N-{2-[(phenylcarbamoyl)amino]phenyl}benzenesulfonamide

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

There was no evidence of adverse effects on parental reproductive indices following repeated oral exposure of the test substance, with the NOAEL set as 1000 mg/kg/day in parental animals under the conditions of the test.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Specific details on test material used for the study:

Lot no.: 140220

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc. (Hino Breeding Centre).
- Age at study initiation: (P) 9 wks
- Weight at study initiation: (P) Males: 320.9-367.7 g; Females: 214.9-240.8 g
- Housing: Males and females (except during the gestation and lactation periods); stainless steel cages. For females during the gestation and lactation periods; polymethylpentane cages.
- Diet (e.g. ad libitum): Ad libitum (except during the urine collection and during measurement of motor activity) autoclaved sterilized pellet diet (CRF-1, oriental yeast co. Ltd.). Animals were fasted on the day before scheduled necropsy.
- Water (e.g. ad libitum): Ad libitum (except during the measurement of motor activity) well water.
- Acclimation period: 6 Days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 to 24.3
- Humidity (%): 48.6 to 69.8
- Air changes (per hr): 10 to 20
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark

Route of administration:

oral: gavage

Vehicle:

other: 0.5% w/v% methyl cellulose aqueous solution

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The dosing preparations were prepared once every 4-8 days (based on the results of previous stability and homogeniety assessments). The test substance was initially ground in an agate pestle and mortar, and a few drops of vehicle added. This suspension was then transferred into a measuring cylinder. The appropriate volumes of the vehicle was added to the measuring cyclinder and mixed to achieve the required concentrations which were then added into brown glass vials and stored refrigerated until use.

VEHICLE
- Concentration in vehicle: 0, 11, 33 and 100 mg/mL
- Amount of vehicle (if gavage): Dose volume of 10 mL/kg/day
- Lot/batch no. (if required): PDG1792

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: Until evidence of mating was apparent (upto 6 days).
- Proof of pregnancy: vaginal plug and/or sperm in vaginal smear referred to as day 0 of gestation.
- After successful mating each pregnant female was caged individually.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

At the initial test substance preparation, 15 mL of each of the analytical samples were tested for homogeneity and analytical determination. The measured concentrations were within acceptable ranges (actual values 98.4 to 100.3% of nominal).

Duration of treatment / exposure:

Males: From 14 days before mating (Days 1 to 15) until the day before the necropsy through the mating period (42 days in total).
Females: From 14 days before mating (Days 1 to 15) until Day 4 of lactation (day of delivery was Day 0 of lactation) through the mating and pregnancy periods and delivery. One female not which did not deliver was kept until day before the necropsy.

Frequency of treatment:

Parental males and females daily by oral gavage.

Details on study schedule:

- Age at mating of the mated animals in the study: approximately 11 weeks

Remarks:

Doses / Concentrations:
0, 110, 330 and 1000 mg/kg/day.
Basis:
nominal conc.

No. of animals per sex per dose:

- A total of 106 rats were used on study (48 males and 58 females).
- Each test group consisted of 12 males (including 5 males for the control and 1000 mg/kg groups in the recovery test) and 12 females (5 females
each were added for the control and 1000 mg/kg groups in the recovery test).

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: A preceeding 14 Day repeated dose oral study with the test substance to rats, indicated that at a high dose of 1000 mg/kg/day would be expected to provide obvious toxicological changes.

Parental animals: Observations and examinations:

See Chapter 7.5.1, record Repeated dose toxicity: oral.001

Oestrous cyclicity (parental animals):

Vaginal smears were collected with swabs from all test females in the morning (same time every day) from the initial dosing day to the day of mating or end of the mating period to confirm the estrous cycle. The obtained smears were collected on a plate for each animal, and stained with Giemsa.The estrous cycle was classified into diestrus (D), proestrus (P), estrus (E), and metestrus (M). The mean estrous cycle (number of
days from the estrous period to the next estrous period) and the number of the estrous period during the test period were calculated.

Sperm parameters (parental animals):

Not examined

Litter observations:

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

GROSS EXAMINATION OF DEAD PUPS:
Yes, for external abnormalities.

Postmortem examinations (parental animals):

Necropsy and histopathology procedures are described in Chapter 7.5.1, record Repeated dose toxicity: oral.001

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring were sacrificed on Day 4 of lactation.
- These animals were subjected to postmortem macroscopic examination (external anomalies).

GROSS NECROPSY
- Gross necropsy consisted of a check for external anomalies, including that in the oral cavity.

Statistics:

Histopathological findings were analysed using Fisher's exact probability test. Copulation index, fertility index, gestation index, delivery index and sex ratio were analysed using the Chi-squared test. Implantation index, stillborn rate, external anomaly index, external anomaly typing index, live birth index, and viability index were analysed using Wilcoxon's rank sum test.

Number of rearing, grip strength, motor activity, body weights, food consumption, urinalysis (reagent strip), hematology, blood chemistry, absolute and relative organ weight, estrous cycle, number of estrus, days until copulation, gestation length, number of corpora lutea, number of implantations, number of delivered offspring, number of live offspring, body weight of offspring (both sexes) were initially analysed using Bartlett's test. When the data was homogenous the Dunnet's multiple comparison test was applied, and when not homogenous, Steel's multiple comparison test was applied for the control group and each test group. For the urinalysis reagent strip, Steel's test was performed.

Reproductive indices:

Compulation index (%), fertility index (%), gestation length, implantation index (%), gestation index (%) and delivery index (%) were the determined reproductive indices.

Offspring viability indices:

Live birth index (%), stillborn rate (%), Viability index on Day 4 (%), Sex ration (%), external anomaly index (%), and external anomaly typing index (%), were the determined offspring viability indices.

Clinical signs:

not examined

Description (incidence and severity):

Examined in section 7.5.1

Body weight and weight changes:

not examined

Description (incidence and severity):

Bodyweight examined in section 7.5.1

Food consumption and compound intake (if feeding study):

not examined

Description (incidence and severity):

Bodyweight examined in section 7.5.1

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

not examined

Description (incidence and severity):

Examined in section 7.5.1

Other effects:

not examined

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Reproductive performance
There were no treatment-related differences in:
- Estrus cycling
- precoital time
- the number of pregnant females
- duration of gestation
- mean numbers of corpora lutae
- number of total, pre-implantation and post-implantation sites

Selected results by dose level (0, 110, 330 or 1000 mg/kg bw/day).
- females mated: 12/12, 12/12, 12/12, 12/12
- females pregnant: 12/12, 12/12, 12/12, 11/12
- females with live pups: 12/12, 12/12, 12/12, 11/12
- Copulation indices (%): 100, 100, 100, 100
- fertility Indices (%): 100, 100, 100, 91.7

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: No adverse effects at highest dose tested

Clinical signs:

not examined

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:

no effects observed

Histopathological findings:

not examined

VIABILITY (OFFSPRING)
No statistically significant difference was noted in number of live offspring at birth, number of stillborn at birth, sex ratio, stillborn rate, or viability indexon Day 4 between the control group and any treated group. In addition, a high value of the birth index was observed in the 1000 mg/kg group; however, this change was judged to be not related to the test substance treatment because the post-implantation losses were a few.

BODY WEIGHT (OFFSPRING)
No statistically significant difference was observed between the control group and any treated group in body weight on days 0 and 4 of postpartum.

GROSS PATHOLOGY (OFFSPRING)
No external anomalies were observed in any offspring.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: No adverse effects at highest dose tested

Key result

Reproductive effects observed:

not specified

Conclusions:

There was no evidence of adverse effects on parental reproductive indices and litter data following repeated oral exposure of the test substance, with the NOAEL set as 1000 mg/kg/day in parental animals and offspring under the conditions of the test.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Effects on developmental toxicity

Description of key information

There was no evidence of adverse effects on litter data following repeated oral exposure of the test substance, with the NOAEL set as 1000 mg/kg/day in offspring under the conditions of the test.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

A reliable combined repeated dose and reproductive screening study was conducted which indicated that there was no evidence of adverse effects on parental reproductive indices and litter data following repeated oral exposure of the test substance, with the NOAEL set as 1000 mg/kg/day in parental animals and offspring under the conditions of the test. The test material was not classified for reproductive effects based on the available information under the CLP Regulation (EC) No 1272/2008, as amended for the twelfth time in Regulation (EC) No 2019/521.