4-[[5-(anilino)carbonyl-2-methoxyphenyl]azo]-3-hydroxy-N-(3-nitrophenyl)naphthalene-2-carboxamide

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, NOAEL for test material was considered to be 1000 mg/kg bw/day for reproductive toxicity, when male and female rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data from various test chemicals

Justification for type of information:

Weight of evidence approach based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on reproductive toxicity studies on rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data avaiable

Species:

rat

Strain:

Crj: CD(SD)

Details on species / strain selection:

No data avaiable

Sex:

male/female

Details on test animals or test system and environmental conditions:

1.TEST ANIMALS
- Age at study initiation:9 weeks old

2.TEST ANIMALS
- Source: Charles River Japan Co., Ltd
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Age at study initiation: 9 weeks
- Weight at study initiation: male : 309 to 355 g female: 206 to 232 g
- Fasting period before study:
- Housing: a cage made of stainless steel hanging type wire mesh is excluded for the period excluding pregnancy / nursing period, and made of polycarbonate with a floor covering for laboratory animals (beta chip, Nippon Charles River Co., Ltd.)
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum): tap water irradiated with ultraviolet rays after autoclave sterilized solid feed for laboratory animals (CRF-1, Oriental Yeast Industry Co., Ltd.) and filtering with a pore size of 5 μm.
- Acclimation period:6 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 ° C.,
- Humidity (%):55 ± 15%
- Air changes (per hr): ventilation at about 12 times / hour,
- Photoperiod (hrs dark / hrs light): lighting 12 hours / day (7: 00-19: 00)
IN-LIFE DATES: From: To:

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: 1 w/v% Tween 80 solution 2.0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80

Details on exposure:

1.PREPARATION OF DOSING SOLUTIONS: Test material dissolved in 1 w/v% Tween 80 solution

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0, 40, 200, 1000 mg/kg
- Amount of vehicle (if gavage):
- Lot/batch no. (if required):

2.PREPARATION OF DOSING SOLUTIONS: test material dissolved in 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80
DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water): 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80
- Concentration in vehicle: 0,100, 300, 1000 mg/kg
- Amount of vehicle (if gavage): 10ml/kg
- Lot/batch no. (if required):
- Purity:
- Purity:

Details on mating procedure:

2.- M/F ratio per cage:1:1
- Length of cohabitation: together day and night on the 14th day
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: vaginal plug formation or sperm was observed in vaginal plaque specimens was taken as copulation formation, and that day was taken as the 0th day of pregnancy
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- After successful mating each pregnant female was caged (how):
- Any other deviations from standard protocol:

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data avaiable

Duration of treatment / exposure:

1.Male: 42 days
Female: 42 – 48 days (from 14 days before mating to day 4 of lactation)
2.Males, 37 days
Females, from 14 days before mating to day 4 of lactation

Frequency of treatment:

Daily

Details on study schedule:

Recovery period: Males, 14 days
Females (satellite), 14 days

Sacrifice : Males, day 43 of treatment and day 15 of recovery
Females, day 5 of lactation
Females (satellite), day 15 of recovery

Remarks:

1.Doses / Concentrations:
0, 40, 200, 1000 mg/kg
2.0 (vehicle), 100, 300, 1000 mg/kg

No. of animals per sex per dose:

1.
No of animals Total: 116
0 mg/kg bw/day:12 male , 12 female
40 mg/kg bw/day:12 male , 12 female
200 mg/kg bw/day:12 male , 12 female
1000 mg/kg bw/day:12 male , 12 female

Recovery:
0 mg/kg bw/day: 5 male , 5 female
1000 mg/kg bw/day: 5 male , 5 female
2.Total: 96
0 mg/kg bw/day: 12 male, 12 female
100 mg/kg bw/day: 12 male, 12 female
300 mg/kg bw/day: 12 male, 12 female
1000 mg/kg bw/day: 12 male, 12 female

Control animals:

yes, concurrent vehicle

Details on study design:

No data avaialble

Positive control:

No data avaialble

Parental animals: Observations and examinations:

Clinical signs, FOB, Body weight, Food consumption, Urinalysis, Hematology and clinical chemistry were observed.

Oestrous cyclicity (parental animals):

Estrous cycle, numbers of corpora lutea or Implantations were observed

Sperm parameters (parental animals):

No data avaialble

Litter observations:

Viability, Clinical signs and Body weight were observed

Postmortem examinations (parental animals):

Gross pathology and Histopathology were observed

Postmortem examinations (offspring):

Gross pathology and Histopathology were observed

Statistics:

Regarding the weighing data, parametric data was tested for equal variance by the Bartlett method, and one-way ANOVA was performed when variance was equal. Where the variances are not equal and nonparametric data were tested by Kruskal-Wallis. When significant difference was observed between groups, multiple comparisons of Dunnett method or Dunnett type were performed. Pathological findings in counting data were examined by χ 2 for a × b . When significant difference was observed , comparison was made between control group and each test substance administered group by Armitage χ 2 test. Other counting data were tested by Fisher's direct stochastic method. The significance level of each test was 5%. For the data on newborn babies, the average value calculated for each mother was set as a sample unit

Reproductive indices:

Copulation index, fertility index, delivery index, gestation length, implantation index and gestation index were observed

Offspring viability indices:

Viability on day 4 were observed.

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

No effects on mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behavior of treated rats were observed.

Clinical signs: No clinical signs of toxicity was observed in treated rats.

Body weight: No effect on body weight of treated rats was observed as compared to control.

Reproductive function: estrous cycle: No effects estrous cycle, numbers of corpora lutea or Implantations were observed in treated rats as compared to control.

Reproductive performance: No effects on copulation index, fertility index, delivery index, gestation length, implantation index and gestation index were observed in treated rats as compared to control.

Organ weights : No effect on organ weight of treated rats were in treated rats as compared to control.

Gross pathology: No gross pathological changes were observed in treatd rats as compared to control.

Histopathology: No histopathological changes were observed in treatd rats as compared to control.

other findings No effect on food consumption, hematology, blood chemistry, urinalysis of treated rats were observed as compared to contorl.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect on Clinical signs, FOB, Body weight, Food consumption, Urinalysis, Hematology, Blood chemistry, organ weights, reproductive performance and Histopathology

Remarks on result:

other: overall no effects on reproductive parameters

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

no effects observed

Description (incidence and severity):

1.No clinical signs of toxicity were observed in offspring as compared to cotnrol.

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

2.No significant differences in numbers of offspring or live offspring were observed as compared to cotnrol.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

2.No effect on body weight of offspring were observed as compared to cotnrol.

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Description (incidence and severity):

2. No gorss pathological changes were observed in offspring as compared to cotnrol.

Histopathological findings:

no effects observed

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Viability: No effect on viability on day 4 were observed in offsprings to treated rats.

Clinical signs: No clinical signs of toxicity was observed in treated rats

Body weight: No effect on body weight of treated rats was observed in offsprings to treated rats as compared to control.

Gross pathology: No gross pathological changes were observed in offsprings of treated rats as compared to control.

Histopathology: No histopathological changes were observed in offsprings of treated rats as compared to control.

other findings: No effect on number of offspring or live offspring at birth, sex ratio and live birth index of offsprings of treated rats as compared to control.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect on number of offspring or live offspring at birth, sex ratio, live birth index, or viability index , gross pathology and Histopathology

Remarks on result:

other: overall no effects on developmental parameters

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Reproductive effects observed:

not specified

Treatment related:

not specified

Relation to other toxic effects:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Conclusions:

NOAEL was considered to be 1000 mg/kg bw/day for P and F1 generation when Crl:CD (SD) male and female rats were treated with test chemical orally by gavage for 42 days.

Executive summary:

Data available from different studies were reviewed to determine the reproductive toxicity of test chemical. The studies are as mentioned below:

Study 1

In combined repeated dose toxicity study with reproduction and developmental screening, Crj:CD(SD)IGS male and female rats were treated with test chemical in the concentration of 0, 40, 200 and 1000 mg/kg bw/day 1 w/v% Tween 80 solution orally by gavage for male 42 days and females from 14 days before mating to day 4 of lactation (Females (satellite), 42 days).All animals were observed for clinical sign , body weights, food consumption. Males were sacrificed on day 43 of treatment and day 15 of recovery while Females were sacrificed on day 5 of lactation Females (satellite), day 15 of recovery and Offspring on day 4 after birth.

No effect on clinical sign, body weights, food consumption, haematology, clinical chemistry, urinalysis, organ weights, gross pathology and histopathological of treated rats as compared to control. Similarly no effect on estrous cycle, copulation index, fertility index, delivery index, gestation length, numbers of corpora lutea or implantations, implantation index, gestation index, number of offspring or live offspring at birth, sex ratio, live birth index, or viability index on day 4 of offspring were observed as compared to control. In addition, no effect on clinical signs, body weights, gross pathology and histopathology of treated offspings were observed as compared to control. Therfore, NOAEL was considered to be 1000 mg/kg bw/day for P and F1 generation when  Crl:CD (SD) male and female rats were treated with test chemical orally by gavage for 42 days.

Study 2

In a combined repeated dose toxicity study with reproduction and developmental screening, Crj:CD(SD)IGS male and female rats were treated with test chemical in the concentration of 0, 100, 300, 1000 mg/kg bw/day in 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80 orally by gavage.The dosing period is from 42 to 47 days for a total of 37 days before mating by mating and 4 days before breeding and the day before necropsy, for females for 14 days before mating, a mating period, a gestation period and delivery until 4 nursing days did. The non-delivering animal was taken until the day before necropsy. At the time of administration, it was administered by gavage once a day using the teflon stomach probe, in the morning. The volume of the solution to be administered was 10 mL / kg, and it was calculated based on the body weight measured on the nearest day. Female vaginal plaque was collected daily in the morning from the start of administration to the start of the mating, the sexual cycle was examined, and the mean cycle duration was calculated. After the administration period before mating, pairs of males and females 1 were established within each group and allowed to live together day and night on the 14th day. Mating confirmation was done every morning, and cases where vaginal plug formation or sperm was observed in vaginal plaque specimens was taken as copulation formation, and that day was taken as the 0th day of pregnancy. Mating pairs were separated from male and female and submitted for further examination. Body weight was measured on both the male and female on the administration start date, 3, 7 and 14 days after the start of administration, once a week for females, females with mating completed on 0, 7, 14, 20 gestation days and 0 and 4 days of pregnancy It was measured

No effect on clinical sign, body weight, food consumption, haematology and clinical chemistry of treated rats as compared to control. Similarly, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour were observed in treated rats. Increase in liver weight was observed in male and female rats. No gross pathological and histopathological changes were observed in treated rats. In addition, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour and numbers of offspring or live offspring, sex ratio, live birth index, viability index or body weights of offspring were observed. No gross pathological changes were observed in offspring as compared to control. Therefore, NOAEL was considered to be 1000 mg/kg bw/day when Crj:CD(SD)IGS male and female rats were treated with test chemical orally by gavage.

Based on the data available from different studies, test material did not showed reproductive toxicity at dose concentration 1000 mg/kg body weight/day, when male and female rats were treated with test material orally, Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

 

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

Quality of whole database:

Data is Klimicsh 2 and from authoritative database

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive toxicity study

Data available from different studies were reviewed to determine the reproductive toxicity of test chemical. The studies are as mentioned below:

Study 1

In combined repeated dose toxicity study with reproduction and developmental screening, Crj:CD(SD)IGS male and female rats were treated with test chemical in the concentration of 0, 40, 200 and 1000 mg/kg bw/day 1 w/v% Tween 80 solution orally by gavage for male 42 days and females from 14 days before mating to day 4 of lactation (Females (satellite), 42 days).All animals were observed for clinical sign , body weights, food consumption. Males were sacrificed on day 43 of treatment and day 15 of recovery while Females were sacrificed on day 5 of lactation Females (satellite), day 15 of recovery and Offspring on day 4 after birth.

No effect on clinical sign, body weights, food consumption, haematology, clinical chemistry, urinalysis, organ weights, gross pathology and histopathological of treated rats as compared to control. Similarly no effect on estrous cycle, copulation index, fertility index, delivery index, gestation length, numbers of corpora lutea or implantations, implantation index, gestation index, number of offspring or live offspring at birth, sex ratio, live birth index, or viability index on day 4 of offspring were observed as compared to control. In addition, no effect on clinical signs, body weights, gross pathology and histopathology of treated offspings were observed as compared to control. Therfore, NOAEL was considered to be 1000 mg/kg bw/day for P and F1 generation when  Crl:CD (SD) male and female rats were treated with test chemical orally by gavage for 42 days.

Study 2

In a combined repeated dose toxicity study with reproduction and developmental screening, Crj:CD(SD)IGS male and female rats were treated with test chemical in the concentration of 0, 100, 300, 1000 mg/kg bw/day in 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80 orally by gavage.The dosing period is from 42 to 47 days for a total of 37 days before mating by mating and 4 days before breeding and the day before necropsy, for females for 14 days before mating, a mating period, a gestation period and delivery until 4 nursing days did. The non-delivering animal was taken until the day before necropsy. At the time of administration, it was administered by gavage once a day using the teflon stomach probe, in the morning. The volume of the solution to be administered was 10 mL / kg, and it was calculated based on the body weight measured on the nearest day. Female vaginal plaque was collected daily in the morning from the start of administration to the start of the mating, the sexual cycle was examined, and the mean cycle duration was calculated. After the administration period before mating, pairs of males and females 1 were established within each group and allowed to live together day and night on the 14th day. Mating confirmation was done every morning, and cases where vaginal plug formation or sperm was observed in vaginal plaque specimens was taken as copulation formation, and that day was taken as the 0th day of pregnancy. Mating pairs were separated from male and female and submitted for further examination. Body weight was measured on both the male and female on the administration start date, 3, 7 and 14 days after the start of administration, once a week for females, females with mating completed on 0, 7, 14, 20 gestation days and 0 and 4 days of pregnancy It was measured

No effect on clinical sign, body weight, food consumption, haematology and clinical chemistry of treated rats as compared to control. Similarly, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour were observed in treated rats. Increase in liver weight was observed in male and female rats. No gross pathological and histopathological changes were observed in treated rats. In addition, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour and numbers of offspring or live offspring, sex ratio, live birth index, viability index or body weights of offspring were observed. No gross pathological changes were observed in offspring as compared to control. Therefore, NOAEL was considered to be 1000 mg/kg bw/day when Crj:CD(SD)IGS male and female rats were treated with test chemical orally by gavage.

Based on the data available from different studies, test material did not showed reproductive toxicity at dose concentration 1000 mg/kg body weight/day, when male and female rats were treated with test material orally, Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

 

Effects on developmental toxicity

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

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

Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.