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`52-week dog:
`Pure-bred beagles. 4/sex/dose, age 5.75 to 7.25 months (mean 6.25’) at study initiation, weight range 8.5-
`11.1 kg m and 6.9-10.9 kg f.
`‘these dogs seem to be fairly youn . in eneral mean a e is 9
`Study No. DS-93-011/Laboratory:H
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`Route: oral gelatin capsules.
`Doses: 10, 20, 40 mg/kg/d for 1 yr. The control group received similar number of empty gelatin capsules.
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`onths.
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`lnltlal weight (g) and weight change after 7 8. 5m
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`Results:
`QumLSJgns; motor excitability in 2 HD fwk1 and 1 HD f wk3 of dosing.
`Modality. none
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`'
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`' body weight gain significantly decreased in all doses and both
`sexes starting at end of treatment wk1 onward (Table below from sponsor). The decrease was greater for
`MD and HD males and HD females compared with the controls during wks 1-7 (dogs were fed 400 g/d
`diet). Increasing the food to 500 g/d during wks 8-52 did not reverse the weght loss. When the sexes
`were combined, the mean gain remained significantly reduced for mid and high dosed dogs compared
`with the control group. MDF had no effect on food consumption. MD and HD dogs (m+f) drank on the
`average significantly more water than the control dogs during wks 1-4, 21-49. These animals showed
`some tendency to drink more water during the pre-dosing period relative to the control.
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`MW no drug-related findings.
`Hematology; Hb, PCV, and RBC count were slightly but significantly reduced in mid and high dosed dogs
`relative to the control (613%). Total WBC count, lymphocytes count was slightly reduced in mid and high
`dosed dogs (21-26%), eosinophil count was reduced 63% in MD but not in HD dogs. and platelet count
`was increased in HD. There was interanlmal variation and the levels for RBC, Hb, and PCV were low
`during pre-dose in drug groups relative to the control. These effects make it difficult to conclude with
`certainty that these changes are drug-induced.
`It can be assumed that the changes in HD animals are
`drug-related.
`Mlinahfiifi; no drug-related findings.
`W serum alkaline phosphatase (SAP) level was significantly increased in HD wks 6 and 12
`(1 .7-1 .8 fold) relative to the control but no significance was noted on wk 50. On wk 25, SAP levels were
`elevated (insignificantly) in all three groups. Also on wk 50, Na level was significantly decreased in all
`treated groups. creatinine significantly decreased in males of MD and HD. and a slight insignificant
`increase in total protein of HD dogs.
`~ W5; increase in liver weight in 7/8 HD and 5/8 MD dogs over the 4% of body weight normal
`upper limit. Liver weight was significantly increased in all three groups and relative kidney weight
`increased significantly in mid and high dose dogs (see Table). The absolute and relative mean weights of
`prostate in MD and HD treated dogs are reduced relative to those of the control (139 control, 6 and 79 MD
`and HD respectively for the absolute wt and the corresponding values for the relative wts are 0.09. 0.05
`and 0.069). The absolute thymus weight was reduced in HD relative to the control (8.8 and 139
`respectively. or 32%).
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`Organ wts-results of statistical analysis for male and female rats combined
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`' Slums-n or the 5‘ Incl of milk, (William m)
`" Si'u‘fimn in Ii Incl of Milky (Willi-fl II!)
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`W no drug-related changes.
`filstgpamojpgy; 4/4 HD male dogs had slightly increased hemosiden'n deposition in hepatic Kupffer cells
`relative to the control. However. no other changes noted that could account for the weight change or
`hematology and clinical chemistry effects. Minimal reactive hyperplasia of lymph nodes was found in 1/4
`each fand m MD and HD respectively and none noted in the control. An extracapsular nodule of cortical
`tissue in adrenals and focal vacuolation of the zona glomerulosa were found in 2/4 LD. 2/4 MD. and 2/4
`HD male dogs and to a lesser extent in female dogs.
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`Oral administration of 10. 20. or 40 mg/kg MDF to dogs for 52 wks did not cause any deaths or clinical
`signs of toxicity except some motor excitation in high doses. Body weight gain was reduced although
`appetite seem to be normal. water intake was increased in HD relative to the control the toxicological
`significance of which is unclear. RBC, Hb. and PCV values were slightly but significantly reduced in high
`doses. On wk 50. Na level was significantly decreased in all treated groups. Liver, and kidney weights
`were significantly increased in high doses and mean weight of prostate in MD and HD treated dogs were
`reduced relative to the control. There were no macroscopic findings. and histopathology revealed slightly
`increased incidence of hemosiderin deposition in hepatic Kupffer cells relative to the control in high doses.
`An extracapsular nodule of cortical tissue in adrenals and focal vacuolation of the zona glomerulosa were
`found in one half of all treated male groups and to a lesser extent in female dogs. A NOEL could not be
`determined. 10 mg/kg is the LOEL based on body weight loss. clinical chemistry, and histopathology
`findings.
`
`Summary and Conclusions of Chronic Studies:
`Oral administration of modafinil to rats and dogs was well tolerated upto 52wks of repeate dosing. No
`mortality was seen in rats upto 60mg/kg dosed for 52wks and in dogs at 40mg/kg dosed for 52wks.
`In the
`rat 6mo study, one male rat dosed 200mglkg was killed on wk23 due to 229 loss in wt and necropsy
`showed enlarged spleen and small flaccid testes. another male in this dose gr was found dead on wk1
`and prior to death it had severe respiratory distress, convulsions, and salivation; necrospy did not reveal
`any sig finding except for a dark liver. Also in the 6mo rat study. one f dosed 200mglkg was found dead
`and necropsy showed enlarged and cystic kidneys with subcapsular discoloration. one m closed the low
`dose of 20mg/kg died on wk2 and found to have enlarged liver. The sponsor did not related any of these
`4 deaths to modafinil. The following findings were recorded:
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`decr in wt gain in rat dosed z 50mg/kg and dogs dosed 20&40mg/kg.
`decr in Hb, PCV. RBC in female rats dosed 200mglkg for 6mo and dogs dosed 20&40mg/kg for
`1yr. Also in the dog, the total WBC & lymphocyte count was decr.
`cholesterol level was incr in the rat dosed 200mglkg for 6mo and rats dosed 30&60mg/kg for 1yr.
`ALP level was incr in the dog dosed 40mg/kg during mid study, serum Na deer in all 3 drug grs.
`total protein incr in dogs dosed 40mg/kg.
`liver wt (absol and/or rel) was incr in rats dosed _>_30mglkg dosed for 6mo and 1yr and in dogs the
`incr was dose-dependent in all 3 drug grs.
`weight of spleen and kidneys was incr in 200mglkg rats closed for 6mo and incr in rats dosed
`60mg/kg; kidney wt was also incr but only in rats dosed 60mg/kg for 1yr.
`the wt of the thymus was decr in dogs dosed 40mglkg and the prostate wt was deer in
`20&40mg/kg dose gr.
`there was an incr in hemosiderin deposition in hepatic Kupffer cells in dogs dosed 40mg/kg.
`NOEL could not be determined in the rat 6mo study or the dog 1yr, in the rat 1yr the NOEL was
`6mg/kg.
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`REPRODUCTIVE AND DEVELOPMENTAL STUDIES
`
`Seg l rat fertility study/Study conducted in 1984—198_on~6LP/study# 03-93-014.
` .
`
`Modafinil was administered by oral gavage to Sparague-Dawley rats (24mlgr and 27flgr) at 20. 50. or
`100mglkg/d. The cont gr was administered the vehicle. Modafinil was prepared as a suspension in 0.5%
`CMC and shaken well before administration. Rats were 9wks old at study initiation and weighed 174-2249
`m and 194-2619 f. Males were treated for M prior to mating and females for 2wks prior to mating.
`Dosing continued throughout mating period for both sexes (mating period was 11d) and gestation period
`for the f. One half of females were killed at end of gestation (gd20) for teratology study and. the remaining
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`half was allowed to give birth to F1 generation and lactate. The latter half of the females did not meet the
`requirement of having a minimum of 10 pregnant f by end of study, therefore. 3 additional confirmed-
`pregnant f (therefore. the total of 27flgr noted above) was added. These females were, treated
`throughout gestation. birth, and lactation until weaning of F1 generation at which time they were killed. At
`weaning. 1-2 males and 1-2 females from each litter of F1 rats. were retained and mated when they
`reached 13wks of age. These F1 females were allowed to give birth spontaneously and feed their young
`(F2 generation). The remaining F1 rats were kept and killed on ppd21-23 for autopsy and organ wts; no
`histopath was done. F2 rats (4/sexllitter) were observed till weaning.
`
`The following parameters were assessed:
`F0 generation:
`Growth. signs. B.wt, food intake, repro parameters for females included: mating
`index, mating interval. fertility index. repro index. wt.gain and food intake during
`gestation. ForMW embryotox, no. of corpora lutea.
`implantation sites, fetal viability. sex ratio. resorptions, fetal exam for skeletal and
`visceral anomalies. ForW no. of pregnant females,
`gestation length. B.wt. food intake during lactation, gestation index. resorption
`index. no. and sex ratio upto ppd21, viability index. lactation index. and necropsy.
`
`F1 generation:
`
`mommating; physico-behavioral development were examined: righting reflex
`(pde). startle response (ppd8), traction reflex (ppd3). reaction to noise (ppd9),
`openning of eye lids. eruption of incisors, etc.. Mating; B.wt, food intake,
`signs, repro index. gestation index, no. and morphology of implantation sites. no.,
`wt, and sex ratio of F2 generation up to ppd21, and. viability and lactation index.
`
`Results;
`There were no drug related findings on the assessed parameters except as noted. Most of the findings
`occurred at comparable rates between the cont and drug grs. Mating interval for F0 (the only drug treated
`generation) was 2.54d in control and 2.04-2.22d in drug grs indicating no drug effect Resorption index
`(RI) was incr in HDFO f at 12% vs. 6% in the cont, the RI was similar in LD&MDf to that of the cont. RI
`was also incr in HDF1 fat 15% vs. 9% in cont and again. similar incidence in LD&MD to the cont. Also in
`F1 females. one MDf had 9 stillboms but the incidence in all the rest in this gr and in LD&HDf was similar
`to that in the cont. Therefore. this finding was not considered drug related due to its randomness.
`
`It is concluded that oral administration of modafinil to rats had no effect on male or female fertility
`or any other effects on repro parameters including F1 and F2 generations except for 1.7-2% incr in
`resorptions in HDFO&F1 compared with the cont. This incr is small and unclear if it is drug
`related, more info are needed to verify this finding. Therefore, the NOEL for fertility and repro in
`this study is 100mglkgld.
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`Seg ll teratology study in rats/Study concluded at end of 1981_LP/Stlldy# IDS-93'
` .
`015.
`‘
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`Modafinil was administered by oral gavage to pregnant Sparague-Dawley rats (25flgr) at 50. 100.
`200mg/kg/d during gd6~15. Pregnant females were killed on gd 20. The cont gr was administered the
`vehicle. Modafinil was prepared as a suspension in 10% gum arabic. B.wt at study initiation was not
`provided.
`
`The following parameters were assessed:
`During gestation: mortality, signs. B.wt and food intake. The following repro parameters were assessed
`on pregnant females killed on gd20: no. of corpora lutea. implantation sites, and early and late resorptions.
`The fetuses were divided into 2 grs, one examined for skeletal and the other gr for visceral anomalies.
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`Results;
`There was an incr in resorptions in HD: the number of females with resorptions was 19 vs. 10 in cont,
`and. the total no. of resorptions was 42 vs. 28 in cont These parameters in LD&MD were comparable to
`the cont. However, the mean no. of gestating females at term was similar among the drug grs and the
`cont Also. the no. of resorptions per female presented with resorptions. was not different:
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`50 100contDose:Resorptions: 200
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`mean no.1s.d
`of gestating f
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`no. fpresenting
`resorptions
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`mean no. of resorp.
`per f presenting with
`resorptions
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`1.17
`1.33
`12.15 11.81
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`1.68
`1.43
`11.40 11.63
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`10
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`13
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`19
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`2.55
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`2.15
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`2.30
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`2.20
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`For the fetal visceral exam. there was an incr in hydronephrosis (HS) in fetuses from HD was as follows:
`Cont
`HD
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`Left HS
`L&R HS
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`4/126(3.2%)
`3/126(2.4%)
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`9/150(6%)
`12/150(8%)
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`No difference in LD&MD.
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`The only skeletal effect was an incr in incomp ossification (head bones) in fetuses from LD&HD rel to the
`cont (28/150(18.5%). 27I150(18%) vs. 8/120(6.7%) in cont, respectively).
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`These skeletal and visceral changes are relatively small and may not be drug related.
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`It is concluded that oral administration of modafinil to pregnant rats during organogenesis did not
`cause maternal toxicity and was not teratogenic upto 200mglkg. There was some embryotoxicity
`noted in the absence of maternal toxicity, however, it may not be of a true biological significance.
`The total no. of resorptions In HDf was incr and, litter hydronephrosis and, incomp ossification in
`fetuses from HDf were also incr. The incr in number of resorptions might have been due to the
`higher no. of implantations noted in this gr relative to the cont and other drug grs as noted in the
`above table from the sponsor. Although, an incr In resorptions was also seen in the above study
`at 100mglkg (non-GLP). Because of the weak signal of embryotoxicity, the NOEL for the maternal
`repro parameters. embryotoxicity, and teratogenicity is 200mglkg.
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`Seg n teratology study in rabbits/Study conducted in early 1982-non-GLPIstudy# DS-93-
` .
`016.
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`Modafinil was administered by oral gavage to pregnant New Zealand white rabbits (21flgr. age 16-19wks)
`at 25. 50. and 100mg/kg/d during gd6-18. The cont gr was administered the vehicle. Pregnant females
`were killed on gd 29. Modafinil was prepared as a suspension in 10% gum arabic. B.wt at study initiation
`was not provided.
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`The following parameters were assessed:
`During gestation: mortality, signs, B.wt and food intake. The following repro parameters were assessed
`on pregnant females killed on gd29: no. of corpora lutea, implantation sites, resorptions. The fetuses
`were divided into 2 grs, one examined for skeletal and the other gr for visceral anomalies.
`
`Results;
`Mean wt gain during treatment (gd6—18) was reduced in HDf rel to the cont (2.4 vs. 7% cont). Mean wt
`gain in HDf was minimally decr throughout the study (gd0—29) rel to the cont (12.8% vs. 13.3% cont).
`Decline in food intake accompanied the deer in mean wt gain in HD during treatment (1479 vs. 1609 cont).
`An incr in no. of aborting females (those fertilized but did not have fetuses at c-section) noted in MD (3/21)
`vs. cont (0/20), also the no. of resorptions per female was elevated in MD at 37/21 (mean1sd 2.2132) vs.
`the cont at 13/20 (mean1sd 0911.2). The sponsor indicated that this was contributed to 2 MDf that
`together had a total of 17 resorptions. The number of live fetuses (total per f) was slightly reduced in HDf
`at 78 vs. 117 in cont. There were no drug effects on fetal skeletal or visceral parameters.
`
`It is concluded that oral administration of modafinil to pregnant rabbits during organogenesis,
`caused a small decr in mean wt gain and food intake In HD. Only in MDf. there was an incr in no.
`of resorptions and no. of aborting females. These latter findings may not be drug related since
`they were non-dose dependent. The NOEL for maternal repro tox is 50mglkg and that for
`embrytox and teratogenicity is 100mglkg.
`
`Seg lll peri- and post-natal study in rat/Study conducted at end of 1984—GLP/study# DS-
` .
`93-017.
`
`Modafinil was administered by oral gavage to pregnant Sprague-Dawley rats (25flgr, age 8—10wks) at 20,
`50, and 100mg/kg/d from gd15 and through lactation (ppd21). Females were killed on ppd21. The cont gr
`was administered the vehicle. Modafinil was prepared as a suspension in 5% CMC. B.wt at study
`initiation was 192-2909.
`
`The following parameters were assessed: mortality, signs, B.wt, food intake, water intake, nursing
`behavior, resorption index, implantation sites. and other repro parameters. For the F1 generation: viability
`and growth including standard clinical tests such as righting reflex, startle response, response to
`noise...etc. Necropsy was done on all females that did not give birth, gestating females, and their litter.
`The females that did not give birth and gestating females were sacrificed between ppd21-23. From each
`litter, 2/sex were necropsied with the following organs weighed and examined: thymus, heart, liver, spleen,
`kidneys, adrenals, and lungs; (no histopath was done but these organs were preserved in 10% formalin).
`
`The sponsor indicated that the drug conc prepared at the start of lactation were lower than the proposed
`theoratical conc specially at the HD. This was due to the poor homogenization of the suspension
`immediately preceeding sample withdnwal. However, the sponsor stated that these samples were stirred
`constantly throughout intubation.
`It is unclear if this affected the study results.
`
`Results;
`maternal food intake was reduced in all grs including the cont dun'ng gd18-20, at 56-649 compared to
`140-1489 at gd0-6, but no changes during lactation. There was no drug effect on gestating or lactating
`females re. B.wt or water intake and no effect on gestation duration or nursing behavior. Resorption index
`was incr dose-dependently in MD&HDf rel to the cont (12.7 and 13.4% vs. 10% in cont). However, the
`sponsor indicated that these values were within historical data for SD rats at 10.8119 and therefore, may
`not be drug related. At birth, the no. of live young per gestating female was reduced in all 3 drug grs non-
`dose dependently with moderate deer in MD (mean1sd: 14.3124, 13612.5, 11715.8, and 12.514 in cont,
`LD, MD, and HD respectively).
`In addition, the no. of stillboms was elevated in MD&HD rel to the cont at
`1.5138 MD, 1.112.6 HD vs. 0310.6 in cont The stillborn rate (stillbomlyoung born) was: MD 32/290
`(11%), HD 26/326 (8%), cont 7/349 (2%). Among the physico-anatomical changes, the startle reflex was
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`slightly delayed in MD&HD rel to the cont but was earlier in LD.; no other effects. There were no drug
`effects on organ wts or macroscopic exam.
`
`it is concluded that oral administration of modafinil to female rats caused no maternal tox upto
`100mglkg. There was an lncr in RI that was dose-dependent but stated to be within historical data
`for this strain of rats. The no. of live fetuses per female was reduced in all 3 drug grs but the
`pattern was non-dose dependent and the stillboms and stillborn rate were incr in MD&HD rel to
`the cont parameters. Fetal and litter wt were not affected by drug treatment however, growth
`seemed to be slightly retarded as indicated by the delay in startle response in MD&HD. The
`maternal NOEL is 100mglkg whereas, that for embryotoxicity is 20mglkg.
`
`Se lll
`eri- and
`-natal study in rat with functional and behavioral evaluation/Study conducted in 1995/
`H- PA/GLPlstudy# 03-95-022
` .
`
`Modafinil was administered by oral gavage to pregnant Sprague-Dawley rats (30flgr‘) at 50. 100, and
`200mg/kg/d from gd7 through ppd20. All survivng pregnant females were killed on
`d 21. The cont gr
`was administered the vehicle. Modafinil was prepared as a suspension inflwhite viscous liquid).
` ., .
`B.wt at study initiation was 220-2719.
`
`' Blood was collected from 8 mated rats pre-dose on gd6, 5 of these rats were assigned to a postpartum
`satellite gr for collection of milk and blood (dosing from gd7-11 of lactation). Blood was collected on
`ppd3&9 from 5 rats pre-dose and pre-administration of oxytocin for collection of milk. Blood was also
`collected on ppd5&11 at 2hr postdose. Blood was sampled from the jugular or tail vein. Milk was
`collected pre-dose on ppd3&9 and 2hr postdose on ppd4&10. Oxytocin was injected 5min before milk
`sampling (see above).
`
`The following parameters were assessed: mortality, signs, B.wt, food intake, mating behavior, gestation
`period, implantation sites, litter size, pup viability. and nursing behavior. Gross necropsy was done on rats
`that did not deliver by gd25. any gestating rats that sacrificed moribund or found dead, all surviving
`females on ppd21, and, all pups on ppd21 not selected for continued observation. Statistical analyses
`were done on given parameters.
`
`
`
`There were no drug related deaths or clinical signs. Mean wt gain was sig reduced in HDf (40%) during
`the 7-10 gestation period; no difference at later periods. However, this early decr affected the
`cummulative periods of 7-20 and 0-20 (p50.01 at 128.8% respectively). Mean wt was comparable in HD
`rel to the cont except a sig decr recorded on gd14&15. During lactation period, a deer in mean wt (3-5%
`of cont) noted in HD during pp days 2-5 87-8 (reaching significant level p50.05/0.01 at pp days 3-4&7-8);
`no effect on wt gain. Absolute & rel food intake in MD&HD was sig decr during the period 7-10&7-20. The
`deer was between 4-7% of the cont except during gd7-10 it was 20% in HD relative to cont (this was the
`period of deer in wt gain of 40% in HD). No effect on food during lactation period. No deaths occurred in
`any gr. There were fewer pregnancies in LD rel to the cont (20/25 vs. 25/25 in vehicle), however. this was
`considered not drug related since it was not seen at the higher doses and dosing was initiated post gd7
`which is after implantation. No drug effect on gestation period, duration of delivery per pup per litter,
`duration of parturifion per pup, or on implantation sites. An incr in incidence of stillborn noted in LD
`(p_<_0.01) and HD (non-sig)(1.4, 4.6. & 3% in cont. LD. 8. HD respectively). This finding was considered by
`the sponsor to be drug unrelated since it was not dose-dependent
`In the HD satellite gr, the no. of pups
`found dead or cannibalized was incr (p_<_0.01. at 14.5% vs. 4.4% in cont). Also in this gr. lactation index
`(no. of live pups on ppd11/live pups on ppd7) was sig (p50.01) decr (84%) rel to the cont (94%). The
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`sponsor considered these findings non drug related because they were not observed in the main study
`(more rats than the 5/gr satellite). Milk was absent from stomach on ppd1 of 4/20LD or 80% (p50.01) rel
`to 1/23 or 12.5% cont. The sponsor indicated that this finding was not dose dependent therefore. not drug
`related. There were no other drug effects on any of the assessed parameters.
`
`It is concluded that oral administration of modafinil to rats during pregnancy and through lactation
`dayzo was not teratogenic upto 200mglkg. There was a 40% deer in wt gain of rats dosed
`200mglkg during gestation period 7-10; values were comparable thereafter. This decr affected the
`cummulative wt during gd7-20&0-20. Mean wts were slightly but sig deer in HD during lactation.
`Decrease in food intake accompanied the deer in wt. Decrease in food intake noted In MD&HD
`during gd7-10&7-20. There were no drug effects on the several repro parameters measured in this
`study except for few sporadic effects. The affected parameters Included an Increase in incidence
`of stillborn. lactation index. no. of pups found dead or cannibalized. These findings were small,
`occured either in LB or were non-dose-dependent, or noted only in the 5 rats satellite gr and not in
`the main study. Modafinil therefore, was not embryotoxic upto 200mglkg. The NOEL for maternal
`toxicity is 100mglkg.
`
`SUMMARY AND CONCLUSIONS) FOR THE REPRODUCTIVE STUDIES:
`
`Modafinil effect on reproductive and/or developmental parameters was tested in the following studies:
`1.
`Seg l in rats/doses: 20. 50, 100mglkg/Non-GLP
`2.
`Seg II in rats/doses: 50, 100. 200mglkg/d/Non-GLP
`3.
`Seg ll in rabbits/doses: 25. 50. and 100mglkg/Non-GLP
`4.
`Seg III in rats/doses: 20, 50. 100mglkg/GLP
`5.
`Seg III in rats/doses: 50. 100, 200mglkg/GLP
`The NOEL values are as follows:
`NOEL (mg/kg)
`Study
`End—point
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`fertility
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`Seg ll rat
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`Seg Ill rat
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`maternal tax
`teratogenicity
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`maternal tax
`teratogenicity
`embryotox
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`maternal tox
`teratogenicity
`embryotox
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`100
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`200
`200
`200
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`50
`100
`100
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`100
`100
`20
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`Seg lll rat
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`maternal tax
`100
`teratogenicity
`200
`embryotox
`200
`Modafinil administered orally to rats and rabbits upto 200mglkg (this dose is 30x the max anticipated
`human dose of 400mg/d). was not teratogenic. Male and female fertility in rats was not affected upto
`100mglkg. Embryotox seen as incr resorptions, occurred at 100mg/kg in F0&F1 rats.
`In absence of
`maternal toxicity, modafinil in a Seg ll rat study incr total no. of resorptions in fdosed 200mglkg, in
`addition, there was an incr in litter hydronephrosis and incomp ossification in fetuses from these females.
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`These embryo-fetal toxicities may not be of real biological significance because the number of resorptions
`per female was not affected and the visceral and skeletal effects are relatively small. The NOEL for
`embryotoxicity in this study is therefore. 200mglkg. Rabbits dosed modatinil during organogenesis period.
`showed a small decr in mean wt gain and food intake at 100mg/kg. The NOEL in rabbits was 50mg/kg for
`maternal tox and 100mg/kg for embryotox and teratogenicity.
`In a pre- and post-natal Seg lll rat study.
`modafinil was dosed during pregnancy and lactation. There was a dose-dependent incr in RI but stated by
`the sponsor to be within historical data for this rat strain. Though dose-independent. number of live
`fetuses per female was decr in all 3 drug grs and the stillborn rate and number of stillboms were incr in
`females dosed 50&100mg/kg. The only effect on growth was a delay in startle response in the
`50&100mglkg dosed groups. The maternal NOEL is 100mg/kg whereas. that for embryotoxicity is
`20mg/kg.
`In a more recent Seg III study in rats, modafinil was not teratogenic upto 200mglkg but caused
`a 40% decr in wt gain in females in this gr during gestation period 7-1 0; no effect thereafter. This decr
`however. affected the cummulative wt during gestation periods 7-20&0-20. Mean wt was slightly but sig
`decr during lactation period. Accompanying the decr in wt was a 7-20% decr in food intake in the
`100&200mglkg dosed grs during gestation periods 7-10&7-20.
`In contrast to the previous Seg Ill study
`where the NOEL for embryotoxicity was 20mglkg. in this repeate study. modafinil was not embryotoxic
`upto 200mg/kg; the maternal NOEL was 100mg/kg similar to that in the previous study.
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`Labelling;
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`Genetic Toxicology
`_conducted in 1979 the following assays (08-93-018) all of which were non-GLP:
` .
`-
`Ames bacterial reverse gene mutation assay Plate incorporation— labs— conc
` .
` .
`tested were 10, 100. 1000, and 10.000ug/plate in 4+ 89. An incr in no. of revertants was seen in +89 in
`TA98 and TA1538 (the former is a derivative of the latter) when the assay repeated 2x; the findings were
`negative in -SQ. The incr was 2.2o3x negative cont in TA1538 and 2-3x negative cont in TA98. The
`sponsor concluded that modafinil at 5000 and 10.000ug/plate in +59 was mutagenic in TA98 and TA1538.
`
`_labs repeated Ames test with oonc upto 5000ug/plate using the 5 standard strains. each strain
` .
`tested in 3 plates per conc in 4+ 89 from Aroclor induced rat livers. Modafinil in DMSO was non-toxic
`upto 5000ug/plate which was the conc used in the main assay. Modafinil was not mutagenic in this assay
`upto 5000ug/plate in either 4+ 89 including TA98 & TA1538.
`
`A complementary 3rd Ames was awning to OECD guidelines but still non-GLP in
` .
`1982 to check the results found by
`above: only TA98 & TA1538 were examined in
` .
`4+ 89 at 500-10,000ug/plate modafinil. The results were negative and modafinil was found non-
`mutagenic in these 2 strains.
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`A more recent Ames test was done by—1995 under GLP (StUdW 03-95-
` .
`001) and sponsored by Cephalon. This study was done because the other 3 studies were non-GLP and a
`positive response was seen in strains TA98 and TA1538 in +89. This study was well done. the
`appropriate positive controls and their responses were valid and a cytotox assay was done prior to the
`main assay using TA100 and WP2uvRA (modafinil conc ranged between 5~5000uglplate and it was not
`cytotoxic upto 5000uglplate which is the conc used in the main assay). Modafinil similar to the above 3
`assays was dissolved in DMSO so did the positive cont except for 2 (NaAz & MMS) were dissolved in
`water. The strains tested were TA98, TA100, TA1535, TA1537, and E.coli WP2er Both the plate
`incorporation and pre-incubation methods were tested in -I+SQ. Cone of modafinil tested in the main
`assay were 250. 500. 1000. 2500, and 5000ug/plate in -I+SQ. The results were clearly negative in -/+SQ in
`the plate incorporation and pro-incubation assays upto 5000ug/plate modafinil conc. therefore. modafinil is
`considered non-mutagenic in the Ames bacterial reverse mutation assay.
`
`-
`In vitro Human lymphocyte assay/GLP—1986
` .
`Cytotox was assessed using modafinil conc at 30, 100, 300, 1000. and 3000ug/ml in DMSO and mitotic
`index (MI) was determined. The highest conc used in the main assay was 300uglrnl due to limit by
`solubility as stated by the sponsor. Modafinil cone in the main assay were 30. 100. 300uglml in +I-SQ at 1
`8. 24hr exposure respectively. Modafinil was not clastogenic in human lymphocyte assay upto 300ug/ml
`cone in -/+ $9.
`
`In vitro gene mutation of Chinese hamster V79 lung fibroblasts at the HPRT locus/Lafon
`-
`labs/followed OECD guidelines but not GLP (in the spirit of GLP as stated by the sponsor)l
`Exposure to test sub was 1&3hrs (OECD recommends 3-6hr and sometimes upto 1.5x cell cycle).
`Modafinil conc was given in mM and upto 10‘mM in +l- $9. The results were negative and modafinil was
`not mutagenic in this system.
`
`-
`In vivo bone marrow MN in Chinese hamste_on6LP
` .
`Modafinil was orally dosed to 6/gr Chinese hamsters for 5d at 200&1000mg/kgld and chromosomes
`checked for abnormalities. This assay failed to meet several of me OECD criteria such as no. of cells
`used per animal (recommended minimum of 100 here only 50 were used). no. of doses usually one dose
`and other designs should be justified. no justification was given for dosing for 5days. MI is determined in at
`least 1000cellslanimal in this study only 50 cells were used. A priliminary tox study showed 2/4 animals
`died at 2000mg/kg but no deaths at 1000mglkg dose. The results showed that modafinil was not
`clastogenic in Chinese hamster bone marrow MN in vivo assay upto 1000mglkg dose.
`
`As stated, it is the opinion of the reviewer that the above assays were non-GLP and some did not follow
`OECD giodelines and therefore. firm conclusions can not be made based on the data submitted.
`
`In vitro fonivard gene mutation at TK locus in L5178 MLP cells—3396-
`-
` .
`003/1996/GLP.
`Modafinil was dissolved in DMSO. Conc were based on cytotox assay and solubility characteristics. The
`drug remained in sol upto 1500ug/ml but after 4hr treatment period a precipitate was noted at zi100ug/ml.
`Note that the dmg did not alter the pH in culture medium.
`In the dose range finding assay. both untreated
`and vehicle controls were used, exposure was for 4hr at 37C atterwhich, cells were washed and
`resuspended in growth medium. Cell count was made after 24hr to measure reduction in cell growth
`relative to the cont The appropriate positive controls were used: MMS in -89 and MCA in +89.
`Modafinil in dose range finding was tested between 3.12-1451ug/ml in 4+89, it was cytotoxic at 1451 with
`65-11% cytotoxicity relative to negative controls.
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`«89:
`There were 4 assays in -89. #2 was terminated without mutant selection because there was a shift in
`cytotox causing too few doses to clone and proper evaluation of the drug; trial 3 was unacceptable
`because cloning efficincies were very high and contamination