Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
a off-white powdery solid, without irritating odor

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
The animals were housed in TECNIPLAST cages 1500U from the Tecniplast Company, Italy. The cages have high density polypropylene body, measuring 480 x 375 x 210 mm - floor area 1500 cm2. The cages, cage racks and other equipment were sanitized according to standard operation procedures.
On arrival, animals were placed into the cages, 5 rats per cage.
48-hours after arrival animals were weighted and kept in their cages until the start of the study to allow for acclimation to the animal room conditions, which are identical as defined for the experimental part of the study. After randomisation were placed three and two rats of the same sex.
The study was carried out in the experimental animal house of the Test Site with the pressure air-condition system. The animals were housed in one room under the defined laboratory conditions. The temperature and relative humidity in the animal room were recorded daily (dataloger No.: TH 464); mean values of temperature were 22.35±0.83°C at mean relative humidity 48.16±6.04% (mean±SD).
The artificial lighting was setting in a 12-hour light and 12-hour dark photoperiod.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Duration of treatment / exposure:
The animals were dosed daily for 7 days a week.
Frequency of treatment:
Once per day.
Females were treated during:
• 14-day pre-mating,
• 14-day mating (maximum)
• 22-day gestation (approximately)
• 13-day lactation
Females with no evidence of copulation and that failed to deliver were dosed for a total of 53 doses.
Males were treated during:
• 14-day pre-mating,
• 14-day mating (maximum)
Doses / concentrationsopen allclose all
Dose / conc.:
15 mg/kg bw/day
Remarks:
Low
Dose / conc.:
50 mg/kg bw/day
Remarks:
Mid
Dose / conc.:
150 mg/kg bw/day
Remarks:
High
Control animals:
yes, concurrent vehicle

Examinations

Statistics:
Individual quantitative data were summarized and analysed using descriptive statistics. This was done separately for males and females. The differences between the component groups were assessed statistically using appropriate parametric and/or non-parametric methods. The categorical data were evaluated applying contingency tables.
Significance level of 0.05 was considered to make relevant statistical conclusions.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
No changes in gait, posture and response to handling as well as the presence of clonic or tonic movements or bizarre behaviour (self-mutilation, walking backwards) were observed
Mortality:
no mortality observed
Description (incidence):
No moribund animals were observed, and no test item-related mortality was recorded during the study.
Description (incidence and severity):
Significantly increased birth weights of male and female pups in the mid-dose group cannot be attributed to account of the extended intrauterine development, because no significant differences in gestation length were observed.
Under defined test conditions, the test item, specifically in the mid-dose group, induced alteration of physical development presented consequently as decrease of pup body weight in the lactation period dose- and gender dependently.
Statistically significant changes in offspring body weights might be considered as adverse in the sense of non-specified modulation effect of test item.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
The food consumption was not observed during period of mating.
In summary, decreased food consumption in significant extent in high dose group of female rats were not accompanied with changes in the body weights and was not considered to be of toxicological relevance.
Organ weight findings including organ / body weight ratios:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
ca. 750 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
reproductive function (oestrous cycle)

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality 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
Gross pathological findings:
no effects observed

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
ca. 750 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: reproduction toxicity

Overall reproductive toxicity

Reproductive effects observed:
no
Lowest effective dose / conc.:
750 mg/kg bw/day (nominal)
Treatment related:
no

Applicant's summary and conclusion

Conclusions:
The test item 1-(9,9-Dibutyl-9H-fluoren-2-yl)-2-methyl-2-morpholin-4-yl-propan-1-one
- did not cause mortality of animals
- had no visible toxic effect on animals
- did not cause test item related alterations of organ and tissues. Macroscopic findings observed during gross necropsy (being sporadic incidence and without test item dose dependence) after histopathological evaluations were considered not to be test item related in the sense of the prevalence, severity and character.
- did not cause histopathological changes on examined reproduction organs
Statistically significant changes of relative organ weights observed in the testes, epididymis (right), thyroid glands and uteri including cervix based on the histological examinations revealed no serious morphological alterations and statistically significant differences were considered to be findings without biological/toxicological significance.
- did not show any effect on the reproductive performance of the exposed animals nor endocrine disrupting potential.
- caused dose-dependent decrease (both in offspring males and females) at 0.05 significance level of the body weight variable measured at Day 13 post-partum
- caused statistically significant decrease of body weight (both in offspring males and females) at the high dose (750 mg/kg) at Day 13 post-partum.
The statistically significant gender- and dose specific changes observed in the body weights of offspring might be considered as adverse in the sense of non-specified modulation effect of test item.
No-observed-adverse-effect level (NOAEL)
Based on the reproduction toxicity
• NOAEL for females and males was concluded to be 750 mg/kg