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T- 4q 28 Day Percutaneous AbsorPtiOn Study with FC-98 in Albino Rabbits L-44-) Experiment No.: Conducted At: Dates Conducted: Conducted By: 0979ABO630 Safety Evaluation Laboratory Riker Laboratories, Inc. St. Paul, Minnesota October 25, 1979 to December 17, 1979 K. D. O'Malley6 BS Advanced Toxicologist Study Director Date Reviewed By: dc: M. T. Case K. L. Ebbens F. D. Griffith W. C. McCormick K. L. Ebbens, BS Date Supervisor, Acute Toxicology Summary A 28 day percutaneous absorption study with FC-98 was conducted from October 25, 1979 to December 17, 1979 at Rikez Laboratories, Inc., St. Paul, Minnesota using male and female albino rabbits ranging in body weights from 1.88 to 2.30 kg. The test article was administered by de-ml application to ten male and ten female rabbits each a dosage level of 5,000 mg/kg body weight for a 24 hour exposure period. No mortalities, untoward behavorial reactions or body weight losses were noted during the 28 day study. Necropsies were performed on all animals upon termination of the study with no visible lesions noted. Preliminary serum analysis (see Appendix V) indicates dermal absorption of FC-98 in albino rabbits, however, due to the limited number of samples analyzed, no concrete conclusion may be drawn. Introduction The objective b of this stud.Y determine the percutaneous absorption potential of FC-98 in male and female albino rabbits. The study, which was initiated at Riker Laboratories, Inc., St. Paul, Minnesota on October 25, 1979 and completed on December 17, 1979, was not conducted to support a government submission or marketing permit and is therefore not regulated by the Good La.boratory Practice Regulation of 1978. The raw data generated by the Study Director and the final report are stored in the conducting laboratory's archives. .E A preliminary rangefinder study was conducted to determinp the appropriate dosage level to be used in this study. b Riker Toxicity Experiment No.: 0979ABO630, Test Method 699 a Young adult albino rabbits of the New Zealand breed-@-were used in this Lu@;L. Ali aiiiinzil@-w;ere JiL.-Iucnider 4LIuarantinefor several days prior to testing wi.Liiotily iiiiiitziLw:l;iicii to be Ln cjoodhc.,altahnd suitable as test aiiiinai!@a;L the iniLiaLion of the study used. The rabbits were housed individ- 11;1-Lli.lyl L;tcCl,wire-bottomcd caegas and maintained on a standard .L,ti)orLiLora-LYi(.)1i)iw.i,lli food and Water availablu ad libitum. An initial rangefinding study was conducted using two male and two female rabi-)iL:f@or Llac;icilo!@;ayluevel. The trunk of each animal was clipped free of ii.LLiliL!L@.;.-aifLic.Lujilac(.-oan Lho surface of the intact skin which covered aidproximately40!ttotal body surface area. After administration of thL,Lost article, a flexible plastic collar was fitted on each animal;and ttiutrunk wrapped with impervious plastic sheeting which will occlude the test article. The animals were returned to their cages for a 24 hour period aft(2rwhich time the test article was removed from the dermal surface of the animals. The animals were observed for pharmacotoxic reactions both during the exposure period (immediately Post dose administration, one and two hours) and after removal of the test article (daily for 14 days following dose administralion)with all ruaclions recorded (Table 3). initial and final body weiglit5were also recorded (Table 1). The information derived from the initial rangefinder was used in determining the dosage level for the 28 day percutaneous study. Preparation of 10 male and 10 female animals for dosing and application of the test article were conducted in the same manner as the rangefinder study with the exception of the collection of blood samples from the orbital sinus plexus prior to application and again-on days 1, 7, 14 and 28 after initiation of the study for serum which was frozen for sponsor analysis. After the 24 hour exposure ab Pel Freez, Inc., Rogers, AR i-lurinai@abbitChuw, lialstonPurina, St. Louis, MO I)L!riocLllic LLSL arlicle was removed from the dermal surface of the animals and the animals returned to their cages for the following 28 days. Initial, '/, Lilaiill2L$d.Ay UL)(lywLiytiLs wurl- rul;c)rcie(dTable 2) as were any phar-micoLOX.ic:!@igns noted during the 28 day observation period (Table 4). A gross necropsy was conducted on all animals sacrificed on day 28 and all findings recorded (Table 2). The protocol, principal personnel involved in the SLudy, coml)osition characteristics, and Quality Assurance statement are contained in Appendices I - IV. 4. ACUTE F)ERMAL RANGZRINDER TOXICITY STUDY - ALBINO RABBITS with FC-98 Mortality and Body Weight Data Dosea Sex (mg/kg) Animal Number individual Body Wcights Test Day Number 0 14 (kg) Number Dead Number Tested Percent Dead 5000 m 9B2599 m gB2602 F 9B2617 F 9B2614 2.37 2.41 2.42 2.15 2.53 2.62 2.31 2.52 0/4 0 1000 m gB2605 m 9B2608 r 9B2618 F 9B2621 2.22 2.47 2.25 2.35 2.51 2.55 2.16 2.45 0/4 0 a Test article was dosed as a suspension in water ACIJTI-:PERCUTANI-'O(JS A13SORPTION TOXICITY STUDY - ALBINO Mortality with FC-98 and Body Weight Data RABBITS a Dose Scx (mg/kg) Animal Number Individual Test 7 Body Weights Day Number 14 (kg) 28 Number Dead Number Tested Percent Dead 5000 m 9B3041 2.16 2.58 2.47 2.63 0/10 -'0 m 9B3047 2.30 2.03 2.27 2.68 m 1)],,'3053 2. 1(") 2.13 2.45 2.79 m 9B3059 2.15 2.10 m ()11304-) 2. 3rl 2.15 2.33 --1.47 2.54 2 m 9B3049 2.11 1.96 2.11 2.37 m 1)133055 2.19 2.22 2.47 2.75 m 9B3061 2.18 2.22 2.31 2.46 m 9B3045 2.30 2.33 2.52 2.79 m 9B3051 2.22 2.15 2.38 2.70 5000 F 9B2921 2.25 2.03 2.25 2.66 1/10 10 F 9B2927 2.12 2.00 2.29 2.78 F 9B2933 2.18 1.87 2.01 2.37 r 9B2939 1.99 1.72 2.09 2.42 F 9B2923 2.08 1.99 2.30 2.65 F 9B2929 1.88 1.72 1.90 2.21 F 9B2935 2.11 2.14 2.33 2.85 F 9B2941 2.02 2.13 2.25 2.48 1.' 9132925 2.12 1.98 1.70 b F 9B2931 1.94 1.94 2.14 2.37 a Test article was dosed as a suspension in water Animal sacrificed on day 14 due to accidental back day 7 bleeding period injury inflicted during the @!@O D S@ Necropsies performed on all animals upon termination visible lesions. Animal 9B2925 which was sacrificed paralysis, also revealed no visible lesions. of the study revealed no on day 14 due to posterior U) E@ C94 u tn 0 El I u C-) 0 t .-4 0 0 0 tn U) a) tn r 0 u tn 0 (a 2 -@4 tn 4J fa r- 44 .-f @) IA tn el 4J L4 -C.) C4 4) 4j 4j 4.) 4J 4J 4.J 4J a0 0 a 0c 0 u fa a r_ 0 a0 c u u -@4 0 -.1 tn C 44 r. .u,q En -@u4 U) 44 C q&4 r- 0 0 .,q 0 0 a% t3) 41 tn 4.j 4.) U -,4 L) .@4 0 @4 0 @4 z z 0 )@4 0 @4 z x 4) rA4 U) 0 2E L0n 0 .C U) 41 0 0r tn 0 2 0 .@4 tn C0 faAi f0w 1f9c cn tn (D lm in a) wr 4J w0 C4 EU) E- 12 th 0 44 tn 0 .@4 0 1-4 P-4 o 0 E- (n U) 14 a) 0 0 r- to CO .@4 cn tg-4 C i,@4 r_ r_ 0 o 4j tn 41 4J to 0 @4 0 @4 z rk 0 2 Ln l@.kk u i Alli'rNDEX T r.LjiiL:iiL Nw OLi (iH L:5C@D Singie .1@]IONSOf@: 3M CONDUCTED BY: 'I'ES'IA'RTICLE: Dose 28 Day Percutaneous Absorption tc-@Q Safety Evaluation Laboratory, Riker Study Laboratories, Division Inc., St. Paul, Minnesota (-nt@'I'r,0A1r,TICT.r.: 1)A'I'L0'1;'STUDY: SYSrl'1.71A,N1I) SC)LJI@CL: Now ZL:zil.and White Albino Rabbits i'L:i Inc. , f(ocjers,Arkansas Sex: Number: 4- t:@ 104-10 Weight Range: r,@--5k4t UL,JI-C'I'IVL: 'I'lioLb,jective of this study will be to determine T)O',:Cjlt:iozfliLilu tce;t article in albino rabbits. the percutaneous absorption Rabbits were selacted as the LUSL system for their historical use in de.-mal absorption handling and general availability. studies, ease of from a lar(jlr colony by hcaltli 4ancl L--xly w(--igilL,will Lic r,iiicioinllyiou.,@udin standard wire-mesh cages in temperature and humidity con- trolled rooms witi, food2@and water offered ad libitum. Each an4-:nal ',Q us..;igncda numuered ear tag, which will correspond to a card affixed to the Outside of ttic cage. The trunk of each animal will be clipped free of hair arid tiic test article applied as a single dosage of !51)()C).@ng/kg to intact skin coverinq anproximately 10% total body surface aiea. A flexible plastic collarb will I)c fitted on each animal and the trunk wrapped with impervious t)l,islicslioctinq, which will occlude the test article. -The animals will then be returned to their cages for a 24 hour exposure period after which the test article will be removed. Prior to the'application, blood samples will be collected from the orbital sinus plexus and again on days 1, 7, 14, and 28 after initiation of the study for serum which will be frozen for sponsor analysis. A gross necropsy will be conducted on all animals which may die during the conduct of the study as well as all animals sacrificed on day 28. All gross findings will be recorded and tissue samples of liver, spleen, brain, kidney and bone possible future weights will be marrow (sternum) will be fixed in 10% microscopic examination. initia 1, 7, recorded as well as any pharmacoto' xic buffered fo=alin for 14, and 28 day body signs noted during the conduct of the study. All raw data, other than the blood analysis data which will be the responsibility of the sponsor, and the final report will be stored in the Riker Laboratory's Archives, St. Paul, Mi-nne*ota. a Purina Rabbit Chow, Ralston Purina, St. Louis, Missouri The collar will be worn for the duration of the study to reduce oral ingestion of residual test article. Sponsor n Study Directo@.- klkt--ij'.xl)4L,rLlilk:NIui.L APPI-:NDTX T (CotitLniiecl) ACU L,--i),.:i,urii 'I'OXC.;L.LLY Ian S P'ONSOR: 3M CONDUCTED BY: Safety @)4a@uation TES7' AR'I'ICLL: Q I rliLlIgS L,-fiJy Laboratory, Riker Laboratories, Divi Inc., St. Paul, Minne @,"I'A)('I'INC;/CuMI,Ll@-*,I,IIJUAN'1'1U-l.-'' S'I'LJL)Y. 'i*l-:S'SIY'S'1'1:'MAN[) SOUf@Ci,-': N(iw Zealand white A Pul Inc., Rogers, je7 o Rabbits Arkansa4 a.5 S(--X: N )er Bod weight Range: C)LIJI-'C:'I'IVE:I'I'LO-bjective of this study will be to approximate the acute dlrnial.toxici ul' Llit:Lu!;L jrLiclu in alliino r;AljljiL,!;. liabUiLL; werv selected iAs the Lust system for their sensitivity of response, historical data, ease of handlin and general availability. ml;,I,Ilul-); 'i'liuaii.Liii.AIL;,:;L:IucLcLl frujit iA larcjcr colony by health and wc:iylit,will L)v randomly housed in standard wire-mesh cages in temperature and humiditv controlled rooms with foH6d2@and water offerui ad libitum. E:ach animal wil be assigned a numbered ear tag, which will correspond to a card affixed to the outside of the cage. The trunk of each animal will be clipped frftc of hair and the tet article placed on the surface of the intact skin at sing dosages of,," mg/kg, however, if these dosage levels c riot adequatuly haracterize the toxicity of the test article, additional animals will be administered the test article at supplemental dosage level! Any additional dosage levels will be documented and filed with this protocc The test article will be administered to the animals in the form received from the sl)ons r. AFter admini$tration of the test article, a flexible I)Iastic collar- will@ be fitted on each animal and the trunk wrapped with itrirj(--rvi1o.u)sIasticsliceting whicit will occlude the test article. The aniii will be returned to their cages for a 24 hour exposure period after which Lime the test article will be removed from the dermal surface of the animal 'I'liacnimals will be observed for pharmacotoxic reactions both during the ex 1.)osurcperiod (inunediately 1)ost dose administration, one and two hours) and after removal of the test article (daily for 14 days following dose adminis tration) with all reactions being recorded. Initial and final body weights will also be recorded. The acute median lethal dose (LD50) of the test article will be approximated. -kll raw ddta and the final report will be sti in the Riker Laboratories Archives, St. Paul, Minnesota. Purina Rabbit Chow, Ralston Purina, St. Louis, Missouri The collar will be worn for the duration of the study to reduce'oral ingestion of residual test article. Sponsor Date ///;Ixl@@le, Riker Experiment No.; APPENDIX I (Concluded) Amendment to.Protocol 10. "Doc 2. A^A Study Directo& study Director Date v,r r,C) c Study Director Date 4. study A.AA 0- Dir@ectov 0 AA(@ u ) n Study Director Date Date study Director Date 7. Study Director Date Study Director Date AL"1)1-.'NDIXIl Principal Participating Personnel Involved in the Study NjiintK. i.. I-l'bi)(!n1.3-S;, Function Supervisor, Acute Toxicology O'Malley, ES Dr. V. Pothapragada C. C. Pecore Advanced Toxicologist Study Director Commercial Chemicals Chemist Supervisor Animal Laboratory APPENDli 111 12. Composition Characteristics This study is not regulated by the Good Laboratory Practice Regulation of 1978 and therefore information pertaining to composition characteristics is not applicable for inclusion in this study. 13. APPENDIX IV ouality Assurance Statement This study is not regulated by the Good Laboratory Practice Regulation of 1978 and therefore a statement signed and prepared by the Quality Assurance group is not applicable. This study was, however, audited by the Quality Assurancc groul). In addition to the data audit, different significant phases for studies underway in the Toxicology Laboratory are inspected weekly on a recurring cycle, ana tne tacilities are examined by Laboratory Quality Assurance on a three month schedule. LI I A. Ui)cl 22o-21: 14. APPENDIX 5 L. L-'BBL-NS - lilKL-'kSAI-L-'TYE-VAI,UATION LAB W. C. McC:ORNIICK - MI:I)ICAL I)EPI*.- TOXICOLOGY 203-1 SERVICR-.S - 220-21-: SKLN ABSOIIP*RION S-FUDII:S ON 1-*(:-128,I:C-129 aild I:c-98 -JUNF: 27, 1980 FC-143, FC-95, FC-99, FC-134, FC-13S, ]'leaseconsi(lei-t.ilis.11a,uthorization for your laboratory to releasu LIIC(.Iennaltoxicity/skin inentioned compounds. absorption studies conducted on the above It is understood that the studies are being issued in an incomplete Conn iiisofaras the fluorochomical analysis of the serum samples have not been completed and will not be included in the report. I)i-eliminary serum sami3le analysis indicates absorption of the comj)ounds. The serum data analysis arc not sufficient enough to draw any concrete conclusions concerning comparitive toxicity. However, the animal data you have generated addresses this matter in a broader L:Olltext. It is not certain wliejtihe remaining samples will be analyzed and their completion should not hold up your report any longor. 'I'liaiyioku for your paticnce in this matter. WC@i: k 1li A 1). 15. Ai@i'L'NL).LX*5 (Conciuded) COMKERCIAL CHEMICALS DlVlSIUN ANAT.YTICAL I.AB REII'ORI'#1.lo(j I(j W. C. MCCORMICK 220-2E-02 ii(ifn V. POTHAPRAGAI)A AND V. BUNNELLE 236-3A I(IKER SKIN ABSURI'TION STUDY L.@jitl- June 9, 1980 JO Reference: Commercial #15669 Chemicals I)ivislots An&.Lytic-al Request For lack analyzed. of time, only a selected out of aurum awnplt--s was T(YRAL F, ppm Coinpound FC-129 I-'C-134 FC-128 FC-98 FC-135 FC-95 FC-99 I)a 7 lay -- 1.0.1 Females Day 1 I)ay 28 14ales Day 1 I)ay 28 26.1 69.6 11.4 23.3 0.2 4.4 18.1 16.5 18.8 1. () 2.'J.9 it)..5 226.4 93.1 271.9 94.3 6.9 20.8 2.3 7.6 0.9 128.0 10.3 130.2 42.5 53.1 Ul.5 U9. 8 129.1 72.7 73.5 66.6 Females Malet; Day 14 _U_@8 .1@tL_7 pay. .14 1);Iy 28 12.1 3.5 5.4 0.8 /1. M--i Iii)tf i)l* Ais:tl-lo&ltt: ()XY)-.t!ok D. F. Hageti, Anal. Bluchein; Its-lltils8-/, 545, isia.1 19@8). V. A. Bunnelle VAB/hc I r) . V. Putliapragada@ Read by and RevJt-w@--,; 1) Wiiitcr ? -/ .1 .0oo@ %.a.- 4-