N-[3-(dimethylamino)propyl]oleamide

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in accordance with OECD guideline 407 and GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N-[3-(dimethylamino)propyl]oleamide

EC Number:

203-661-5

EC Name:

N-[3-(dimethylamino)propyl]oleamide

Cas Number:

109-28-4

Molecular formula:

C23H46N2O

IUPAC Name:

(9Z)-N-[3-(dimethylamino)propyl]octadec-9-enamide

Test material form:

other: liquid

Details on test material:

Color: reddish
Batch: R401/57
Purity: 78.2 area-%

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Supplier: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, 97633 Sulzfeld
Age at beginning of administration period: 42+/-1 days

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

drinking water

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analyses of the test-substance preparations were carried out as a separate study at the test facility GKA Competence Center Analytics, BASF SE, Ludwigshafen, Germany under the responsibility of the Study Director of this facility. The study was carried out in compliance with the Principles of GLP. The stability of the test substance in drinking water for a period of 7 days was proven before the start of the administration period.

Duration of treatment / exposure:

28 days

Frequency of treatment:

Once daily.

Doses / concentrations

Remarks:

Doses / Concentrations:
10, 50 and 150 mg/kg body weight
Basis:

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

Rationale for selection of doses: In a pre-test, the test substance was administered to Wistar rats at dose levels of 1000 and 300 mg/kg body weight/day by gavage for a period of 2 weeks. Several severe clinical findings were observed even at 300 mg/kg including premature death. Therefore, 150 mg/kg body weight/day was selected as the highest dose in the present study.

Examinations

Observations and examinations performed and frequency:

- Mortality: twice daily on working days, once daily on Saturdays, Sundays and public holidays
- Clinical observations: daily before and after administration
- Detailed clinical observations: performed in all animals prior to the administration period and thereafter at weekly intervals. Parameters examined: abnormal behavior during “handling”, fur, skin, posture, salivation, respiration, activity/arousal level, tremors, convulsions, abnormal movements, impairment of gait, lacrimation, palpebral closure, exophthalmus, feces (appearance/consistency), urine, pupil size.
- Food consumption: determined weekly over a period of 1 day
- Water consumption: determined weekly over a period of 4 days
- Body weight: determined before start of administration period, at start of administration period and weekly thereafter
- Food efficiency: calculated based upon individual values for body weight and food consumption
- Functional observational battery: performed in all animals at the end of the administration period
- Motor activity assessment: performed in all animals towards the end of the administration period
- Ophthalmoscopy: performed in all animals prior to the start of the administration period and in the control and high dose animals on day 28

Sacrifice and pathology:

Parameters examined in clinical pathology:
- Hematology: Leukocyte count, Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet count, differential blood count, reticulocytes, prothrombin time
- Clinical chemistry: ALT, AST, ALP, GGT, sodium, potassium, chloride, inorganic phsophate, calcium, urea, creatinine, glucose, total bilirubin, total protein, albumin, globulins, triglycerides, cholesterol, magnesium
- Urinalysis: pH, protein, glucose, ketones, urobilinogen, bilirubin, blood, specific gravity, sedinment, color/turbidity, volume

Organs/parameters examined in pathology:
- Weights: anestzetized animals, liver, kidneys, adrenal glands, testes, epididymides, ovaries, uterus, spleen, brain, heart, thymus, thyroid glands
- Preserved organs/tissues: all gross lesions, brain, pituitary gland, thyroid glands, parathyroid glands, thymus, esophagus, salivary glands (mandibular gland, sublingual gland), trachea, lungs, pharynx, larynx, nose (nasal cavity), aorta, heart, liver, pancreas, spleen, kidneys, adrenal glands, testes, ovaries with oviducts, uterus, vagina, epididymides, prostate, seminal vesicles, stomach (fore- and glandular stomach), duodenum, jejunum, ileum, cecum, colon, rectum, urinary bladder, lymph nodes (mesenteric and axillary lymph node), sciatic nerve, bone marrow (femur), eyes, extraorbital lacrimal glands, skin, female mammary glands, skin, female mammary gland, spinal cord (cervical, thoracic and lumbar cord), sternum with marrow, femur with knee joint, skeletal muscle.
- Histopathology: all gross lesions, salivary glands (mandibular and sublingual), brain, pituitary gland, eyes, esophagus, thyroid glands, parathyroid glands, thymus, trachea, lungs, aorta, heart, liver, pancreas, spleen, kidneys, adrenal glands, testes, ovaries with oviducts, uterus, vagina, female mammary gland, epididymides, prostate, seminal vesicles, stomach (fore- and glandular stomach), duodenum, jejunum (with Peyer's plaques), ileum, cecum, colon, rectum, urinary bladder, lymph nodes (mesenteric and axillary lymph node), sciatic nerve, skin, bone marrow (femur), spinal cord (cervical, thoracic and lumbar cord)

Statistics:

Means and standard deviations of each test group were calculated for the following parameters:
- Body weight, body weight change (comparison of each group with the control group was performed using a two-sided DUNNETT's test for the hypothesis of equal means)
- Feces, rearing, grip strength length forelimbs, grip strength length hindlimbs, footsplay test, motor activity (Non-parametric oneway analysis using a two-sided KRUSKAL-WALLIS test. If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using a two-sided Wilcoxon-test for the equal medians.)

Statistics of pathology:
Weight parameters were analyzed with a non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pair wise comparison of each dose group with the control group was performed using the WILCOXON test for the hypothesis of equal medians.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

No premature mortality occured. Salivation was observed predominantly after treatment in all animals of either sex of dose group 3 (150 mg/kg bw/d) on few days between study day 11 and 28. Besides, respiration sounds were observed predominantly after treatment in 3 male and 4 female animals of dose group 3 (150 mg/kg bw/d) on several days between study day 7 and 28. These findings were assessed as adverse and related to the test article.

Mortality:

mortality observed, treatment-related

Description (incidence):

No premature mortality occured. Salivation was observed predominantly after treatment in all animals of either sex of dose group 3 (150 mg/kg bw/d) on few days between study day 11 and 28. Besides, respiration sounds were observed predominantly after treatment in 3 male and 4 female animals of dose group 3 (150 mg/kg bw/d) on several days between study day 7 and 28. These findings were assessed as adverse and related to the test article.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

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

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

BODY WEIGHT AND WEIGHT GAIN:
Body weight was significantly increased (+10.6%) in male animals of the 10 mg/kg bw/d dose group on day 28. This single occurence was without any correlative to food consumption or other clinical oarameters. It was therefore considered as not treatment-related.

HAEMATOLOGY:
Some deviations in the differential blood cell counts without any change of the total white blood cell count were found in the female rats of the 10 and the 150 mg/kg bw/d dose groups: an increase of the absolute and relative neutrophil counts as well as relative basophil counts and a decrease of the relative lymphocyte counts. The statistically significant changes in the dosed rats were due to low neutrophil and basophil and high lymphocyte counts in the control rats of this study. The deviations were not dose-dependent. All values were within, or very slightly out of the borders of the historical control ranges.

CLINICAL CHEMISTRY:
In the male rats of the 150 mg/kg bw/d dose group, the calcium concentration was increased compared to the controls. This was the only deviated clinical chemistry parameter in these animals. Therefore it was regarded as non adverse, although it cannot be excluded that this change was treatment related.

URINALYSIS:
In the female rats of the 50 mg/kg bw/d dose group, the specific gravity of the urine was statistically significantly decreased. This change was not dose-dependent or accompanied by any other deviations in the urine. It was therefore considered not adverse.

NEUROBEHAVIOUR

ORGAN WEIGHTS:
Kidney weights were significantly changed in the female rats of the 10 mg/kg bw/d dose group as compared to the control group. As not significant deviations from the control animals were observed in the higher dose groups, this finding was considered not treatment-related.
The relative brain weights were changed in all treated female rats as compared to the control group. No dose-response relationship or histopathological correlate ws observed, and therefore this finding was considered to be not treatment-related.

GROSS PATHOLOGY
All gross lesions occured singly or were biologically equally distributed between control and treatment groups.

HISTOPATHOLOGY:
All findings noted were either single observations or they were biologically equally distributed between control and treatment group. All of them are considered to be incidental or spontaneous in origin and without any relation to treatment.

Effect levels

Dose descriptor:

NOAEL

Effect level:

> 50 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: Salivation and respiration sounds in male and female animals in the 150 mg/kg dose group

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The oral administration of 3-Dimethyl-aminopropyl-ölsäureamide by gavage over a period of 28 days at dose levels of 10, 50 and 150 mg/kg bw/day caused slight toxicity in animals in the high dose group of either sex. Therefore, the NOAEL under the conditions of the present study was 50 mg/kg body weight/day in both male and female rats.

Executive summary:

3-Dimethyl-aminopropyl-ölsäureamide was administered to groups of 5 male and 5 female Wistar rats at doses of 10, 50 and 150 mg/kg bw/day with a control group concurrently treated with vehicle (drinking water) over a period of 28 days by gavage. Food and water consumption and body weight were determined weekly. The animals were examined for signs of toxicity and mortality at least once a day. Additionally, further clinical examinations were carried out daily. Detailed clinical examinations in an open field were conducted prior to the start of the administration period and weekly thereafter. Ophthalmological examinations were conducted before the beginning and towards the end of the administration period. Additionally, a functional observational battery (FOB) as well as measurement of motor activity was carried out at the end of the study. Clinicochemical and hematological examinations as well as urinalyses were performed towards the end of the administration period. All animals were assessed by gross pathology, followed by histopathological examinations.

No substance-related adverse findings were obtained in the 10 and the 50 mg/kg bw/day dose groups. In the 150 mg/kg bw/day dose group, salivation was observed after treatment in all male and female animals on few days between study day 11 and 28. Furthermore, Respiration sounds were observed after treatment in 3 male and 4 female rats on several days between study day 7 and 28. Based on the findings in the high dose group, a NOAEL of 50 mg/kg bw/day was drived for both male and female rats.