Document N278Ep5rLRdG7ZnybR9zRLYnV

C)--aTleratology *-tudyof T-299SCoC in P@r-a-@ Mcoeriment No.: Conducted At. Dosing Period: Study Director: 0681TRO110 Safety Evaluation laboratory Riker Laboratories, Inc. St. Paul, Minnesota April 6, 1981 through April 16, 1981 E. G. Gortner E. G. Gortner Date Senior Research 'i*chno7-ogist Animal Teretology Reproducl--ion E- G. Lampreclit, DVM, PhE) Date RP-search Veterinary Pathologist T. Case, DVM, PhD Date ?4anager, Pe.@jl,)logy-Toxic@;logy Safety Evaluation !Ab,,)ratcxy summary Oral administration of T-299SCoC at doses of 0, 150, 50, 1.5 and 0.05 mg/kg/day to pregnant Spracjue-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 dA-r-. The compound did not cause abnormal gross, internal, or skeletal malformations of the fetuses. T-299SCoC was not teratogenic in the rat. T-299SCoC 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. 2. Introduction This teratology studya in rats was conducted to evalgate the embryotoxic and teratogenic effects of orally administered T-299SCoC-. 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 11 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. Foo,(]Fa-nd 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-299SCoC dissolved in distilled water daily at 0, 150, 5, 1.5 or'0.05 mg/kg/day. T-299SCoC 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-299SCoC analysis was provided by 3K 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 Bouin'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. a Riker Experiment No. 0681TRO110 i@ FC-143 c Purina Laboratory Chow, Ralston Purina Co., St. Louis, MO 3. Results alndDiscussion 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-299SCoC 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-299BCoC dose groups were not significantly different from the control (Table 2, Appendix VI). T-299SCoC did not cause compound-related abnormal gross fetal findings (Table 3), nor did T-299SCoC treatment produce fetal skeletal malformations (Table 4, Appendix VII). A significanthigher 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 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-299SCoC in utero exposed fetuses did not have compound-related changes in their lenses. Table 1 oral Teratology Study of T-299SCoC in Rats Mean 14&terial Body Weights With Standard Deviations Dose Groups 0 mg/kg/day 150 mg/kg/day 5 mg/kg/day 1.5 mgAg/day 0.05 mgAg/day :12 :it, 20 ------------------------------------------------ MEAt4 :196 -qTAt-4. DEV :16. :1 2 2 251 27t----3..:It-l24E;C-i 4 21. 4 20. 6 22. 27. 6 MERN ;2C'cn_'22.'? 2:ZE-S STRt-4. [.,-EV14. 0 1 ti.1'. 5 2C,. t@; MEFit-@ 2C'l 2-7't- -STRN. E.,E%.--'t 1 :1 ? :14. :1 :kl Li MEAt4 196 213, 245 21':-@'-; STRI-4. E@E%o' :1!:7-. J.4. P---:.1 5 15. t, 9 2 E..i- MEFit-,'1'-; 249 2 :I'i.,. 2 7.-'@l, 1. Ci 2 C.1 -.2 4 a significantly lower than the control group (Dunnett's t test p 4r.0.05) Table 2 Oral Teratology Study of T-299SCoC in Rats Mean Litter Data and Fetus Weights with Standard Deviations-a Dose Groups 0 mgAg/day 150 mg/kg/day 5 mgAg/day 1.5 g/kg/day 0.05 mgAg/day GPP t-IC'."-IF'I.-'If-IF:FL.F.E-1 E. p: *1t@"IHi.- si Al.4 E@EV 1. 1. STAt4 C.-E-. @t. 5. 1 1 1 2. .:I--:. STFit-I L',E',,@ 1-4 T Flf-IL@EV !t7o.2 2. @l 1(:'. 4 1. 7 4. 2. 5 E' 21 .F.'TFit-LJ,EV 5. 1 4. 1. '.? 2. 1 Cl. C-1 I E.:'='. I I-IF,LHt-4THT'@lit-I '=.ITES ci ci. ci l@l. fLi ci (-iC.-i ci. cl 2 Cl. 0. II Cl. 4 1. 2 11. 0 8 1 Cl. !T. 2. @:1 Treatment groups were not significantly different from the control group (Dunnett's t te 7. Table 3 Oral Teratology Study of T-299SCoC in Rats Number of Fetuses with Gross FindingsS Findings Total Fetuses Examined Runted Small Umbilical hernia Total Normal Fetuses Total Abnormal Fetuses 0 mg/kg/day 196 1 ----195 1 iso mg/kg/day 140 -----140 0 5mgAg/day 219 ------219 0 1.5 mg/kg/day 162 --- 2 --160 2 0.05 mgAg/day 212 ----- 1 210 1 Treatment groups were not significantly different from the control group (Chi-square p < 0.05) 8. Table 4 Oral Teratology Study of T-299SCoC in Rats Number and Percent of Fetuses with Skeleton Findings Skeleton Findings Fontanelle not closed Hole in frontal Frontals not ossified Parietals not ossified Interparietals not ossified Sternebrae not ossified Sternebrae bipartite Sternebrae asymmetrical One sternebrae missing Two sternebrae missing Four sternebrae 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 0 mgAg/day 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) iso mgAg/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 97 88 (92) 9 (8) 5 mg/kg/day 17 (11) 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) 150b 136 14 (91) (9) 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) 112 b 93 (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 21 (86) (14) a Significantly higher than the control b. Results from one fetus.are missing percent of total examined group (Chi-square p < 0.05) 9. Table 5 Oral Teratolocjy Study of T-2998CoC in Rats Number and Percent of Fetuses with Internal Findings internal Findings 0 mg/kg/day Fetuses with lens 5 findings dark streak in the lens of one eye dark streak & cleft in the lens of one eye cleft in the lens of 5 one eye 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 11 (26)@@ 2 (5) 3 (7) 5 (12) 1 (2) 1 (2)1 3 (7) 2 (3) 1 (1) 1 (1) 1 (1) 1 (1) 4 (6)1 3 (4) 1.5 mg/kg/day 6 (12) 6 (12) 9 (18) 1 (2) 0.05 mg/kg/day 5 (8) 2 (3) 2 (3) 1 (2) 10 (16) 3 (5) 30 (70) 13 (30) 43 59 (87) 9 (13) 68 37 (76) 12 (24) 49 46 (73) 17 (27) 63 2@Significantly different from the control group (Chi-square p -,0.05) percent of total examined lo. TITLE: Appendix I Protocol for Oral Teratology Study of T-299SCoCA -in Pats (Piker Experiment Number 0681TRO110). OBJECTIVE: A teratology study will be used to evaluate the embryotoxic and teratogenlc effects of orally administered T-299SCoC 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 1976 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 wlre 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. I>urina 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 computergenerated random numbers table. CONTROL ARTICLE: Corn oil. TEST ARTICLE: T-299SCoC. 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 Dose Level Group Size 150 mg/kg/day 22 5 mg/kg/day 22 1.5 mg/kg/day 22 0.05 mg/kg/day 22 0 mg/kg/day 22 The oral route of administration will be used because metabolism studies showed radiolabeled T-299SCoC 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 REI>ORT: 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 The control article for Experiment Number 0681TRO110 (oral teratology study of T-299SCoC 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-299SCoC 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. 12. Appendix 11 List of Principal Participating Personnel NAME Edwin G. Gortner Elden G. Lamprecht Gary C. Pecore Vinkateswa Pothapragoda r,oren 0. Wiseth FUNCTION Study Director Veterinary Pathologist Supervisor - Animal Care Commercial Chemical - Analytical Technician Appendix 111 13. STATEMENT OF QUALITY ASSURANCI@ STUDY NUMBER: TITLE: 0681 TROI 10 -oral Teratology Study of T 2998CoC in Rats 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 April 1981 20 April 1981 20 August 1981 7 December 1981 14 December 1981 21 April 1981 21 April 1981 28 August 1981 14 December 1981 14 December 1981 @C-oam@p ia@nce@Au Riker Laboratories, Inc. Date 14. Appendix IV Prestudy and,Poststudy Analysis ofaT-299SCoC Distilled Water Solutions- Dose Level Expected Analysis EEEEtudya_ oststudy 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 Appendix V Oral Teratolocjy Study of T-299SCoC in Rats individual Body Weights (g) and Mean Body weights With Standard Deviations For Pregnant Rats -------------------------------------------- C, 14:IF'. 2 4 2E.4 2:?@--' 2 L'i 4 -A N:LF'. 2 9'-;E. :194 ;21'-;, ;247 ;-281 7-1@5 -7'. t4lP 3000 ISE, 214 24a 261 2 9 C--*3!!C7, NIP 3001 199 222 252 288 325 414 141P 2 0 0 "-:' 1'-71--@: 216 2,9 261 2'-=11 -7'56 t4lP 170 22"@;: 2t:--l ;2-'t-;o-, 4:1!@, t-i:IF' CiCi4 I t--- :144 1 -S"=-@ .21,,i: 2T.4 -@@'41 NIF' --t'0O5 1 -S. 2 1 C-@'51 2'i@E, 12 NIP ClCiE. :IE:!f, L, C-1 4 4 2E-.El NIF;' r@C,7 17-.-' 21:1 2 NIP 105@7' 1'-;1 220 252 NIP 5.@; 21.i, 25@;@ 2 -1 2 5,-L 2,-; 271, ---,14 4 C,1 7 E. NIP 04 6 261 Z@C, 4 NIP' 0 2 5 ZE-7 .2,-741 7 9,-4: NIP 56 20 2@@ 2tE8 2 9.:-; 2 - NIP 2051@ 201- 217 t--, t 325 7 't41 F-..' -;@C-15, =-' -IE44 2 2 ;2 4 *-7@, 2 Z-":-; -7@5: 4 NIP :<C,5.P 191 NIP 2061 204 N I F,.' --<C,t 191 2:IE. 27 .'zo 214 212 2 E. 2 t--. Z,5:1 2,-=. 2E.1 2:E.E. Ci 2Et-. 45 4 Ci 7@41 MEFit-4 196 22 STAt-4. E-E%.'16.1 21. 4 251 276 310 380 1. 4 20. E. 2 E. I-JC'tP-FJEGt-4Flt-4FTiNItIFiL@---. NIP' 299'-:y 196 21,-:@ N:IP. :LE.E: 225 9-;.'4E2.5:i: 2SCi 4-;,:@;5 16. Appendix V (Continued) Oral Teratology study of T-299SCoC in Rats 3:ndividualBody Weights (g) and Mean Body Weights With Standard Deviations For Pregnant Rats 7: :LZ, :15 2 ci -------------------------------------------- C:LF, C,Cl: CIIF.' "-@CIlI O:LF.' OIR 3012 OIP 0 15 OIF' I OIP '@::1.O OIF., C.I, E: OIF: C, Clip. cifE4. OIF' C,t-5. C iI F-.., --<CiE. OIF, C- C,IF., ri7 C-i C-IIF* -@zCl I OIF, C, 14 2 E. IE:5 215 :19"; Z:';.(:.'1 19$ 2--<l 1 r--@ 2-75 :1-ci4 ;224 1 2 ci 2 1,-;t 4-:1,14 4 1 2C-11.-:; L'2 254 1,-; 2:1,-:4 CiCi 22,=-: I I-5c4 -:-Ci 2iE.4 1,-:; 2 1 -;5'E. 2t@l 2'-f:"; 1-47-q 25 -Z4;*4 2 5'-4; 271 e@-46 267 280 201 26-:1 :<Ol 2 7-"-;1 2!f-2 271 181 ci r-i 2 C.,,-; 25C, 270 2152 2p-z',@ C,C1. 2 29E. 259 279 -@Cl4 'e--, 2!@-ii. 25- 2 2 --z, 2E,4 2'-@:5 2Cc,.-2i11 2 -@%'4 217@2 294 --'.,17 ci t@-i 7 27@7 3.-;L7 346 '5 ci 9 4 C, 7--.4 ---,cL-l::7 ci MEFit-4 202 229 226 2 5,-zl 282 ---:61 S.TFit-4[.@E'-,14@. Cl 1 1 2'-1=.5 2 C'.E. 2 5 2 e.. f-ICit-4F,F*EGt-lRf-jATNIr-IFiL-=.' OIR OIP OIF' OIR OIP OIF' 09 ZOICI I -4 3CI66 306f30-1@'2 1 @-4; IE"z'i 1 18 192 19'-; 220 174 2Ci,:4 17-:@ 2Ci. 2C'5 201 15--,. 22@z:- 22e 215 207 2229 2-k7 Cl 22-@224 242 25 Ci 2--@:9 2 7@*6 2 Cl 251 25 AninAl died 17. Appendix V (Continued) oral Teratology Study of T-2998COC in Rats individual Body weights (9) and Mean Body Weights with Standard Deviations For Pregnant Rats ---------------------------------------------- 5 PiR PIF' PIR P:LF.' P:IF' F-IR PIF* PIR PIF' PIF, PIF.' PIF* PIR P:LF, PIF' PIF.' PIF,' PIF., PIP PI.F' PIP 205 225 241 264 aO2O 191' 215 2 4'@@ 268 30 --'" 18 *.:-@ 205 2 --5< 260 3022 182 211 245 274 2 1'-1;'i: 2 1 25-@@. 2 7 :,,024 20E@ 2-7.2 2p--.8 2 8'-@ 3-.025 214 241 2 6 '0 aoc-@16 18-Ci Ci I'@t 2:1@. --C<lL.9 2-00 -7*@C, --1-4 2C,@:. 2ot-@ -='I4 2-7 ;2@.1 22@:- 25 2.L",:: 254 259 2E.6 2E 27E., 2"179 2075 214 241 25E: 20 :1 -q --@, 2Ci-:- 20*@'@7 191 219 241 266 3 7 E.' 2 1 9 I 2 4 a-.' 2 -c@ 7-,' -.-02 246@ 2i---.Cl .7';1 L,2,-: 252 28.2 Ci,,=,-,-@: 19'17 E.4, 2 f.'C@,l 24'@ 2 7 t--- 3Cit. 2:IC@ 2 -.1 2 4'-; 2:-:.'4 ;2!@,:l 271@l 29(@ :1"1 294 3 Ci --1< @,'2 2'-:; 29" 12 271 7-@4 2*1@@5 29-.P -85 410 4--.4 4 3 C.i 290 29-@. .1@.'4!T. --<45 -:zl2 4 272 :,;@--ti--. @-75 45--=-41"7: 445 324 le-4 MEFit-4 201 225 2 5C-L,27-i@ 312 STFIN. E@E'-.@ :1 :1 :L4. 6 17. 1 2 390 Cl. i.-@l NON PRErjNRt4T P:LF: 2C,9 25Ci 21---.42E.9 27E., APpendix V (Continued) Dral Teratology Study of T-299SCoC in Rats Individual Body Weights (g) and Mean Body weights With Standard Deviations For Pregnant Rats :12 :L5 2 C, --------------------------------------------- 1. 5 MG.,-'i-'G.-,'[*@f@ C!IF. QIF' OIR OIF' C!IF, C,IF::l OIF' QIF.' C!IF' QIR QIF' OIF: F-., QIF., C-!IF' C!IP 0 2.'0 -@-'O :I 30-@2 3022 207@4 O'@7:6: C'-@'7 0 10@9 :,0: 4 C-1 30E!6 --'OSE: 3091 3 0,-:4 3 C"-: --<C'94 -7':095 17E: 202 IE:- 2:LC' 212 227 185 211 190 21-@. 1 *.,@r,-; 2 19C' 21:=2 C, 21-@ 1 184 216 2 20 20--Li 1 214 221 17@: 1-;14 2C'4 229 :.'5:L 221 244 201 222 2.'@-'4 2.7*E;. 25-:@ 241 2 4.:-6@' 2 --4< 241 2 25621i-I 12 261 271 2 5. 25 22'-; 24 25 2-; 24-:1 2:1 275 2--@I 249 2-1 264 2tFl 4 j. 254 29 2 e. 274 2:?;l E. 2 251@ 415 291 360 3Ci6 C.1 2 S-t--. 5 Ci 2 9.:-@ ZA 1. 29 4Clt4 Ci 271 -@4:=- ;Zel.5 5 E' 2 2 211 !8t, MEFIN 196 STFIN. DE%., :It Z'l3 245 26:? 7.01 7:6:@ :14.6 :1E..5 J.5. 5 :1 27 NON PREGNFit-4T Flr-JIr-IFIL-c QIF' 307z5 166 2--,'.2752 265 QIR 308"@ 186 20@ 2aO 2-:5 249 261 C41F' 3090 191 212 2Z,5 244 242 255 19. Appendix V (Concluded) Oral Teratology Study of T-299SCoC in Rats 3:ndividual Body Weights (g) and ?4ean Body Weights with Standard Deviations for Pregnant Rats :Ito 2 ci ---------------------------------------------- Ci. C,to MG.,'@--*"G,,'E.- RIR 3041 RIF, 2042 F,:lF., 3Cl4l, F.'IF., 2C145 RIF' -7,:064 F,IF* a04,@ RIF, F,IP. F.I, F, F.I* F.* PIP P:LF, PIR RIF, PIP F'IR F,IF, P:LF* PIF: F*IF. 3049 '-@C.--5" C--'l -:-C5'1 :,0 -a@t-'. a@9i" Cl,@.::--, 0,-.94 3100 -7@'It.I-L, 3 1 ri I C, 3:Lrj4 3 1 tL5-1 -,'106 212 20--, 1?8 20 I"@ 18-7' 1?,. :1 191 I "C@"@:"I-L 18Et 2 C,k'-i 189 18-7. 18S 207 179 205 21L' 241 2 --@2: 222 2:,.'7 205 214 27-,l 2 4-1;, 2:LZ, :-@@'5Cl -L*.f*-!C2 0 C-. 2:12 212 204 242 2!f. 2 r:@ 24 2 E. 26a 2 9'L.-@'3 2 40@@. 251 27E'. 304 164 237 2E.7 299 370 267 2:?8 327 4:L'-@. 225 24t- 2 0 5 8 2E.Cl C-I 25C, 2:?.Ci -i-Z, @-1 4 i.7fE,. 2517 280 al4 2S-17 2 --1,7E'. 2 5 t-. 2 *17,-:4 '-@'45 4 -7'1c 2E.9 292 414 226 2 4,-@4 281 2 -: - 254 28.'@ 5'--: 27@.5 252 274 .14 255 2'7@,17 245 ;2,i,. 5 29--,. 5 4 L-5i 2 295 2 4 Ci Z,27 247 2 E.1-@l29Ci ;274 'aS 3 2 :-z' 4 264 2E:7 320 406 MEFit-i 19 STAt-4. E)E'-.:-L' 225 249 27a :1E.. 2 :1 ;2:1.C, 280 2 4 NC'ti PP.E'Bt-IFlt-iATt4lMAL'---'. RIP -@C144 18-@' 211 2LI2 2-@.1 24E, 20. Appendix Vi Oral Teratology Study of T-299SCoC in Rats individual Litter Data with Mean Fetus Weights AN I MFIL 0 mg/kg/day VIFIBLE FETUSES m F TOTAL CEFICFETUSES NIP NIP NIP NIP 14IR NIP NIP N:IF' NIP tqlp 14IR N:IF' NIP N:LR NIP N:IF' N:IF, NIP NIP NIP NIP NIP 299-(@' 299:E: 299-q 3 C'C,0 3C'iDi 3002 3003 3CIC14 3CIO5 3000---. --:r<@ 0 -(,@ --<052 3 Cl5 :<Cl5 4 3r-155 3056 205f@ 305E' --<t-;159 3060 3r@161 3 CA 6 3 9 4 6 10 NOT PFEGNFir-JT 5 5 :Lo E. 6 12 4 6 lci i@; 5 1:1 E. 4 1 ci 5 4 9 41 E. 5 :1 1 !!, 8 4 7 4 7 IE. ll 12 :L7. i:l :li 4 NOT 7 2 4 a PRECjt4At4T 5 12 8 :10 MEFIN STFIN. DEV. 4. 9 1. 8 4. -q 9.8 1. 8 2. 5 ci ci ci ci 0 ci ci 0 &--I ci Pi0.0 0.0 RESOF' IMPLRN PTION TFITION ';ITEE. SITES C:(-JFF'FFi MEFit-4FETLI,-=.W*l-'G'..l LUTER FIVG m F r_-i 9 :1 0 io 0 12 0 io 0 :L:i :17. 2 :11 :1C-i 11 1. 1 :12 14 12 1 14 1 12 ci 11 9 0 6 C-1 E.' 0 :L2 ci io 0. 5 0.8 :1El2.5 9 14 14 1 ci :li I ri :1L, :L(:l :1:1.4 1.6 4. 9 3. 4. 9 so 4. 9 a. E. 4. 2 4. 5 4. 2 4. 8 4. 0 4. E.' 4. c-;* 4. 5 4. 4 5. 4 4. C, 1. 9 4. 5 4. 9 4. 6 4. 4. 5 4. 6 4. 7-: 4. 7 4. Cl 4. 9 4. 2 4. 4 4. !5 5. 5 4. ri 4. 0 4. E. 4. 9 4. 7 4. 4 4. 1 4. 4 4.1 4. 4. 0 4. E. 4. 1 4. E. 4. 5. 4. C, 3. 8 4. 4 4. 8 4. 5 -@<. 9 4. 4 4. 0 4. 5 4. 2 4. 2 3. 4. 4 0.4 21. Appendix VI (continued) oral Teratology Study of T-299SCoC in Rats individual Litter Data with Mean Fetus Weights ANIMAL 150 mg/kg/day VIFIBLE FETUSES m F TOTAL DEFIC.FETUSES O:LR 3clos 4 7 :L:l ci OIR 3009 NOT PF'EGNAR-JT O:IF.' 3010 NOT PREGf4Flt4T OIF.' 3011 !fl i 8 0 OIR 301:;, 5 5 :Lo 0 OIR 3013 7 9 0 O:LR 3014 NOT PF.*EGNFINT 0 1 F..' 3C'15 4 7 :L:l 0 OIR 3016 5 8 0 CI:IR C-17 CjEFil.@ 0 1 F,,. O:IF' :<C4 Z'. 4 7 El 8 :14 ci clip 3064 E. 0 OIR 3065 1-0 5 5 OIR 306t:@ C-EFiE., O:IF: 3 CiE.'17 NOT PF..'El:it-4Fit-JT OIF.' 3CItES' 5 12 c;i 4 8 :12 0 OIP 3070 IE. 9 0 OIR 3071 1 1 2 0 OIR 3C472 NOT PREGt4At4T OIR 2073 E)EAL, MEAN 4. 9 5. 1 :1 Ei Cl. 0 ';TFIN.DEV. 2. 1 2. 2 3 0. 0 REE-OF' IMF'LFIN COF-F-F@rl MEFir-4 FETI-I-- PTION TATION LLITER FIVG m F SITES SITEE. 2 :L3 :12 9 4. Ei :L 9 0 :1ci 0 9 1 12 2 :1C-i :1 r@i ci 14 0 12 0 15 ci 12 0 C-1 9 C, 2 !i@ :Iri :1 @-i :1, :1 C-1 9 14 14 14 :Is. 11 1 El 9 4. 4 4. 5 4. 4. 1 4. 2 4. 2 4. 1 9 4. 4 4. 5 4. c 4. 5 4. 7-. 4. 5 4. 7 4. 4. C-t E: 8 4. 5 :E: E 4. C, 4. 0 4. 4. 1 9 4. 4 4. 0 Z%. @A 7 4. 0. 16 1. ci 1 l:l. .1 1.9 4. 2 0. 3 22. Appendix VI (Continued) Oral Teratology Study of T-2998coC in Rats Individual Litter Data with Mean Fetus Weights Fit-M4F1IL 5 mg/kg/day VIRE-LE FETUSES DEFi[@, m F TOTFIL FETUSES RE-qOF., IMPLFIN F-TION TFITION SITES SITES. C:OF.-PF,.-FMlERIN LUTEH Fi%,-'G FETUS m I-IT<G.. F F*:LF: CiI --4; F,I.f-@, C.i Ci F'IF' 21 C- ci C-@.,@ - PIF.- 0 :-4:, PIF' F-'IF: 0 C-L.'5 F,IF, F,IF-: Ci.;-@ F,lr-: C,1 PIF. C, 4 F'!F' 2C,17@t- F,:IF: Cii@E. Ci F*IR 07E: F'IF: 0 7,-: P:IF: C.I a-:Ci 2081 F':IF: F'IF' C.I P:If@l.: '-*=4 2 I:, 4 5 7 E. 7 5 5 :< !? 9 5 7 2 8 2 4 5 4 E. 7v 5 4 11 5 E: 9 9 io 1 1 C-1 :12 14 9 1 ci 9 I ci :12 :lz :12 :1-n 5 4 9 NCIT PF.EGt4At-4T t-, 1 7 4 E. I ci r-IEFit-i 5. 2 2. 2 5. 2 1 Ci.4 2. 4 :1.9 0 C-1 0 0 1:-i 0 0. 0 0 0 0 ci 0 ci ci ci 0 ci 0 Ci.ci 0. 0 1 1 0 2 :12 0 17. :10 ci 1 :13 0 14 2 0 0 0 ci 12 0 12 :1 12 C-i I :12 2 ? ci :io Cl.5 0.8 :1:10. 1. 7 1 1 ci :14 12 11 14 14 10 1 c@l :1 11 :1-@, 10 11 :12 I ci 11. -7: 1. 4 4. :1 5. C' 4. 2 4. 4 4. Z 4. 6 4. 5 4. 5 4. 6 4. 4. 5 4. 0 4. 4. 4. 5 4. 4. 5 4. 4 5. 4. 2 4. 4 4. 2 4. 4. 4. 5 4. 7 4. 2 4. 4. El 4. !t' 4. 4. 6 4. *7* 4. 6 4 3. E' 4. 0 4. 8 4. 2 4. 5 4.2 4. E. 4. 1 4. 4 4. 5 4. 1 4. 1 4. 0 4. 4. 4. 4 4. 2 4. Z,. 4. 4. 4 4. 4. 4. t- 4. a 4. 0. 3 23. APPendix Vi (Continued) Oral Teratology Study of T-299SCc>C in Rats Individual Litter Data with Mean Fetus Weights Fit-I4tlAL 3.-S mg/kg/day VIFIE.LE FETUSES CAEFil- m F TOTFIL FETUSES C!:LF, 2,Ci 17j C!IF.' C!:Ip: C,I, F: Q:l F,' C!:lF, C!IF, C!IF' ci C. -Cl.4 5 cl,-EZ.: 07 Ci C-i C,4 C, C!:LF, C!IF-. C!:If@, C@:IF, C@:IFI c@1F. C!IF, C!lp; C!IR C!IF., C, ,i; C-1 Cia-: 3 ci C, ci 1 C-1 2 ci @;:0'-;14 ci -:?@-I 8 1:1 45 9 5 a :1-:@ 5 6 :1:1 5 5 1 Cl NCIT F'F.EriNFlP-JT 2 7 9 24 E. 5 5 1 Ci 4 :117-1 :1 :L 2 5 5 1 ci :1 t-41-IT F*F.,Ei-jNFit-4T 5 E. l:l 7: :1 4 Ni:il-PPE'31-JFit-JT 54 9 1 8 9 1 2 --< 5 5 1 C-1 E. e. 14 MEFit-4 2. '='IFiN. I)E@,.'. 1. E: 4. e. S. 5 2. 5 0 0 0 0 0 0 0 0 0 0 ci ci ci 0 ci 0 0.0 0. El FEE-OF.' IMPLFit4 C:OF.'PRF1 PTION TFi7IOt4 LUTEA SITES SITES t-IEFit@FETUS FIVG t-1 WT<G.'.$ F 1 :12 :1 :1ci (1 :1-,.: ci :L:L 0 :1ci ci 9 2 e. :1 :1 5 7 :12 5 E. l:i i:l :12 :1:1 :11. 9 9 :1 1 ci E. :14 E., cl I 1 1. 5 5 14 9. 6 2. E: E: 11 9 4 I ci 14 10. 1 5 4. Ci 4. 4 4. E. 4. 5 4. E. 4. 1 4. 0 4. t---.4. 4. 5 4. 7 4. 4. 4 4. 7 4. 5 4. 1 3. 7 5. 1 4. 4. E. 4. 4 4. 2 3. 5 5. 4 5. 1 5. :1 4. !!1 4 4. :1 2. E., 4. 9 4. E. 4. 4. Cl. ci 4. 5 4. :1 4. 5 8 4. E. 4. r-I 4. 1 4. 2 4. 4 4. C, 4. 7 4. 4 4. 5 4. IL-1 4. 2 E. 4. 1 Z-:. 7 4. 2 8 4. 'z: 0. 4 24. Appendix Vi (Concluded) -Oral Teratology Study of T-299SCoC in Rats Individual Litter Data with Mean Fetus Weights Fit-Ii1-1@fL '-.:iHL-;LFE'ETLISE:i. [.,EFiL,, 1-i F Tt--IT'AFLETI-ISE:I 0-05 mg/kg/day F.'EE-Cifl-P.l,PLFit4 F,TIC't-4TFITILt-i LU-i'Ll@ SITEA. :@,ITES Fl',,,'i@ F:-1 F-:I i;: @i .4 - F.@LF.. F:iF;. 4 F,-*iL;: F.iF: :@Ci4E. F,.'F:.l' C,4 F: F:i C-i F-I, r;. SI'Fit-4. 4 E. :1 ci NC,T F-F.,EGt.4Ht.iT I P---. I:L E. 4 1 C-1 4 ci C., 5 ci 5 :L:l C, C, 4 C, C, :L -4. 11-i. C-1 1. 2. E. 0. 2 :1C, ci 10 ci I ci C, 5 ci 0 5 C, I ci Li.4 Li. :Li I ci :iC, :L 1. ci 5. Ci 4. 5 4. 4. E. 4. 7 4. 4. 1 4. 4 4. 5 4. 4 4. 4. t- 4. 1 4. 'E. 4. E. 4. 4. 41 4. 4. 1 .4. ri 4. :1 4. "z. -4. 4. @4. 4 4. 4 4. 4. 4 11. C, 5 4, 4. 4 4. 2 4. 4. E. 4, 4. 5 4. E. I. C--i 4. E. .-4 ci 4. @l. 2 4. 0 m9Ag/day Dam Number Total Number Fetuses Fontanalle not closed Prontals not ossified Parietals not ossified Interparietale not ossified Holes in pareitals Sternebra* not ossified oternebree any@ trical one sternebras missing TWo aternabras missing 13 ribs 13 ribs spurred Navy ribs 1 Protrusion on ribs one body vertebrae bipartite TWo bodies vertebrae bipartite 7bree bodies vertebrae bipartite Total Abnormal Petumos Total Normal Fetuses Appendix VII Oral Teratology Study of T-299OCoC in Rats Number of Fetuses by Dam with Skeletal Findings 2997 2998 3000 3001 3002 3003 3004 3005 3006 3007 3052 3053 3054 3055 305 6 7 7 a 7 a 7 6 9 6 1 9 8 1 3 1 1 2 1 1 1 3 2 3 2 1 3 1 1 2 4 2 2 1 3 1 1 2 4 2 2 1 3 1 1 2 4 2 4 5 3 6 5 7 4 1 a 2 (7 1 2 1 1 2 2 2 1 2 3 2 1 2 1 2 1 2 7 5 5 1 1 2 4 1 1 1 I I I 1 1 1 1 1 1 1 3 3 4 1 2 5 1 1 I 1 1 2 I 2 2 2 1 2 1 2 I 5 7 7 8 7 7 7 4 9 5 9 0 9 8 a 1 0 0 0 0 1 0 2 0 1 0 1 0 0 0 150 mg/kg/day Dam Number Total Number Fetuses rontanelle not closed lprontalsnot ossified Parietals not ossified InterparLetals not ossified Sternebrae not ossified sternebrae bipartite Sternebrae asy@ trical One sternobrae stinsing Two sternebras missing 13 ribs 13 ribs spurred Navy ribs Protrusion on ribs One body vertebrae bipartite 7wo bodies vertebrae bipartite One body of vertebrae missing Total Abnormal Fetuses Total Normal Fetuses Appendix Vil (Continued) Oral Teratology Study of T-299SCoC in Rate Number of Fetuses by Dam with Skeletal Findings 3008 3011 3012 3013 3015 3016 3018 3063 3064 3065 3068 3069 3070 3071 a 6 7 6 2 1 1 1 1 a 6 5 10 a 10 a 8 6 1 1 2 2 2 4 2 2 5 5 1 2 5 5 1 2 5 5 1 1 1 2 6 4 3 6 4 9 6 6 3 a 5 1 I 3 2 1 2 1 1 1 2 1 1 5 1 10 3 3 3 1 1 5 2 1 4 2 1 2 1 2 2 1 1 4 1 3 1 1 I 1 1 3 1 2 2 2 I I 5 4 7 6 6 6 4 10 7 10 8 a 6 1 3 2 0 0 2 0 1 0 1 0 0 0 0 0 Appendix VII (Continued) Oral Teratology Study of T-299SCoC In Rate Number of Fetuses by Dam with Skeletal Findings 5 ag/kg/day Dam Number 3019 3020 3021 3022 3023 3024 3025 3026 3027 3028 3029 3074 3075 3076 3077 3078 Total Number Fetuses Fontanelle not closed Frontals not ossified Parietals not onfifled Interparistain not ossified 6f- 6 7 9 7 a 10 6 6 7 6 7 a 8 a 9 3 1 1 3 1 2 2 I Sternebrae not ossified Stenrebrao say@ trical On* sternobrae missing Two sternabrae sinning 2 5 3 5 3 6 6 4 4 6 4 7 7 5 4 a 1 1 1 1 2 1 1 2 2 4 2 1 2 5 1 1 3 4 1 1 3 1 2 13 ribs I I 13 ribs spurred 1 2 Wavy ribs Protrusion on ribs 3 1 One body vertebrae bipartite 2 2 1 1 3 1 2 3 3 2 2 Two bodies vertebrae bipartite 2 2 2 1 Three bodies vertebrae bipartite I I I I 2 I I I Total Abnormal Fetuses Total Normal retuses 5 6 7 6 6 7 10 4 6 6 6 7 7 a 8 1 0 0 3 1 1 0 2 0 1 0 0 1 0 0 1 Peoults from one fetus are missing 1.5 agAg/day Dam Number Total Number Fetuses Tontanelle not closed Prontals not ossified Parietals not ossified Intorperietals not ossified Hole in frontal Sternabrae not ossified Sternabrae asymmetrical One sternabrao missing Two sternebrae missing Four sternabrae 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-299BCoC In Rats Number of Fetuses by Dam with Skeletal Findings 3030 3031 3032 3033 3034 3036 3037 3038 3039 3040 3095 3086 3088 3089 3091 7S 6 9 8 7 6 4 7 7 1 7 1 a 3 6 3 1 1 2 1 1 2 1 1 1 1 1 2 1 1 1 1 1 2 1 1 1 I 1 5 9 3 4 1 3 5 4 1 6 1 6 1 3 I 1 1 2 2 2 1 3 3 1 1 1 2 1 3 1 I I 1 1 2 1 2 1 1 3 1 1 4 2 2 2 3 2 1 1 2 6 7 5 3 4 6 6 1 7 1 a r 4 5 0 2 1 2 3 0 1 1 0 a 0 0 1 2 Results from one fetuo'!'Are missing Appendix VII (Conq4tuded) Oral Teratology Study of T-2998coc in Rate Number of Fetuses by Dan with Skeletal Findings 0.05 mg/kg/d&y Dam Number Total Number Fetuses rontansile not closed Frontals not ossified Parietals not ossified Interparietals not ossified 3041 3042 3043 3045 3046 3047 3049 3049 3050 3051 3096 3097 3098 3099 3100 3101 0 6 7 a 7 5 a 7 4 a 9 8 6 1 6 a 1 1 3 1 1 1 2 3 I 1 1 1 1 1 1 1 1 1 I I sternebrao not ossified Sternabree bipartite Stermsbrae any@trical One sternabrae nkinsing Two sternebras missing 3 4 5 2 3 4 5 4 4 3 6 6 5 1 3 5 I I I 2 1 1 1 2 1 1 1 1 3 1 1 2 1 2 2 2 I 1 1 13 ribs 13 ribs spurred Navy ribe 2 1 1 1 1 1 2 1 1 Protrusion ribs One body vertebrae bipartite 1 2 1 Two bodies vertebrae bipartite 2 1 Three bodies vertebrae bipartite I 2 2 2 1 1 1 1 1 2 1 I Total Abnormal Fetus*@ Total Normal Petumon 7 5 6 7 7 4 a 6 4 3 7 6 7 1 4 7 1 1 1 1 0 1 0 1 0 s 2 2 1 0 2 1 0 mg/kg/day Dm Number Total Number of Fetuses A cleft in the Ions of one oye gn]Larg*d renal polvim Abdominal cavity full of blood Total Abnorml Fetuses Total I#orsal Fetuses Appendix Vill Oral Teratology Study of T-299SCoC in Rats Number of Fetuses by Dan With Internal Findings 2997 2998 3000 3001 3002 3003 3004 3005 3006 3007 3052 3053 3054 305S 3056 3057 3 3 3 4 3 3 3 3 4 3 3 1 a 3 3 3 I I I I 1 4 2 1 3 1 2 1 0 1 0 4 0 0 0 2 1 3 1 1 1 1 2 2 3 2 3 0 3 3 3 1 3 0 2 0 3 2 1 1 Appendix Vill (Continued) Oral Toratology Study of T-299SCoC in Rats Number of Fetuses by Dam With Internal Findings ISO mg/kg/day Dan Number 3008 3011 3012 3013 3015 3016 3018 3063 3064 3065 3060 3069 3070 3071 Total Number retumen 3 2 3 3 3 2 2 4 4 5 4 4 3 1 A dark streak in the Ions 1 1 of one eye A dark streak a cleft in the Ions of one eye 2 1 A cleft in the Ions of on* eye 1 2 1 1 A cleft in the Ions of I both eyes gniarged renal pelvis I Abdominal cavity full of blood I I I Total Abnormal Fetuses Total Normal Fetuses I 1 0 2 0 0 1 0 3 1 1 1 2 0 2 1 3 1 3 2 1., 4 1 4 3 3 1 1 5 mg/kg/daY Dm Number Total Number Votumen A cleft in the Ions of one eye A Cleft in the lens of both eyes Hydronsphroxis Rnlar"d renal pelvis Abdominal cavity full of blood Total Abnormal Fetuses Total Normal Fetuses Appendix Vill (Continued) Oral Teratology Study of T-299SCoC in Rats Number of Fetuses by Dam With internal Findings 3019 3020 3021 3022 3023 3024 3025 3026 3027 3028 3029 3074 3075 3076 3077 3079 2 3 3 4 3 4 4 2 3 3 3 3 4 4 4 4 I I I I I I I I I 0 1 1 0 0 0 2 0 1 0 0 0 1 0 0 2 2 2 2 4 3 4 2 2 2 3 3 3 3 4 4 2 Appendix Vill (Continued) Oral Teratology Study of T-2990CoC in Rats Number of Fetuses by Dam With Internal Findings 1.5 mg/kg/day Dm Nu abor Total Number Fetuses h cleft in the Lens of one eye Sal " ad renal pelvis 3030 3031 3032 3033 3034 3036 3037 3038 3039 3040 3085 3086 3088 3089 3091 3092 3 3 4 3 3 3 2 3 3 1 3 0 3 1 3 3 1 2 1 1 1 1 1 1 1 1 2 Abdominal cavity full of blood Total Abnormal iretuess Tbtal Normal Fetuses 0 0 1 2 1 1 1 1 0 1 0 3 3 3 1 2 2 1 2 3 0 3 0 0 2 0 3 1 1 3 Appendix VIII (Concluded) Oral Teratology Study of T-299SCoC in RAts Number of Fetuses by Dm With Internal Findings 0. OS mg/kcj/day Dm Number 3041 3042 3043 3045 3046 3047 3048 3049 3050 30SI 3096 3097 3098 3099 3100 3101 Total Number Fetuses dark streak in the lens of one eye cleft in the lens of one eye cleft In the Ions of both eyes Enlarged ronal pelvis 3 2 3 3 3 2 4 3 1 3 4 4 3 1 2 4 I I I I I I 1 1 2 Abdominal cavity full of blood I I Thtal Abnormal Fetuses Total Normal Fetuses 1 1 2 1 0 1 0 1 0 1 3 1 0 0 0 0 2 1 1 2 3 1 4 2 1 2 1 3 3 1 3 4 DISTRUMON -LIST J4. C. @(@aaa EI.. .<;.. ..C@iT@o@e x -P.-Zeller (QA @files) recht 2. :4..44yt&ch R@ 4L 1442Aqn -R. :P. -.10ber -W. C. McC=mick -P-.F.D. Grif-fithta) G -R. Stef fen J. D. PP-Dder.spp -Y,. '.M3b=s 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 -o-P.D. Griffith (2) W. H. Pearlson G. R. Steffen J. D. Henderson L. Ebbens AmencTment4io the Final Report"ok t@4ioral t'eratology Study of T-299t .8coc in Rats Issued 12/15/81 Please insert the amended page 3 to the above report.,..F'iveword changes were ..iL made in the last two paragraphs. The study:cohtlbb odi i:@W n67t ch-a:-ngedby this endment to the results and discussion secti6h @of the i@'epo@ri'. L E. G. Gortner Senior Research.-Technologist Animal ileratology Reproduction Date E. G. Lamprecht, DVM, PhD Date Research Veterinary Pathologist M. T. Case, DVi4, PhD -Manager-),-Pa tho-l-ogy-Toxicology Safety Evaluation Laboratory Date Amended page 3 to the Oral Teratology Study of T-299SCoC in Rats - Experiment No. 0681TRO110 Results and Discussion T-299BCoC 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 tlirouyhout 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-299SCoC 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 Cc: M. T. CASO W. C. 14CCoratiek E. G. Gortner (original +1) R. A. Nelson I?.Keller R. A. Prokop E. G. Lamprecht L. 0. Wiseth TLTLE: Protocol for Oral Teratology Study of T299SCoC! in Rats (Riker Experiment Number 0681TROIIO). OBJECTIVE: A teratology study will be used to evaluate the embryotoxic and teratogenic effects of orally administered T299SCoC 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 Usew). 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. RANDOt4IZATION: The animals will be assigned cages according to a computer-generated random numbers table. CONTROL ARTICLE: Corn oil. TEST ARTICLE: T299SCoC. 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 Dose Level 150 mg/kg/day 5 mg/kg/day 1.5 mg/kg/day 0.05 mg/kg/day 0 mg/kg/day Gro2e Size 22 22 22 22 22 The oral route of administration will be used because metabolism studies showed radiolaboled T299BCoC 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 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 abnormlities. The proposed date for the final report is 2-3 months after detailed pup examinations have been completed (approximately third quarter, 1981). E. G. Gortner Date Senior Research Technologist Animal Teratology-Reproduction Study Director 'llr@ -Z eg4J- M'. T. Case, DVM, PhD Date Manager, Pathology-Toxicology Safety Evaluation Laboratory E. G. Lamprecht, DtM, PhD Date Research Veterinary Pathologist W. C. McCormick, MS Toxicologist Sponsor Representative Da@--el