Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Waiver - Toxicity to reproduction screening
Waiver - Toxicity to reproduction extended one-generation study

Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Toxicity to reproduction - fertility (screening)

According to Regulation (EC) No 1907/2006, Annex VIII, 8.7.1, column 2, testing for effects on fertility by a screening test is not required as a test for prenatal developmental toxicity (Annex IX, 8.7.2) is available for the test substance.

Toxicity to reproduction - fertility (extended one-generation study)

According to Regulation (EC) No 1907/2006, Annex IX, 8.7.3, column 1, an extended one-generation reproductive toxicity study is not indicated as the available subchronic oral (90-day) toxicity study in male and female rats conducted with the target substance does not indicate any adverse effects on reproductive organs or tissues as assessed by gross and histopathological examination. Therefore, on behalf of animal welfare, the conduct of animal studies on reproduction toxicity with the target substance by any route of exposure is considered scientifically unjustified.



Effects on developmental toxicity

Description of key information

Oral (OECD 414), rat: NOAELteratogenicity ≥ 1000 mg/kg bw/day

Oral (OECD 414), rat: NOAELmaternal toxicity ≥ 1000 mg/kg bw/day

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 May - 22 Jun 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
22 Jan 2001
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
OGYÉI National Institute of Pharmacy and Nutrition, Budapest, Hungary
Limit test:
no
Species:
rat
Strain:
other: Hannover Wistar rat (CRL:WI(Han))
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sulzfeld, Germany
- Age at study initiation: at least 11 weeks
- Weight at study initiation: 188-252 g
- Housing: individually in Type II polycarbonate cages with Lignocel® bedding were used during mating and gestation period; Arbocel crinklets were used as nesting material
- Diet: ssniff® SM Autoclavable Complete Diet for Rats/Mice – Breeding and Maintenance (Ssniff Spezialdiäten GmbH, Soest, Germany); ad libitum
- Water: tap water; ad libitum
- Acclimation period: at least 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.0-24.2
- Humidity (%): 34-68
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
other: 1% aequeous methyl cellulose solution
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Formulations were prepared daily as follows. On the previous afternoon, the necessary amount of the test item (powdered if necessary) was weighed into a calibrated glass
container and the required amount of vehicle was added and stirred overnight at room temperature by a magnetic stirrer to obtain homogenous formulations. On the next morning formulations were transported for administration to animals. Formulations (off-white suspensions) were kept at room temperature, and were stirred continuously
by magnetic stirrer until the end of the treatment.
Stability of the test item in the vehicle was assessed in the conditions employed on the study during the analytical method validation. In this study, test item formulation samples in the 10-220 mg/mL concentration range (using 1% methyl cellulose as vehicle) were proven to be stable for at least 17 days when stored at room temperature (20±5°C).

VEHICLE
- Justification for use and choice of vehicle: Based on the available information of a previous rodent studies and the results of the DRF study 1% aqueous methyl cellulose solution was selcted as vehicle for this study in agreement with the Sponsor.
- Amount of vehicle: 5 mL/kg bw
- Lot/batch no.: Methyl cellulose: 5115851
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis of test item formulations for concentration and/or homogeneity was performed at the test site using a validated GC-FID (Gas Chromatography - Flame Ionization Detection) method. Top, middle and bottom samples were taken from the test item formulations three times during the study (during the first week of the treatment and two times during the second half of the treatment period including a sample on the last week of treatment). Samples were taken in duplicate (2 mL/each*), one set to analyse and one set as a back-up, if required for any confirmatory analyses. Similarly, duplicate samples were taken from the middle of the vehicle control formulation for concentration measurement. Acceptance criterion of the concentration analysis was set according to the analytical method validation, expected to be at 100 ± 15% of the nominal concentration. Acceptance criteria of the homogeneity was that the CV of replicates (top, middle and bottom of test item formulations) had to be less than 10%. The mean concentration of all formulations were found to be in the range of 97.0 - 106.2% of their nominal concentrations (20, 60 and 200 mg/mL) and were found to be homogenous. No test item was detected in the vehicle control formulations. Based on these results, test item formulations were considered suitable for the study purposes.
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1
- Length of cohabitation: 2-3 h
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
14 days (GD 6 - 19)
Frequency of treatment:
daily, 7 days/week
Duration of test:
20 days
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
27 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
The dose levels were set by the Sponsor in consultation with the Study Director based on available data, including the results of an acute oral toxicity study in rats according to the OECD guideline 401, where the LD50 >2400 mg/kg bw was estimated, and the results of an acute dermal study in rats according to the OECD guideline 402, where the LD50 >2000 mg/kg bw was estimated. Furthermore, in a 13-week repeated dose toxicity study in rats (data on file at the Sponsor), NOAEL level of 1000 mg/kg bw/day was concluded. In the Dose Range Finding (DRF) study in pregnant rats, conducted under same conditions as the main study, no signs of maternal toxicity, embryotoxicity or foetotoxicity were observed up to and including 1000 mg/kg bw/day.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations included: morbidity and mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at the onset of treatment (GD 6) and weekly afterwards
- Observations included: behavioural changes and signs of toxicity including mortality, changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), bizarre behaviour (e.g. selfmutilation, walking backwards), tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma
On GD 13 and/or 14, the sperm positive females were examined for the presence of vaginal bleeding or “placental sign” (intrauterine extravasation of blood as an early sign of pregnancy in rat, which is considered to confirm implantation).

BODY WEIGHT: Yes
- Time schedule for examinations: on GD 0, 3, 6, 8, 10, 12, 14, 16, 18 and 20

FOOD CONSUMPTION: Yes
- Time schedule: on GD 0, 3, 6, 8, 10, 12, 14, 16, 18 and 20.
Food consumption was calculated for each interval, including GD 0-6, GD 6-20 and GD 0-20.


WATER CONSUMPTION AND COMPOUND INTAKE: No


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: Dams vicera were examined macroscopically for any structural abnormalities or pathological changes.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: early and late embryonic or foetal deaths and for the number of live foetuses
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: No
Statistics:
see "any information on materials and methods incl. tables"
Indices:
Preimplantation loss: %, group mean
Number of corpora lutea-Number of implantations x100/Number of corpora lutea
- Postimplantation loss: %, group mean
Number of implantations-Number of live foetuses x100/ Number of implantations

Foetal data:
Sex distribution: %, group mean
Number of male (female) foetuses x100/Number of foetuses
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related effects or systemic clinical signs were noted in the treated animals of the low (100 mg/kg bw/day), mid (300 mg/kg bw/day) or high (1000 mg/kg bw/day) dose groups.
Nodule (approx. 1 cm) on the neck ventral area was detected for a high dose animal on GD 6. The nodule was growing rapidly, by GD 12 it reached the 3-4 cm
size. Due its large size it pressed the trachea and made the breathing difficult for this animal, furthermore it interfered with the treatment procedure, thus this animal was pre-terminally euthanized on GD 12. However, this fact was considered not being a treatment related effect.
Thin fur / alopecia on some areas was recorded for one animals of mid dose group and for 3 animals of high dose group on some days, but those findings were not considered as a test item related effect.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
No mortality was recorded for any animals in the study.
One animnals was pre-terminally sacrificed due to nodule on the neck ventral area (see "clinical signs").
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No test item related effect on body weight was observed in the low, mid and high dose groups (100, 300 and 1000 mg/kg bw/day) when compared to control. No toxicologically significant changes were observed in the mean body weights of the low, mid or high dose groups when compared to control. Similarly, no statistically significant or biologically relevant differences were seen in the body weight gain, corrected body weight gain or net body weight gain values during the treatment period (GD 6-20) or entire study (GD 0-20) compared to the control value, the only exception (statistically significant increase in the GD 16-18 body weight gain of the low dose group) was considered as irrelevant finding based on the lack of dose response and biological significance.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No biologically relevant changes were observed in the mean daily food consumption of dams in the low, mid and high dose groups (100, 300 and 1000 mg/kg bw/day, respectively) compared to the control value.
The food consumption of the low, mid and high dose groups was comparable with the control group, the difference in the daily mean food consumption (calculated for the entire period of the study) was not larger than 5.1% in any dose groups when compared to the control. There was no statistical significance and dose dependence, furthermore based on the absolute values, so any minor differences were considered being non-test item related background variations.
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
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment related observations were recorded for any evaluated animals in the study.
Thin fur was detected for three animals at necropsy, these are with correlation in the findings recorded during the in life phase. However, these facts were considered as biologically not relevant findings and not related to the test item administration.
For the pre-terminally euthanized high dose animal, a subcutaneous dark red nodule (on the ventral cervical area) was recorded. Gritty; yellow/green material and dark red liquid (blood) was detected on the cut surface. This fact was not related to the test item administration.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Number of abortions:
no effects observed
Description (incidence and severity):
There was no statistically significant difference in foetal death in any test item treated groups compared to the control.
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
No significant differences were noted in preimplantation loss or number of implantations of the test item treated groups when compared to the control, minor changes in the high dose group were considered as biological variability, not related to the test item treatment.
There was no statistically significant difference in the postimplantation loss or total intrauterine mortality between the test item treated and control groups.
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
The early and the late embryonic loss values of the test item treated groups were comparable with control. There were no dead foetuses in any groups of the study.
Early or late resorptions:
no effects observed
Description (incidence and severity):
The early and the late embryonic loss values of the test item treated groups were comparable with control.
Dead fetuses:
no effects observed
Description (incidence and severity):
There were no dead foetuses in any groups of the study.
There was no statistically significant difference in the postimplantation loss or total intrauterine mortality between the test item treated and control groups.
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
One hundred and eight females (27* in each group) were mated in the study. The number of confirmed pregnant, evaluated dams was 23 in the control group, 26 in the low dose group (100 mg/kg bw/day), 22 in the mid dose group (300 mg/kg bw/day) and 26 in the high dose group (1000 mg/kg bw/day).

*Note: Originally, mating of 24 animals per groups was planned. However, as placental sign evaluation on GD13/14 showed higher than usual ratio of non-pregnant females, three extra animals were mated for each group to ensure the required number of pregnant, evaluated animals in the study.
Other effects:
no effects observed
Description (incidence and severity):
No abnormalities were observed on the placentas of any animals in the control, low (100 mg/kg bw/day), mid (300 mg/kg bw/day) or high (1000 mg/kg bw/day) dose groups.
Key result
Dose descriptor:
NOAEL
Remarks:
maternal toxicity
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: No adverse effects on maternal toxicity observed up to the highest dose tested.
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Description (incidence and severity):
The mean foetal weight per litter in the test item did not differ significantly from the control mean value. Minor increases (without statistical significance) were seen in this parameter, but based on the biological relevance those differences were considered as biological variability.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
The mean number of viable foetuses was comparable with the control mean in all test item treated groups.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
There was no toxicologically significant difference in the sex distribution of foetuses between the control and treatment groups.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
The mean number of viable foetuses was comparable with the control mean in all test item treated groups.
Changes in postnatal survival:
not examined
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
The total number of retarded foetuses (runts) as well as the number of affected litters was similar in all test item treated group as in the control group, indicating no effect on this parameter.
Based on the external findings the number of malformed / variant / intact foetuses were comparable with the control in the low, mid and high dose groups (100, 300 and 1000 mg/kg bw/day, respectively).
In case of short or absent mandible malformation, one case was recorded in the mid and high dose groups. Although the calculated incidence was higher than the relevant historical control mean value and no similar case was observed in the study control, but based on the low absolute values (only one foetus in one litter was affected in both dose groups) and the lack of statistical significance, it was considered as incidental finding, unrelated to test item administration.
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
All of the skeletal findings correspond with the current historical control (HC) or the concurrent study control data, or were considered to be incidental findings without dose response.
Based on the skeletal findings the number of malformed / variant / intact foetuses were comparable with the control in the low, mid and high dose groups (100, 300 and 1000 mg/kg bw/day, respectively), as the observed statistically significant differences (negative trends) had no biological relevance.

A malformation (short, fused mandible) was recorded for one foetus in the mid dose group and another in the high dose group (this finding corresponded the mandible abnormality noted at external evaluation. The affected high dose foetus was identified as a runt.). The observed incidence was higher than the relevant historical control data and no such a malformation was detected in the study control. However, based on the isolated occurrence and due to lack of other malformations / variations connected to facial bones, these findings were considered as an incidental background finding, unrelated to treatment.

In case of some skeletal variations (incomplete ossification of skull (3 or more bones), ossified sternebra (2 or less), dumbbell / asymmetrically ossified vertebras and wavy ribs), the foetal or litter based incidence in the test item treated groups was comparable with the current study control or historical control values. Therefore, they were considered as biologically not relevant findings and not related to the test item treatment.
Incomplete ossification of hyoid body was observed only in two foetuses of the control group; similarly, bipartite ossification of vertebra was detected only in one control foetus. These findings were considered as irrelevant findings from the study point of view.
Some of the observed variations (misaligned sternum, vertebra (sacral 3 or more unossified), and ossified tarsus (≤ 2.5)) were considered incidental findings based on the isolated occurrence (recorded only in one foetus), these were not related to the treatment.
The incidence of Carpal, ossified ≤ 2.5 was higher in the low and mid dose groups than in the relevant HC data, however, there was no dose response (the high dose group did not show this variation). Similar case was observed for large fontanelle, interrupted ribs and for unossified/incompletely ossified pubis, where the foetal or litter based incidence in the low dose group was higher than the study control or relevant historical control, but there was no dose response (the mid and low dose groups showed the variation comparable manner to study control or they did not even contain the variation). Furthermore, none of these differences (without dose response) were statistically significant, thus these findings were considered not being treatment related.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
All of the visceral findings (malformations or variations) were consistent in general nature and incidence with the concurrent study control data / existing historical control data or showed incidental occurrence, therefore considered as incidental findings.
The incidence of abnormal lung lobation malformation was higher in the mid dose group than the relevant historical control data (and no similar case was recorded in the study control group), however, there was no dose response (the low and high dose groups showed no such a malformation) and there was only one isolated occurrence. Therefore, this finding was considered not being treatment related.
In case of Situs inversus malformation, increased incidences were observed in the mid and high dose groups compared to the historical control values (and no similar case was recorded in the study control group). However, there was no clear dose response and there was only one isolated occurrence in both dose groups, thus these findings were considered as not being treatment related.
In case of several visceral variations (short/absent brachiocephalic trunk, convoluted/dilated ureter and small renal papilla), the foetal or litter based incidence in the test item treated groups was comparable with the study control and/or historical control values. Therefore, they were considered as biologically not relevant findings and not related to the test item treatment.
Thymic cord variation was observed only in one foetus of the control group. This finding was considered as irrelevant findings from the study point of view.
In case of dilated renal pelvis variation, recorded only in the low dose group, no similar observation was recorded in the study control or historical control database. But based on the low absolute value (only one foetus was affected), it was considered as incidental finding, unrelated to test item administration.
Key result
Dose descriptor:
NOAEL
Remarks:
teratogenicity
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects on developmental toxicity observed up to the highest dose tested.
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no
Conclusions:
In this study, no signs of maternal toxicity were noted and there were no toxicologically relevant adverse effect on embryos or foetuses in any test item treated groups. Therefore, the NOAEL for maternal toxicity and teratogenicity was 1000 mg/kg bw/day, respectively.
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 1) and consistent studies, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.7, of Regulation (EC) No 1907/2006.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Developmental toxicity/ teratogenicity

The teratogenicity of Oleyl Palmitamide was assessed in a prenatal developmental toxicity study in Hannover Wistar rats according to OECD Guideline 414 and under GLP conditions (Hargitai, 2017). The dams were treated daily by oral gavage, at concentrations of 100, 300 and 1000 mg/kg bw/day from gestation day 6 (GD 6) up to and including gestation day 19 (GD 19), where sperm positive day was counted as day 0 of pregnancy (GD 0). Control dams were treated with the vehicle (1% aqueous methyl cellulose solution) only. Caesarean sections, necropsy of dams and examination of uterine contents were performed on GD 20.

Dose levels were chosen based on a Dose Range Finding study in pregnant rats, were no signs of maternal toxicity, embryotoxicity or foetotoxicity were observed up to and including 1000 mg/kg bw/day.

Parameters monitored during the main study included mortality and clinical observations, body weight, body weight gain and individual food consumption. Maternal reproductive parameters associated with uterine examination were evaluated, and the foetuses were weighed and examined for external, visceral and skeletal abnormalities. Placentas were examined macroscopically. The number of confirmed pregnant, evaluated dams was 23 in the Control, 26 in the low (100 mg/kg bw/day), 22 in the mid (300 mg/kg bw/day) and 26 in the high (1000 mg/kg bw/day) dose groups, respectively. All test item formulations were within the range of 97.0-106.2% of nominal concentration and were found to be homogenous. No test item was detected in the vehicle control samples. Based on these results, test item formulations were considered suitable for the study purposes. There were no mortalities or clinical signs in the study related to the test item treatment (pre-terminal euthanasia of a high dose female was not related to test item administration). There was no test item related adverse effect on maternal body weight / body weight gain, maternal corrected body weight / body weight gain, and on food intake in the low, mid and high dose groups (100, 300 and 1000 mg/kg bw/day, respectively). There were no toxicologically significant differences, or test item related-changes in intrauterine development examined up to and including 1000 mg/kg bw/day. The mean number of viable foetuses in all test item treated groups as well as their sex distribution were comparable with the control mean. No test item related effect was observed on the mean foetal body weight per litter. No remarkable internal or external observations related to test item treatment were recorded for any pregnant animals during necropsy. No remarkable abnormalities were observed on the placentas in any examined groups. There were no statistically significant effects of the test item on external, visceral and/or skeletal development of foetuses in the study. A low incidence of an unusual malformation was reviewed and considered as not being a test item related effect.

In conclusion, no signs of maternal toxicity were noted and there were no toxicologically relevant adverse effect on embryos or foetuses in any test item treated groups in this study. Therefore, the NOAEL for maternal toxicity and teratogenicity was 1000 mg/kg bw/day, respectively.

Justification for classification or non-classification

The available data on toxicity to reproduction of the test substance do not meet the criteria for classification according to Regulation (EC) No 1272/2008, and are therefore conclusive but not sufficient for classification. Available subchronic oral (90-day) toxicity study in male and female rats conducted with the test substance does not indicate any adverse effects on reproductive organs or tissues as assessed by gross and histopathological examination. Therefore, on behalf of animal welfare, the conduct of an extended one-generation reproductive toxicity study with the target substance by any route of exposure is considered scientifically unjustified. Under these circumstances, the health hazard ‘reproductive toxicity’ as defined in Section 2.1 GHS and the reason for no classification (conclusive but not sufficient for classification) is considered to cover both the reproduction and developmental toxicity.