NDA 21-372
`
`Page 131
`
`4. Hematologv Changes: A decrease of eosinophils counts were seen in treatment related
`manner in animals included in palonosetron treatment groups (eosinophil counts were 0.17 to
`0.22x109/1 in males and were 0.12, 0.14, 0.13, 0.14 and 0.06x109/1 in females belonging 2
`control and 3 treatment study groups. The differences were only of statistical importance.
`
`5. Toxicokinetic/Plasma concentration Estimation: The peak plasma concentrations on day 1
`were less than the concentrations seen on week 26, 52, 78 and 104. The steady state was not
`reached on day 1. The increase in peak plasma concentrations of the compound was linear but
`non-dose proportional. The plasma concentrations of the compound were more in females than
`males. On week 26, the plasma concentrations (AUC values) were 136.9 and 308.2 times the
`AUC value in man reported by sponsor in a study at the suggested clinical dose of 5 ug/kg. The
`pharrnacokjnetic data of the study animals are shown in the following tables. The half-lives of
`the palonosetron in males included in high dose treatment group were estimated to be 1.5, 2.2
`and 2.1 hr on study weeks 26, 52 and 78 (not calculated for study day 1 and week 104). The
`half-life ofthe compound in females was more variable as it was 0.8 to 4.2 hr on study week 26,
`0.8 to 2.4 hr on week 52 and 78. The plasma concentrations (Cmax and AUC values) of the
`compound in male and female rats are shown below (sponsor’s table)
`
`‘
`
`TABLE 21
`
`Pharmacokinetic parameters of Palonosetron on Day 1 and during Weeks 26, 52, 78 and 104
`01’104-Week Rat Carcinogenicity Study1
`
`a. Cmax (ny'ml)
`Sex
`Doses
`
`Cmax
`
`M
`
`F
`
`(mg/kg/day)
`15
`30
`60
`
`'
`
`Day 1 W6 Wk 52 Wk 78 WK 104
`19.48
`253.2
`99.8
`119.6
`120.3
`153.4
`593.3
`212.8
`557.8
`276.9
`427.3
`1744.7
`1293.7
`1521.2
`1117.5
`
`15
`45
`90
`
`25.90
`141.2
`702.8
`
`392.7
`119.7
`1224.6
`718.7
`1645.2 2349.7
`
`181.4
`350.6
`1180.7 662.8
`1288.0 2014.0
`
`13. AUC(9_14N) (ng.h/m1)
`
`Sex
`M
`
`F
`
`Doses
`(mg/kg/day)
`15
`-
`30
`60
`
`_
`
`'
`
`AUCwmn (ng.h/ml)
`Day 1 W6 Wk 52 Wk 78 WK 104
`38.8
`195.0
`215.3
`462.6
`311.5'
`361.7
`372.1
`1088.5' 1606.6
`1766.8
`1402.0 4078.4 '6139.5 6294.8 4968.0"
`
`15
`45
`90
`
`275.0‘
`624.8
`662.4
`211.3
`39.3
`2758.8 3308.4 3391.4 4162.3'
`479.6
`2511.2 9185.3
`14933.3 6441.7 95372“
`
`

`

`NDA 21-372
`
`Page 132
`
`Cmax values calculated from 3 animals; except ‘2 animals; AUCWMQ values calculated fi'om 18 samples, except
`'17 samples, t’15 samples, c = samples taken during week 103; I = 65 of rats/group
`
`At the peak levels, the metabolite RS-17825-007 levels were present in a linear but in a non-dose
`proportional manner within 0.25 to 1 hr post dosing of palonosetron in mid and high dose
`treatment groups. AUC values of the metabolite from study week 26 to 104 were similar
`indicating that the compound and metabolite were not acsumulated after the repeat
`administration of the compound. The metabolite to compound ratio was higher after repeat
`dosing than the single dose ofthe compound and the halflife ofthe compound at different
`intervals were not estimated from the data. The pharmacokinetic profile ofthe metabolite is
`tabulated below (tables taken from sponsor submission):
`
`TABLE 22
`
`
`Pharmacokinetic parameters of Rs-17825-007 on Day 1 and during Week 26, 52, 78 and 104 of 104-Week
`carcinogenicity Study in Ratsl
`Sex
`Doses
`Cmax (ng/ml)
`(mg/kg/day)
`Day 1 W6 Wk 52 Wk 78 WK 104
`15
`BLQ
`BLQ
`11.5
`14.0
`BLQ
`30
`BLQ
`43.1
`43.9
`55.4
`18.7
`60
`32.4
`117.8
`55.5
`60.5
`53.2
`
`M
`
`F
`
`BLQ
`18.4
`22.4
`BLQ
`BLQ
`15
`38.4
`91.2
`77.3
`53.1
`BLQ
`45
`90
`56.33
`93.8
`96.3
`55.9
`59.3
`
`
`M
`
`b. AUCtO-Z‘hr) (ng.h/ml)
`Sex
`Doses
`(mg/kg/day)
`15
`30
`60
`15
`45
`90
`
`1-'
`
`AUCMM (ng.h/m1)
`Day 1 W6 Wk 52 Wk 78 WK 104
`a
`a
`a
`a‘
`a
`19A
`2346
`2088
`805
`50]
`90.2
`669.3
`353.4
`360.4
`258.3
`a
`a
`a
`a
`a
`15.5b
`200.7
`157.4
`292.9
`160.2”
`298.6
`630.9
`671.7
`305.4
`337.4“
`
`—_'_—_'—._'——.‘-.—‘fif_—._——~I_
`Cmax values calculated from 3 animals; except 2 animals;
`sample taken during week 103,
`AUQMW) values
`not calculated, AUCWNM values calculated from 18 samples, exceptbl 7 samples, c = samples taken during week
`103; I = 65 ofrats/group
`
`Orally administered palonosetron produced a dose related but non-proportional plasma peak
`concentration from low to high doses of the study. The estimated half lives was short, i.e., from
`1.5 to 2.2 hr in males and 0.8 to 4.2 hr in females during the Study as shown in the table above.
`. There was no evidence of accumulation of the compound during the study. The study indicated
`that the exposure of the compound in female was higher than male animals during the study.
`
`

`

`NDA 21-372
`
`Page 133
`
`6. Organ Weights Changes: The absolute weight of liver was increased by 24.0 and 23.0% in
`males and females and, adrenal weight was increased by 37.5% and 30.9% in males and females
`included in high dose treatment group. The mean absolute weight oflung/bronchj and spleen
`were increased by 8.4 and 46.0% among males of high dose treatment group. The absolute
`weight of spleen of the animals of high dose treatment group was increased by 33.1 and 21.3% in
`males and females, respectively. The absolute weights of salivary glands and testes in males of
`high dose treatment group testes were lower than control animals by 12.2 and 56.3%,
`respectively. The absolute weight of ovaries was decreased in a treatment related manner, i.e.,
`70.5, 72.5 and 71.6% among females oflow, mid and high dose treatment groups, respectively.
`The mean relative weight (in relation to body weight) ofliver was increased by 36.8 and 56.1%
`in male and female of high dose treatment group and, uterus + cervix relative weight was
`reduced by 1.7, 2.32 and 2.5 times than the weight of the tissues ofthe control group animals.
`
`7. thsical Examination/Ophthalmoscopic Changes: These changes were not observed
`during the study.
`
`8. Gross Patholoox,' Changes: At sacrifice, an increase in the incidences of dark areas in
`adrenals was seen in 2, 0, 1, 4 and 8 males and, 9, 11,15, 19 and 15 females included in 0, 0, low,
`mid and high dose treatment groups, respectively. Enlarged adrenal was seen in 2 males of high
`dose treatment group. Small epididymides was seen in O, 0, 2, 1 and 11 males of control 1,
`control 2, low, mid and high dose treatment groups, respectively. The enlarged kidneys were
`reported in only 2, 0, O, 1 and 5 males of control 1, control 2, low, mid and high dose treatment
`groups, respectively. The flaccid testes were seen in 7, 10, 6, 5 and 42 males, testicular fluid in
`subscapsular region in 2, O, 0, l and l 1 males, small testes in 0, 0, 3, 0 and 18 in males and, blue
`colored testes in O, 0, 2, 3 and 9 males were reported in of control 1, control 2, low, mid and high
`dose treatment groups, respectively. Enlarged and swollen liver was seen 15 males and 16
`females of high dose treatment group. Only a trend of an increase of mammary masses wee seen
`in both sees ofanimals. These and the other pathological findings noted in the animals are
`shown in the following table.
`
`TABLE: Patholo-ical Chan-es in Or-ans/Tissues of Rats"
`
`Dark Adrenal
`
`F
`
`
`
`Enlarged Adrenal M
`F
`.E-idid 'mides small M
`Eididvmides swollen M
`
`Enlar. Kidneys
`
`Liver Swollen
`
`

`

`NDA 21-372
`
`Page 134
`
`Testes Flaccid
`
`Testes Subscao. Fluid
`
`II-
`AmeLymthodeMass _
`
`
`
`
`
`
`
`Pancreat Lymph Node M _l-—__
`Masses
`F __I—__
`Skin Masses Present
`l-
`__
`
`M F
`
`‘= 65/sex/group
`
`9. Histopathological Changes:
`
`.Noon-Neoglastic Changes.
`
`a. Males: Epididymal hypospermia, the absence of colloid from
`
`seminal vesicle and testicular tubular germinal epithelial degeneration were seen in higher
`incidences (p<0.05 — 0.005) than in control group animals. A significant increase in the
`incidences ofprogressive senile renal nephropathy (p<0.05) were seen in males of high dose
`treatment group (see table). Sinus erythrocytes and erythrophagocytosis in mesenteric lymph
`node were present in slightly higher incidences in males of the high dose group than control
`group. A dose related increase in the incidents of accumulation of alveolar macrophages of lung
`was seen in treatment groups males. The degeneration of testicular tubular germinal epithelium
`was noted in high dose treatment group (60 Vs 14-16 in control group). The incidences of
`epithelial hyperplasia ofthymus and thymus cysts were high among males of high dose group
`animals. The epidermal hyperplasia of slight to moderate nature was present in mid and high
`dose treatment groups (see table below).
`
`b. Females: Increased incidences of adrenal cortical vacuolation were seen in females of mid
`
`and high dose treatment groups animals, i.e., 6 males in each ofmid and high dose treatment
`group, respectively (Vs 2 in control group). The incidences of adrenal medullary hyperplasia,
`chronic renal nephropathy (senile) were higher than control in females of high dose treatment
`group. The focal medullary hyperplasia was noted in 8 and 1] males and females ofhigh dose
`teatment group.
`In mesenteric lymph nodes, sinus erythrocytes and erythrophagocytosis were
`reported in higher incidences in high dose treated animals than control females. The
`accumulation of alveolar macrophages in lung was noted in higher incidences, i.e., 26 females in
`high dose treatment group animals. The islet cell hyperplasia ofpancreas was noted in 0, l, l, l
`and 3 females ofstudy groups. The diffirse hyperplasia ofpituitary was found in 0, l, 0, l and 5
`females. The incidences of scabs on tail were 7, 6, 10, 23, and 30 females and the observed
`epidermal hyperplasia was of slight to moderate nature in animals (see table below). Clear cell
`foci in liver were seen in 9, l3, 2], 20 and 25 females. Follicular abscess was in a dose-related
`rnannen
`
`

`

`NDA 21-372
`Page 135
`
`TABLE: Incidencee of Non-Neoplastic Lesions in Control and Treatment groups Ratsl
`
`
`
`
`
`
`
`
`
`
`
`Histopathological Lesion
`
`Cortical Vacuolation
`
`M
`
`Medullary l-lyperplasia Focal MI‘1
`
`
`Treatment Grou-s (N = 65/sex/ on—-5—
`
`——II—
`
`N
`
`Male Reproductive Or_ans:
`l-lI——__Il-_
`—_——I-_
`——I_l_lI-II_E_
`progressive senne
`M _——I__
`Nepmopamy
`r __—I_I_
`Liver
`
`
`
`
`
`
`Focal Congestion
`
`I!
`
`._. N
`
`_. DJ
`
`Il-
`-
`Lungs
`Accumul. Alveolar Macroph. M _
`F ——_——
`Pen‘vasc- Lymphoid Aggreg- M —_-__
`Lungs
`F _I—___
`
`Mammary Gland
`--___I_
`35
`F
`_!_II-_- 18
`__——_—-
`Myofibre Degeneration
`M __I___
`F __I-I-I-
`S-leen
`
`__—I—__
`F __I-II_I_
`M _____
`F
`
`IE-
`
`
`
`Hemosiderosis
`
`Thymus
`
`______
`F _——:—n-
`M ____I-
`F
`
`Thymus Cysts
`
`—————_
`Eidermal H -e -lasia
`M —_—_I_
`_-—__I_
`———II__I_
`—___II__
`—__-_I_
`
`
`
`
`___—__
`
`= 65 of rats/group
`
`

`

`NDA 21-372
`
`Page 136
`
`b. Neoplastic Changes:
`lncreased incidences of the benign pheochromocytoma were seen in a treatment related manner
`in 9, 13, 16, 18 and 27 males of 0, 0, 15, 30 and 60 mg/kg/day treatment groups. Among
`females, the incidences of benign pheochromocytoma were 1, 0, 2, 4 and 7 in 0, 0, 15, 45 and 90
`mg/kg/day treatment groups. The incidences were 28.7 and 41.5% in mid and high dose
`treatment groups males. The background control data range provided by sponsor for males was
`10.9 to 25%. The incidence of the tumor was 10.8% in females, which was more than the
`background range of 2 to 10%. The pairwise p values were 0.0004 for 60 mg/kg/day treatment
`group male 0.0004 for females of 90 mg/kg/day treatment group (pairwise comparison with
`pooled control). The p values with control 1 and 2 were 0.0007 and 0.0132 for high dose males.
`These were not significant for control 2. The combined incidences of benign and malignant
`pheochromocytoma were significantly increased for males of 60 mg/kg/day treatment group
`(trend test p < 0.001). The pairwise p test with pooled controls for the combined benign and
`malignant pheochromocytoma was not significant in these animals. The increase in the
`incidences ofislet cell adenoma (p<0.05 and pairwise p<0.029) and combined incidences ofislet
`cell adenoma and carcinoma (trend p<0.005) was significant in study males. The pairwise
`comparison showed that'tl'ie combined incidences were significantly higher in all the treatment
`groups study animals (p<0.01). The incidences ofrare benign hepatocellular adenoma were
`increased in males and females of high dose treatment group. The increase in incidences was
`significant (trend test p<0.0002) in high dose treatment group females and incidences were
`significant by pairwise testing (p<0.0012) and the incidences were more than the historical
`control range ofO to 5.3. The thyroid C-cell adenoma incidences (common) were increased in
`high dose females (exact p>0.0001; pairwise p with pooled control p<0.0053). The mammary
`adeno-carcinoma incidences in females of45 and 90 mg/kg/day treatment groups were more
`than control 1 but not when compared with control 2 or pooled control animals. The combined
`incidences of mammary adeno-carcinoma and benign fibroadenoma were within sponsor
`background range of 1.4 to 6.0%. The significant increase in the incidences of benign adenoma
`pars distalis (common) were seen in male rats included in low, mid and high dose treatment
`groups (pair wise p values 'were 0.0001, 0.0005 and 0.0005 in animals of 3 treatment groups).
`The incidences were greater than the historical control data of sponsor laboratory.
`
`TAB LB
`
`lncidences of Neo -
`
`
`Histopathological Lesion
`
`
`
`
`
`
`
`lastic Lesions in Control and Treatment _rou - s Rats
`’
`Treatment Gro us
`
`_—
`Cont.l Cont.2
`Low
`Mid
`High
`Trend Tes: Pairwise
`Dose
`Dose
`Dose
`(p value)
`p (pooled
`control
`Adrenals:
`
`
`
`Benignpheocmmocwm
`M __-_—I_I_
`
`F
`I-
`__l--
`Malignant Pheochromocytoma M
`l-___ 0.158 _
`I-
`F
`__l__ 0658 —
`
`l4
`17
`22
`29
`_
`'"Z
`
`Combined lncidences
`
`10
`
`
`
`
`
`
`
`

`

`NDA 21—372
`
`Page 137
`
`:
`
`
`
`MemmmmmmeeMee
`M -_I-___—
`Carcinoma (MFG)
`F ___I-_—-
`Hepatocellular
`M nun—“—
`WW F __- _- °'°°”
`Combined Incidences of
`M ——_____
`MHC +BHA
`F __——I-_-
`—-—__--—
`_——-—__—
`F _____flfl-_
`|AcinarCeHAdenocarcinoma M
`[_E-I-E-I-_—
`
`-
`F _l-I-___—
`M _———-__
`eeememe ea or
`Adm" AFFFFFFFCFFFW F ____I_—-
`Islet Cell Adenoma
`M
`-——I_E-I-II_
`F --__I-—
`
`-' "ml-—
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`—_—__—_—
`F _------
`———l-_I_n-ma-
`
`Tememmnmemmn
`Adenoma
`
`M _-——-I-_
`F _l-l- -=g
`
`c-CeHCeeee-mme
`
`Benign Adenoma-ParDistalis M
`r
`
`-
`
`
`
`Mammarv Gland
`
`Adenocarcinoma
`
`M ___I___-
`F _m— 2
`—
`
`F
`
`Commeememen mm
`MFFFMF
`
`M _l-_
`F
`
`
`
`
`
`Thyroid C-CellAdenoma M __l_ m 0.0136
`F
`-II__I_-——
`
`
`M
`I-_I-l-l_——
`
`
`F __I-_I_——
`
`
`Cemeeeeec-Ceumemme
`M _n-n-n-u-__
`
`
`FC-CF'FFFFFFFF
`F “mm-—
`
`
`
`M ‘0
`<0.0001
`0.005
`III-hWU!
`MAO‘b-I
`WAebb!
`
`<O.69
`
`
`
`
`
`—.___
`[_l__
`
`Benign fibroadenoma
`M _ _—_
`F
`III——
`
`

`

`NDA 21-372
`
`Page 138
`
`In summary, the study was conducted in adequate number of animals (male/female) at the oral
`gavage doses of 0, 0, 15, 30 and 60 mg/kg/day in males and, 0, O, 15, 45 and 90 mg/kgday in
`females. The administered compound attained a non-dose proportional plasma concentrations of
`the parent compound and the metabolite RS-l7825 (major human metabolite). The plasma
`concentrations in females were more than males of the treatment groups during the study. On
`study week 26, the exposure ofthe compound in male and female animals was 136.9 and 308.2
`times the plasma concentration (AUC values) achieved after the suggested clinical dose of5
`ug/kg in man. Increased incidences of benign pheochromocytoma in male and females were
`seen. Increased incidences of combined benign and malignant pheochromocytoma and,
`pancreatic islet cell adenoma and combined incidences ofislet cell adenoma and carcinoma were
`seen in all treatment group animals. The adenoma ofpar-distalis incidences were significantly
`high in males of treatment groups. The incidences of hepatocellular adenoma and thyroid C-cell
`adenoma were increased in females of high dose treatment group. The increased incidences of
`tail squamus cell papilloma in high dose males were reported.
`'
`
`REPRODUCTIVE TOXICITY STUDIES:
`
`1. Intravenous male fertilitv studv with RS-25259-197 in rats: (Report. No. 6267)
`
`Testing Laboratories: Syntex Discovery Research, Palo Alto, CA
`
`Studv Started: April 6, 1992
`
`Studv Completed: December 16, 1992
`
`GLP Reguirements: A Statement of compliance with the GLP regulations and quality
`assurance unit was included.
`
`Animals: Crl: CDR (SD)BR VAF/Plus strain Rats; Males approximately 15 weeks old, weighing
`427-577 g; females approx. 4-5 months old, weighing 262-404 g (post mating day l weights for
`females).
`.
`
`Drug Batch No.: PA15303-51/11902, 11903, and 11904
`
`Methods: RS-25259-197 was intravenously administered (via the tail vein) to 4 groups of 10
`male rats were at doses of0 (vehicle), 1, 3, or 10 mg/kg/day for a period of2 weeks prior to
`mating with untreated females and continuously for 4 weeks thereafter: The vehicle used in the
`present study was a sodium phosphate buffer solution which consisted of 0.087 g% sodium
`chloride, USP; 0.020 g% monobasic sodium phosphate monohydrate, USP; and 0.058 g%
`dibasic sodium phosphate anhydrous, USP in purified water, USP. RST2S259-197 and vehicle
`were each administered in total volumes of 1 ml/kg. Dose selection was based on the results of a
`1 month i.v. toxicity study (Syntex 89-R—91 , AT 5962), wherein rats given a 10 mg/kg daily
`dose exhibited transient clinical signs, but no clinical or anatomic pathological changes. Male
`
`

`

`NDA 21-372
`Page 139
`
`rats were observed daily for general condition and weighed weekly and terminally. Female body
`weights were determined on day 1 of pregnancy or at the end of the cohabitation period
`(nonpregnant females) weekly thereafter and terminally. During the 4 day cohabitation period
`females were examined daily for the presence ofa copulatory plug or- sperm in the vaginal
`lavage. Females with evidence of mating were euthanized on gestation day 14, whereas
`nonpregnant females were euthanized at 13 days after the end of the mating period. All females
`were subjected to partial necrOpsies in which the whole reproductive tract was examined. The
`number of corpora lutea, live fetuses (viable implants) resorptions (non-viable implants) and
`abnormal conditions were also determined in pregnant females. Following review of the
`gestational data, males were euthanized and discarded, with no gross examinations conducted.
`
`
`Results: Male rats which received the 10 mg/kg dose exhibited vocalization immediately after
`dosing for up to 45 sec on various days. Males in all other groups appeared normal with no
`mortality observed in any group. Males in all treated groups had non dose-dependent
`suppression of body weight gains (-23 to -34%) compared to gains in control groups. All dams
`survived to the scheduled sacrifice, with no gross pathologic changes observed. No treatment-
`related effects on mating performance: [proportion (%) of males mating with at least 1 female;
`proportion (%) ofmales with at least 1 female pregnant and the proportion (%) of males with at
`least 1 female pregnant among those with evidence ofmating] were observed. Table 6, on the
`following page presents a summary of the gestational indices of female rats examined at mid
`gestation (Reproduced from Sponsor's Summary Table 4, vol 1, pgs. 82-83).
`
`Table 6.
`
`Summarv of Re roductive Indices in Female Rats Examined at Mid
`
`Gestation (Reproduced from Sponsor's Summary Table 4, vol 1., gs. 82—83)
`
`1
`
`3
`
`1
`
`Variable
`# Animals
`
`# Preg. (%)
`# w/ Resorp. (%)
`
`# Corpora lutea
`# Implantations
`Live litter size
`# Total resorptions
`Implantation index
`Reso tion index
`
`
`
`20
`
`13 (65%)
`7 (53%)
`
`16713.7
`13.1 i 4.2
`12.114.1‘
`0.9 i 0.9
`77.6 i 17.6
`8.3 i 10.5
`
`l
`'
`
`20
`
`15 (75%)
`6 (40%)
`
`'
`
`18.1 :21
`15.8 i 1.9
`15.2: 1.7
`0.7 i 0.8
`88.2 i 10.7
`3.9 :43
`
`20
`
`16 (80%)
`4 (25%)
`
`17.8:26
`14.2 i 3.2
`13.9:34
`0.3 i 0.6
`79.6 i 16.4
`3.4 i 7.4
`
`20
`
`14 (70%)
`9 (64%)
`
`17.6128
`15.9 i 0.9
`14811.1
`1.1 i 0.7
`91.8 i 8.4
`6.6 :43
`
`,
`* Significantly different from control values P < 0.05
`IMPLANTATION INDEX =(1MPLANTATIONS/CORPORA LUTEA) X 100; RESORPTION
`INDEX = (TOTAL RESORPTION/IMPLANTATIONS) X 100
`
`Briefly, the data in Table 6 (above) show that treatment of male rats with RS-25259-197 at doses
`up to 10 mg/kg for 2 weeks prior to and during mating had no effect on the No. ofcorpora lutea,
`
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`NDA 21-372
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`
`implantations, and resorptions. A significant reduction in the live litter size was observed,
`however it's relation to treatment is not apparent as no other gestational parameters showed
`treatment-related changes.
`
`In conclusion, intravenous administration of RS—25259 to male rats at doses of l, 3, or 10
`mg/kg/day for 2 weeks prior to mating, produced nansient vocalizations at the high doses and
`suppressed body weight gain at all doses, but had no effegts on mating performance or fertility.
`However, the 2-week dosing schedule used currently is not in accordance with the recommended
`duration for a normal Segment 1 Fertility and Reproductive Toxicity Study in male rats. Thus,
`the aforementioned study cannot be used in lieu ofa normal Segment I study in male rats.
`
`2. Segment 1. Fertilitv and Reproductive Performance Toxicitv of Orallv Administered
`RS-25259—007 in Male Rats: (Study # 4l-R-94/ AT 6700)
`
`Testing Laboratogy: Syntex Discovery Research, Palo Alto, CA
`
`Dates of Start and Completion of Study: August 19, ]993 and June 27, 1994
`
`CL? and (ZAU Reguirements: Statement was included.
`
`Animals: Sprague Dawley female and male rats (Crl: CDR (SD)BR VAF/Plus strain) with an
`average weight of 260 g (females) and of non-specified weights (males) were used in the study.
`
`Lot No. of RS-25259-007: Lot # 2422, 24 and 25.
`
`Methods: One hundred and twenty female and 60 male rats were divided in 4 groups (15 males
`and 30 females/group). Male rats were administered a single oral dose of either 0, 18, 60 or 120
`mg of RS—25259—OO7/kg/day (in a volume of 5 ml/kg/day adjusted daily according to daily body
`weight for at least 6] to 63 days prior to cohabitation and were mated with untreated females.
`The treatment of male rats continued during mating period and these animals were weighed and
`observed for physical changes weekly. The dose selection of the study was based on the results
`of l-month oral toxicity study in rats (Study # l6-R-92, AT 6329). In this study, a dose of 60
`mg/kg/day was a 'no effect dose' and a dose of 180 mg/kg/day produced a reduction in the body
`weight gain and abnormal histopathological changes in liver, testes and thymus. In the present
`study, ]/2 of the female rats included in each of the control and treatment groups were
`necropsied at midgestation and the other half were allowed a natural delivery (day 25/26 of
`gestation). The female rats were observed daily for clinical observations, weighed on day 1 of
`pregnancy, weekly and at termination of the study. Those rats who did not litter were necropsied
`3 to 9 days after the expected day of parturition. These were not examined histopathologically.
`The numbers of pups born (live/dead) were recorded. On postpartum day 4, litter sizes were
`standardized to 8 pups/litter (1:1/MsF). The general condition of the pups, body weight, sex and
`number of live pups were recorded on postpartum day 4, 7, l4 and 21 of the study. At
`
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`NDA 21-372
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`
`midgestation, females were killed and mammary chain and reproductive tract were separated.
`The number of implantations, corpora lutea, distribution of live and dead fetuses/uterine horn
`determined and number of empty implantation and resorbed sites were noted. Pups died within 4
`days of parturition were preserved in 70 % ethanol for examining extemal abnormalities.
`
`Results:
`
`1. General Observational Effects on Dams: R525259 up to a dose of 120 mg/kg/day did not
`produce any effect in male rats. Two ofthe pregnant females developed an external
`encrustation. One female was killed during the study as it delivered all dead pups. A treatment
`related retardation in the body weight gain of 10.3 and 9.06 % were seen in groups of animals
`necropsied at midgestation and belonging to 18 and 60 mg/kg/day treatment groups. Since none
`of the mated female rat included in 120 mg/kyday treatment group was pregnant, sponsor did
`not include the data of body weight change of females belonging to this group.
`
`2. General Reproductive and Fertilitv Data of Dams: A treatment related reduction in the
`number of matings among males included in 120 mg/kg/day treatment group was seen. The
`percent of matings were 90, 96.7, 93.3, 73.3 % in control, 18, 60 and 120 mg/kg/day treatment
`groups, respectively. Sponsor reported incorrect mating of 87 % among animals treated with 120
`mg/kg/day treatment group in Summary table # 4 of mating performance and fertility. None of
`the females those mated with a male belonging to 120 mg/kg/day treatment group was pregnant,
`thus RS-25259 at this dose was toxic to rats.
`
`Implantation sites, live litter size, total resorptions and resorption index were not affected among
`'18 and 60 mg/kg/day treatment groups (Table ). But implantation index was dose dependently
`reduced, this was 91.4, 87.9 and 77.4 % among animals included in control, 18 and 60
`mg/kg/day treatment groups respectively.
`
`3. Effect on Litter Data: The mean number oflive fetuses/litter among animals included in 18
`and 60 mg/kg/day of RS-25259 treatment groups was not statistically different from those of
`control group (14.4, 15.0 and 11.7 in control, 18 and 60 mg/kg/day treatment groups
`respectively). The percent survival index of F1 rats included in 18 and 60 mg/kg/day treatment
`groups was not statistically different fiom the control group animals and these were 100 % in all
`of these treatment groups (Table). The body weight of the pups born to dams included in 18 and
`60 mg/kg/day treatment group was also comparable with the control group.
`
`Segment 1. Reproductive Performance Study ofRS 25259 in Male Rats
`
`Table 1
`
`Parameter Measured
`
`Control
`0
`
`Low dose
`18
`
`Mid Dose
`60
`
`High Dose
`120
`
`Dams Data (FD)
`
`

`

`NDA 21-372
`
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`
`Total # Dams used .
`
`i
`
`Copulation Rate
`
`Pregnancy Rate
`
`Observations (C-Section)
`
`30
`
`90
`
`100
`
`Mean number of Corpora lutea 18.2
`Mean Implantation
`16.4
`Mean Liner Size
`15.1
`Mean Total Resorptions
`1.3
`% Resorption Index
`8.2
`
`Neonatal Obsrvations:
`
`Mean Gestation Period
`# Born Alive
`Live Pup Body Weight
`Day 4
`Male
`Female
`Dav 17
`Male
`Female
`Day 14
`
`Male
`Female
`
`Day 21
`
`21.5
`14.4
`
`10.0
`9.2
`
`16.7
`15.7
`
`34.9
`33.6
`
`:
`
`3o
`
`96.7
`
`100
`
`7.8
`15.8
`15.1
`0.7
`4.3
`
`21.3
`15.0
`
`10.0
`9.3
`
`16.8
`15.8
`
`35.7
`34.5
`
`30
`
`93.3
`
`93
`
`18.1
`13.8
`13.3
`0.4
`3.1
`
`21.8
`11.7
`
`10.9
`10.1
`
`17.7
`16.9
`
`36.5
`35.3
`
`Male
`Female
`
`55.0
`52.9
`
`56.9
`54.9
`
`58.4
`56.0
`
`3o
`
`73.3
`
`0
`
`-
`-
`-
`-
`-
`
`-
`-
`
`-
`~
`
`—
`-
`
`-
`-
`
`-
`
`- S
`
`ix pups (Two pups from 2 control group dams # 0132 and 0144 and, 4 pups 01'4 dams # 328, 332, 342, 350 and
`356 of60 mg/kg/day treatment group) were found dead during the study. Only pup # 144 (control group) had wavy
`thoracic rib. All of other dead pups were comparable with control group pups.
`
`In conclusion, R825259 produced a dose dependent decrease of the implantation index in
`animals treated with low and mid doses of the study. The highest dose of the study, 120
`mg/kg/day was toxic in animals and the treated animals were infertile as the rate of copulation
`was markedly reduced and none of the female got pregnant. No effects on the reproductive and
`fertility of animals treated with 18 and 60 mg/kg/day dose were noted.
`
`3. Segment 1. Fertility and Reproductive Performance Toxicity of Orally Administered
`RS-25259-007 in Female Rats: (Study # 78-R-94/AT 6750)
`
`Testing Laboratorv: Syntex Discovery Research, Palo Alto, CA.
`
`GLP & QAU Reguirements: The statement of compliance was included.
`
`

`

`NDA 21-372
`
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`
`Date of start and Comp;letion of Studv: March 15, 1994 and September 9, 1994.
`
`Animals Used: One hundred and twenty female Crl:CD-lR (lCR)BRVAF/Plus strain rats
`approximately 3 to 4 months of age were used in the study.
`
`Lot No. of RS-25259—007: Lot # PA 17555—40
`
`Methods: These animals were divided in 4 groups (30/group) and were administered a single
`oral dose of either 0 (acetic acid and sod. acetate solution pH 5.0), 9, 30 or 60 mg of RS-25259-
`197/RQday (in a volume of5 ml/kg/day adjusted daily according to daily body weight) for 2
`weeks prior to cohabitation with untreated males (1 male with 2 females). The treatment of
`female rats continued through gestation period 9. Females without evidence of mating were
`treated through 9 days after the end of the mating period. The dose selection ofthe study was
`based on the results of 1-month oral toxicity study in rats (Study # l6-R—92, AT 6329).
`In this
`study, a dose of 60 mg/kg/day was a 'no effect dose' and a dose of 180 mg/kg/day produced
`retardation in the body weight gain and abnormal histopathological changes in liver and thymus
`and ovarian cyst in females. The day 1 ofpregnancy was an evidence of sperm plug in vaginal
`lavage. On day 15 of gestation, females with mating evidence were necropsied. Females with
`no evidence of pregnancy were necropsied 15 days after cohabitation period and number of
`implantation and corpora lutea counted. 1n the present study, female rats were observed daily for
`clinical observations, weighed on day l ofpregnancy, twice weekly thereafter and at termination
`of the study. All ofthe surviving and rats died during the study were necropsied to determine the
`cause oftheir death. The number of implantation, corpora lutea, distribution of viable and
`resorbed implants/uterine horn determined and any abnormal conditions were recorded.
`
`
`Results:
`
`1. General Observational Effects on Dams: RS-25,259 up to a dose of 60 mg/kg/day did not
`produce any effect in dams. No significant retardation in the body weight gain was seen in
`animals belonging to (l, 9, 30 and 60 mg/kg/day treatment groups respectively. Three animals,
`i.e., 1 and 2 animals belonging to control and 9 mg/kg/day treatment groups died during the
`study. The percent of animals mated were 93, 86, 83 and 73 % in 0, 9, 30 and 60 mg/kg/day
`treatment groups respectively and the number of conceived were 85, 95, 83 and 73 % in animals
`belonging to 0, 9, 30 and 60 mg/kg/day treatment groups respectively. A trend of a decrease of
`number of matings and number of animals conceived was seen among animals included in 60
`mg/kg/day treatment group. This was comparable with the animals included in the control
`group.
`
`2. General Reproductive and Fertilig Data of Dams: The number of corpora lutea and
`implantation were not affected by RS-25259 treatment (Table ). The live mean litter size was
`15.6, 13.1 and 13.6 in 9, 30 and 60 mg/kg/day treatment groups respectively vs 14.0 in the
`control group. The numbers of early resorptions were 0.8, 1.0, 1.] and 1 in 0, 9, 30 and 60
`
`

`

`NDA 21-372
`
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`
`mg/kg/day treatment groups respectively. The data is shOwn in the following table (Table taken
`from sponsor's submission at page 020.
`
`Table 1
`Segment 1. Reproductive Performance Study of RS 25259 in Female Rats
`
`. C
`
`Parameter Measured
`
`Control
`0
`
`Low dose
`9
`
`Mid Dose -
`30
`
`High Dose
`60
`
`Total # Dams used
`
`Copulation Rate
`
`Pregnancy Rate
`
`Observation at C-Section
`
`Mean number of Corpora lutea
`Mean Implantation
`Mean Litter Size
`
`Mean Early Resorptions
`% Resorption Index
`Implantation Index
`
`29
`
`93
`
`85
`
`18.1
`14.7
`14.0
`
`0.8
`4.8
`18.21
`
`28
`
`86
`
`95
`
`19.8
`16.6
`15.6
`
`1.0
`5.7
`12.15
`
`30
`
`83
`
`83
`
`17.5
`14.2
`13.1
`1.1
`9.6
`20.39
`
`30
`
`73.3
`
`73
`
`18.2
`14.6
`13.6
`1.0
`6.9
`24.57
`
`In conclusion, R525259 produced no effect on the fertility of female rats up to a dose of 60
`mg/kg/day dose. Only a trend of a decrease of number of matings and conceived were seen at a
`dose of 60 mg/kg/day dose. The dose selection of the study appears to be not appropriate as it
`was a no effect dose in l-month toxicity study. A dose of180 mg/kyday was toxic dose in the
`same study, therefore, MTD was not identified.
`
`4. Oral Segment II Teratologv Studv with RS-25259-l 97 in Rats
`(Report No. 67-R-94-25259-l97—PO-7T)
`
`Testing Laboratorie : Syntex, Palo Alto, CA
`
`Studv Started: January 18, 1994
`
`Studv Completed: October 3, 1994
`
`GLP Reguirements: A Statement of compliance with the GLP regulations and quality
`assurance unit was included.
`
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`

`NDA 21-372
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`
`Animals: Sexually mature Cr1:CD7BR VAF/Plus Sprague Dawley Rats, Females:
`approximately 15 weeks of age, 209-290 g in weight; Males: approximately 5 months of age,
`weight not indicated.
`
`Drug Batch No.: PAl7555-40ML
`
`Methods: Three groups of female CD7Rats (25-26/group) were orally administered (by gavage)
`RS-25259-197 at doses of 18, 60, and 120 mg/kg (dissolved in vehicle and administered at a
`volume of2 ml/kg) during day 7 through 1.6 of gestation. A forth group of 24 female rats were
`given 2 ml/kg ofvehicle (13.2 mg/ml glacial acetic acid + sodium acetate 53.08 mg/ml in 1 ml of
`distilled water) alone and served as controls. Dose selection was based on an oral range-finding
`teratology study in rats where RS-25259-197 was administered at doses of 30, 90, or 180 mg/kg/
`day. The 180 mg/kg dose produced reduced maternal weight gain and increased numbe