3.20 3.12 3.45 3.34 3.44 2.97 3.59 2.86 3.11 DOSAGE GROUP II 1 MG/KG/DAY 10609-- 7/11 MA FA MA FA MA MA MA i/ M A FA MA MA MA FA MA FA MA 3.39 3.15 3.29 3.64 3.45 3.73 3.07 3.27 3.17 3.28 3.43 3.29 3.29 3.23 3.18 3.68 10610 NOT PREGNANT 10611 9/ 9 10612 9/ 9 10613 10/ 7 10614 5/ 7 10615 5/ 9 10616 6/ 7 FA FA FA FA FA MA FA FA / FA FA FA E FA FA FA MA 3.84 3.60 3.39 3.82 3.56 4.06 3.59 3.93 3.47 3.50 3.46 3.82 3.68 3.70 3.96 MA FA FA MA MA FA FA ,/ M A FA FA FA FA MA FA 3.23 3.50 3.44 3.55 3.86 3.72 3.23 3.27 3.45 3.48 3.45 3.43 3.74 3.62 FA MA FA FA MA FA MA FA MA / MA MA MA MA MA FA E 3.18 3.44 3.62 3.36 3.52 3.31 3.44 3.41 3.52 3.39 3.35 3.38 3.59 3.87 3.47 FA /F FA FA FA FA MA FA 2.95 2.94 2.77 3.06 3.22 3.31 2.84 MA FA FA FA / MA MA FA MA MA FA FA MA MA 3.59 3.21 3.61 3.50 3.50 3.64 3.57 3.96 3.80 3.31 3.45 3.63 3.71 FA MA MA FA FA MA / FA MA MA MA FA MA FA 3.33 3.56 3.66 3.41 3.06 3.39 3.39 3.70 3.36 3.33 3.49 3.54 3.30 M = MALE F = FEMALE A ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G) . 418-011: PAGE F-55 003804 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE -RADGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 15 (PAGE 2) : FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA FETUS 8 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 16 19 20 21 22 23 DOSAGE GROUP III 5 MG/KG/DAY RAT 8 CLs 10617 7/ 7 10618 7/10 10619 10/ 7 10620 9/ 9 10621 10/ 6 10622 7/ 8 10623 4/ 9 10624 6/12 MA MA MA FA FA MA MA / FA MA MA FA FA E FA 3.39 3.35 3.99 3.36 3.39 3.87 3.74 3.01 3.40 3.69 3.07 3.54 3.56 MA E FA FA FA / FA FA FA FA MA MA FA E FA FA 3.67 3.29 3.14 3.54 2.97 3.07 3.11 3.20 3.26 3.26 3.14 3.22 3.26 MA FA MA FA MA MA FA FA MA FA / FA MA FA MA FA FA 3.36 3.38 3.25 2.96 3.12 3.17 3.11 2.63 2.89 2.60 2.99 3.29 2.56 3.15 2.80 3.20 FA MA E E MA FA MA E FA / FA MA FA MA FA MA MA MA 3.28 4.00 3.52 3.51 3.28 3.40 3.38 3.26 3.43 3.31 3.07 2.72 3.49 3.43 FA MA MA MA MA MA FA MA FA / FA FA MA MA FA MA 2.91 3.55 3.57 3.56 3.59 3.72 3.32 3.55 3.56 3.08 3.47 3.67 3.50 3.40 3.83 MA FA MA MA FA FA MA / FA FA MA MA MA FA FA FA 3.25 3.20 3.10 3.18 3.07 2.85 2.54 2.95 3.10 3.04 3.32 2.72 3.38 3.16 2.86 FA MA MA MA / FA MA MA FA FA FA MA MA 2.96 3.28 3.40 3.34 3.24 3.81 3.41 3.02 3.13 3.04 3.40 3.46 FA FA MA MA FA MA / MA MA FA MA MA MA FA MA MA MA FA 3.20 3.11 3.23 3.44 3.40 3.32 3.17 3.27 3.20 3.36 3.40 3.35 3.06 3.10 3.59 3.22 3.28 FA 3.46 DOSAGE GROUP IV 10 MG/KG/DAY 10625 13/ 8 10626 3/ 9 10627 9/ 5 10628 6/10 10629 8/ 7 10630 10/ 5 10631 12/ 6 10632 8/ 9 MA MA FA FA MA E FA FA FA FA FA MA / FA FA MA FA 2.64 3.09 2.76 3.49 3.34 3.33 3.52 3.44 3.16 3.45 3.13 3.26 3.59 3.20 2.98 FA MA MA /' FL MA FA MA MA MA FA MA 3.71 4.02 3.74 1.68 3.78 3.42 3.57 3.76 3.51 2.44 3.72 FA FA FA MA MA MA FA FA FA /1FA MA E MA MA 3.25 3.30 3.17 3.77 3.49 3.50 3.56 3.40 3.17 3.47 3.74 3.40 2.64 FA FA MA MA FA / FA MA FA MA FA FA MA MA MA MA 3.24 3.66 3.59 3.63 3.30 3.05 3.41 3.48 3.28 3.43 3.50 3.52 3.44 3.24 3.65 FA MA FA FA FA FA FA FA / MA MA FA MA FA MA MA 3.13 3.60 3.39 3.65 3.31 2.94 3.50 2.85 3.51 3.22 3.34 3.50 3.22 3.46 3.75 MA FA FA MA MA MA MA FA FA FA ,/ MA MA MA 3.08 3.10 3.26 3.52 3.33 3.40 3.33 3.08 3.26 3.00 3.51 3.62 3.61 MA FA MA FA FA FA FA FA MA MA FA / FA FA MA FA 3.32 3.61 3.30 3.52 3.23 3.20 3.39 3.30 3.44 3.63 3.52 3.54 3.48 3.47 3.71 FA FA MA E MA FA FA E / FA FA FA MA MA FA FA MA 3.08 3.02 3.47 3.38 3.15 1.49 3.01 3.12 3.10 2.63 3.40 3.26 3.07 3.48 = MALE F S FEMALE A = ALIVE E = EARLY RESORPTIONI L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G) . 00 U > m0 CJI 01 PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-63I6.7) TABLE 15 (PAGE 3): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA 003805 FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 IB 19 20 21 22 23 DOSAGE GROUP V 20 MG/KG/DAY RAT # 10633 10634 10635 10636 10637 10638 10639 10640 CLs 4/11 8/ 8 7/ 5 8/ 8 9/ 6 6/ 8 8/ 9 8/11 MA FA MA MA / FA MA FA FA MA MA FA MA MA MA 3.79 3.65 3.70 3.65 2.88 3.58 3.03 3.34 3.56 3.34 3.29 3.53 3.42 3.25 FA MA FA MA MA MA MA MA / FA FA FA FA MA MA MA FA 2.80 3.11 3.18 3.43 3.07 3.22 3.43 3.16 2.63 3.26 3.24 3.24 3.43 3.32 3.18 2.95 FA MA FA FA FA MA FA / FA MA MA MA E 3.29 3.48 3.19 3.31 3.14 3.15 2.81 2.93 3.19 3.15 3.29 MA FA MA E FA FA FA FA / FA FA MA MA MA MA MA MA 3.35 3.26 3.64 3.50 3.45 3.35 3.57 3.00 3.46 3.53 3.59 3.61 3.71 3.48 3.44 FA FA MA MA MA MA FA FA FA /r FA MA FA MA MA MA 2.58 2.89 3.02 3.12 3.15 3.12 2.94 3.21 2.87 2.94 3.19 2.99 3.36 3.14 3.23 MA FA FA MA FA FA / FA MA FA MA FA MA FA MA 3.50 3.09 3.53 3.49 3.06 3.31 3.09 3.16 3.22 3.33 2.84 3.69 3.41 3.82 MA MA MA FA FA MA FA MA / MA E MA FA FA FA MA FA MA 3.34 3.52 3.75 3.34 3.40 3.24 2.71 3.58 3.09 3.25 3.19 3.37 3.25 3.34 2.83 2.83 MA MA MA MA FA MA MA ,/ M A FA MA FA FA MA MA FA MA MA 3.21 3.21 3.43 3.19 3.05 3.04 2.99 3.16 3.00 3.28 2.99 3.14 3.13 3.06 2.87 2.77 3.04 DOSAGE GROUP VI 25 MG/KG/DAY 10641 7/ 6 10642 7/10 10643 9/ 8 10644 9/11 10645 8/ 8 10646 7/ 8 10647 3/ 8 10648 11/ 7 FA FA MA FA FA FA MA ,/ MA FA FA FA FA MA 3.49 3-42 3.69 3.47 3.14 3.22 3.21 3.53 3.11 3.30 3.24 3.38 3.40 FA FA MA MA MA FA MA ,/ F A MA FA MA MA FA MA FA MA 3.05 2.68 3.17 3.07 3.07 3.02 3.26 2.90 3.15 2.88 3.06 3.24 2.92 3.20 2.76 2.89 MA FA FA FA MA MA MA FA FA /' MA FA FA MA MA MA 3.44 3.33 3.41 3.44 3.68 3.52 3.47 3.33 3.25 3.51 3.17 3.51 3.22 3.25 3.51 MA MA FA FA FA E MA MA MA /' MA MA E FA MA MA MA MA FA MA MA 2.92 2.64 2.79 2.81 2.83 3.10 2.97 3.00 3.01 2.74 2.66 2.89 3.02 2.81 2.95 2.90 2.89 2.73 FA FA MA MA MA FA / MA FA FA MA FA MA MA FA i3.05 3.16 3.26 3.26 3.05 3.06 3.02 3.01 3.16 3.34 2.97 3.13 3.09 2.76 MA FA FA MA MA , MA FA MA FA FA MA MA FA 3.07 3.04 3.16 3.02 2.97 2.95 3.02 3.13 2.77 3.15 3.12 3.07 2.98 FA E FA / MA FA FA MA FA FA FA MA 3.25 3.18 3.37 3.18 2.66 3.61 2.84 2.92 3.12 3.32 MA FA MA MA FA FA MA FA FA MA / MA MA MA FA FA FA FA 2.99 2.90 3.12 2.77 3.12 2.73 3.15 2.95 2.75 3.26 3.12 3.19 3.17 3.03 2.76 2.81 2.59 M = MALE F * F&4ALS A = ALIVE E EARLY RESORPTION L = LATE RESORPTION /" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G) . s-tfc ? O -a > 0 5m m0 -n 1 01 Ol PROTOCOL 418-011P: ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS (SPONSOR'S STUDY NUMBER: T-6316.7) TABLE 15 (PAGE 4) : FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA FETUS ft 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 DOSAGE GROUP VII 35 MG/KG/DAY RAT ft CLs 10649 7/ 6 10650a 8/ 9 10651 10/ 8 10652 7/ a 10653 9/ 5 10654 9/13 10655 5/ 7 10656 9/10 FA E FA MA FA MA MA / FA E FA FA FA FA 2.36 2.61 2.85 2.44 2.80 2.75 2.35 2.61 2.30 2.57 3.08 FA FA FA E E FA MA / E MA MA FA FA MA FA FA MA 2.01 1.75 2.08 2.20 2.15 2.04 2.12 1.66 1.14 1.64 2.02 1.95 1.95 FA MA FA FA FA FA MA MA MA FA /' MA MA MA MA FA HA FA 3.04 3.34 3.03 3.13 2.96 3.35 3.25 3.03 3.14 3.22 2.98 3.19 3.35 3.20 3.10 3.66 3.46 MA MA HA FA HA MA FA / FA MA FA MA FA MA FA 3.07 3.29 3.51 3.07 3.27 3.00 2.88 3.19 3.21 3.20 3.45 2.72 3.04 2.59 FA MA HA FA FA MA MA FA E / FA FA MA MA FA 3.04 3.20 3.09 2.71 2.90 3.46 2.93 3.22 3.27 2.94 3.29 3.55 3.17 FA MA FA FA HA MA FA FA FA / MA FA E FA MA MA MA FA FA 2.76 3.03 3.06 3.20 2.97 3.22 2.86 2.80 3.12 3.21 2.98 2.81 3.46 3.37 3.24 3.01 2.87 MA MA MA FA FA / FA MA MA MA MA MA MA 3.23 3.36 3.40 3.17 3.23 2.82 3.53 3.32 3.03 3.21 3.10 3.52 MA FA MA FA FA MA MA MA MA / MA FA FA MA FA MA MA MA 3.37 3.30 3.12 2.85 3.22 3.35 3.33 3.52 3.13 3.22 3.11 3.36 3.45 2.56 3.21 2.84 3.25 M = MALE F - FEMALE A ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G) . a. Fetuses 10650-1 and 10650-10 had cleft palate and an edematous body, and fetuses 10650-2, 10650-3, 10650-6, 10650-7, 10650-9, 10650-11, 10650-12, 10650-13, 10650-14, 10650-15 and 10650-16 had cleft palate. 418-011 P A G E F-57 003806 4k. ooo "0 TJ > n m Ul -I 418-011 .PAGE F-58 418-011P:PAGE 58 ATTACHMENT 1 PROTOCOL AND AMENDMENT 003807 418-011: PAGE F-59 418-011P:PAGE 59 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044 T: (215)443-8710 F: (215) 443-8587 PROTOCOL 418-011P SPONSOR'S STUDY NUMBER: T-6316.7 STUDY TITLE: PURPOSE: TESTING FACILITY: STUDY DIRECTOR: SPONSOR: STUDY MONITOR: ALTERNATE STUDY MONITOR: Oral (Gavage) Dosage-Range Developmental Toxicity Study of N-EtFOSE in Rats The purpose of this study is to provide information for the selection of dosages to be used in the developmental toxicity (embryo-fetal toxicity and teratogenic potential) study of N-EtFOSE administered orally via gavage to Crl:CDBR VAF/Plus presumed pregnant female rats. Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Telephone: (215)443-8710 Telefax: (215)443-8587 Raymond G. York, Ph.D., DABT Associate Director of Research 3M Toxicology Services 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 Marvin T. Case, D.V.M., Ph.D. Telephone: (612)733-5180 Telefax: (612)733-1773 Andrew M. Seacat, Ph.D. Telephone: (612) 575-3161 Telefax: (612) 733-1773 003808 418-011 PAGE F-60 418-011 P:PAGE 60 Protocol 418-011P Page 2 REGULATORY CITATIONS: U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guideline on detection of toxicity to reproduction for medicinal products. Fed eral Register, September 22,1994, Vol. 59, No. 183. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). G ood Laboratory Practice Standard for S afety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. European Economic Community (1989). Council decision on 2 8 July 1989 on the acceptance by the European Economic Community o f an O EC D decision/recom- m endation on com pliance with principles o f goo d laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. REGULATORY COMPLIANCE: This study will be conducted in the spirit of the Good Laboratory Practice (GLP) regulations cited above in that the Testing Facility personnel will adhere to the Standard Operating Procedures for laboratory operations and data collection. The Testing Facility Quality Assurance Unit (QAU) will not audit the protocol, the raw data, the reports or the critical phases of the study. All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintained with the protocol. STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. 003809 418-011 .PAGE F-61 418-011P:PAGE 61 Protocol 418-011P Page 3 TEST ARTICLE AND VEHICLE: Identification: Test Article: Name: Physical Description: Lot/Batch Number: Specific Gravity: Purity: Expiration Date: N-EtFOSE. Waxy solid. FM-3929 (30035, 30037, 30039). -1.7. 99.1%. May, 2000. Information on the identity, composition, strength and purity of the test article is on file with the Sponsor. Vehicle: 2% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). Supplier and lot identification of Tween 80 to be documented in the raw data. Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the vehicle that would interfere with the results of this study. Therefore, no analyses other than those mentioned in this protocol will be conducted. Safety Precautions: Gloves, mask, appropriate eye protection and a uniform/lab coat are to be worn during formulation preparation and administration. The Material Safety Data Sheet (MSDS) is attached to the protocol (ATTACHMENT 2). Storage: Bulk Test Article: Vehicle Components: Prepared Vehicle: Prepared Formulations: Room temperature. Room temperature. Room temperature. Frozen (-20C). All test article shipments to the Testing Facility should be addressed to the attention of Julian Gulbinski, Manager of Formulations, at the previously cited address and telephone number. Shipments should include information concerning storage conditions and shipping cartons should be labeled appropriately. The recipient should be notified in advance of shipment 003810 418-011:PAGE F-62 418-01 IP :P A G E 62 Protocol 418-011P Page 4 FORMULATION: Frequency of Preparation: Formulations (suspensions) will be prepared daily at the Testing Facility. Vehicle will be prepared weekly at the Testing Facility. Detailed preparation procedures are attached to this protocol (ATTACHMENT 3). Adjustment for Purltv: The test article will be considered 100% pure for the purpose of dosage calculations. Testing Facility Reserve Samples: The Sponsor will reserve a sample (1 g) of each lot of the bulk test article used during the course of this study. The Testing Facility will reserve a sample (5 mL) of each lot of the vehicle components used during the course of this study. Samples will be stored under the previously cited conditions. ANALYSES: Samples additional to those described below may be taken if deemed necessary during the course of the study. Bulk Test Article Sampling: No analyses of the bulk test article will be conducted during the course of this study. Information on the stability of the bulk test article is on file with the Sponsor. Analyses of Prepared Formulations: At the request of the Sponsor, no analyses of prepared test article formulations will be conducted during the course of the study. However, records will be maintained to document how the test article formulations were prepared. DISPOSITION: Prepared formulations will be discarded at the Testing Facility. All remaining bulk test article will be returned to the Study Monitor at the previously cited address. 003811 418-011: PAGE F-63 418-011 P:PAGE 63 Protocol 418-011P Page 5 TEST SYSTEM: Soeeies/Strain and Reason for Selection: The Crl:CDBR VAF/Plus (Sprague-Dawley) rat was selected as the Test System because: 1) it is one mammalian species accepted and widely used throughout industry for nonclinical studies of developmental toxicity (embryo-fetal toxicity/teratogenicity); 2) this strain has been demonstrated to be sensitive to developmental toxins; 3) historical data and experience exist at the Testing Facility0*3'; and 4) the test article is pharmacologically active in the species and strain. Number: Initial population acclimated: 75 virgin female rats. Population selected for study: 56 mated female rats (8 per dosage group). Body Weight and Age: Female rats will be ordered to have body weights of 200 g to 225 g each at receipt, at which time they will be expected to be at least 60 days of age. Actual body weights will be recorded the day after receipt and will be documented in the raw data. Sex: Female rats will be given the test article. Male rats of the same source and strain will be used only as breeders and are not considered part of the Test System. Source: Charles River Laboratories, Inc. The rats will be shipped in filtered cartons by air freight and/or truck from Charles River Laboratories, Inc., to the Testing Facility. identification: Rats are permanently identified using Monel self-piercing ear tags (Gey Band and Tag Co., Inc., No. MSPT 20101). Male rats are given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats are assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study on the basis of day 0 of presumed gestation body weights. 003812 418-011 PAG E F-64 418-011 P:PAGE 64 Protocol 418-011P Page 6 ANIMAL HUSBANDRY: All cage sizes and housing conditions are in compliance with the Guide for the Care and Use o f Laboratory A nim ate. Housing: The rats will be individually housed in stainless steel, wire-bottomed cages except during the cohabitation period. During cohabitation, each pair of rats will be housed in the male rat's cage. No nesting materials will be supplied because the female rats will be sacrificed before parturition is expected. Room Air. Temperature and Humidity: The animal room is independently supplied with at least ten changes per hour of 100% fresh air that has been passed through 99.97% HEPA filters (Airo Clean room). Room temperature will be maintained at 64F (18C) to 79F (26C) and monitored constantly. Room humidity will also be monitored constantly and maintained at 30% to 70%. Light: An automatically controlled 12-hour light:12-hour dark fluorescent light cycle will be maintained. Each dark period will begin at 1900 hours EST. Diet: Rats will be given Certified Rodent Diet #5002 (PMI Nutrition International) available ad libitum from individual feeders. Water: Water will be available ad libitum from individual bottles attached to the cages or from an automatic watering access system. All water will be from a local source and passed through a reverse osmosis membrane before use. Chlorine will be added to the processed water as a bacteriostat; processed water is expected to contain no more than 1.2 ppm chlorine at the time of analysis. Water is analyzed monthly for possible bacterial contamination and twice annually for possible chemical contamination. Contaminants: Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the certified diet or in the drinking water at levels that would interfere with the results of this study. Therefore, no analyses other than those routinely performed by the feed supplier or those mentioned in this protocol will be conducted. O03813 418-011:PAGE F-65 418-011 P:PAGE 65 Protocol 418-011P Page 7 RANDOMIZATION AND COHABITATION: Upon arrival, male and female rats will be assigned to individual housing on the basis of computer-generated random units. After acclimation, virgin female rats will be cohabited with breeder male rats, one male rat per female rat. The cohabitation period will consist of a maximum of five days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ will be considered to be at day 0 of presumed gestation and assigned to individual housing. Healthy mated female rats will be assigned to dosage groups based on computer generated (weight-ordered) randomization procedures. ADMINISTRATION: Route and Reason for Choice: The oral (gavage) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately administered; and 2) it is one of the possible routes of human exposure. Method and Frequency: Female rats will be given the test article once daily on days 6 through 17 of presumed gestation, the period of organogenesis. Dosages will be adjusted for the most recently recorded body weight and given at approximately the same time each day. Rationale for Dosage Selection: Dosages will be selected by the Sponsor on the basis of previous studies conducted with the test article. 003814 418-011: PAGE F-66 418-011P:PAGE 66 Protocol 418-011P Page 8 Dosage Levels. Concentrations and Volumes: Dosage Group Number of Rats Dosage (mg/kg/day) Concentration (mg/mL) Dosage Volume (mL/kg) Argus Batch Number 1 8 0 (Vehicle) 0 5 B-418-011 P-A(Dav.Month.Year) II 8 1 0.2 5 B-418-011P-B(Dav.Month. Year) III 8 5 1 5 B-418-011 P-C(Day.Month.Year) IV 8 10 2 5 B-418-011 P-D(Day.Month.Year) V8 20 4 5 B-418-011 P-E(Day.Month.Year) VI 8 25 5 5 B-418-011-P-F(Dav.Month. Year) 35 7 5 B-418-01 l-P-G(Dav.Month.Year) The test article will be considered 100% pure for the purpose of dosage calculations. TESTS. ANALYSES AND MEASUREMENTS: Viability: All Periods: At least twice daily. Clinical Observations and/or General Appearance: Acclimation Period: Weekly. Predosage Period: Day 0 of presumed gestation. Dosage Period: Twice daily. Onceapproximately one hour postdosage and then four to six hours later. Postdosage Period: Once daily. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or Study Monitor. Body Weights: Acclimation Period: Weekly. Predosage Period: Days 0 and 4 of presumed gestation. Dosage Period: Daily. Postdosage Period: Daily. 003815 418-011: PAGE F-67 418-011P:PAGE 67 Protocol 418-011P Page 9 Feed Consumption Values (recorded and tabulated): Predosage Period: Days 0 and 4 of presumed gestation. Dosage Period: Days 6, 8,10, 12,14 and 16 of presumed gestation. Postdosage Period: Days 18 and 20 of presumed gestation. Feed consumption values may be recorded more frequently if it is necessary to replenish the feed. These intervals will not be tabulated. Mating Performance: Mating will be evaluated daily during the cohabitation period and confirmed by observation of spermatozoa in a smear of the vaginal contents and/or a copulatory plug observed in situ. Caesarean-Sectioning Observations: Rats will be Caesarean-sectioned on day 20 of presumed gestation. The fetuses will be removed from the uterus and placed in individual containers. The rats will be examined for number and distribution of: Corpora Ltea. Implantation Sites. [Placentae that appear abnormal (size, color or shape) will be noted in the raw data.] Live and Dead Fetuses. (A live fetus is defined as one that responds to stimuli; a dead fetus is defined as a term fetus that does not respond to stimuli and that is not markedly autolyzed; dead fetuses demonstrating marked to extreme autolysis are considered to be late resorptions.) Early and Late Resorptions. (A conceptus is defined as a late resorption if it is grossly evident that organogenesis has occurred; if this is not the case, the conceptus is defined as an early resorption.) 003816 I 418-011: PAGE F-68 418-011P:PAGE 68 Protocol 418-011P Page 10 Fetal Observations: Fetuses will be examined for sex and for gross external alterations. Late resorptions and dead fetuses will be examined for gross external alterations to the extent possible. The body weight of each fetus will be recorded. Only body weights of live fetuses will be used to determine litter fetal body weight averages. Fetuses with gross external alterations will be fixed in Bouin's solution; all other fetuses will be discarded. Representative photographs of fetal gross external alterations will be taken. METHOD OF SACRIFICE: Rats will be sacrificed by carbon dioxide asphyxiation. Live fetuses will be sacrificed by an intraperitoneal injection of euthanasia solution (Beuthanasia-D Special, manufactured by Schering-Plough Animal Health). NECROPSY: Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation (a table of random units will be used to select one control group rat from which ail tissues examined at necropsy will be retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions). Unless specifically cited below, all other tissues will be discarded. Scheduled Sacrifice: On day 20 of presumed gestation, female rats will be Caesarean-sectioned, and a gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. Uteri of apparently nonpregnant rats will be stained with 10% ammonium sulfide to confirm the absence of implantation sites<5). Rats Found Dead or Moribund: Rats that die or are sacrificed because of moribund condition, abortion or premature delivery will be examined for the cause of death or moribund condition on the day the observation is made. The rats will be examined for gross lesions. Pregnancy status and uterine contents of female rats will be recorded. Aborted fetuses and/or delivered pups will be examined to the extent possible, using the same methods described for fetuses. Uteri of apparently nonpregnant rats will be stained with 10% ammonium sulfide to confirm the absence of implantation sites(5). 003817 418-011 PAGE F-69 418-011 pipa g e 69 Protocol 418-011P Page 11 STATISTICAL EVALUATION: Averages and percentages will be calculated. Litter values will be used where appropriate. Additional procedures and/or analyses may be performed if deemed appropriate. DATA ACQUISITION. VERIFICATION AND STORAGE: Data will be hand- and/or computer-recorded. Records will be reviewed by the Study Director and/or appropriate management personnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All original data will be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the draft final report, after which time the Sponsor will be contacted to determine the disposition of these materials. RECORDS TO BE MAINTAINED: Protocol and Amendments. Test Article, Vehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (if prescribed by Staff Veterinarian). General Comments. Clinical Observations and/or General Appearance. Body Weights. Feed Consumption Values. Caesarean-Sectioning and Fetal Observations. Gross Necropsy Observations. Organ Weights (if required). Photographs (if required). Study Maintenance (room and environmental records). Feed and Water Analyses. Packing and/or Shipment Lists. 003818 418-011P:PAGE 70 Protocol 418-011P Page 12 KEY PERSONNEL: Executive Director of Research: Mildred S. Christian, Ph.D., ATS Director of Research: Alan M. Hoberman, Ph.D., DABT Associate Director of Research and Study Director: Raymond G. York, Ph.D., DABT Director of Laboratory Operations: John F. Barnett, B.S. Manager of Study Coordination: Valerie A. Sharper, M.S. Manager of Animal Operations and Member, Institutional Animal Care and Use Committee: Dena C. Lebo, V.M.D. Manager of Regulatory Compliance: Kathleen A. Moran, M.S. Consultant, Veterinary Pathology: W. Ray Brown, D.V.M., Ph.D., ACVP REPORT: A letter report for the purpose of dosage selection for the full study will be prepared immediately following completion of the in-life phase. A summary report will be prepared including: abstract, summaries of the methods, results and conclusion: table of contents; copy of the protocol: amendments: summary and individual tables; and reports of supporting data (if appropriate). The report will be included as an appendix to the full study report. The Sponsor will receive one copy of the draft report and two copies of the final report. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE STATEMENT: The procedures described in this protocol have been reviewed by the Testing Facility's Institutional Animal Care and Use Committee. All procedures described in this protocol that involve study animals will be conducted in a manner to avoid or minimize discomfort, distress or pain to the animals. The Sponsor's signature below documents the fact that information concerning the necessity for conducting this study and the fact that this is not an unnecessarily duplicative study may be obtained from the Sponsor. No alternative (in vitro) procedures were available for meeting the stated purposes of the study. 003819 418-011:PAGE F-71 418-011P:PAGE 71 Protocol 418-011P Page 13 REFERENCES: 1. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 2. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November?, 1983), J. Clin. Psychiat. 45(9):7-10. 3. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River Crt:CDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 4. Institute of Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski, E. (1964). FSrbemethode zum makroskopischen Nachweis von Impiantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. 003820 PROTOCOL APPROVAL: FOR THE TESTING FACILITY George^. Deariove, Ph.D., DABT Associate Director of Research 418-011 .PAGE F-72 418-011P:PAGE 72 Protocol 418-011P Page 14 /'-ft/Zl/'*? P Date / o - Z L / v - r)Sr Date Dena C. Lebo, V.M.D. Member, Institutional Animal Care and Use Committee FOR THE SPONSOR T Q <.LC. Marvin T. Case, D.V.M., Ph.D. Study Monitor Date Z . ____I L ^ a ! 1fff Date 003821 418-011: PAGE F-73 418-011P:PAGE 73 ATTACHMENT 1 SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE 003822 ATTACHMENT 1 418-011: PAGE F-74 418-011P:PAGE 74 Protocol 418-011P Page 1 of 2 STUDY SCHEMATIC DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY" Sta rt of Dos age C o h a tstation F em ale Rats Day 6 of Pres Limed G e s t ation CaesareanSectioning Emj of Dos age Day 17 of Pres jm ed Gestation Day 20 of Presumed Gestation Dosage Period. a. For additional details see "Tests, Analyses and Measurements'' section of the protocol. b. Fetal evaluations (all fetuses - external examinations). 003823 ATTACHMENT 1 418-011: PAGE F-75 418-011P.PAGE 75 Protocol 418-011P Page 2 of 2 SCHEDULE* 16 JUN 98 22 JUN 98 PM - 27 JUN 98 AM 23 JUN 98 - 27 JUN 98 29 JUN 9 8 -1 4 JUL 98 13 JUL 9 8 -1 7 JUL 98 24 JUL 98 29 SEP 98 Arrival Date - Acclimation Begins. Cohabitation Period. Day 0 of Presumed Gestation. Dosage Period (Days 6 through 17 of presumed gestation). Caesarean-Sectioning Period (Day 20 of presumed gestation). Letter Report. Summary Report. a. The study initiation date is the date the Study Director signs the protocol. 003824 418-011: PAGE F-76 418-011P:PAGE 76 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 003825 418-011:PAGE F-77 M ATERIAL SAFETY DATA SHEET 3M N - +FOSC 418-011P:PAGE 77 3M C e n te r S t . P a u l, M in n eso ta 5 5 1 4 4 -1 0 0 0 1 -8 0 0 -3 6 4 -3 5 7 7 o r (612) 737-6501 (24 h ou rs) C o p y rig h t, 1998, M in n eso ta M in in g and M a n u fa ctu rin g Com pany. A ll r ig h ts r e se r v e d . C o p yin g an d /or d ow n load in g o f t h is in fo r m a t io n f o r t h e p u r p o s e o f p r o p e r ly u t i l i z i n g 3M p r o d u c ts is allo w ed p ro vid ed th a t: 1) th e in fo rm a tio n is co p ied in f u l l w ith no chan ges u n le ss p r io r agreem en t i s o b ta in e d from 3M , and 2) n e ith e r th e copy nor th e o r ig in a l is re so ld or o th erw ise d is tr ib u te d w ith th e in te n tio n o f e a rn in g a p r o fit th e re o n . D IV IS IO N : 3M C H E M IC A L S TRADE NAME: FC-1 0 FLUORAD Brand F lu o ro c h e m ic a l A lc o h o l ID NUMBER/ U .P .C .: 9 8 -0 2 1 1 -1 1 1 3 -7 0 0 -5 1135-09495-2 9 8 -0 2 1 1 -1 1 8 3 -0 9 8 -0 2 1 1 -1 5 7 5 -7 0 0 -5 1135-02145-3 9 8 -0 2 1 1 -6 6 2 0 -6 Z F -0002-0572-2 ISSU ED : Ja n u a ry 2 9 , 1998 SUPERSEDES: Novem ber 05, 1997 DOCUMENT: 1 0-3778-7 0 0 -5 1 1 3 5 -0 9 5 4 2 -3 0 0 -5 1 1 3 5 -1 0 4 3 9 -2 1 . INGRED IENT C .A .S . NO. PERCENT PERFLUOROOCTANESULFONAM IDO ALCO H O L. PERFLUOROHEXANESULFONAM IDO ALCOH OL. PERFLUOROHEPTANESULFONAM IDO ALCOHOL PERFLUOROBUTANESULFONAM IDO ALCO H O L. PERFLUOROPENTANESULFONAM IDO ALCOHOL 1 6 9 1 -9 9 -2 3 4 4 5 5 -0 3 -3 6 8555-73-7 3 4449-89-3 6 8 555-72-6 8 0 .0 3 .0 2 .0 2 .0 1 .0 - 9 0 .0 - 7 .0 - 6 .0 - 6 .0 - 3 .0 2 . PH YSICAL DATA B O IL IN G P O IN T :............. VAPOR P R E S S U R E :........... VAPOR D E N S IT Y :............. EVAPORATION R A T E :... SO LU B ILIT Y IN WATER: S P E C IF IC G R A V IT Y :. . . PERCENT V O L A T IL E :. . . P H :.......................................... V I S C O S I T Y :........................ M ELTING P O I N T : . . . . . . c a . 118 C 6 1 mm H g < 1 0 mmHg C a lc 6 20 C > 1 . 0 A ir= 1 C a lc 6 20 C . < 1 . 0 BuOAc=1 n e g lig . c a . 1 .7 W ater=i (o f m elt) 0h N/A N/D N/D 003826 Ahhi-.vi.rinn? N/D . Not Determined N/A - Not Applicable CA * Approximately 418-011: PAGE F-78 M SDS: FC-1 0 FLUORAD B rand F lu o ro c h e m ic a l A lc o h o l Ja n u a ry 2 9 , 1998 418-011P:PAGE 78 PAGE 2 2 . PH YSICAL DATA (co n tin u ed ) APPEARANCE AND OOOR: Amber waxy s o lid 3 . F IR E AND EXPLOSION HAZARD DATA FLA SH P O IN T :.............................................. > 148 C S e t a f l a s h FLAMMABLE L IM IT S - L E L :.................... N /A FLAMMABLE L IM IT S -U E L :.................... N /A A U T O IG N IT IO N T EM P E R A T U R E :............. N /A EX T IN G U ISH IN G M EDIA: H a te r , Carbon d io x id e , D ry c h e m ic a l, Foam SP E C IA L FIR E FIGH TING PROCEDURES: Hear f u l l p r o te c tiv e c lo th in g , in c lu d in g h elm et, s e lf-c o n ta in e d , p o s itiv e p re ssu re or p re ssu re demand b re a th in g a p p a ra tu s, bunker co a t and p a n ts, bands around arm s, w a ist and le g s , fa c e m ask, and p ro te c tiv e co verin g fo r exposed areas o f th e head. UNUSUAL FIR E AND EXPLOSION HAZARDS: See H azardous D ecom p osition s e c tio n fo r p ro d u cts o f com b u stion . 4 . R EA CTIVIT Y DATA ST A B ILIT Y : S ta b le IN CO M PA T IB ILIT Y - M ATERIALS/COND ITIO NS TO AVO ID : Not a p p lica b le . HAZARDOUS POLYM ERIZATION: H azard o u s p o ly m e r iz a tio n w ill n o t o c c u r . HAZARDOUS DECOM POSITION PRODUCTS: C arb on M onoxide and C arb on D io x id e , O x id e s o f N itr o g e n , O x id e s o f S u lfu r , Hydrogen F lu o rid e , T o xic V ap o rs, G ases or P a r tic u la te s . 5 . ENVIRONM ENTAL INFORM ATION SP ILL RESPONSE: R e fe r to o th e r s e c tio n s o f t h is MSDS fo r in fo rm a tio n re g a rd in g p h y s ic a l and h e a lth h a za rd s, r e s p ir a to r y p r o te c tio n , v e n tila tio n , and p e rso n a l p r o te c tiv e eq u ip m en t. C o lle c t s p ille d m a te r ia l. C le a n up r e sid u e . P la ce in a U . S . D O T-approved c o n ta in e r . 003827 Akh w/n . not-prmined N/A - Not Applicable CA - Approximately 418-011:PAGE F-79 MSDS: FC-10 FLU0RA0 Brand Fluoroeheaical Alcohol Ja n u a ry 2 9 , 1998 5 . ENVIRONM ENTAL INFORM ATION (co n tin u ed ) 418-011P:PAGE 79 PAGE 3 RECOMMENDED D ISP O SA L: In c in e r a te in a p erm itted hazardous w aste in c in e r a to r in th e p resen ce o f a co m b u stib le m a te r ia l. Com bu stion p ro d u cts w ill in c lu d e H F. D isp o se o f w aste p rod u ct in a f a c i l i t y p erm itted to a cce p t ch em ical w aste. ENVIRONM ENTAL DATA: L a b o r a to r y t e s t s show ed no b io d e g r a d a tio n . 9 6 -H r . LD50 F a th e a d M innow (P im ep h ales p rom elas) - No m o r ta lity a t w ater s a tu r a tio n . No s t a t is t ic a lly s ig n ific a n t e ffe c t on % h a tch , % s u r v iv a l, w e ig h t, and le n g th in 30 d ay F a th e a d M innow e g g f r y s tu d y . Lab t e s t s show ed 200 fo ld b io c o n c e n tra tio n o f FC-1 0 in to m u scle f i l l e t s o f ch a n n e l c a t fis h . REGULATORY INFORM ATION: V o la tile O rgan ic Com pounds: N/A. VOC L ess H20 & Exem pt S o lv e n ts : N/A. T h is p ro d u ct co m p lies w ith th e ch em ica l r e g is tr a tio n req u irem en ts o f TSCA, EIN ECS, CD SL, A ICS and K orea. EPCRA HAZARD CLA SS: FIR E HAZARD: No PRESSURE: No R EA CT IV IT Y : No ACUTE: Y es CH RONIC: Y es 6 . SUGGESTED FIR ST AID EYE CONTACT: Im m ed ia tely flu s h ey es w ith la r g e am ounts o f w a te r. G et im m ediate m ed ical a tte n tio n . SK IN CONTACT: Im m ed ia tely w ash s k in w ith so ap and la r g e am ounts o f w a te r. Remove co n tam in ated c lo t h in g . I f sign s/sym p tom s o c c u r , c a ll a p h y s ic ia n . Wash co n ta m in a te d c lo t h in g b e fo r e re u se and d is p o s e o f c o n ta m in a te d shoes. INH ALATION: I f sign s/sym p tom s o c c u r , rem ove p erso n to fr e s h a ir . I f sign s/sym ptom s c o n tin u e , c a ll a p h y s ic ia n . IF SWALLOWED: C a ll a p h y s ic ia n IM M ED IATELY. I f sw a llo w e d , in d u ce v o m itin g im m ed iately as d ir e c te d by m e d ica l p e rso n n e l. N ever g iv e a n y th in g m outh to an u n c o n sc io u s p e r s o n . by 00382S Il /rt 'ni!!nori N/A - Not Aoolicable CA - Approximately 418-011: PAGE F-80 M SDS: FC-1 0 FLUORAD Brand F lu o ro ch e m ica l A lc o h o l Ja n u a ry 2 9 , 199B 418-011 P .P A G E 80 PAGE 4 7 . PRECAUTIONARY INFORM ATION EYE PROTECTION: A void eye c o n ta c t. Wear s a fe ty g la s s e s w ith s id e s h ie ld s . SK IN PROTECTION: A v o id sk in c o n ta c t. N ear a p p ro p ria te g lo v e s when h a n d lin g t h is m a te r ia l. A p a ir o f g lo v e s made from th e fo llo w in g m a te r ia l(s ) recom m ended: b u ty l ru b b e r. U se one o r m ore o f th e fo llo w in g p erso n a l p ro te c tio n item s as n e ce ssa ry to p reven t sk in c o n ta c t: co v e ra lls. are RECOMMENDED V EN T ILA T IO N : U se w ith a p p ro p ria te lo c a l e x h a u st v e n tila tio n . P ro v id e s u ffic ie n t v e n t ila t io n to m a in ta in e m issio n s below recom m ended ex p o su re lim it s . I f exh au st v e n tila tio n is not ad eq u ate, use a p p ro p ria te re sp ira to ry p ro te ctio n . RESPIRATORY PROTECTION: A void b re a th in g o f a irb o rn e m a te r ia l. S e le c t one o f th e fo llo w in g NIOSH ap proved r e s p ir a to r s b ased on a ir b o r n e c o n c e n tr a tio n o f co n tam in an ts and in acco rd an ce w ith OSHA r e g u la tio n s : h a lf-m a sk d u st r e s p ir a to r , f u ll-fa c e su p p lie d a ir r e s p ir a to r . PREVENTION OF ACCID ENTAL IN G EST IO N : Do not e a t, d rin k or sm oke when u sin g areas th o ro u gh ly w ith soap and w a te r. b efore e a tin g . th is p ro d u ct. Nash exposed Nash hands a fte r h an d lin g and RECOMMENDED STORAGE: S to r e away from h e a t. Keep c o n ta in e r c lo s e d when n o t in u se . F IR E AND EXPLOSION AVOID ANCE: N on flam m ab le. OTHER PRECAUTIONARY INFORM ATION: No sm o k in g: Sm oking w h ile u s in g t h i s p ro d u ct can r e s u lt in co n ta m in a tio n o f th e to b a cco an d /or sm oke and le a d to th e fo rm a tio n o f th e h azard ou s d e co m p o sitio n p ro d u cts m ention ed in s e c tio n 4 o f th is MSDS. H M IS HAZARD R A T IN G S: H EALTH : 1 FLA M M A B ILITY: 1 R E A C T IV IT Y : 0 PERSONAL PROTECTION: X (See p r e c a u tio n s , s e c tio n 7 .) INGRED IEN T EXPOSURE LIM IT S VALUE UNIT 003823 TYPE AUTH SK IN PERFLUOROOCTANESULFONAM IDO ALCOHOI_____ 0 .1 MG/M3 PERFLUOROHEXANESULFONAM IDO ALCOHOI_____ 0 .1 MG/ M3 PERFLUOROHEPTANESULFONAM IDO A LC O H O L...................................................................... 0 .1 MG/ M3 TNA 3M TN A 3M TNA 3M Y Y Y si m n o / i N/A . Not Anolicable CA - Approximately 418-011: PAGE F-81 M SDS: FC -1 0 FLUORAD Brand F lu o r o c h e n ic a l A lc o h o l Ja n u a ry 2 9 , 1998 418-Q 11P:PA G E 81 PAGE 5 EXPOSURE LIM IT S (co n tin u ed ) INGRED IENT VALUE UNIT TYPE AUTH SKIN ' PERFLUOROBUTANESULFONAM IDO ALCOHOI----- 0 .1 MG/M3 PERFLUOROPENTANESULFONAM IDO A LCO H O L..................................................................... 0 .1 MG/M3 TWA 3M TWA 3M Y Y * SK IN NOTATION: L is t e d s u b s ta n c e s in d ic a te d w ith 'Y ' u n d er SK IN r e fe r to th e p o te n tia l co n trib u tio n to th e o v e r a ll exposure by th e cutaneous rou te in c lu d in g m ucous membrane and e y e , e ith e r by a ir b o r n e o r , m ore p a r t ic u la r ly , by d ir e c t c o n ta c t w ith th e su b sta n ce . V e h ic le s can a lte r sk in a b so rp tio n . SOURCE OF EXPOSURE LIM IT DATA: - 3M: 3M Recom m ended E x p o s u r e G u id e lin e s 8 . HEALTH HAZARD DATA EYE CONTACT: No adverse h e a lth e f f e c t s a re e x p e cte d from eye c o n ta c t. SK IN CONTACT: Product is not expected to be ir r ita tin g to th e sk in . May be absorb ed th ro u gh th e sk in and p e r s is t in th e body fo r an extended tim e. INH ALATION: May be ab sorb ed by in h a la tio n and p e r s is t in th e body fo r an exten ded tim e. IF SWALLOWED: In g e stio n is not a lik e ly route o f exposure to th is p rod u ct. I lln e s s may o cc u r a f t e r a s in g le sw a llo w in g o f r e la t i v e ly la r g e q u a n titie s o f th is m a te ria l. M U TAGENICITY: Not m u tagen ic in in -v it r o a s s a y s . REPRODUCTIVE/DEVELOPM ENTAL T O X IN S: S u b sta n ce was n o t te r a to g e n ic in th e r a t a t d o ses as h igh as 30 m illig ra m s per k ilo gram per day v ia o r a l r o u te . OTHER HEALTH HAZARD INFO RM ATIO N: T h is p ro d u ct i s n o t known to c o n ta in any su b sta n ce s r e g u la te d under C a lifo r n ia P ro p o sitio n 6 5 . A P ro d u ct T o x ic it y Sum m ary S h e e t i s a v a ila b le . 00383O Il/A AfmitpaKi P.A - Annroximatlv I 418-011: PAGE F-82 M SDS: FC-1 0 FLUORAD Brand F lu o r o e h e a ic a l A lc o h o l Jan u ary 2 9 , 1998 418-011P:PAGE 82 PAGE 6 SECTIO N CHANGE DATES H E A D IN G SECTIO N CHANGED SIN CE Novem ber 0 5 , 1997 ISSU E A b b r e v ia tio n s : N/D - N ot D eterm in ed N/A - N ot A p p lic a b le CA A p p ro x im a te ly T he in fo r m a tio n in t h i s M a t e r ia l S a fe t y D a ta S h e e t (M SDS) i s b e lie v e d to be c o r r e c t a s o f th e d a te is s u e d . 3M MAKES NO W ARRANTIES, EXPR ESSED OR IM P L IE D , IN C LU D IN G , BUT NOT LIM IT ED TO , ANY IM P LIED WARRANTY OF M ER CH A N T A B ILIT Y OR F IT N E S S FOR A P A R T ICU LA R PURPO SE OR CO URSE OF PERFORMANCE OR USAGE OF TRAD E. U se r i s r e s p o n s ib le fo r d e te rm in in g w h e th e r th e 3M p r o d u c t i s f i t f o r a p a r t i c u l a r p u r p o s e and s u i t a b l e f o r u s e r ' s m ethod o f u se o r a p p lic a t io n . G iv e n th e v a r ie t y o f f a c t o r s th a t ca n a f f e c t th e u se and a p p lic a t io n o f a 3M p r o d u c t, som e o f w h ich a r e u n iq u e ly w ith in th e u s e r ' s know led ge and c o n t r o l, i t i s e s s e n t ia l th a t th e u s e r e v a lu a t e th e 3M p r o d u c t t o d e te r m in e w h e th e r i t i s f i t f o r a p a r tic u la r p u rp o se and s u it a b le fo r u s e r 's m ethod o f u se o r a p p lic a tio n . 3M p r o v id e s in fo r m a t io n in e l e c t r o n i c fo r m a s a s e r v i c e t o i t s c u s to m e r s . Due to th e rem ote p o s s i b i l i t y t h a t e le c t r o n ic t r a n s fe r may have r e s u lte d in e r r o r s , o m is s io n s o r a l t e r a t i o n s in t h i s in fo r m a t io n , 3M m akes no re p re se n ta tio n s as to i t s co m p leten ess or a c cu ra c y . In a d d itio n , in fo rm a tio n o b ta in e d from a d a ta b a se may n o t be a s c u r re n t a s th e in fo rm a tio n in th e MSDS a v a ila b le d ir e c t ly from 3M. 003831 418-011:PAGE F-83 418-011P:PAGE 83 ATTACHMENT 3 TEST ARTICLE AND CONTROL ARTICLE PREPARATION PROCEDURE 003832 418-011: PAGE F-84 418-011P:PAGE 84 ATTACHMENT 3 Protocol 418-011P Version: 4 1 8 -0 1 1 P (09 J U N 981 Page 1 of 3 TEST ARTICLE AND CONTROL ARTICLE PREPARATION PROCEDURE Test Article: N-EtFOSE Vehicle: 2% Tween 80, in R.O. Water A. Purpose: The purpose of this procedure is to provide a method for the preparation of dosage suspensions of N-EtFOSE and the control article for oral administration to rats on Argus Study 418-011P. B. General Information: 1. All suspension containers will be labeled and color coded. Each label will specify the protocol number, test article identification, Argus batch number, concentration, dosage level, preparation date, expiration date and storage conditions. 2a. Suspensions will be prepared: X Daily __ Weekly For days of use Vehicle will be prepared: Daily X Weekly __For__ __ days of use 3. Suspensions will be prepared at a final dosage volume of 5 ml_/kg. 4. Safety: X Gloves, lab coat, goggles or safety glasses and faceshield X Dust-Mist Respirator __ Half-Face Respirator __ Full-Face Respirator/Positive Pressure Hood __ Tyvek Suit/Apron 5. Dosage solutions adjusted for Free base and % Purity. __ Yes X No (Calculations based on 100%) ___ Free Base ___ Purity 6. Sampling requirements: Cited in protocol. 7. Storage: Cited in protocol. 003833 418-011:PAGE F-85 418-011P.PAGE 85 ATTACHMENT 3 Protocol 418-011P V ersion: 4 1 8 -0 1 1 P (09 J U N 981 Page 2 of 3 TEST ARTICLE AND CONTROL ARTICLE PREPARATION PROCEDURE NOTE: Test article will be prepared as a serial dilution from the high dosage to the low dosage. Once the final volumes are achieved, stir bars are to be added to the containers; mixing should occur during sampling and/or administration. C. Preparation of Vehicle 1. Add the required amount of R.O. deionized water to an appropriately labeled container. Heat the water to 50C, 5C, add the required amount of Tween 80 and mix until uniform (See TEST ARTICLE CALCULATIONS). D. Test Article Suspension Preparation: 1. To prepare the 7-mg/mL, Group VII suspension, add the required amount of test article (See TEST ARTICLE CALCULATIONS) into an appropriately sized, labeled container. Add the required amount of vehicle and heat the mixture to 80C, 5C for approximately 30 minutes. 2. Once the test article has dissolved; spin over night while the solution cools. (Be sure there is a visible vortex, this will achieve the desired emulsion.) 3. To prepare the 5-mg/mL, Group VI suspension, remove the required amount of stock suspension (Group VII) (See TEST ARTICLE CALCULATIONS), add the required amount of vehicle and mix. 4. To prepare the 4-mg/mL, Group V suspension, remove the required amount of stock suspension (Group VI) (See TEST ARTICLE CALCULATIONS), add the required amount of vehicle and mix. 5. To prepare the 2-mg/mL, Group IV suspension, remove the required amount of stock suspension (Group V) (See TEST ARTICLE CALCULATIONS), add the required amount of vehicle and mix. 003834 418-011: PAGE F-86 418-011P:PAGE 86 ATTACHMENT 3 Protocol 418-011P Version: 418-011P (09 J U N 981 Page 3 of 3 TEST ARTICLE AND CONTROL ARTICLE PREPARATION PROCEDURE 6. To prepare the 1-mg/mL, Group III suspension, remove the required amount of stock suspension (Group IV) (See TEST ARTICLE CALCULATIONS), add the required amount of vehicle and mix. 7. To prepare the 0.2-mg/mL, Group II suspension, remove the required amount of stock suspension (Group III) (See TEST ARTICLE CALCULATIONS), add the required amount of vehicle and mix. Written by: Approved by: Clarification: j/ j No ___Yes (See attached clarification form.) Initials/Date : -?/?? 0383S 418-011:PAGE F-87 418-011P.PAGE 87 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044 T: (215) 443-8710 F: (215) 443-8587 PROTOCOL 418-011P ORAL (GAVAGE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS SPONSOR'S STUDY NUMBER: T-6316.7 Amendment 1 - June 18,1998 1. Clinical Observations and/or General Appearance (page 8 of the protocol): Clinical observations during the dosage period will be taken twice daily, prior to dosage administration and once approximately one hour postdosage, rather than once approximately one hour postdosage and then four to six hours later. Reason for Change: This change was made at the request of the Sponsor to match the time frames of the other reproductive/developmental toxicity studies using the same test grticle. / George E^earlove, Ph.D., DABT Date RaymncbG. York, Fh.CL DABT Associate Director of Research Associate Director of Research and Study Director J z M tfZ Date J'-miJic'T-______/f\JtUrh fF Dena C. Lebo, V.M.D. Date Member, Institutional Animal Care and Use Committee O*- Marvin T. Case, D.V.M., Ph.D. Study Monitor 003836 Date APPENDIX G HISTORICAL CONTROL DATA 003S37 418-011: PAGE G-1 SUMMARY OF REPRODUCTIVE INDICES CD RAT PERIOD JUNE 1995 -J U N E 1997 NUMBER OF STUDIES 97 NUMBER OF RATS: TESTED PREGNANT FOUND DEAD ABORTED DELIVERED 2132 1967 3 0 0 NUMBER OF RATS PREGNANT AT CAESAREAN-SECTIONING 1957 NUMBER OF RATS WITH SINGLE CONCEPTUS LITTER: LIVE RESORBED ABORTED 5 0 0 MEAN o r PREGNANT 92.8 AVERAGE # CORPORA LUTEA 1 7 .3 AVERAGE # IMPLANTATIONS 1 5 .6 AVERAGE LITTER SIZE AVERAGE # LIVE FETUSES 14.8 AVERAGE # DEAD FETUSES 0 .1 AVERAGE * RESORPTIONS 0.8 AVERAGE * EARLY RESORPTIONS 0 .7 AVERAGE t LATE RESORPTIONS 0.0 RANGE/STUDY MEAN o r % (64.0-100) (15.2-21.0 ) (12 .9 -18 .0 ) (11.8 -17.0 ) (0 -1.1) (0 -1.6) (0 -1.6) (0 -0 .2 ) 003838 418-011 .PAGE G-2 SUMMARY OF REPRODUCTIVE INDICES CD RAT MEAN o r AVERAGE % DAMS WITH ANY RESORPTIONS 48.4 AVERAGE DAMS WITH ALL CONCEPTUSES RESORBED 0.1 AVERAGE DAMS WITH ONE OR MORE LIVE FETUSES 99.9 AVERAGE SEX RATIO, {% MALES/LITTER) 50.2 AVERAGE FETAL BODY WEIGHT (G) 3 .4 7 AVERAGE FOR MALES (G) 3.56 AVERAGE FOR FEMALES (G) 3.37 AVERAGE % DEAD OR RESORBED CONCEPTUSES/LITTER o in RANGE/STUDY MEAN o r % (0 -7 5 .0 ) (0 -4 .5 ) (9 5 .4 -10 0 ) (42.1-57.0 ) (3 .10 -3 .7 8 ) (3.17-3.90 ) (2 .9 8 -3 .6 6 ) (0 -10 .9 ) 003839 SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT PERIOD JUNE 1995 - # STUDIES # RATS TESTED # RATS PREGNANT # RATS DIED # RATS ABORTED # RATS DELIVERED # RATS WITH 100% RESORPTION JUNE 1997 120 2579 2387 4 0 439 1 EXTERNAL OBSERVATIONS Red su b s ta n ce around noae and /or mouth In ciso rs m isaligned broken and/or m issing Chromodacryorrhea A lo p e c ia White substance p re se n t in a n te r io r chamber o f eyes MEAN RANGE /STUDY N% N 1 0.04 0 -1 (0 -3 .3 ) 2 0.08 0-2 (0 -6 .7 ) 2 0.08 0-2 (0 -6 .7 ) 3 0 .12 0-3 (0 -10.0) 1 0.04 0 -1 (0 -4 .2 ) GROSS LESIONS SUBMANDIBULAR LYMPH NODES Dark red JUGULAR VEIN Distended w ith blood CHEST Subcutaneous f a t , dark red on v e n t r a l s id e THORACIC CAVITY F ille d w ith lig h t red flu id THYMUS D iscolored areas Large AXILLA Mass p re se n t 1 0.04 0 -1 (0 -3 .3 ) 1 0.04 0 -1 (0 -3 .3 ) 1 0.04 0 -1 (0 -3 .3 ) 1 0.04 0 -1 (0 -4 .0 ) 2 0.08 0-2 (0 -6 .7 ) 1 0.04 0 -1 (0 -4.0 ) 1 0.04 0 -1 (0 -4 .0 ) 003840 418-011 .PAGE G-4 SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT LIVER GROSS LESIONS Adhesion between l e f t l a t e r a l lo b e and l e f t l a t e r a l abdominal w a ll MEAN RANGE! /STUDY N N 1 0.04 0 -1 (0 -4.0) STOMACH/INTESTINE Stomach, f l u i d - f i l l e d Stomach and in t e s t in e s distended w ith gas I n t e s t in e s , empty Cecum c o n ta in e d a b la c k substance 1 0.04 0 -1 (0 -4.0) 1 0.04 0 -1 (0 -4.0) 1 0.04 0 -1 (0 -4.0) 1 0.04 0 -1 (0-4.0) BACK Spine, p ro tru sio n o f bone on d o rsa l th o r a c ic p o r tio n 1 0.04 0 -1 (0-4.0) KIDNEY(S ) P e lv is , slight/m oderate d ila tio n w ith or w ithout flu id P e lv is , marked/extreme d ilatio n M ottled Large Sm all Cortex p itte d L e ft; p e lv is contained yellow flu id 17 0.66 0-2 (0 -12 .5 ) 5 0.19 0 -1 (0-4.0) 2 0.08 0 -1 (0 -4.0) 2 0.08 0 -1 (0-4.0) 1 0.04 0 -1 (0-4.0) 1 0.04 0 -1 (0 -3.4 ) 1 0.04 0 -1 (0-4.0) ABDOMINAL CAVITY F ille d w ith lig h t red flu id 1 0.04 0 -1 (0 -4.0 ) SPLEEN Two w h it e a r e a s on serosal surface 1 0.04 0 -1 (0 -4.0) BLADDER F lu id -fille d W all th ic k , contained one c a lc u li 1 0.04 0 -1 (0 -4.0) 1 0.04 0 -1 (0 -12 .5 ) 003841 418-011 PAGE G-5 SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT GROSS LESIONS URETERS Distended w ith c le a r flu id MEAN RANGE /STUDY N% N % 1 0.04 0 -1 (0 -4 .0 ) VAGINA/CERVIX C ervix distended w ith flu id Cervix contained a th ick , brown su bstan ce C ervix contained a dark red, gelatinous substance Cervix contained green, viscous flu id V agina con tain ed brown, viscous flu id 1 0.04 0 -1 1 0.04 0 -1 1 0.04 0 -1 1 0.04 0 -1 1 0.04 H O (0 -4 .0 ) (0 -4 .0 ) (0 -4 .0 ) (0 -4 .0 ) (0-2 .1 ) UTERUS Contained red-brown f lu id and one dead, brown fe tu s R ight horn, absent R ight horn, th rea d lik e L e f t horn, lumen absen t on c e r v ic a l end; ovarian end, distended w ith cle a r flu id R ight horn, c le a r masses. contained gelatin o u s substance 1 0.04 0 -1 2 0.08 0 -1 1 0.04 0 -1 1 0.04 0 -1 1 0.04 10 H (0 -4 .0 ) (0 -16.7) (0-2 .1 ) (0 -4 .0 ) (0 -4 .0 ) FORELIMB Lesion present 1 0.04 0 -1 (0 -3 .3 ) 003842 418-011: PAGE G-6 SUMMARY 07 FETAL EXTERNAL ALTERATIONS CD RAT PERIOD JUNE 1995 - JUNE 1997 * STUDIES INCLUDED * LITTERS EXAMINED * LIVE FETUSES EXAMINED 86 1745 25613 ALTERATION HEAD Exencephaly Hematoma M icrocephaly EYES Eye bulges depressed L id s open Microphthalm ia EARS Low s e t SNOUT Short TONGUE Absent PALATE C left RANGE /STUDY N %N % L 5 0.29 0 -1 (0-4.5) F 5 0.02 0 -1 (0-0.4) L 1 0.06 0 -1 (0 -2.2) F 1 0.00 0 -1 (0-0.2) L 1 0.06 0 -1 (0-4.0) F 1 0.00 0 -1 (0-0.3) L 14 0.80 0-2 (0 -12 .5 ) F 15 0.06 0-2 (0-0.9) L 1 0.06 0 -1 (0 -4.2) F 1 0.00 0 -1 (0-0.3) L 1 0.06 0 -1 (0-4.0) F 1 0.00 0 -1 (0-0.3) L 3 0 .17 0-1 (0 -12.5) F 3 0.01 0 -1 (0-0.9) L 2 0 .11 0 -1 (0-4.2) F 2 0.01 0 -1 (0-0.3) L 1 0.06 0 -1 (0-4.0) F 1 0.00 0 -1 (0-0.3) L 4 0.23 0 -1 (0-4.5) F 4 0.02 0 -1 (0-0.4) L: LITTER INCIDENCE P: FETAL INCIDENCE 003843 418-011: PAGE G-7 SUMMARY OF FETAL EXTERNAL ALTERATIONS CD RAT ALTERATION JAWS M icrognathia A g n a th ia L F L F BODY Edema L F U m bilical hernia L F G astrosch isis L F Trunk sh ort L F Spina b ifid a L F E xtra limb pro L tr u d in g from back F Hematoma L F PLACENTA Enlarged L F FORELIMBS Two d i g i t s p r e se n t on forepaw L F HINDLIMBS R o tated L F ANUS No op en in g p r e s e n t L F RANGE /STUDY N %N 5 0.29 0 -1 (0 -4 .5 ) 5 0.02 0 -1 (0 -0 .3) 1 0.06 0 -1 (0 -12 .5 ) 1 0.00 0 -1 (0 -0 .9) 2 0 .11 0 -1 (0-4.5) 2 0.01 0 -1 (0-0.4) 7 0.40 0 -1 (0 -16 .7 ) 7 0.03 0 -1 ( 0 -1 .1 ) 1 0.06 0 -1 (0 -4.0 ) 1 0.00 0 -1 (0 -0 .2) 3 0 .17 0-2 (0 -9 .1) 3 0.01 0-2 (0 -0 .7) 1 0.06 0 -1 (0 -4.0) 1 0.00 0 -1 (0 -0 .2) 1 0.06 0 -1 (0 -4 .5 ) 1 0.00 0 -1 (0 -0 .4 ) 1 0.06 0 -1 (0 -2 .4 ) 1 0.00 0 -1 (0 -0 .2 ) 1 0.06 0 -1 (0 -16 .7 ) 1 0.00 0 -1 ( 0 -1 .1 ) 1 0.06 0 -1 (0 -4 .3) 1 0.00 0 -1 (0 -0 .3) 1 0.06 0 -1 (0 -4.0) 1 0.00 0 -1 (0 -0 .2 ) 2 0 .11 0-1 (0-4.5) 2 0.01 0-1 (0-0.4) L: LITTER INCIDENCE F: FETAL INCIDENCE 003844 418-011:PAGE G-8 SUMMARY OF FETAL EXTERNAL ALTERATIONS CO RAT ALTERATION TAIL T h r e a d lik e A g e n e sis S p lit Short C o n str ic te d RANGE /STUDY N %N L 5 0.29 0 -1 (0 -5 .3 ) F 5 0.02 0 -1 (0 -0 .4 ) L 3 0.17 0-1 (0-4.5) F 3 0.01 0 -1 (0-0.3) L 1 0.06 0 -1 (0 -4 .5 ) F 1 0.00 0 -1 (0 -0 .4 ) L 2 0 .11 0-1 (0-4.3) F 2 0.01 0 -1 (0 -0.3) L 1 0.06 0 -1 (0 -2 .4 ) F 1 0.00 0 -1 (0-0 .2 ) L: LITTER INCIDENCE Fs FETAL INCIDENCE 003845 418-011:PAGE G-9 SUMMARY OF FETAL SOFT TISSUE ALTERATIONS CD RAT PERIOD JUNE 1995 # STUDIES INCLUDED # LITTERS EXAMINED # FETUSES EXAMINED JUNE 1997 36 845 6091 BRAIN ALTERATION Lateral ven tricles, moderate d ila tio n Lateral ven tricles, marked d ila t io n Third v e n tr ic le , marked d i la t io n L a te r a l and t h ir d v e n t r ic le s , irregu larly shaped L F L F L F L F RANGE/STUDY N% N 1 0.12 0 -1 (0 -3.4) 1 0.02 0 -1 (0 -0 .6 ) 1 0.12 0 -1 (0 -4 .2) 1 0.02 0 -1 (0 -0 .6 ) 1 0.12 0 -1 (0 -4 .2) 1 0.02 0 -1 (0 -0 .6 ) 1 0.12 0 -1 (0 -4 .2) 1 0.02 0 -1 (0 -0 .6 ) BYES Microphthalm ia L 3 0.36 0 -1 (0 -4.0 ) F 3 0.05 0 -1 (0 -0 .6 ) PALATE C left L 2 0.24 0 -1 (0 -4 .0 ) F 2 0.03 0 -1 (0 -0 .6 ) TONGUE Sm all Absent L 1 0.12 0 -1 (0 -4.3) F 1 0.02 0 -1 (0 -0 .6 ) L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .6 ) JAW M icrognathia L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .6 ) HEART Septal defect L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .6 ) L: LITTER INCIDENCE F: FETAL INCIDENCE 003846 418-011: PAGE G-10 SUMMARY OF FETAL SOFT TISSUE ALTERATIONS CD RAT ALTERATION RANGE/STUDY N N VESSELS Innominate, absent L 8 0.95 0-2 (0-8.0) F 8 0 .13 0-2 (0 -1.2 ) Innom inate, a r i s e s on le ft Subclavian a r te ry , absent Ductus a r te r io s u s , absent U m bilical a r te r y , d isp la c e d Situs inversus L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0-0.6) L 1 0.12 0 -1 (0-3.4) F 1 0.02 0 -1 (0 -0 .6) L 1 0.12 0 -1 (0-3.4) F 1 0.02 0 -1 (0 -0 .6 ) L 10 1 .1 8 0-2 (0 -8 .3) F 1 1 0.18 0-2 (0 -1.2 ) L 1 0.12 0 -1 (0-4.2) F 1 0.02 0 -1 (0 -0 .6) Aorta, descends to rig h t L F 1 0.12 0 -1 (0-4.0) 1 0.02 0 -1 (0 -0 .6 ) Pulmonary a r t e r y , descends to r ig h t behind aorta L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .6) LUNGS R ight a p ic a l, card iac and d ia p h ra g m a tic ' lo b es appear as one Interm ediate lo b e. absent L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .6 ) L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .6) BODY Edema L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .6 ) ABDOMINAL CAVITY S itu s inversus o f liv e r , in te s t in e s , stomach, sp le en , pancreas and kidneys L F 1 0.12 0 -1 (0-4.0) 1 0.02 0 -1 (0 -0 .6 ) P elvis, slig h t d ilatio n L F 1 0.12 0 -1 (0 -3.7) 1 0.02 0 -1 (0 -0.6) L: LITTER INCIDENCE F: FETAL INCIDENCE 003847 418-011: PAGE G-11 SUMMARY OF FETAL SOFT TISSUE ALTERATIONS CD RAT ALTERATION SPLEEN Absent URETERS D isten d e d RANGE/STUDY N% N % L 1 0.12 0-1 (0-4.0) F 1 0.02 0 -1 (0 -0 .5) L 2 0.24 0-2 (0 -8 .7) F 3 0.05 0-3 (0 -1 .7 ) L: LITTER INCIDENCE F: FETAL INCIDENCE 003848 418-011:PAGE G-12 SUMMARY OF FETAL SKELETAL ALTERATIONS CD RAT PERIOD * STUDIES * LITTERS * FETUSES JUNE 1995 INCLUDED EXAMINED EXAMINED JUNE 1997 35 820 6318 RANGE/STUDY ALTERATION Nt N SKULL F r o n ta l(8 )s in co m p letely or L 2 0.24 0 -1 (0 -4.2) not o ssified F 2 0.03 0 -1 (0 -0.6) P arietal(s): not ossified L 2 0.24 0 -1 (0 -4.2) F 2 0.03 0 -1 (0-0.6) N asal(s): short L 3 0.36 0 -1 (0 -4.2) F 3 0.05 0 -1 (0-0.6) BaBisphenoid: in com pletely L 1 0.12 0 -1 (0-4.0) o ssified F 1 0.02 0 -1 (0 -0 .5) Sphenoid: ir r e g u la r ly shaped L 1 0 .12 0 -1 (0-4.0) --- t n v jt ---4---- Braft1x"x1---_-___-_--___ ....---_ _ F 1 0.02 0 -1 (0 -0 .5) L-- - . 0 - 1 (0--4 .0 )-- F 2 0.03 0 -1 (0 -0.6) M a x illa e and P rem a xilla e: L 3 0.36 0 -1 (0 -4.2) Bhort F 3 0.05 0 -1 (0-0.6) S k u ll: incom pletely or L 2 0.24 0 -1 (0 -4.2) not o ssifie d F 3 0.05 0-2 (0 -1.2 ) S k u ll: fused L 1 0.12 0-1 (0-4.0) F 1 0.02 0 -1 (0 -0 .5) VERTEBRAE c e r v ic a l: Arch, open : Fused L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .5) L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .5) T h o ra cic: Centrum, b i f i d L 71 8.66 F 77 1.22 : C en tra , u n ila te r a l L 8 0.98 o ssificatio n F 8 0.13 : Centrum, incom pletely or L 3 0.36 not o ssified F 4 0.06 0-8 (0-29.6) 0-9 (0 -4 .7 ) 0-2 (0-8.0) 0-2 (0 -1.0 ) 0-2 (0-8.3) 0-3 (0 -1.8 ) L: LITTER INCIDENCE F: FETAL INCIDENCE 003849 418-011: PAGE G-13 SUMMARY OF FETAL SKELETAL ALTERATIONS CO RAT ALTERATION VERTEBRAE (CONT.) Thoracic (co n t.) C e n tr a , fu se d A r c h , open A r c h , sm all RANGE/STUDY Nt N L 1 0.12 0 -1 (0 -4.2) F 1 0.02 0 -1 (0 -0 .5 ) L 1 0.12 0 -1 (0-4.0) F 1 0.02 0 -1 (0 -0 .5 ) L 1 0.12 0 -1 (0 -3.4) F 1 0.02 0 -1 (0 -0 .5 ) Lumbar: Centrum, b i f i d L 2 0.24 F 2 0.03 Centra, fused L 1 0.12 F 1 0.02 Centrum, in c o m p le te ly L 2 0.24 or not o ssified F 3 0.05 A r c h e s , in c o m p le te ly L 15 1.8 3 or not ossified F 21 0.33 1, present L 1 0.12 A r c h , open F 1 0.02 L 1 0.12 F 1 0.02 C e n tr a , u n i l a t e r a l L 2 0.24 o ssificatio n F 2 0.03 0-1 (0-3.8) 0-1 (0-0.5) 0-1 (0-4.3) 0 -1 (0-0.6) 0-2 (0 -8 .3) 0-3 (0 -1.8 ) 0-3 (0 -12 .0 ) 0-4 (0 -2 .1) 0-1 (0-4.3) 0 -1 (0-0.6) 0 -1 (0-4.0) 0-1 (0-0.5) 0-1 (0-4.2) 0 -1 (0-0.6) S a c r a l: None, present A rch, open L 1 0.12 0-1 (0 -4.3) F 1 0.02 0 -1 (0 -0 .6 ) L 1 0.12 0-1 (0-4.0) F 1 0.02 0 -1 (0 -0 .5 ) C a u d a l: None p resen t 1 , p r e s e n t L 1 0.12 0 -1 (0 -4.3) F 1 0.02 0 -1 (0 -0 .6 ) L 1 0.12 0 -1 (0 -4.3) F 1 0.02 0 -1 (0 -0 .6 ) RIBS C ervical R ib (s ) present One o r more, wavy One or more, in co m p le te ly o s sifie d (hypoplastic). or not o s s ifie d L 31 3.78 0-5 (0-20.0) F 32 0 .5 1 0-5 ( 0 - 2 .7 ) L 44 5.36 0 -7 ( 0 -3 1 .8 ) F 73 1 .1 6 0 -15 (0 -8 .3 ) L 33 4.02 F 53 0.84 0-5 (0 -22.7) 0-9 (0 -5 .0 ) L: LITTER INCIDENCE F: FETAL INCIDENCE 038So 418-011: PAGE G-14 SUMMARY OF FETAL SKELETAL ALTERATIONS CD RAT ALTERATION RIBS (CONT. ) Fused S p lit Two segm ents RANGE/STUDY N% N L 3 0.36 0 -1 (0 -4 .3 ) F 3 0.05 0 -1 (0 -0 .6 ) L 2 0.24 0 -1 (0 -4 .0 ) F 2 0.03 0 -1 (0 -0 .5 ) L 1 0.12 0 -1 (0 -3 .7) F 1 0.02 0 -1 (0 -0 .5 ) MANUBRIUM D uplicated L 1 0.12 F 1 0.02 0-1 (0-3.8) 0-1 (0-0.4) STERNEBRAE One o r more in c o m p le te ly o s sifie d or not o ssifie d D uplicated Fused A sym m etric L 114 F 164 L1 F1 L1 F1 L1 F1 13.90 2.60 0 .12 0.02 0.12 0.02 0.12 0.02 0-7 0 -11 0 -1 0 -1 0 -1 0 -1 0 -1 0 -1 (0 -3 3 .3 ) (0 -6 .2 ) (0 -3 .8 ) (0 -0 .4 ) (0 -3.7) (0 -0 .5 ) (0 -4 .2 ) (0 -0 .5 ) PELVIS Pubis(es) and/or Ischium (a): incom pletely or not o s s ifie d P u b is(es): incom pletely o s s ifie d Pubis(e8): not o s s ifie d Ischium (a): incom pletely or not o ssifie d L 139 F 224 L 114 F 185 L8 F8 L 50 F 70 16.95 3.54 13.90 2.93 0.98 0.13 6.10 1 .1 1 0-9 0-16 0-9 0-16 0-3 0-3 0-4 0-9 (0 -39.1) (0 -9 .0 ) (0 -39.1) (0 -8 .9 ) (0 -13.6) (0 -1.6 ) (0 -16.7) (0 -4 .7 ) FORELIMBS K etacarpals: 1, present : 2, present F o red igits: 2, present Forephalanges: 1 , present L 1 0.12 0 -1 (0 -4.3) F 1 0.02 0 -1 (0 -0 .6 ) L 1 0.12 0-1 (0 -4 .3) F 1 0.02 0 -1 (0 -0 .6 ) L 1 0.12 0-1 (0 -4 .3) F 1 0.02 0 -1 (0 -0 .6 ) L 1 0.12 0 -1 (0 -4 .3) F 1 0.02 0 -1 (0 -0 .6 ) L: LITTER INCIDENCE F: FETAL INCIDENCE 003851 418-011: PAGE G-15 SUMMARY OF FETAL OSSIFICATION SITES SKELETAL AVERAGES CD RAT (CAESAREAN-SECTIONED DAY 20 GESTATION) PERIOD: JUNE 1995 - JUNE 1997 # STUDIES INCLUDED 33 # LITTERS EXAMINED 772 # FETUSES EXAMINED 5944 SKELETAL AVERAGES HYOID VERTEBRAE CERVICAL THORACIC LUMBAR SACRAL CAUDAL RIBS (p a ir a ) STERNUM MANUBRIUM STERNAL CENTERS XIPHOID FOREPAWS ( C a lc u la te d a s average per limb) CARPALS METACARPALS DIGITS PHALANGES HINDPAWS ( C a lc u la te d as average per limb) TARSALS METATARSALS DIGITS PHALANGES FETUS/LITTER MEAN RANGE/STUDY 0.84 (0 .6 9 -0 .9 5 ) 7.00 13.03 5.97 3.00 4.83 13.02 -- (1 2 .9 9 -1 3 .1 5 ) (5 .8 5 -6 .0 0 ) (2 .9 6 -3 .0 0 ) (4 .3 5 -5 .2 1) (12 .9 9 -13 .0 8 ) 1.00 3.57 0.99 (0 .9 8 -1.0 0 ) (3 .2 6 -3 .8 5 ) (0 .9 4 -1 .0 0 ) 0.00 3.49 5.00 5.05 -- (3 .3 3 -3 .6 2 ) -- (4 .9 0 -5 .2 7 ) 0.00 3.99 5.00 4.96 -- (3 .9 3 -4 .0 4 ) -- (4 .77-5.13) 003852 418-011: PAGE G-16 SUMMARY OF FETAL OSSIFICATION SITES SKELETAL AVERAGES CD RAT (CAESAREAN-SECTIONED DAY 2 1 GESTATION) PERIOD: JUNE 1995 - JUNE 1997 # STUDIES INCLUDED 2 # LITTERS EXAMINED 48 # FETUSES EXAMINED 374 SKELETAL AVERAGES HYOID VERTEBRAE CERVICAL THORACIC LUMBAR SACRAL CAUDAL RIBS ( p a ir s ) STERNUM MANUBRIUM STERNAL CENTERS XIPHOID FOREPAWS (C a lc u la t e d a s average per limb) CARPALS METACARPALS DIGITS PHALANGES HINDPAWS ( C a lc u la t e d as average per limb) TARSALS METATARSALS DIGITS PHALANGES FETUS/LITTER MEAN RANGE/STUDY 0 .9 7 (0 .9 6 -0 .9 7 ) 7.00 13 .0 4 5.96 3.00 7 .4 6 13 .0 3 -- (13 .0 2 -13 .0 6 ) (5 .9 4 -5 .9 8 ) -- (7 .2 3 -7 .6 7 ) (13 .0 1-13 .0 5 ) 1.0 0 3.98 1.0 0 -- (3 .9 7 -3 .9 8 ) -- 0.00 3.98 5.00 7.64 -- (3 .9 8 -3 .9 9 ) -- (7 .5 3 -7 .7 5 ) 0.02 4 .6 1 5.00 5.84 -- (4 .6 0 -4 .6 2 ) (5 .7 8 -5 .8 9 ) 003853 APPENDIX H STATEMENT OF THE STUDY DIRECTOR 003854 OPrimedica 418-011 P A G E H-1 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215) 443-8587 PROTOCOL 418-011: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF N-EtFOSE IN RATS SPONSOR'S STUDY NUMBER: T-6316.7 STATEMENT OF THE STUDY DIRECTOR This final report accurately reflects the raw data obtained during the performance of the study. No significant deviations from the U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations; Final Rule3, the Japanese Ministry of Health and Welfare (MHW) Good Laboratory Practice Standard fo r Safety Studies on Drugsband the European Economic Community (EEC) Council decision on 28 July 1989 on the acceptance by the European Economic Community o f an OECD decision/recommendation on compliance with principles o f good laboratory practicecoccurred that affected the quality or integrity of the study. Z7.rJ>r >nd G. York, pG i.f DABT Associate Director of R isearch and Study Director Date a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. b. Japanese Ministry of Health and Welfare (1988). G ood Laboratory Practice Standard fo r Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. c. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community o f an OECD decision/recommendation on compliance with principles o f good laboratory practice. Official Journal of the European Communities: Legislation. 32(No. L 315; 28 October): 1-17. 003855 APPENDIX I QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT 003856 O P rimedica 418-011 P A G E 1-1 Argus Research Laboratories, Inc. 9 05 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 4 4 3 -8 7 1 0 Telefax: (215) 4 43 -8 5 8 7 QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT Study Director: Raymond G. York, Ph.D., DABT Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Protocol 418-011: Oral (Gavage) Developmental Toxicity Study of N-EtFOSE in Rats Sponsor's Study Number: T-6316.7 The draft protocol for this study was audited for adherence to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice on 13 JUL 98. Critical phases of this study were inspected five times; study information and raw data were audited twice (see tables 1 and 2 for dates and phases/data). The draft final report and the raw data for this study [except for Appendix F, the Pilot Report, which was conducted in the spirit of Good Laboratory Practice (GLP)] were compared and audited for accuracy, for adherence to protocol requirements, and for adherence to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice between 12 NOV 98 and 02 DEC 98, and for revisions requested by the Sponsor on 10 DEC 98 and 17 DEC 98. 003857 418-011 P A G E I-2 This study was conducted according to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice. and Compliance Date Heather L. F o, M.S. Date Quality Assurance Supervisor and Principal Auditor 003858 TABLE 1 CRITICAL PHASES INSPECTED 418-011 P A G E I-3 Cohabitation Date of inspection: 21 AUG 98 Date results reported to the Study Director and Management: 21 AUG 98 Test Article Preparation Date of inspection: 02 SEP 98 Date results reported to the Study Director and Management: 28 SEP 98 Test Article Administration - Gavaae Date of inspection: 03 SEP 98 Date results reported to the Study Director and Management: 28 SEP 98 Blood Collection Date of inspection: 10 SEP 98 Date results reported to the Study Director and Management: 10 SEP 98 Caesarean-Sectioning Date of inspection: 10 SEP 98 Date results reported to the Study Director and Management: 28 SEP 98 003859 418-011:PAGE I-4 TABLE 2 RAW DATA AUDIT(S) The following study information and raw data were audited on 08 OCT 98, and 15 OCT 98 to 17 OCT 98: Protocol. Protocol amendments. List of personnel and computer operator codes. Error codes and codes for clinical sign observations. Animal receipt, randomization, physical examination and acclimation. In-life transaction record. Feed consumption. Cohabitation. Caesarean-sectioning. Maternal gross observations. Fetal gross observations. Fetal fixative assignment. Fetal visceral examination. Fetal skeletal examination. Necropsy. Tissue packing lists. Male breeder colony records. General comments. Study maintenance records. Temperature and relative humidity reports. Feed and water analyses. Edit requests. Dosage volumes. Deviations. Data review page. Key for test facility computer back-up record abbreviations. Blood collection data and packing lists. The results of this audit were reported to the Study Director and Management on 21 OCT 98. 003860 418-011 PAG E The following study information and raw data were audited on 08 OCT 98 ancMO OCT 98: Vehicle receipt, preparation and use. Test article receipt, preparation and use. Test article packing lists. The results of this audit were reported to the Study Director and Management on 21 OCT 98. 003861 Summary N-EtFOSE Rat Teratology Study Numbers: 3M T -6316.7, Argus 418-011 Compound & Lot: N -EtFO SE (N-Ethyl-Perfluoroocyanesulfonam ido Ethanol) - Lot FM3923 (30035, 30037, 30039) 98.2% pure (SM D Analytical Request 52489) Analytical Documentation filed along with final report. Note - same lot as used in two-year rat carcinogenicity study (T-6316, Covance 6329-212). Study Title: Oral (Gavage) Developmental Toxicity Study o f N -EtFOSE in Rats Report Date: 17 December 1998 Study M onitor Summary: (written by M arvin T. Case) Pregnant rat fem ales, 25 per group, were given daily oral (intubation) o f N -E tFO SE at dose levels o f 0, 1, 5, 10, and 20 mg/kg/day on days 6 through 17 o f gestation. The rat fetuses w hich had been exposed in utero during organogensis were collected at day 20 o f gestation. Approxim ately one-half o f the fetuses w ere examined for soft tissue (visceral) changes and other h alf were examined for skeletal changes. There were no compound related clinical signs o f toxicity in the dosed rat dams. A body w eight effect, reduced body w eight gains, occurred in the pregnant fem ales in highest two dose levels - 10 & 20 mg/kg. This body w eight effect correlated with reduced food consumption in the same two groups. The maternal toxicity at 10 & 20 mg/kg w as reflected in the pups as reduced pup weights. There was no effect on other litter parameters (corpora lutea, implantations, litter size, number o f live fetus, early or late resorptions, pup sex ratio) at any dose level. Increased delayed ossification sites were found in 10 & 20 mg/kg fetuses another reflection o f maternal toxicity at these dose levels. No compound-related soft tissue or skeletal malformations were found at any dose level. Thus, the compound w as not found to be teratogenic in the rat. The maternal toxic N O E L was 5 mg/kg. The fetal toxic N O E L w as also 5 mg/kg. Teratogenic N O E L in the rat w as > 20 mg/kg. N O T E : Dose preparation samples were collected tw ice during the study and these samples were sent to 3M Evirom m ental Analytical laboratory in B ld g 2. Likew ise, samples were collected from extra fem ales assigned as toxicokinetic satellite animals (3/group except control and high dose 5/group). These animals were killed on gestation day 18 (day after last dose) and the follow ing samples were collected: serum and liver from pregnant dams, fetuses and placentas (pooled by litter) from uterus. These samples were frozen and sent to 3M Evirom m ental A nalytical laboratory in B ld g 2. A t time o f this summary, January 1999, analysis o f these samples had not been done; therefore, these analytical results were not included in the final report nor in this summary. ' 003862