Registration Dossier

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-11-11 to 2016-01-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: EPA OPPTS 870.3550 (Reproduction/Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study report): JNJ-119717-AAA (T001036)
- Physical state: solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: A14JB3442
- Expiration date of the lot/batch: 2016-09-30 (retest date)
- Purity test date: 2015-03-04

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Solubility and stability of the test substance in the solvent/vehicle: Stability of formulations over 6 hours at room temperature under normal laboratory light conditions (concentration range 1-200 mg/mL) was confirmed as part of the analytical method development and validation study (Test Facility Study no. 509775).

FORM AS APPLIED IN THE TEST (if different from that of starting material): Solution (groups 2, 3 and 4)

OTHER: correction factor = 1.07

Test animals

Species:
rat
Strain:
Wistar
Details on species / strain selection:
This species and strain of rat has been recognized as appropriate for reproduction toxicity studies. Charles River Den Bosch has general and reproduction/developmental historical data in this species from the same strain and source. This animal model has been proven to be susceptible to the effects of reproductive toxicants.
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: approx. 10 weeks (start pretest); approx. 12 weeks (start treatment) (females); approx. 11 weeks (start treatment) (males)
- Weight at study initiation: 191-235 g (females), 303-339 g (males)
- Fasting period before study: no
- Housing:
Pretest: Females were housed in groups of 5 females/cage in Macrolon plastic cages (MIV type, height 18 cm).
Pre-mating: Animals were housed in groups of 5 animals/sex/cage in Macrolon plastic cages (MIV type, height 18 cm).
Mating: Females were caged together with males on a one-to-onebasis in Macrolon plastic cages (MIII type, height 18 cm).
Post-mating: Males were housed in their home cage (Macrolon plastic cages, MIV type, height 18 cm) with a maximum of 5 animals/cage. Females were individually housed in Macrolon plastic cages (MIII type, height 18 cm).
Lactation: Pups were kept with the dam until termination in Macrolon plastic cages (MIII type, height 18 cm).
- Diet (e.g. ad libitum): Free access to pelleted rodent diet
- Water (e.g. ad libitum): Free access to tap-water
- Acclimation period: At least 5 days prior to start of pretest (females) or treatment (males).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24°C
- Humidity (%): 40-70%
- Air changes (per hr): at least 10 room air changes/hour
- Photoperiod (hrs dark / hrs light): 12h/12h licht/dark

IN-LIFE DATES:
From: 2015-11-11 (females; start pretest); 2015-11-25 (males; start treatment); 2015-12-31 and 2016-12-01/02/03/04/05/12/14 (pups; delivery of litters = PND 1)
To: 2015-12-24 (males; scheduled necropsy); 2016-01-12/13/14/15/18/19/26 (pups; scheduled necropsy on PND 13-15); 2016-01-13/14/15/18/19/20/27 (females; scheduled necropsy on PND 14-16)

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Remarks:
specific gravity 1.036
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
- Formulations (w/w) were prepared daily within 6 hours prior to dosing and were homogenized to a visually acceptable levle. Adjustment was made for specific gravity of the vehicle (1.036). A correction was made for the purity/composition of the test item. A correction factor of 1 was used.
- Formulations were placed on a magnetic stirrer during dosing.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at Charles River Den Bosch.
- Concentration in vehicle: 0 mg/mL (group 1), 3 mg/mL (group 2), 10 mg/mL (group 3), 30 mg/mL (group 4)
- Amount of vehicle (if gavage): 5 mL/kg body weight. Actual dose volumes were calculated according to the latest body weight.
- Lot/batch no. (if required): no data
- Purity: no data
Details on mating procedure:
- M/F ratio per cage: 1 animal/sex/cage
- Length of cohabitation: Following a minimum of 14 days of treatment for the males and femlaes, until detection of mating was confirmed
- Proof of pregnancy: Detection of mating was confirmed by evidence of sperm in the vaginal lavage or by the appearance of an intravaginal copulatory plug. This day was designated Day 0 post-coitum. Once mating was confirmed, the males and females were seperated. A maximum of 14 days was allowed for mating, after which female no. 53 (Group 2) who had not shown evidence of mating was seperated from her male.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): Females were individually housed in Macrolon plastic cages (MIII type, height 18 cm).
- Any other deviations from standard protocol: no
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Chemical analyses were conducted on a single occasion during the treatment phase (2015-12-02), according to a validated method (Test Facility Study no. 509775). Three sets of duplicate samples were collected. Two sets of duplicate samples were stored as reserve samples. After the analytical results were approved in the raw data by the Principal Scientist, the reserve samples were destroyed. Samples of formulations were analyzed for homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations).
In addition to the criteria mentioned in the validated analytical method, each calibration curve was accepted if the average of the retention times and response factors of the data points used to construct the calibration line were within a range of ±10% compared to those obtained during the method validation.
The accuracy of preparation is considered acceptable if the mean measured concentrations are 90-110% of the target concentration. Homogeneity is demonstrated if the coefficient of variation is ≤ 10%.
Stability of the test item under test conditions was performed as part of the method validation study (Test Facility Study no. 509775).
Duration of treatment / exposure:
29 days (males), 49-63 days (females that delivered), 47 and 54 days(females that failed to deliver)
Pups were not dosed directly but were potentially exposed to the test item in utero, via maternal milk or from exposure to maternal urine/faeces.
Frequency of treatment:
Once daily for 7 days per week, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Group 1 (control)
Dose / conc.:
15 mg/kg bw/day (nominal)
Remarks:
14 mg eq/kg bw/day, Group 2
Dose / conc.:
50 mg/kg bw/day (nominal)
Remarks:
47 mg eq/kg bw/day, Group 3
Dose / conc.:
150 mg/kg bw/day (nominal)
Remarks:
140 mg eq/kg bw/day, Group 4
No. of animals per sex per dose:
10 animals/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
Dose levels were based on the results of an acute oral toxicity study in the rat (Test Facility Study no. 509004) and a 28-day repeated toxicity (gavage) study in the rat (RCC B65125; data on file at Sponsor). In the acute oral toxicity study, JNJ-119717-AAA (T001036) was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure two additional groups of three females were dosed at 300 mg/kg body weight. Treatment at 2000 mg/kg resulted in severe toxicity: 2 out of 3 females had to be euthanized for humane reasons on Day 2. Clinical signs included lethargy, flat posture, hunched posture, slow breathing, shallow respiration, piloerection, watery discharge from the eyes and/or ptosis were noted for the animals on Days 1 and/or 2. At necropsy, dark red contents of the urinary bladder were found in the two animals sacrificed preterm. The surviving animal showed reduced body weight gain between Days 8 and 15. At 300 mg/kg, lethargy, hunched posture, uncoordinated movements, piloerection and/or ptosis were noted on Days 1 and/or 2. There were no treatmentrelated effects on the other parameters measured (mortality, body weight/body weight gain, macroscopic examination at necropsy). Based on these data, the oral LD50 value of JNJ-119717-AAA (T001036) in Wistar rats was established to be within the range of 300-2000 mg/kg body weight. In a subsequent 28-day repeated toxicity (gavage) study in the rat (RCC B65125; data on file at Sponsor) dose levels of 30, 100 and 300 mg/kg/day were tested. In the absence of any adverse effect, the NOAEL was initially established at 300 mg/kg/day by the Study Director, but subsequently put on 100 mg/kg/day in an Expert Report. Reason for this was that some rats at 300 mg/kg/day were noted with sedation and ataxia during the first week of treatment. Based on these data, dose levels of 15, 50 and 150 mg/kg/day for the current reproductive toxicity screening study were selected in consultation with the Sponsor. However, due to an error in the correction factor for purity/composition of the test item the actual doses given to the animals were by a factor of 1.07 lower, i.e. 14, 47 and 140 mg eq/kg/day.
- Rationale for animal assignment (if not random): randomized
Positive control:
no

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least twice daily (early in the morning and close to the end of the working day).

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily from treatment onwards up to the day prior to necropsy, detailed clinical observations were made in all animals immediately after dosing.

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of treatment and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on PND 1, 4, 7 and 13. Body weight and calculated body weight gain were reported.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Weekly during treatment, except for males and females which were housed together for mating and for females without evidence of mating. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on PND 1, 4, 7 and 13. Both absolute food consumption and food consumption relative to body weight were reported.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not applicable

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Time schedule for examinations: Subjective appraisal was maintained during the study, but no quantitative investigation was introduced as no treatment related effect was suspected.
Oestrous cyclicity (parental animals):
Daily vaginal lavage was performed to determine the stage of estrous beginning 14 days prior to treatment (pretest), the first 14 days of treatment and during mating until evidence of copulation was observed. Vaginal lavage continued for those females with no evidence of copulation until termination of the mating period. During pretest, this was done for 48 females. At the end of the pretest phase, 40 females with at least two regular estrous cycles were selected at random2 and further used in the study. The remaining females were removed from the study, and estrous cycle results were not reported but kept in the raw data. On the day of necropsy, a vaginal lavage was taken to determine the stage of estrous.
Sperm parameters (parental animals):
Parameters examined in F0 male parental generations: additional slides of the testes (to examine staging of spermatogenesis), testis weight
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no, on PND 1, all pups were individually identified by means of subcutaneous injection of Indian ink. When general hair growth blurs the identification, the pups were identified by tattoo on the feet.
- Maximum of 8 pups/litter (4/sex/litter as nearly as possible); Blood samples were collected from two of the surplus pups; excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
- Mortality / Viability: The numbers of live and dead pups were determined on PND 1 and daily thereafter. If possible, defects or cause of death were evaluated.
- Clinical signs: At least once daily, detailed clinical observations were made for all animals.
- Body weights: Live pups were weighed on PND 1, 4, 7 and 13.
- Sex: Sex was determined for all pups on PND 1 and 4.
- Anogenital distance: Anogenital distance (AGD) was measured for all live pups on PND 1. The AGD was normalized to the cube root of body weight.
- Areola/nipple retention: On PND 13, all males in each litter were examined for the number of areola/nipples.

GROSS EXAMINATION OF DEAD PUPS:
Yes, for external and internal abnormalities; If possible, defects or cause of death were evaluated.

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: no

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: no
Postmortem examinations (parental animals):
SACRIFICE
All animals surviving to the end of the observation period were deeply anaesthetized using isoflurane and subsequently exsanguinated. Necropsy was conducted on the following days:
- Male animals: Following completion of the mating period (a minimum of 28 days of dose administration).
- Female animals: On PND 14-16 (females which delivered); On Post-coitum Days 27-29 (female nos. 49 and 50 with evidence of mating) or 26 days after the last day of the mating period (female no. 53 without evidence of mating).

GROSS NECROPSY
- After sacrifice, all animals were subjected to a limited necropsy, with special attention being paid to the reproductive organs. Descriptions of all macroscopic abnormalities were recorded.
- The number of implantation sites were recorded for all paired females.
- Samples of the following tissues and organs were collected and fixed in 10% buffered formalin (neutral phosphate buffered 4% formaldehyde solution): Cervix (F), Clitoral gland (F), Coagulation gland (M), (Cowper’s glands) (M), Epididymides (M), Mammary gland area (M/F), (Glans penis) (M), (Levator ani plus bulbocavernosus muscle complex (LABC)) (M), Ovaries (F), Preputial gland (M), Pituitary gland (M/F), Prostate gland (M), Seminal vesicles (M), Testes (M), Thyroid including parathyroid if detectable (M/F), Uterus (F), Vagina (F), All gross lesions (M/F), Identification marks: not processed (M/F)
- Tissues/organs mentioned in parentheses were not examined by the pathologist, since no signs of toxicity were noted at macroscopic examination.

ORGAN WEIGHTS
- The following organ weights and terminal body weight were recorded from the surviving males on the scheduled day of necropsy: Cowper’s glands, Epididymides, Glans penis, Levator ani plus bulbocavernosus muscle complex (LABC), Testes, Thyroid
- Also for all F0-females thyroid organ weight and terminal body weight were recorded on the scheduled day of necropsy.
- Absolute organ weights and organ to body weight ratios were reported.

HISTOPATHOLOGY
- All organ and tissue samples, as defined under Histopathology, were processed, embedded and cut at a thickness of 2-4 micrometers. These slides were stained with haematoxylin and eosin. The additional slides of the testes (to examine staging of spermatogenesis) were stained with PAS/haematoxylin.
- The following slides were examined by a pathologist:
1) The ovaries, testes and epididymides of the animals of Groups 1 and 4.
2) The additional slides of the testes of the males of Groups 1 and 4, and all males suspected to be infertile, to examine staging of spermatogenesis.
3) All gross lesions of all animals (all dose groups).
4) Thyroid gland of Groups 1 and 4 males and females of Groups 1 and 4.
5) Thyroid gland of Groups 2 and 3 males and females, based on (possible) treatment-related changes in this organ in Group 4.
6) The reproductive organs of all males that failed to sire and all females that failed to deliver healthy pups (all because not pregnant): Female/Male nos. 49/09 and 50/10 (group 1) and nos. 53/13 (group 2)
- All abnormalities were described and included in the report. An attempt was made to correlate gross observations with microscopic findings.
- A peer review on the histopathology data was performed by a second pathologist.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 4 days of age (culling) or at 13-15 days of age (terminal sacrifice).
At culling on PND 4, from 2 pups per litter blood samples (0.4 mL in total) were collected by decapitation between 7.00 and 10.30 a.m
At terminal sacrifice on PND13-15, from 2 pups per litter (if possible from one male and one female) blood samples were collected at planned necropsy. As part of the necropsy procedure, blood samples (0.9 mL) were drawn by aorta puncture under anaesthesia using isoflurane, between 7.00 and 10.30 a.m..
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- All pups were sexed both externally and internally, and descriptions of all external abnormalities were recorded. Particular attention was paid to the external reproductive genitals to examine signs of altered development.
- At terminal sacrifice (PND 13-15), the thyroid from 2 pups per litter, i.e. the same pups as selected for blood sampling (see also sections 7.10 and 7.12.1), was collected and preserved in 10% buffered formalin.
- The stomach of pups not surviving to the scheduled necropsy date was examined for the presence of milk, if possible. If possible, defects or cause of death were evaluated.

HISTOPATHOLOGY / ORGAN WEIGTHS
not examined
Statistics:
The following statistical methods were used to analyze the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-to-one rank test) was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test was applied to frequency data.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.
Reproductive indices:
For each group, the following calculations were performed:
- Mating index (%) = (Number of females mated/Number of females paired) x 100
- Fertility index (%) = (Number of pregnant females/ Number of females paired) x 100
- Gestation index (%) = (Number of females bearing live pups/Number of pregnant females) x 100
- Duration of gestation = Number of days between confirmation of mating and the beginning of parturition
Offspring viability indices:
Survival indices:
- Post-implantation survival index (%) = (Total number of offspring born/ Total number of uterine impl antation sites) x 100
- Post-implantation survival index was expressed as 100% when the number of offspring exceeded the number of implantation sites recorded.
- Live birth index (%) = (Number of live offspring on Day 1 after littering/Total number of offspring born) x 100
- Viability index (%) = (Number of live offspring on Day 4 before culling/Number live offspring on Day 1 after littering) x 100
- Lactation index (%) = Number of live offspring on Day 13 after littering/Number live offspring on Day 4 (after culling)) x 100
- Group mean values were calculated from individual litter values
- Sex ratio (percentage males) = (Number of males in litter/Total number of offspring in litter) x 100

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Slight salivation occurred after dosing in both sexes at 50 and 150 mg/kg/day. The incidence increased dose-dependently. This salivation was considered to be a physiological response rather than a sign of systemic toxicity considering the nature and minor severity of the effect and its time of occurrence (i.e. after dosing). This sign may be related to taste of the test item.
Incidental findings that were noted among control and/or treated animals included alopecia, scales, scabbing or a wound on different parts of the body and rales. These findings occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study. At the incidence observed, these were considered signs of no toxicological relevance.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
There were no premature decedents in this study.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weights and body weight gain of treated animals remained in the same range as controls over the treatment period.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No treatment-related changes in food consumption before or after allowance for body weight were noted.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
The serum concentration of the thyroid hormone T4 (total T4), measured in males at scheduled sacrifice, was not affected by treatment up to 150 mg/kg/day.
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
Organ weights and organ to body weight ratios were not affected by treatment.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Macroscopic observations at necropsy did not reveal any alterations that were considered to have arisen as a result of treatment with the test item.
Incidental findings among control and treated animals were within the background range of findings that are encountered among rats of this age and strain, and did not show a dose-related trend. These necropsy findings were therefore considered to be unrelated to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Test item-related thyroid gland findings consisted of a slightly increased incidence and/or severity of follicular cell hypertrophy in males and females treated at 150 mg/kg/day. The incidences and severities recorded for the remaining dose groups including controls were within background pathology of rats of this age and strain.
The remainder of the recorded microscopic findings were within the range of background pathology encountered in rats of this age and strain. There was no test item-related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.
There were two couples of the control group (male 9/female 49, male 10/female 50) and one couple at 15 mg/kg/day (male 13/female 53) which had no offspring. No abnormalities were seen in the reproductive organs, which could account for their lack of offspring. Based on the fact that all couples at 50 and 150 mg/kg/day had offspring, the infertile couple at 15 mg/kg/day was considered to be unrelated to the treatment. The minor increase was regarded to be an adaptive change and considered non-adverse.
There were no morphological findings in the reproductive organs of either sex which could be attributed to the test item and spermatogenic staging profiles were normal for all males examined.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
Length and normality of the estrous cycle were not affected by treatment up to 150 mg/kg/day.
Most females had regular cycles of 4 days. Extended di-estrus occurred in one control female and two females at 15 mg/kg/day. One of the two low dose females (no. 53) could not be mated. As no extended di-estrus was noted at the higher doses of 50 and 150 mg/kg/day, it was considered to be a chance finding, rather than treatment-related. Irregular cycles occurred in two females at 150 mg/kg/day. As both females had normal litters, it was considered a sign of no toxicological relevance.
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
spermatogenic staging profiles were normal for all males examined.
Reproductive performance:
no effects observed
Description (incidence and severity):
REPRODUCTION DATA
- No treatment-related changes in reproductive parameters were noted up to 150 mg/kg/day.
- Except for one female (no. 53) at 15 mg/kg/day, all females showed evidence of mating. Two control females (nos. 49 and 50) were not pregnant. Mating andfertility indices, precoital time, and number of implantation sites were not considered to have been affected by treatment.
- For females no. 44 (control), 51, 59 (15 mg/kg/day), 64, 66, 67 (50 mg/kg/day) and 72 (150 mg/kg/day), the number of pups born was slightly higher than the number of implantations. This was considered to be due to the normal resorption of these areas as these enumerations were performed on Days 14 or 16 of lactation.

DEVELOPMENTAL DATA
The numbers of females with living pups on Day of 1 of lactation were 8, 9, 10 and 10 at 0, 15, 50 and 150 mg/kg/day, respectively.
No toxicologically relevant effects on gestation index and duration, parturition, maternal care and early postnatal pup development (postnatal endpoints evaluated are given below) were observed up to 150 m/kg/day.
- Gestation: The gestation index and duration of gestation were unaffected by treatment up to 150 mg/kg/day.
- Parturition/maternal care: No signs of difficult or prolonged parturition were noted among the pregnant females. Examination of cage debris of pregnant females revealed no signs of abortion or premature birth. No deficiencies in maternal care were observed.
- Post-implantation survival index: Post-implantation survival index was unaffected by treatment. All values remained within the normal range of biological variation.
- Early postnatal pup development : At dose levels up to 150 mg/kg/day, no treatment-related changes were noted in the live birth, viability and lactation indices, sex ratio, anogenital distance, areola/nipple retention, clinical signs, body weights, external macroscopy and serum concentration of the thyroid hormone T4 (PND 13-15).

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Remarks:
parental
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Dose descriptor:
NOAEL
Remarks:
Reproduction
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance

Target system / organ toxicity (P0)

Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Description (incidence and severity):
Pups that went missing showed no clinical signs, except for the pup at 50 mg/kg/day that had no milk in the stomach at the first litter check. Incidental clinical signs in surviving pups consisted of dragging of the left hind leg (in a single control pup), blue spot(s) on neck or back, swelling in the neck, a black nodule on neck or shoulder due to excess of ink, lean appearance, alopecia, or scales on the neck. The nature and incidence of these clinical signs remained within the range considered normal for pups of this age, and were therefore considered to be of no toxicological relevance.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
At the first litter check, one pup at 50 mg/kg/day was found dead. During lactation, one pup of the control group, one pup at 15 mg/kg/day and one pup at 50 mg/kg/day were missing, most likely due to cannibalism. This incidental mortality was not related to treatment.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weights of pups showed no toxicologically relevant changes.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Description (incidence and severity):
The serum level of the thyroid hormone T4 in male and female pups was not affected by treatment up to 150 mg/kg/day.
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The pup at 50 mg/kg/day that was found dead at first litter check showed beginning autolysis. Macroscopic findings in surviving pups were limited to dragging of the left hind leg (in a single control pup), and a black nodule on neck or shoulder due to excess of ink in two pups at 15 mg/kg/day. These incidental findings were not related to treatment.
Histopathological findings:
not examined
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
SEX RATIO: Sex ratio was not affected by treatment up to 150 mg/kg/day.
At 15 mg/kg/day, sex ratio indicated statistically significantly more female than male pups (62% female pups versus 43% in the control group) at first litter check. This finding was considered not to be related to treatment because sex ratio was normal at the higher dose levels (i.e. 50% female pups at 50 and 150 mg/kg).
ANOGENITAL DISTANCE: Anogenital distance in male and female pups was not affected by treatment up to 150 mg/kg/day.
The statistically significantly higher median anogenital distance of male pups at 50 and 150 mg/kg/day occurred without a dose related-trend and was, therefore, not attributed to treatment.
AEROLA/NIPPLE RETENTION: Treatment up to 150 mg/kg/day had no effect on areola/nipple retention. For none of the examined male pups nipples were observed on PND 13.

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Effect levels (F1)

Dose descriptor:
NOAEL
Remarks:
Developmental
Generation:
F1
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
clinical biochemistry
gross pathology
other: Sex ratio; Anogenital distance; Aerola/nipple retention

Target system / organ toxicity (F1)

Critical effects observed:
no

Overall reproductive toxicity

Key result
Reproductive effects observed:
no

Applicant's summary and conclusion

Conclusions:
In conclusion, treatment with JNJ-119717-AAA (T001036) by oral gavage in male and female Wistar Han rats at dose levels of 15, 50 and 150 mg/kg/day revealed no parental, reproduction and developmental toxicity up to 150 mg/kg/day.
Based on these results, a parental, reproduction and developmental No Observed Adverse Effect Level (NOAEL) of 150 mg/kg/day were derived. This corresponds to an actual dose level of 140 mg eq/kg/day. Therefore, the substance is not classified as a reproductive toxictant according to the CLP regulation.