Registration Dossier

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 December 2013 to 28 January 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with OECD GLP (1997) principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
2 group 2 females and 1 group 4 female received 20-30% higher volume dose Days 1-3. 5 group 1 females and 1 group 4 female no body weight was determined on Day 1 of lactation and no food intake recorded for Days 1-4. No food wght recorded on Day 22.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
- Name of test material (as cited in study report): MTDID 9580
- Molecular formula (if other than submission substance):
- Molecular weight (if other than submission substance):
- Smiles notation (if other than submission substance):
- InChl (if other than submission substance):
- Structural formula attached as image file (if other than submission substance): see Fig.
- Substance type: Non-mixture
- Physical state: Liquid
- Analytical purity: 97.8%
- Impurities (identity and concentrations):
- Composition of test material, percentage of components:
- Isomers composition:
- Purity test date: 6 October 2013
- Lot/batch No.: 113F1114 (E27136)
- Expiration date of the lot/batch:
- Radiochemical purity (if radiolabelling):
- Specific activity (if radiolabelling):
- Locations of the label (if radiolabelling):
- Expiration date of radiochemical substance (if radiolabelling):
- Stability under test conditions:
- Storage condition of test material: At room temperature, in the dark, under nitrogen
- Other:

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: (P) 13 wks
- Weight at study initiation: (P) Male Means by Group: 339-345 g; Female Means by Group: 225-230 g; (F1) Male Means by Group: 6.0-6.4 g; Female Mean by Group: 5.6-6.1 g
- Fasting period before study:
- Housing:
- Diet (e.g. ad libitum): Pelleted rodent diet (SM R/M-Z from SSNIFF Spezialdiaten GmbH, Soest, Germany) ad libitum
- Water (e.g. ad libitum): Tap Water ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24
- Humidity (%): 40-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 10 December 2013 To: 28 January 2014

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Remarks:
Polyethylene glycol 400
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water):Based on trial formulations performed at WIL Research Europe.
- Concentration in vehicle: Doses administered were 150, 450, and 1000 mg/kg/day.
- Amount of vehicle (if gavage): The dose volume was 5 mL/kg body weight
- Lot/batch no. (if required): No data
- Purity: No Data
Details on mating procedure:
- M/F ratio per cage: 1/1 (From the same treatment group)
- Length of cohabitation: 13 days maximum
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): Individually housed in Macrolon plastic cages (MIII type, height 18 cm).
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses were conducted according to WIL Research protocol Project 503544.
Duration of treatment / exposure:
Males were exposed for 28 days (2 weeks prior to mating, during mating and up to the day prior to schedules necropsy). Females were exposed for 40-44 days (2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of lactation up to the day prior to scheduled necropsy)
Frequency of treatment:
Once daily, 7 days per week.
Details on study schedule:
- Age at mating of the mated animals in the study: 15 weeks
Doses / concentrations
Remarks:
Doses / Concentrations:
150, 450, and 1000 mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
10 animals/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Doses were chosedn based on the results of a 7-day dose range finding study.
- Rationale for animal assignment (if not random): Random
Positive control:
Not applicable

Examinations

Parental animals: Observations and examinations:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily from beginning of treatement to the day prior to necropsy. The time of onset, grade and duration of any observed sign was recorded.
- Male number paired with, mating date, confirmation of pregnancy, and delivery date were recorded. Pregnant females were examined to detect signs of difficult or prolonged parturition, and cage debris of pregnant females was examined to detect signs of abortion or premature birth. Any deficiencies in maternal care were examined.

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17, and 20 post coitum and during lactation on Days 1 and 4.
Oestrous cyclicity (parental animals):
No Data
Sperm parameters (parental animals):
No Data
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: No

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: The number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, and physical or behavioural abnormalities.

GROSS EXAMINATION OF DEAD PUPS: Yes, for external abnormalities; possible cause of death was determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals were sacrificed following the completion of the mating period (a minimum of 28 days of dose administration).
- Maternal animals: All surviving females which delivered pups were sacrificed on Lactation Days 5-7. Females which failed to deliver were sacrificed on Post-coitum Day 27 (a female with evidence of mating) and Post-coitum Day 21 (a female without evidence of mating).

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were collected: cervix, clitoral gland, coagulation gland, epididymides, female mammary gland area, ovaries, preputial gland, prostate gland, seminal vesicles, testes, uterus, vagina, and any gross lesions.
-Slides of the following tissues were prepared and examined by a pathologist: The ovaries, testes and epididymides of the animals in the control and 1000 mg/kg/day group. Additional slides of the testes of the males in the control and 1000 mg/kg/day group that were suspected to be infertile were created. Slides of the reproductive organs of all males that failed to sire and all females that failed to deliver healthy pups were created.
Postmortem examinations (offspring):
SACRIFICE
- All F1 offspring were sexed and descriptions of all external abnormalities were recorded. The stomach of pups not surviving to the scheduled necropsy date were examined for the presence of milk. Defects or cause of death were evaluated.

GROSS NECROPSY
-F1 offspring were not subjected to gross necropsy.

HISTOPATHOLOGY / ORGAN WEIGTHS
-Histopathology and organ weights were not recorded for F1 offspring.
Reproductive indices:
For each group, the following calculations were performed: mating index, fertility index, conception index, gestation index, duration of gestation, percentage of live males at first litter check, and percentage of live females at first litter check.
Offspring viability indices:
Percentage of postnatal loss, and viability index were calculated.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS): No mortality occured during the study period. No toxicologically relevant clinical signs were noted.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS): No toxicologically relevant changes in body weights, body weight gain, or food consumption were noted.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS): No toxicologically relevant effects on reproductive parameters were noted.

ORGAN WEIGHTS (PARENTAL ANIMALS): Testes and epididymides weights and terminal body weights of treated males were similar to those of control animals.

GROSS PATHOLOGY (PARENTAL ANIMALS): Necropsy did not reveal any toxicologically relevant alterations.

HISTOPATHOLOGY (PARENTAL ANIMALS): All histopathological changes were considered to be incidental findings. There was no test article-related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Target system / organ toxicity (P0)

Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined

Details on results (F1)

VIABILITY (OFFSPRING): No toxicologically relevant changes in pup viability were observed.

CLINICAL SIGNS (OFFSPRING): To toxicologically relevant clinical signs were noted.

BODY WEIGHT (OFFSPRING): Body weights of pups were considered to have been unaffected by treatment.

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
ca. 1 000 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
gross pathology
other: See 'Remarks'

Target system / organ toxicity (F1)

Critical effects observed:
no

Overall reproductive toxicity

Key result
Reproductive effects observed:
no

Applicant's summary and conclusion

Conclusions:
Based on the results of the study, the parental, reproduction and developmental No Observed Adverse Effect Level (NOAEL) was 1000 mg/kg/day.
Executive summary:

The reproductive and developmental toxicity potential of the test article was evaluated in Wistar Han rats. The study was conducted in compliance with OECD GLP (1997) regulations. The test method was based on OECD 421 (1995) and OPTS 870.3550 Reproduction/Developmental Toxicity Screening Test (2000). The test article in polyethylene glycol 400 was administered via oral gavage at 0 (vehicle), 150, 450, or 1000 mg/kg/day to 10 animals/sex/group at a dose volume of 5 mL/kg body weight. Males were exposed for 2 weeks prior to mating, during mating, and up to termination (for 28 days). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and at least 4 days of lactation (for 40-44 days). For the parental animals, mortality/viability (at least twice daily), clinical signs (daily), body weights (Males: Day 1 of exposure, and weekly thereafter. Females: Days 0, 4, 7, 11, 14, 17, and 20 post coitum and lactation Days 1 and 4) food consumption (weekly), and general reproduction data (at post-coitum day 0 and birthing for mothers) were noted. For pups, mortality/viability (daily), clinical signs (at least once daily), body weights (Day 1 and 4 of lactation), sex determination (Day 1 and 4 of lactation), and external examination (at sacrifice) were noted. Parental animals were subject to gross necropsy and the ovaries, testes, epididymides of animals in the control and 1000 mg/kg/day groups were collected and examined by a histopathologist. In parental animals, no mortality occurred during the study period. No toxicologically relevant clinical signs, changes in body weights or body weight gains, or food consumption were noted during the study. Upon macroscopic examination, no toxicologically relevant alterations were observed. The testes, epididymides weights and terminal body weights of treated males were similar to those of control animals. Upon microscopic examination of the selected tissues, histologic changes were considered to be incidental findings. There was no test article-related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations. No toxicologically relevant effects on reproductive parameters were noted. Mating, fertility and conception indices, precoital time, and number of corpea lutea and implantation sites were unaffected by treatment. Two females at 150 mg/kg/day failed to deliver healthy offspring (these did not mate and/or did not become pregnant). Since the incidence of infertile animals showed not dose-response relationship, the findings were considered incidental. No toxicologically relevant effects on gestation index and duration, parturition, maternal care and early postnatal pup development (mortality, clinical signs, body weight and macroscopy) were observed. Based on the results of the study, the parental, reproduction and developmental No Observed Adverse Effect Level (NOAEL) was 1000 mg/kg/day.