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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-05-02 to 2017-06-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2019

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
- Principle of test:
The study was designed to determine, in a preliminary phase, the maximum tolerated dose (MTD) and, successively, the toxicity of the test item over a period of 14 consecutive days of treatment (main phase).

- Short description of test conditions:
In the preliminary phase (Phase 1), 2 male and 2 female rats/groupwere initially administered at a dose level of 300mg/kg/day (Group 1) and then at 2000mg/kg/day (Group 2), each followed by a 7-day observation period.
In the main phase (Phase 2), 4 animals/sex/dose were administered once daily at 100, 500 and 1000mg/kg/day for at least 14 consecutive days.

- Parameters analysed / observed:
– preliminary phase: clinical signs, body weight, gross macroscopic observations;
– main phase: clinical signs, body weight, food consumption, clinical pathology investigations (including thyroid hormones determination), terminal body weight, organ weights and macroscopic observations.
GLP compliance:
no
Remarks:
However, study was carried out in a GLP compliant facility (DRF for OECD 422 study)
Test type:
other: dose - range finder
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
N,N'-[(methylimino)bis(trimethylene)]bis(stearamide)
EC Number:
236-793-7
EC Name:
N,N'-[(methylimino)bis(trimethylene)]bis(stearamide)
Cas Number:
13483-58-4
Molecular formula:
C43H87N3O2
IUPAC Name:
N-{3-[methyl(3-octadecanamidopropyl)amino]propyl}octadecanamide

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Envigo Holland, Kreuzelweg 53, 5961 NM Horst, Netherlands
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 6-7 weeks
- Weight at study initiation: males: 160 - 183; females: 153 - 181 g
- Fasting period before study: Phase 1: overnight fast prior to each dosing day; Phase 2: the diet was offered ad libitum throughout the study
- Housing: up to 5 of one sex to a cage, in clear polysulfone solid bottomed cages
- Diet: commercially available laboratory rodent diet (4 RF 21, Mucedola
S.r.l.) ad libitum
- Water: drinking water via water bottles ad libitum
- Acclimation period: 7 days phase 1; 4 weeks phase 2

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2 °C
- Humidity (%): 55 +/- 15 %
- Air changes (per hr): 1 5 to 20 air changes per hour
- Photoperiod (hrs dark / hrs light): artificaial light for 12 hours each

IN-LIFE DATES: From: 2017-05-02 to 2017-06-09

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
0.5 % aqueous solution
Details on oral exposure:
VEHICLE
- Concentration in vehicle: Phase 1: 30 and 200 mg/L
Phase 2: 10, 50 and 100 mg/mL
- Amount of vehicle (if gavage): 0.5 % aqueous solution


MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

Doses:
Phase 1: 300 and 2000 mg/kg single oral dose
Phase 2: 0, 100, 500 and 1000mg/kg/day treatment for 14 consecutive days
No. of animals per sex per dose:
Phase 1: 2 animals/sex/group
Phase 2: 4 animals/sex/group
Control animals:
yes
Remarks:
Phase II, control animals received the vehicle alone
Details on study design:
The following investigations were performed:
– preliminary phase: clinical signs, body weight, gross macroscopic observations;

– main phase: clinical signs, body weight, food consumption, clinical pathology investigations
(including thyroid hormones determination), terminal body weight, organ weights and macroscopic observations.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during any phases of the study.
Clinical signs:
Phase 1
Clinical signs observed in the animals of the preliminary phase treated at 300 mg/kg were limited to piloerection only on the day of dosing (at 30 minutes and 4 hours after dosing).
No clinical signs were seen in the remaining observation session on the day of dosing and daily thereafter.
No clinical signs were noted in all animals treated at 2000 mg/kg during the entire observation period.

Phase 2
No signs of toxicological relevance were observed during the phase 2.
Body weight:
No significant changes in body weight were observed during neither phase of the study.
Gross pathology:
Phase 1
Animals of both sexes treated at 300 and 2000mg/kg killed at termination did not show macroscopic changes.

Phase 2
No treatment-related changes were noted in treated animals of both sexes sacrificed at termination, when compared to controls.

Any other information on results incl. tables

Food consumption: Phase 2

No treatment-related changes were observed in food consumption in either sex during the study.

Haematology: Phase 2

No changes of toxicological relevance were observed in haematological parameters.

Coagulation: Phase 2

No changes were recorded.

Clinical chemistry: Phase 2

Glucose was increased in males dosed at 1000 mg/kg/day.

Alterations in transaminases were noted in one high dose female and also in one control female animal, therefore the finding cannot be conclusively attributed to treatment.

Thyroid hormone determination: Phase 2

No statistically significant changes were recorded.

Terminal body weight and organ weights: Phase 2

Terminal body weight

No differences of toxicological significance were noted in body weight between treated animals and controls.

Absolute and relative organ weights

Statistically significant increase in absolute penis weight was noted in high dose animals. In addition, statistically significant increase in absolute and relative prostate gland weights was seen in mid- and high dose groups with no relevant difference between the two groups.

No other relevant differences in organ weights were observed for males.

No changes of toxicological significance were observed in the absolute and relative organ weights of female animals.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
For acute oral toxicity of N,N’[(methylimino)bis(trimethylene)]bis(stearamide) a LD50value of > 2000 mg/kg can be established.
Executive summary:

The toxicity of N,N’[(methylimino)bis(trimethylene)]bis(stearamide) after oral administration was investigated in a study similar to OECD guideline 423 by (gavage) to Sprague Dawley rats.

The study was designed to determine, in a preliminary phase, the maximum tolerated dose (MTD) and, successively, the toxicity of the test item over a period of 14 consecutive days of treatment (main phase) in order to select dose levels for the subsequent OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) in rats.

In the preliminary phase (Phase 1), 2 male and 2 female rats/group were initially administered at a dose level of 300 mg/kg/day (Group 1) and then at 2000 mg/kg/day (Group 2), each followed by a 7-day observation period. No toxicity was observed at each dose level investigated.

On the basis of results obtained in Phase 1, in the main phase (Phase 2), 4 animals/sex/dose were administered once daily at 100, 500 and 1000 mg/kg/day for at least 14 consecutive days.

 

No mortality and no clinical signs were recorded in main phase animals; body weights as well as terminal body weights were not affected by treatment. Increase in prostate gland weight was noted in mid- and high dose males. The changes and fluctuations observed in haematology and clinical chemistry analysis performed in the animals at the end of the treatment period cannot be conclusively attributed to treatment or were considered incidental.

 

No treatment-related findings were reported at post mortem macroscopic observations of all treated animals. On the basis of the results obtained in this preliminary study, it can be concluded that the test itemN,N’[(methylimino)bis(trimethylene)]bis(stearamide) showed no toxic effects when administered daily at dose levels of 100, 500 and 1000 mg/kg/days for at least 14 consecutive days.

 

For acute oral toxicity of N,N’[(methylimino)bis(trimethylene)]bis(stearamide) a LD50value of > 2000 mg/kg can be established.

 

 

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