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jg ttjM U W ttl ARaU-O M G3 'C.'- c_1';-*--T < I\jt\ Oral Teratology Study of T-2998CoC in RAts -T 1 Experiment No.: Conducted At: Dosing Period: Study Director: 0681TR0110 Safety Evaluation laboratory Riker Laboratories, Inc. St. Paul, Minnesota April 6, 1981through April 16, 1981 E. G. Gortner / 2 -s r~ ir/ E. G. Gortner Date Senior Research Technologist Animal Teratology Reproduction A & AvtnwW E. G. Lamprecht, DVM, PhD Date Research Veterinary Pathologist 'f a n * .* T (H L , _ M. T. Case, DVM, PhD Manager, Pathology-Toxicology Safety Evaluation Laboratory Date 003034* 1. Summary Oral administration of T-2998CoC at doses of 0, 150, 50, 1.5 and 0.05 mg/kg/day to pregnant Sprague-Dawley rats during days 6 through 15 of gestation (period of organogenesis) was not embryotoxic and did not affect the ovaries or reproductive tract contents of the dams. Ohe compound did not cause abnormal gross, internal, or skeletal malformations of the fetuses. T-2998CoC was not teratogenic in the rat. T-2998CoC administration was maternally toxic to the 150 mg/kg/day dose group animals. It caused significantly low mean body weights during the dosing interval. Toxic clinical signs and deaths occurred in only the 150 mg/kg/day dose group. 003035 2. Introduction This teratology study- in rats was conducted to evaluate the embryotoxic and teratogenic effects of orally administered T-2998CoC-. The study was sponsored by 3M Commercial Chemical Division, St. Paul, Minnesota and was conducted by the Safety Evaluation Laboratory, Riker Laboratories, Inc., St. Paul, Minnesota. Two sets of compound administration groups were dosed between April 6 and April 16, 1981. The protocol and list of the principal participants and supervisory personnel can be found in Appendices I and II respectively. All portions of this study were conducted according to the Good Laboratory Practice (GLP) regulations and the Safety Evaluation Laboratory Standard Operating Procedures (see Appendix III for Quality Assurance Unit statement). The storage location for specimens, raw data and a copy of the final report is maintained in the Safety Evaluation Laboratory's record archives. Methods Time mated Sprague Dawley derived CD rats were obtained from Charles River Breeding Laboratory, Wilmington, Massachusetts, and assigned cages according to a computer-generated random numbers table. The rats, ranging in weight from 167 to 230 grams, were then divided into four groups of 22 animals each. The rats were housed individually in hanging stainless steel cages with wire mesh floors and fronts in a temperature and humidity controlled room. Food^ and water were available ad libitum. The lights were on a 12 hour light/dark cycle. The animals were observed daily from day 3 through day 20 of gestation for abnormal clinical signs. Body weights were recorded on days 3, 6, 9, 12, 15 and 20 of gestation and the rats dosed accordingly using a constant dose volume of 5 ml/kg of body weight. The five groups were dosed with T-2998CoC dissolved in distilled water daily at 0, 150, 5, 1.5 or 0.05 mg/kg/day. T-2998CoC was administered daily by oral intubation with a syringe equipped with a ball-tipped intubation needle to the rats on days 6 through 15 of gestation (day 0 indicated by sperm-positive vaginal smear). T-2998CoC analysis was provided by 3M Commercial Chemical Division, St. Paul, Minnesota (Appendix IV). All surviving animals were sacrificed on day 20 by cervical dislocation and the ovaries and uterus, including its contents, were examined immediately to determine the following: number of corpora lutea, number of viable fetuses, number of resorption sites, pup weights and sex, and any gross fetal abnormalities. Approximately two-thirds of the fetuses were preserved in alcohol for clearing and staining of the skeleton with alizarin red to detect skeletal abnormalities. Approximately one-third of the fetuses were fixed in Boilin's solution for subsequent free-hand sectioning by the Wilson technique to determine visceral abnormalities. In order to evaluate lens findings seen under the dissecting microscope, all eye sections with findings, plus select eye sections without lens findings were inbedded in paraffin, sectioned at 5-6 microns, stained with hematoxylin and eosin and examined histologically. Riker Experiment No. 0681TR0110 FC-143 Purina Laboratory Chow, Ralston Purina Co., St. Louis, MO 003036 |o |d*|o> 3. Results and Discussion T-2998CoC administered during the period of organogenesis was toxic to the high dose group (150 mg/kg/day) rats in causing low mean body weights during the dosing period. At gestational days 9, 12 and 15 (Table 1, Appendix V), the high dose group rats weighed significantly less than controls (0 mg/kg/day). The mean maternal body weights of the intermediate (5 mg/kg/day), mid (1.5 mg/kg/day), and low (0.05 mg/kg/day) dose groups were not different from the controls throughout the study. Abnormal clinical signs were observed and deaths occurred only in the high dose group. Three rats in the high dose group died. All three of the rats that died were ataxic and two of the rats were pale for one to two days before death. The surviving high dose rats did not have abnormal clinical signs and signs of toxicity did not occur in lower dose animals. T-2998CoC was not embryotoxic and did not affect the ovaries or reproductive tract contents of the dams. The mean number of male, female, total and dead fetuses, the mean number of resorption sites, implantation sites, corpora lutea and mean fetus weights of the four T-2998CoC dose groups were not significantly different from the control (Table 2, Appendix VI). T-2998CoC did not cause compound-related abnormal gross fetal findings (Table 3), nor did T-2998CoC treatment produce fetal skeletal malformations (Table 4, Appendix VII). A significant higher incidence of the skeletal finding of one sternebrae missing occurred in the high dose group. One sternebrae missing is a minor skeletal aberration and was not considered a malformation in this study. Further, the incidence of the finding of one sternebrae missing was not different among the control group and the lower three treatment groups. The incidences of skeletal findings associated with delayed ossification and rib aberrations were not different among the five treatment groups. A fetal lens finding was observed to occur in individual fetuses of all dose groups including the control group. The lens findings were localized to the area of the embryonal nucleus, although a variety of morphological appearances were present within that location. The range of morphological appearances as observed under the dissecting microscope included: a discoloration of the lens near the anteriocentral region extending from beneath the lens epithelium to half-way through the lens posteriorly, a cleft at the anteriocentral lens region or a combination of lens discoloration and the presence of a cleft. The lens findings observed under the dissecting microscope were interpreted histopathologically as either a freehand sectioning artifact of a normal area of primary lens fiber degeneration. The cleft was a space opened up at the vestage of the lens vesicle remnant and consisted of a separation of primary lens fibers of the embryonal nucleus from the lens epithelial cells. The dark streak discoloration of the embryonal nucleus resulted from either the lens being freehand sectioned across the area of normal primary lens fiber degeneration or an artifact being created in the lens during freehand and sectioning accentuating the area of normal primary lens fiber degeneration. The 003037 4. differences in the appearance of the lens artifact in individual fetuses and even among dose groups were largely due to the manner and frequency in which the artifact was created and the limitations inherent in visualizing the artifact under the dissecting microscope. Histologically, the lens artifact was the same in all dose groups regardless of the morphological appearance described under the dissecting microscope. T-2998CoC in utero exposed fetuses did not have compound-related changes in their lenses. 003038 5 BEST COPY AYMIABLE Table 1 Oral Teratology Study of T-2998CoC in Rats Mean Material Body Weights with Standard Deviations Dose Groups 0 mg/kg/day 150 mg/kg/day 5 mg/kg/day 1.5 mg/kg/day 0.05 mg/kg/day DRV 2 Q 12 15 20 MERN 1S*6 251 276 318 3S8 STRN. DEV 16. 1 23. 4 21. 4 28. 6 22. 3 27. 6 MERN 282 229 226 -- c. -T*-- c-C'i. * 361 STRN. DEV 14. 8 16. 1 29. 5 28. 6 23. 5 cl'8 - *9 MERN 281 252 277 312 390 STRN. DEV 12. 1 13. 9 14. 6 17. 1 28. 8 3:0 0 MERN 196 219 245 269 381 369 STRN. DEV 15. 2 14. 6 16. 5 15. 5 17. 9 26. 7* MERN 199 249 2*7*3^ --0 1, 390 STRN. DEV 13. 8 16. 2 IS. 2 21. 8 23. 2 32 4 -- Significantly lower than the control group (Dunnett's t test p < 0.05) 003039 Table 2 Oral Teratology Study of T-2998CoC in Rats Mean Litter Data and Fetus Weights with Standard Deviations Dose Groups 0 mgAg/day GRP NO. OR VIABLE RETUSES ANIMALS IT i 1T Hi. ____ 20 STAN DEV 4. 9 4. 9 1. 8 1. 8 Cl G DEAD R.'fcSORE IIUN IMPLANTATION CORPORfi MEAN HT RET USES SI IES SITES LUTEA f e t u s *:g > 0. 0 0. 0 0. 5 0. 8 10. 2 2 .:> 11. 4 1. 6 4.4 0 .4 150 mgAg/day 14 STAN DEV 4. 9 5. 1 10. 0 2 . 1 cl. cl 0.0 0. 0 0. 6 1. 0 10. 6 3. 1 11. 1 1. 9 4.2 0 .3 5 mgAg/day 21 STAN DEV j. 2 5. 2 10. 4 2. 4 1. 31 0. 0 0. 0 0. 5 0. 8 11. 0 1. 7 11. 2 1. 6 4.2 0.2 1.5 mgAg/day 19 STAN DEV 4. 8 1. 8 2. 5 8. 5 6 0. 0 0. 0 1. 1 1. 5 9. 6 2. 8 10. 1 2. 5 4 .2 0 .4 21 0.05 mgAg/day STAN DEV 5. 1 4. 9 1 0 .0 1. 9 cl. cS. 0. 0 0. 2 0. 4 1. 2 10. 5 d'. cl 11. 0 1. 8 4. 2 8. 2 BEST COPY AVA ll ABLE: 003040 a -- Treatment groups were not significantly different from the control group (Dunnett's t test p < 0.05) 7. Table 3 Oral Teratology Study of T-2998CoC in Rats Number of Fetuses with Gross Findings^. Findings Total Fetuses Examined Runted Small Umbilical hernia Total Normal Fetuses Total Abnormal Fetuses 0 mg/kg/day 196 1 -- -- 195 1 150 mg/kg/day 140 -- -- -- 140 0 5 mg/kg/day 219 -- -- -- 219 0 1.5 mg/kg/day 162 -- 2 -- 160 2 0.05 mgAg/day 211 -- -- 1 210 1 -- Treatment groups were not significantly different from the control group (Chi-square p <0.05) 003041 8 Table 4 Oral Teratology Study of T-2998CoC in Rats Number and Percent of Fetuses with Skeleton Findings Skeleton Findings 0 mg/kg/day Fontanelle not closed Hole in frontal Frontals not ossified Parietals not ossified Interparietals not ossified Stemebrae not ossified Stemebrae bipartite Stemebrae asymmetrical One stemebrae missing Two stemebrae missing Four stemebrae missing 13 ribs 13 ribs spurred Wavy ribs Protrusion on ribs One body vertebrae bipartite Two bodies vertebrae bipartite Three bodies vertebrae bipartite One body of vertebrae missing Total Number of Fetuses Total Abnormal Fetuses Total Normal Fetuses 24 (18) 17 (13) 17 (13) 17 (13) 40 (67) 14 (10) 20 (15) 9 (7) 2 (1) 4 (3) 7 (5) 6 (4) 35 (26) 6 (4) 1 136 126 (93) 10 (7) 150 mg/kg/day 18 (19) 14 (15) 14 (15) 13 (14) 64 (67) 1 10 (10) 30 (31)- 7 (7) 2 (2) 10 (10) 7 (7) 7 (7) 15 (16) 2 (2) 1 5 mg/kg/day 17 (ID . 5 (3) 5 (3) 2 (1) 101 (67) 12 (8) 29 (19) 8 (5) 4 (3) 5 (3) 3 (2) 4 (3) 30 (20) 14 3 (2) 1.5 mgAg/day 15 (13) 1 (1) 9 (8) 9 (8) 6 (5) 71 (63) 13 (12) 19 (17) 2 (2) 1 (1) 1 (1) 6 (5) 6 (5) 4 (4) 17 (15) 2 (2) 97 88 (92) 9 (8) 15( 136 (91) 14 (9) 11293 (83) 19 (17) 0.05 mg/kg/day 20 (14) 9 (6) 9 (6) 3 (2) 95 (64) 5 (3) 18 (12) 22 (15) 3 (2) 6 (4) 6 (4) 3 (2) 3 (2) 25 (17) 6 (4) 2 (1) 148 127 (86) 21 (14) Significantly higher than the control group (Chi-square p <0.05) Results from one fetus' are missing ( ) = percent of total examined Itri. 003042 Table 5 Oral Teratology Study of T-2998CoC in Rats Number and Percent of Fetuses with Internal Findings 9. Internal Findings 0 mgAg/day Fetuses with lens findings 5 A dark streak in the lens of one eye A dark streak & cleft in the lens of one eye A cleft in the lens of 5 one eye A cleft in the lens of both eyes Hydronephrosis Enlarged renal pelvis 17 (8) (8) (29) Abdominal cavity full of blood 2 (3) Total Normal Fetuses Total Abnormal Fetuses Total Fetuses Examined 38 (63) 22 (37) 60 150 mg/kg/day 5 mg/kg/day 1.5 mgAg/day 11 (26) -- 2 (5) 3 (7) 5 (12) 1 (2) 1 (2)-- 3 (7) 2 (3) 1 (1) 1 (1) 1 (1) 1 (1) 4 (6)3 (4) 6 (12) 6 (12) 9 (18) 1 (2) 30 (70) 13 (30) 43 59 (87) 9 (13) 68 37 (76) 12 (24) 49 0.05 mgAg/da\ 5 (8) 2 (3) 2 (3) 1 (2) 10 (16) 3 (5) 46 (73) 17 (27) 63 a -- Significantly different from the control group (Chi-square p <.0.05) ( ) = percent of total examined 003043 10. Appendix I TITLE: Protocol for Oral Teratology Study of T-2998CoC^- in Rats (Riker Experiment Number 0681TR0110). OBJECTIVE: A teratology study will be used to evaluate the embryotoxic and teratogenic effects of orally administered T-2998CoC to pregnant rats during the period of organogenesis. The procedure complies with the general recommendations of the FDA issued in January, 1966 ("Guidelines for Reproduction Studies for Safety Evaluation of Drugs for Human Use"). The study will be conducted according to the 1978 Good Laboratory Practice Regulations and Safety Evaluation Laboratory's Standard Operating Procedures. SPONSOR: 3M Commercial Chemical Division, St. Paul, Minnesota. TESTING FACILITY: Safety Evaluation Laboratory, Riker Laboratories, Inc., St. Paul, Minnesota. STUDY DIRECTOR: E. G. Gortner START OF DOSING: April, 1981. TEST SYSTEM: One hundred and ten sexually mature, time mated Sprague-Dawley derived female rats from Charles River Breeding Laboratory will be housed in hanging stainless steel cages with wire mesh floors and fronts in a temperature and humidity controlled room. This strain of rat will be used because of historical control data and time mated females are readily available. Purina Laboratory Chow and water will be available ad libitum. The lights will be on a 12 hour light/dark cycle. TEST SYSTEM IDENTIFICATION: Each animal will be ear tagged and that number will be indicated on the outside of the cage. RANDOMIZATION: The animals will be assigned cages according to a computer generated random numbers table. CONTROL ARTICLE: Corn oil. TEST ARTICLE: T-2998COC. ANALYTICAL SPECIFICATIONS: The test article, composition and purity will be determined by the Sponsor (3M Commercial Chemical group) prior to the start of the study and at the end of dosing. DOSAGE LEVELS AND EXPERIMENT DESIGN: The test article will be suspended in corn oil daily. The test article suspension and control article will be administered by oral intubation to the rats on days 6 through 15 of gestation according to the following: - FC-143 003044 U. Dose Level Group Size 150 mg/kg/day 5 mg/kg/day 1.5 mg/kg/day 0.05 mg/kg/day 0 mg/kg/day 22 22 22 22 22 The oral route of administration will be used because metabolism studies showed radiolabeled T-2998CoC was well absorbed. No dietary contaminants are known to interfere with the test article. The animals will be observed daily from day 3 through day 20 of gestation for abnormal clinical signs. Body weights will be recorded on days 3/ 6, 9, 12, 15 and 20 of pregnancy and the rats dosed accordingly using a constant dose volume of 5 ml/kg of body weight. The females will be killed on day 20 and the ovaries, uterus and its contents will be examined to determine: number of corpora lutea, number of fetuses (live and dead), number of resorption sites, number of implantation sites, pup weight and gross abnormalities. Approximately one-third of the pups will be fixed in Bouin's solution for subsequent free-hand sectioning by the Wilson technique to determine any visceral abnormalities using a dissecting microscope. Select eye sections can be sent to histopath for microscopic examination as deemed necessary by the study director. The remaining approximately two-thirds of the pups will be fixed in ethyl alcohol for subsequent skeletal examination after clearing and staining with alizarin red. DATA ANALYSIS AND FINAL REPORT: The proposed statistical methods to be used for analysis of the data are: Dunnett's t test for dam and pup weights, number of fetuses, number of resorption sits, number of implantation sites and number of corpora lutea; Chi square for percent abnormalities. The proposed date for the final report is 2-3 months after detailed pup examinations have been completed (approximately third quarter, 1981). Amendment to Protocol dhe control article for Experiment Number 0681TR0110 (oral teratology study of T-2998CoC in rats) will be changed from corn oil to water. The test article will not be suspended in corn oil daily as noted in the protocol, but solutions will be made by dissolving T-2998CoC in water by Dr. V. Pothapragada and the solution for the whole study will be submitted to 3M Commercial Chemical group for clearance before the start of the study and at the end of dosing. 003045 12. Appendix II List of Principal Participating Personnel NAME Edwin G. Gortner Elden G. Lamprecht Gary C. Pecore Vinkateswa Pothapragoda Loren 0. Wiseth FUNCTION Study Director Veterinary Pathologist Supervisor - Animal Care Commercial Chemical - Analytical Technician 003046 Appendix III STATEMENT OF QUALITY ASSURANCE STUDY NUMBER: TITLE: 0681TR0110_______________________ Oral T eratology Study o f T 2998CoC in Rats 13. Audits and/or inspections were performed by the Riker Compliance Audit unit for the above titled study, and reported to the study director and to management as follows : Date Performed Date Reported 9,14,16 A p r il 1981 20 A p r il 1981 20 August 1981 7 December 1981 14 December 1981 21 A p r il 1981 21 A p r il 1981 28 August 1981 14 December 1981 14 December 1981 Compliance Audit Riker Laboratories, Inc, *V / f * f / Date 003047 14. Appendix IV Prestudy and .Poststudy Analysis ofaT-2998CoC Distilled Water Solutions-- Dose Level Expected Analysis Prestudyg. Poststudy 0 mg/kg 150 mg/kg 5 mg/kg 1.5 mg/kg 0.05 mg/kg 0.0 mg/ml 30.0 mg/ml 1.0 mg/ml 0.3 mg/ml 0.01 mg/ml 0.0 ppm 30.328 mg/ml 0.983 mg/ml 0.268 mg/ml 0.0087 mg/ml 0.00 ppm 31.0 mg/ml 0.92 mg/ml 0.33 mg/ml 0.0092 mg/ml Pregnant rats were dosed at 5 ml/kg T-2998CoC 003048 15 BEST COPY AVAII ABLE Appendix V Oral Teratology Study of T-2998CoC in Rats Individual Body Weights (g) and Mean Body Weights With Standard Deviations For Pregnant Rats DRV 2 6 9 12 15 20 0 MG/KG/DfiV NlF: N1R N1F: NlF: NlF: N1R N1R NlF: N1R NlF: N1R NlF: NlF: NlF: NlF: NlF: NlF: NlF: NlF: NlF: 2937 293S 3000 3001 3002 3002 3004 3005 3000 2007 2052 3053 3054 3055 2056 2057 3056 3059 3061 3062 --*" 194 186 199 193 170 167 1S6 185 177 131 216 03 206 207 -134 191 204 191 243 219 214 222 216 . L i- 144 217 206 211 220 253 261 25 1 r' 226 Cl w 216 239 214 264 247 242 252 26.1 188 251 244 6 .'5'2 l.ip 289 267 268 270 '4J- 2S 2 266 226 292 281 261 288 261 290 218 276 266' 259 -?"i &1 1 201 262 291 292 235 267 251 288 261 321 215 292 225 291 231 254 12 297 297 214 327 247 227 -A-*>l *>er 289 286 230 286 402 282 250 414 256 415 241 274 -r3 3-6'2- 401 266 422 293 396 297 254 245 403 241 MERN 196 222 251 276 310 380 STfiH. DEV 16. 1 23. 4 21. 4 20. 6 -v 27. 6 NON F'F:EGNRNT RNIMHLS NlF: 2933 196 213 221 248 258 280 NlF: 2060 188 207 225 235 236 252- 003049 BEST copy AVAUA B U 16 Appendix V (Continued) Oral Teratology Study of T-2998CoC in Rats Individual Body Heights (g) and Mean Body Heights Hith Standard Deviations For Pregnant Rats DAV 6 Q 12 15 26 150 MG.-'KG,-'DA 01R 01R 01R 01R DIR 01R 01R 01R 01R 01R 01R 01ft 01R 01R 01R 01R 3:008 3011 3012 3013 3015 3010 3017 3018 3063 3064 3065 3068 3069 3070 3071 373 214 185 197 198 215 194 197 188 214 208 Z*r-* 193 20 182 230 197 236 215 cl!d!0 231 235 224 23 219 241 dl.2-.J254 219 cl!clO 195 264 218 256 179 244 246 201 '22-3 181 209 252 226 259 d!d!J d!d!3 200 'd7cl 188 d!i1. 297 del*Za 256 259 271 267 280 261 301 252 271 0- 0 250 270 26 ? 300 cl!i'd! 296 279 304 256 253 264 295 211 234 294 317 - 0 d"*"17CO_ J-J-r 337 346 J*CJ- "3 cr IT- 0 33-9 385 3-95 409 345 J*'Z*d` 303' 369 0 MEAN 202 229 226 259 36il STAN . DEV 14. 0 16. 1 29. 5 20. 6 23:. 5 28. 6 HON PREGNANT ANIMALS 01R 3009 194 220 174 223 242 d!6*J- 01R 3010 189 209 173 229 252 269 01ft 3014 183 207 25 207 238 257 01R 3066 187 201 153 0- 0 0 01R 3067 192 223 dd!t* 225 239 251 01R 3072 199 215 26? 224 236 253 Animal died 003050 BEST COPYAM IABLE Appendix V (Continued) Oral Teratology Study of T-2998CoC in Rats Individual Body Weights (g) and Mean Body Weights With Standard Deviations For Pregnant Rats 17 DRY 3 6 8 12 15 5 MG/KG/DAY P1R 201? 205 225 241 264 287 371 P1R 3020 182 215 243 268 301 374 P1R 3021 182 205 235 260 284 375 P1R P1R 3022 3023 182 188 211 221 245 253 274 308 il1*'-- 313 387 ---- eJr P1R 3024 208 268 288 323 410 F'lR 3625 214 241 266 300 3----C* 434 P1R 2026 188 206 233 258 *r-*52 355 P1R 3027 182 214 237 266 287 381 P1R 3028 212 221 257 c-3 322 <.'-*11^. P1R 3028 200 225 257 278 318 288 P1R 2074 206 254 278 308 385 P1R 3075 214 241 258 288 32'0 236 P1R 2076 182 208 37 258 280 26*3 P1R 2077 181 218 241 266 285 2 75 P1R 3078 210 248 3 r3 .302 345 458 P1R 3078 247 280 313 345 413 P1R 2080 188 228 252 282 312 P1R 3081 225 <*4r' 276 385 345 445 P1R 2083 187 210 231 248 3 icL 324 P1R 3084 200 224 251 271 306 384 MEAN 201 225 252 d'i*i' 312 3801 STAN . DEV 12. 1 13. 8 14. 6 17. 1 20. 01 301. 01 NON PREGNANT ANIMALS P1R 3082 208 250 264 268 278 003051 18 Appendix V (Continued) BEST COPY AVAII ABLE Oral Teratology Study of T-2998CoC in Rats Individual Body Weights (g) and Mean Body Weights With Standard Deviations For Pregnant Rats DRV " 6 12 15 20 1. 5 MG/KG/DA GIF: GIF: GIF: Q1R GIF: GIF: GIF: GIF: GIF: GIF: GIF: GIF: GIF: GIF: GIF: GIF: GIF: GIF: G1R 2630 3021 3022 3022 3024 2026 3027 2:02 202 304G 3085 30S6 308 308 201 302 30 2 304 305 17 1S2 iiilci 1S5 190 17 10 20? 212 12 18 184 1S 1S6 172 IS 22 221 202 202 216 227 211 212 20 21 225 226 cicicl 220 20 214 ciL2.il i4 204 ci51 244 222 224 25 226 261 25 272 241 261 242 271 234 ' 252 241 25 26 21 266 23 256 282 242 271 225 261 22 264 241 261 215 241 ciciji- 254 275 -298 273 289 24 274 21 2 2 325 291 306 286 d'S'ci 227 22 29 202 21 29 cif'O 271 2S5 226 321 211 36 357 415 368 290 250 252 412 405 346 273 21 37 221 34 256 372 290 2S5 MEAN 196 21 245 26 201 36 ST FIN. DEV 15. 2 14. 6 16. 5 15. 5 17. 9 26. ? NON PREGNANT ANIMALS GIF: 3025 186 212 227 ci5ci 271 265 GIF: 3087 186 203 230 235 24 261 GIF: 3090 191 212 225 244 242 255 003052 1 S T COPY AVA llABLfc Appendix V (Concluded) Oral Teratology Study of T-2998COC in Rats Individual Body Weights (g) and Mean Body Weights With Standard Deviations for Pregnant Rats DRV 3 6 S' 12 15 2 0. 05 MG/KG/D R1R R1R RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF RIF 3041 3842 3043 3045 3046 3047 3048 3049 3050 3051 3G96 3097 3098 3099 3100 3101 3102 3103 3104 3105 3106 Slid' 241 263 232 198 222 207 237 183 205 197 214 196 ClCl1191 221 180 212 250 221 250 188 206 200 2l3 189 218 183 2l2 188 204 207 242 218 253 179 207 205 234 2lL 236 263 251 237 267 236 250 257 238 279 269 226 233 237 227 268 280 227 260 264 Cl!?cl 276 267 298 245 260 280 280 256 312 292 249 254 252 cl5 5 245 293 295 247 290 287 323 304 299 337 280 296 320 314 286 354 328 281 289 274 286 287 325 274 322 320 403 364 370 413 358 360 406 387 341 424 414 367 355 314 359 352 405 430 338 409 406 MEAN 199 225 249 273 306 380 STAN. DEV 13. 8 16. 2 18. 2 21. 0 23. 2 32. 4 NON PREGNANT ANIMALS R1R 3044 183 196 211 222 231 246 C03053 Appendix VI Oral Teratology Study of T-2998CoC in Rats Individual Litter Data with Mean Fetus Weights 20 ANIMAL 0 mg/kg/day VIABLE FETUSES DEAD M F TOTAL FETUSES RESOR IMPLAN CORPRA PTION TATION LUTEA SITES SITES MEAN FETUS WTO AVG M F N1R N1R N1R N1R N1R N1R N1R N1R NIP. N1R N1R N1R N1R N1R N1R N1R N1R N1R N1R N1R N1R N1R 2997 2998 OCjClQ 3006 3001 3002 3003 3004 3005 300* 3007 3052 3053 3054 3055 3056 3057 3058 3053 3080 3061 3062 63 3 4 6 10 NOT PREGNANT 5 10 6 6 12 4 6 10 6 5 11 6 4 10 54 C| 4 9 13 63 6 5 ii 11 5 8 13 4 l`* 11 4 7* 11 6 Cl J. 3 44 8 NOT PREGNANT 7 3 12 2 8 10 MEAN 4. 9 4. 3 3. 8 STAN. DEV. 1. 8 1. 8 2. 5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0- 0. 0 0. 0 03 1 ii 0 10 0 12 0 10 0 11 13 2 11 0 13 1 10 0 11 l l 14 l 12 0 11 09 0 , 08 0 12 10 0. 5 10- 3 0. 8 2. 5 10 13 11 13 10 12 14 12 13 11 11 C| 14 14 10 11 C| 10 12 10 11. 4 1. 6 4. 9 4. 9 4. 9 3. 8 3. 3 3. 8 4. 3 4. 5 4. 2 4. 8 4. 0 4. 8 4. 2 4. 5 4. 4 5. 4 4. 0 39 4. 5 4. 9 4. 6 4. 2 4. 5 4. 6 4. 3 4. 7 4. 0 4. 9 4. 2 4. 4 4. 5 5. 5 4. 0 4. 0 4. 6 4. 9 4. 7 4. 4 4. 1 4. 4 4. 1 4. 9 4. 0 4. 6 4. 1 4. 6 4. 3 5. 3 4. 0 ft 4. 4 4. 8 4. 5 3; Q 4. 4 4. 5 4. 2 4. 0 4. 2 3. 9 4. 4 0. 4 003054 Appendix VI (Continued) Oral Teratology Study of T-2998CoC in Rats Individual Litter Data with Mean Fetus Weights 21 ANIMAL VIABLE FETUSES DEAD M F TOTAL FETUSES 150 mg/kg/day RESOR IMPLAN CORPRA PTION TATION LUTEA SITES SITES MEAN FETUS WTO AVG M F 01R 01R 01R CUR 01R 01R 01R 01R 01R CUR 01R 01R 01R 01R 01R 01R 01R 01R 01R 01R 01R 01R 3008 30O9 3010 3011 3012 3013 3014 3015 3018 3017 3018 3083 3084 3085 3086 3067 3068 3069 3070 3071 3072 3073- 4 7 11 NOT PREGNANT NOT PREGNANT 53 8 5 5 10 72 9 NOT PREGNANT 4 7 11 35 8 DEAD 34 7 6 8 14 6 6 12 10 5 15 DEAD NOT PREGNANT 5 7 12 4 8 12 63 9 11 2 NOT PREGNANT DEAD MEAN 4. 9 5. 1 10. 0 STAN. DEV. 2. 1 2. 2 3. 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0. 0 0. 0 2 13 12 3. 9 4. 0 3. 3 1 C| 0 10 09 Q 10 10 4. 4 4. 1 4. 2 4. 5 4. 2 4. 3 4. 3 4. 1 3. 9 1 12 12 4. 4 4. 7 4. 3 wo 10 10 4. 5 4. 5 4. 5 10 0 14 0 12 0 15 Cl 4. 7 4. 9 4. 5 14 . o 3- 9 3. 8 14 3. 3 4. 0 *T:` C* 14 -- `c* 3. 8 3. 8 0 12 12 4. 0 3-. 9 4. 0 0 12 11 4. 0 4. 1 3. cj 0 Q 10 y 3. 9 ~ "7 0 Cl 4. 3 4. 4 4. 3 0. 6 10. 6 1. 0 3. 1 11. 1 1. 9 4. 2 0. 3 003055 BESTcopy M i l M i r Appendix VI (Continued) Oral Teratology Study of T-2998COC in Rats Individual Litter Data with Mean Fetus Weights ANIMAL 5 mgAg/day VIABLE FETUSES DEAD M F TOTAL FETUSES RESOR IMPLAN CORPRA PTION TATION LUTEA SITES SITES MEAN FETUS WTCG AVG M F PIR PIP PIR PIP. PiR PiR PIR PIR PiR PIR R1R PIR PIR PIR PiR PiR PiR PIR PIR PIR PiR PiR 2019 226 2021 222 2022 2024 2025 2026 2027 2028 2029 2074 2075 2076 2077 2078 2079 2080 2081 2082 3082 2084 - 6 C| 4 Cl "7 1 0 12 5 5 10 2 C| 1 2 C| 5 14 68 C- 9 8 isl' 1 0 45 C| 4 6 10 r 5 12 4 12 i 11 tr 12 12 6 54 Ci Cl 51 3 12 NOT PREGNANT 61 7 4 6 10 MEAN 5. 2 er * 10. 4 STAN. DEV. 2. 2 2. 4 1. 9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0. 0 0. 0 1 10 0 C| 2 12 0 12 0 10 1 12 0 14 08 cL 1 1 0 10 0Q 0 10 0 12 0 12 1 12 0 12 0s cL 11 0 12 C| 0 10 0. 5 11. 0 0. 8 1. 7 10 10 14 12 11 14 14 C| 11 11 10 10 12 12 11 12 10 11 12 C| 10 11. 2 1. 6 2. 4 2.8 4. 1 5. 0 4. 2 4. 4 4. 2 4. 4. 5 4. 5 4. 6 4. 2 4. 5 4. 0 4. 2 4. 2 4. 5 4. 2 4. 5 2.2 2. 9 4. 4 5. 2 4. 2 4. 4 4. 2 4. 8 4. 6 4. 5 4. 7 4. 2 4. 7 4. 0 4. 5 4. 4 4. 6 4. 2 4. 6 2. 4 2.6 4. 0 4. 8 4. 2 4. 5 4. 2 4. 6 4. 1 4. 4 4. 5 4. 1 4. 1 4. 0 4. 2 4. 2 4. 4 4. 2 4. 2 4. 2 4. 2 4. 2 4. 4 4. 5 4. 2 4. 2 0. 2 003056 BEST COPY AVAII ABLE Appendix VI (Continued) Oral Teratology Study of T-2998COC in Rats Individual Litter Data with Mean Fetus Weights ANIMAL VIABLE FETUSES DEALM F TOTAL FETUSES 1.5 mg/kg/day RESOR IMPLAN CORPRA FT ION TATION LUTEA SITES SITES MEAN FETU AVG M WT<G F QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR QlR 2020 2021 2022 2022 2024 2025 3026 2027 3028 2029 2040 3085 3086 2087 2088 3089 2090 3091 2092 2092 2^894 2095 8 11 45 9 5 8 12 5 6 11 !j 5 10 NOT PREGNANT cl' 7 9 ill 4 6 5 10 ' 4 10 i1 cl 5 5 10 10 1 NOT PREGNANT 5 & 11 14 NOT PREGNANT 54 18 3 C| 12 5 5 10 6 C; 14 MEAN 2. 7 4. 8 8. 5 STAN. DEV. 1. 8 2. 5 2. 6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0. 0 0. 0 1 1 0 0 0 0 2 1 1 5 2 5 0 1 1 0 0 1 0 1. 1 1. 5 12 10 12 11 10 9 8 11 11 7 12 6 11 5 10 9 3 11 14 9. 6 2. 8 11 11 12 11 11 9 Cl 12 10 6* 14 C( 11 z` 11 Cj 4 10 14 10. 1 2. 5 4. 0 4. 4 4. 6 4. 5 4. 6 4. 1 4. 6 4. 5 4. 6 4. 7 4. 0 4. 2 4. 7 4. 4 4. 5 4. 1 2. 7 5. 1 4. 9 4. 6 4. 4 2. 4 4. 2. 5 5. 4 5. 1 5. 1 4. 5 2. 4 4. 1 2. 8 4. 9 4. 6 4. 2 4. 2 0. 0 r 'i V- iv i v iv i v iv i 2 9 4. 1 4. 5 4. 5 4. 0 4. 2 4. 0 4. 4 4. 0 4. 1 4. 4 4. 7 4. 5 4. 2 4. 2 0. 4 003057 BEST COPY AVAtt ABLE 24 Appendix VI (Concluded) JDral Teratology Study of T-2998CoC in Rats Individual Litter Data with Mean Fetus Weights ANIMAL VirtbLE FETUSES DEAD H F TOTAL FETUSES 0.05 mgAg/day RESOR 1MF'LAN CORF RA PT ION T A T ION LUTEA SITES SITES MEAN FE'I US h AVG N R RIF: RIF: RxF. RiF. RiF: RiR R1R RIF. RiR RiR RIR RiR R iR RIR RiR RiF: RiR R:t F: RiR RiR RiR RIR 204i 20h 2 30.42 3.044 i-^'4 L' XfcJH-C' 2047 2046 204 `3 2000 20bx x-0 3*cX 0 's1 202c: 2 33* 2100 216i 2i0>2 2iu'3 3X04 3x60 21018 4 r' 11 5X 8 4 6 10 NOT f r e g n a n t S if li if 4 10 4X r' x 3 ii I x 10 x2 5 t..^-i t, 11 if r' 12 r' tx 12 5 11 ii 2 72 x S 12 l` 4 11 4 4 13 X5 8 if 8 12 r' 0 12 MEAN 0 . i 4. A 10. 0 STAN. d e v . l. A 4. 4 2. 8 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0. 0 0. 01 11 01 s 0 10 0 11 0 10 2s 1- 12 0 10 05 0 11 0i 12 0 12 0 11 5 I1' 0 10 01 12 01 11 01- 12 0 ** 01 12 1 14 0. 4 10. 0 1. 2 4. 4 11 H ii ii .10 C| 10 10 C* ii 12 11 101 12 10 12 11 11 4 12 14 11. 0 1. 8 4. F 0. 01 4. 5 4. 2 4. 2 4. 2 4. 6 4. F 4. 5 4. 1 4. 4 4. 5 4. 4 4. 2 4. 0 4. V 4. 0' 4. 1 4. 2 4. 4 4. 2 4. 0 4. 5 4. 2 4. 6 4. 4 4. 1 4. 1 4. 0 4. 6 4. 1 4. 8 4. 2 4. 8 4. 2 4. 4 4. 2 4. 0 4. 4 4. A 4. 4 4. 1 4 0 4. 2 x. S4 4. 2 A, 1 4. 2 . 4. 6 4. 4 x .if. 4. 2 3-. S 4. 8 4. 2 4. 2 4. 0 4. 0 4. 8 4. 8 4. 2 4. O 4. 2 4. 2 0. 2 003058 Appendix VII Oral Teratology Study of T-2998CoC in Rats Number of Fetuses by Dam with Skeletal Findings 0 ag/kg/day Dam Number Total Number Fetuses Fontanelle not closed Frontals not ossified Parietals not ossified Interparietals not ossified Holes in pareitals 2997 2998 3000 3001 3002 3003 3004 3005 3006 3007 3052 3053 3054 3055 3056 3057 3058 3059 3061 3062 6778 787696819886 4687 131 12 1 11 3232 111 2 1 311 242 1 2 1 311 242 1 2 1 311 242 1 1 Stemebrae not ossified Stemebrae asysmetrlcal One stemebrae missing Two stemebrae missing 4 5 36 574 1820 121 1 2 2 212 3 2 1 212 1 2 755335 7 3 1 12 1 41 112 1 13 ribs 13 ribs spurred Navy ribs 1 Protrusion on ribs One body vertebrae bipartite Two bodies vertebrae bipartite Three bodies vertebrae bipartite 11 111 11 11 111 3341 2 511 1 112 1 22 21 212 1 1 1 2212 1 Total Abnormal Fetuses Total Normal Fetuses 5 7 7 8 7 7 7 4 9 5 8 0 9 88 446 8 5 1000 0 10 20 10 100 0 200 0 2 003059 W tn ISO mgAg/day Dam Number Total Number Fetuses Fontanelle not closed Frontal* not ossified Parietals not ossified Interparietals not ossified Stemebrae not ossified Stemebrae bipartite Stemebrae asymetrical One stemebrae missing Two stemebrae missing 13 ribs 13 ribs spurred Navy ribs Protrusion on ribs One body vertebrae bipartite Two bodies vertebrae bipartite One body of vertebrae missing Total Abnormal Fetuses Total Normal Fetuses Appendix VII (Continued) Oral Teratology Study of T-2998CoC in Rats Number of Fetuses by Dam with Skeletal Findings 3008 3011 3012 3013 301S 3016 3018 3063 3064 3065 3068 3069 3070 3071 86 76 21 1 1 1 8 6 5 10 a 10 8 8 6 1 1 222422 5 512 5 512 5 511 1264 36496 63851 1 321 21 1 1 2 1 1 5 1 10 3 3 3 1 15 2 1 42 1 11 21 2 2 41 1 1 31 1 3 1222 11 1 5 4 7 6 6 6 4 10 7 10 8 8 6 1 32 0 0 20 10 100 0 0 0 C4 m K) Appendix VII (Continued) Oral Teratology Study of T-2998COC in Rats Number of Fetuses by Dam with Skeletal Findings 5 mgAg/day Dam Number 3019 3020 3021 3022 3023 3024 3025 3026 3027 3028 3029 3074 3075 3076 3077 3078 3079 3080 3081 3083 3084 Total Nuaber Fetuses Fontanelle not closed Frontals not ossified Parietals not osfified Interparietals not ossified 6fi. 6 7 9 7 8 10 6 6 7 6 7 8 8 8 9 6 6 8 5 7 31 1 31 2 1322 12 2 12 11 Stemebraa not ossified Stenrebrae asymmetrical One stemebrae missing Two stemebrae missing 25 35366 4464 7754 84 37 35 11 1 12 1 12 11 242 125 1 1 34 4 1 1 31 2 13 ribs 13 ribs spurred Navy ribs 11 12 Protrusion on ribs 31 One body vertebrae bipartite 2 2 1 1 3 1 2 3 32 2 Two bodies vertebrae bipartite 2 2 2 1 Three bodies vertebrae bipartite 1 1 11 2 111 11114 1 222 1 Total Abnormal Fetuses Total Normal Fetuses 5 6 7 6 6 7 10 4 6 6 6 7 7 8 8 8 6 4 8 4 7 100 31 10 20 100 10 0 10 20 10 Results from one fetua are missing Kj -J 003061 1.5 m g A 9 /day Dam Number Total Number Fetuses Fontanelle not closed Frontals not ossified Parietals not ossified Interparietals not ossified Hole in frontal Sternebrae not ossified Stemabrae asymmetrical One sternebrae missing Two sternebrae missing Four sternebrae missing 13 ribs 13 ribs spurred Navy ribs Protrusion on ribs One body vertebrae bipartite Two bodies vertebrae bipartite Total Abnormal Fetuses Total Normal Fetuses Appendix VII (Continued) Oral Teratology Study of T-2998CoC in Rats Humber of Fetuses by Dam with Skeletal Findings 3030 3031 3033 3033 3034 3036 3037 3038 3039 3040 3085 30B6 3088 3089 3091 3092 3093 3094 3095 7-- 6 9 S 7 6 4 7 7 1 7 1 8 3 6 6 2 7 10 3 11 2112 22 2 11 1 11 1 1 2 11 1 11 1 1 2 11 1 1 1 1 59 34 1 354 16 16 1 36 26 8 11 1 222 1 12 33111 2 1 3 1 21 11 1 1 21 211 31 1 4 22 23 2 12 1 1 111 36l 75 3466 1718 t 462 79 50 312 30 1100 0 0 12000 1 28. 003062 Results from one fetus'* are missing Appendix VII (Concluded) Oral Teratology Study of T-299BCOC in Rats Number of Fetuses by Dam with Skeletal Findings 0.05 mgAs/day Dam Number 3041 3042 3043 3045 3046 3047 3048 3049 3050 3051 3096 3097 3098 3099 3100 3101 3102 3103 3104 3105 3106 Total Nuafcer Fetuses Fontanelle not closed Frontals not ossified Parietals not ossified Interparietals not ossified a6 78758 74898 816 8896 89 11 31 112 321 4 1 11111 3 1 11111 3 1 11 Stemebrae not ossified Sternebrae bipartite Stemebrae asymnetrlcal One sternebrae sdssing Two stemebrae missing 34 5234 544 366 5 1 3586 3 78 11 1 11 2 111 211 1 12122 1 311212 22232 11 1 13 ribs 21 11 1 13 ribs spurred Wavy ribs Protrusion ribs 1 1 21 1 1 2 One body vertebrae bipartite 1 2 1 2 2 2 1 1 1 1 2 2 4 1 Two bodies vertebrae bipartite 2 l 1 1 Three bodies vertebrae bipartite 1 1 2 1 2 1 Total Abnormal Fetuses Total Normal Fetuses 756 77486 4 3 76 714 7886 79 111 10 10 10 5 22 10 210 10 10 003063 fo vo 0 mgAq/day Daw Nustoer Total Number of Fetuses A cleft in the lens of one eye Enlarged renal pelvis Abdominal cavity full of blood Total Abnormal Fetuses Total Normal Fetuses Appendix VIII Oral Teratology Study of T-2998CoC in Rata Number of Fetuses by Dam With Internal Findings 2997 2998 3000 3001 3002 3003 3004 3005 3006 3007 3052 3053 3054 3055 3056 3057 3058 3059 3061 3062 3 3 3 4 3 3 3 3 4 3 3 14 3 3 32 24 3 1111 1 14 2 13 1 21 2 ' 11 01040002 32303331 1 3 1 1 11 2 2 1 2 0 0 30 20 32 1 11 043 e 003064 u> o Appendix VIII (Continued) Oral Teratology Study of T-2998CoC in Rats Number of Fetuses by D u With Internal Findings 150 mgAg/day D u N u b e r 3008 3011 3013 3013 3015 3016 3018 3063 3064 3065 3068 3069 3070 3071 Total Huabar Fatusea A dark atraak in the lena of one eye A dark streak C cleft in the lens of one eye A cleft in the lens of one eye A cleft in the lena of both eyes Enlarged renal pelvis 333 1 1 Abdominal cavity full of blood Total Abnormal Fetuses Total Normal Fetuaes 110 313 33334454431 1 31 3 11 1 1 1 11 300 133 10311130 1. 4 1 4 3 3 1 1 003065 u> Appendix VIII (Continued) Oral Teratology Study of T-2998CoC in Rata Number of Fetusea by Dam With internal Findings 5 ig/ki/d/ Nunbcr Total Nuaber Fatuses A cleft in the lena of one eye A cleft in the lena of both ayes Hydronephroaia Enlarged renal pelvis Abdominal cavity full of blood 3019 3020 3021 3022 3023 3024 3025 3026 3027 3028 3029 3074 3075 3076 3077 3078 3079 3080 3081 3083 3084 233 434423333444433423 1 1 11 1 11 1 11 Total Abnormal Fetusea Total Normal Fetusea 011 222 000 434 201 222 000 333 l'OO 344 2 1000 22342 0 3 003066 Lo K> Appendix VIII (Continued) Oral Teratology Study of T-2998CoC In Rats Number of Fetuses by Dam With Internal Findings 1.5 mgAs/day Dam Nuafcer 3030 3031 3032 3033 3034 3036 3037 3038 3039 3040 3085 3086 3088 3089 3091 3092 3093 3094 3095 Total Number Fetuses A cleft in the lens of one eye Enlarged renal pelvis 3 34 3332 3 31 30 31 331 34 1211 1 1 111 1 2 11 Abdominal cavity full of blood 1 Total Abnormal Fetusee Total Normal Fetuses 00 121 1110 10 3331221230 3 00 20 110 31130 24 003067 CUJJ Appendix VIII (Concluded) Oral Teratology Study of T-2998CoC in Rats Number of Fetuses by Dan With Internal Findings 0.05 n g A 9 /day Dan Ntafeer 3041 3042 3043 3045 3046 3047 3048 3049 3050 3051 3096 3097 3098 3099 3100 3101 3102 3103 3104 3105 3106 Total N n b e r Fetuses A dark streak in the lens of one eye A cleft in the lens of one eye A cleft in the lens of both eyes Enlarged renal pelvis 32 33324 31 344 31 34 34 244 11 11 1 1 11 12 11 11 Abdoninal cavity full of blood 11 1 - Total Abnormal Fetuses Total Normal Fetuses 1 1 2 1 0 1 0 1 0 1 3 1 0 0 0 ,0 1 1 0 1 2 2112 314 21 2 13 31 34 2 32 32 * 003068 U> t -DISTRIBUTION BIST 44. jT. Qase E. -G. Gortner (original + 1 ) F . J te lle r (QA f l i e s ) 7E. G. -Laxnprecht E . ;L. p u ts c h R. eleqr.1 -R. iE. Ober ;W. C. Mcparmick .-*F. HD. Griffit& \ f ) p . H. P earlso n G . R. Steffen--* J. D. jgend&rspp K.. '1. Ebbens 003069 M. T. Case E. G. Gortner (original + 1) F. Keller (Q A Files) E. G. Lamprecht E. L. Mutsch + R. A. Nelson R. E. Ober W. C. McCormick -F. D. Griffith (2) W. H. Pearlson G. R. Steffen -*J. D. Henderson -> K. L. Ebbens Amendment^to the Final Report oi th Oral Teratology Study of T-299f8CoC in Rats 'ct'oio Exprimnt Mb ;V 0 6 8 ITRO'fi0s Issued 12/15/81 Please insert the amended page 3 to the above report. Five word changes were made in the last two paragraphs. The study conclusions*&r not changed by this ^amendment to the results and discussion section of the 'report*. L * - -- - E. G. Gortner Date Senior Research Technologist Animal Teratology Reproduction E. G. Lamprecht, DVM, PhD Date Research Veterinary Pathologist ^ 7htoAM *n T * 2 / lf/ M. T. Case, DVM, PhD Date Manager^ -Pathology-Toxicology - Safety Evaluation Laboratory 003070 Amended page 3 to the Oral Teratology Study of T-2998CoC in Rats - Experiment No. 0681TR0110 Results and Discussion T-2998CoC administered during the period of organogenesis was toxic to the high dose group (150 mg/kg/day) rats in causing low mean body weights during the dosing period. At gestational days 9, 12 and 15 (Table 1, Appendix V), the high dose group rats weighed significantly less than controls (0 mg/kg/day). The mean maternal body weights of the intermediate (5 mg/kg/day), mid (1.5 mg/kg/day), and low (0.05 mg/kg/day) dose groups were not different from the controls throughout the study. Abnormal clinical signs were observed and deaths occurred only in the high dose group. Three rats in the high dose group died. All three of the rats that died were ataxic and two of the rats were pale for one to two days before death. The surviving high dose rats did not have abnormal clinical signs and signs of toxicity did not occur in lower dose animals. T-2998CoC was not embryotoxic and did not affect the ovaries or reproductive tract contents of the dams. The mean number of male, female, total and dead fetuses, the mean number of resorption sites, implantation sites, corpora lutea and mean fetus weights of the four T-2998CoC dose groups were not significantly different from the control (Table 2, Appendix VI). T-2998CoC did not cause compound-related abnormal gross fetal findings (Table 3), nor did T-2998CoC treatment produce fetal skeletal malformations (Table 4, Appendix VII). A significant higher incidence of the skeletal finding of one sternebrae missing occurred in the high dose group. One sternebrae missing is a minor skeletal aberration and was not considered a malformation in this study. Further, the incidence of the finding of one sternebrae missing was not different among the control group and the lower three treatment groups. The incidences of skeletal findings associated with delayed ossification and rib aberrations were not different among the five treatment groups. Fetal lens findings were observed to occur in individual fetuses of all dose groups including the control group. The lens findings were localized to the area of the embryonal nucleus, although a variety of morphological appearances were present within that location. The range of morphological appearances as observed under the dissecting microscope included: a discoloration of the lens near the anteriocentral region extending from beneath the lens epithelium to half-way through the lens posteriorly, a cleft at the anteriocentral lens region or a combination of lens discoloration and the presence of a cleft. The lens findings observed under the dissecting microscope were interpreted histopathologically as a freehand sectioning artifact of a normal area of primary lens fiber degeneration. The cleft was a space opened up at the vestage of the lens vesicle remnant and consisted of a separation of primary lens fibers of the embryonal nucleus from the lens epithelial cells. The dark streak discoloration of the embryonal nucleus resulted from either the lens being freehand sectioned across the area of normal primary lens fiber degeneration or an artifact being created in the lens during freehand sectioning accentuating the area of normal primary lens fiber degeneration. The C-03071 CU: M- T.case w. C. McCormick E. G. Gortner(original +1) R. A. Nelson F. Keller R. A. Prokop E. G.Lamprecht L. O. Wiseth ) t TITLE: Protocol for Oral Teratology Study of T2998CoC^ in Rats (Riker Experiment Number 0681TR0110). OBJECTIVE: A teratology study will be used to evaluate the embryotoxic and teratogenic effects of orally administered T2998CoC to pregnant rats during the period of organogenesis. The procedure complies with the general recomendations of the FDA issued in January, 1966 ("Guidelines for Reproduction Studies for Safety Evaluation of Drugs for Human Use"). The study will be conducted according to the 1978 Good Laboratory Practice Regulations and Safety Evaluation Laboratory's Standard Operating Procedures. SPONSOR: 3M Commercial Chemical Division, St. Paul, Minnesota. TESTING FACILITY: Safety Evaluation Laboratory, Riker Laboratories, Inc., St. Paul, Minnesota. STUDY DIRECTOR: E. G. Gortner START OF DOSING: April, 1981. TEST SYSTEM: One hundred and ten sexually mature, time mated Sprague-Dawley derived female rats from Charles River Breeding Laboratory will be housed in hanging stainless steel cages with wire mesh floors and fronts in a temperature and humidity controlled room. This strain of rat will be used because of historical control data and time mated females are readily available. Purina Laboratory Chow and water will be available ad libitum. The lights will be on a 12 hour light/dark cycle. TEST SYSTEM IDENTIFICATION: Each animal will be ear tagged and that number will be indicated on the outside of the cage. RANDOMIZATION: The animals will be assigned cages according to a computer-generated random numbers table. CONTROL ARTICLE: Corn oil. TEST ARTICLE: T2998CoC. ANALYTICAL SPECIFICATIONS: The test article, composition and purity will be determined by the Sponsor (3M Commercial Chemical group) prior to the start of the study and at the end of dosing. DOSAGE LEVELS AND EXPERIMENT DESIGN: The test article will be suspended in corn oil daily. The test article suspension and control article will be administered by oral intubation to the rats on days 6 through 15 of gestation according to the following: - FC-143 C03072  ' f.* Dose Level 150 mg/kg/day 5 mg/kg/day 1.5 mg/kg/day 0.05 mg/kg/day 0 mg/kg/day Group Size 22 22 22 22 22 The oral route of administration will be used because metabolism studies showed radiolabeled T2998CoC was well absorbed. Mo dietary contaminants are known to interfere with the test article. The animals will be observed daily from day 3 through day 20 of gestation for abnormal clinical signs. Body weights will be recorded on days 3, 6, 9, 12, 15 and 20 of pregnancy and the rats dosed accordingly using a constant dose volume of 5 ml/kg of body weight. The females will be killed on day 20 and the ovaries, uterus and its contents will be examined to determine: number of corpora lutea, number of fetuses (live and dead), number of resorption sites, number of implantation sites, pup weight and gross abnormalities. Approximately one-third of the pups will be fixed in Bouin's solution for subsequent free-hand sectioning by the Wilson technique to determine any visceral abnormalities using a dissecting microscope. Select eye sections can be sent to histopath of microscopic examination as deemed necessary by the study director. The remaining approximately two-thirds of the pups will be fixed in ethyl alcohol for subsequent skeletal examination after clearing and staining with alizarin red. DATA ANALYSIS AND FINAL REPORT: The proposed statistical methods to be used for analysis of the data are: Dunnett's t test for dam and pup weights, number of fetuses, number of resorption sites, number of implantation sites and number of corpora lutea; Chi square for percent abnormalities. The proposed date for the final report is 2-3 months after detailed pup examinations have been completed (approximately third quarter, 1981). "v_ J?~ 9 -y / E. G. Gortner Date Senior Research Technologist Animal Teratology-Reproduction Study Director M1.. T. Case, DVM, PhD Date Manager, Pathology-Toxicology Safety Evaluation Laboratory E. G-. Lamprecht, DVM, PhD Date Research Veterinary Pathologist W. C. McCormick, MS Toxicologist Sponsor Representative Dare' 003073