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Administrative data

Description of key information

With regard to the 28-subacute oral toxicity study with male and female Wistar rats a no-observed-adverse-effect-level (NOAEL) of 50 mg/kg body weight was established for an analogue substance, 3-aminopropyldimethylamine.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD 407)
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst Aktiengesellschaft, Kastengrund, SPF breeding colony
- Age at study initiation: approximately 6 weeks
- Weight at study initiation: male: 108 g (mean), female: 114 g (mean)
- Housing: in groups of 5
- Diet: ssniff R/M-H (V 1534) ad libitum, except for the period in which the animals were kept in diuresis cages.
- Water: tap water in plastic bottles ad libitum, except for the period in which the animais were kept in diuresis cages.
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 50 ± 20 %
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
water
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The determination of the test substance was performed by spectrophotometrical detection (λ = 274 nm).
Duration of treatment / exposure:
28 days
Frequency of treatment:
daily (7x week)
Remarks:
Doses / Concentrations:
0, 10, 50, 250 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In a dose range finding study groups of 3 male and 3 female Wistar rats received 3-Dimethylaminopropylamin at dose levels of 50 and 250 mg/kg body weight per day over a period of 14 days. The animals of the 50 mg/kg body weight group showed no clinical signs. The animals of the 250 mg/kg body weight showed sporadically swollen abdomen during the study period.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: the animals were observed once daily.

BODY WEIGHT: Yes
- Time schedule for examinations: The body weights of all animals were determined before the start of the study and then twice weekly throughout the study.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: weekly
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the termination of the study
- Anaesthetic used for blood collection: Yes (intrapenitoneal injection of 50-100 mg Ketamin/kg body weight)
- Animals fasted: No data
- How many animals: all
- Parameters examined: Erythrocyte count, hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), leucocyte count, thrombocyte count, differential leucocyte count and red ceII morphology, reticulocyte count*, heinz bodies*, coagulation time. *These parameters were scored in the control group and high dose group only.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the termination of the study
- Animals fasted: No data
- How many animals: all
- Parameters examined: Sodium, potassium, inorganic phosphorus, uric acid, bilirubin total, creatinine, glucose, urea, calcium, chloride, aspartate aminotransferase (ASAT/GOT), alanine aminotransferase (ALATIGPT), alkaline phosphatase (AP), gamma-glutamyltranspeptidase (GGT), cholesterol, triglycerides, total protein, albumin.

URINALYSIS: Yes
- Time schedule for collection of urine: overnight from day 26 to day 27
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined: Appearance, color, pH-Value, hemoglobin, protein, glucose, ketone bodies, bilirubin, urobilinogen, specific weight, sediment, volume.

OTHERS: The animals were examined weekly for neurological disturbances, damage to the oral mucosa and impairment of dental growth.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes. The autopsy included macroscopic examination of the skin, orifices, eyes, teeth, oral mucosa and internal organs.

HISTOPATHOLOGY: Yes. The necropsy included examination of the heart, stomach, liver, jejunum, kidneys, colon, adrenals, uterus, spleen, ovanies, testes, epididymides, lungs.
Statistics:
The following parameters were compared statistically with the control group values at the level of significance p = 0.05.
Body weights at the designated measurement times, hematological data, clinical chemistry parameters, urine analysis, absolute organ weights and organ to body weight ratios.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
at 250 mg/kg bw/d
Mortality:
mortality observed, treatment-related
Description (incidence):
at 250 mg/kg bw/d
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
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):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
at 250 mg/kg bw/d
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
at 250 mg/kg bw/d
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
Mortality occurred in four females of the 250 mg/kg body weight group. The animals were found dead on days 11, 18, 22 and 25 of the study.
One male animal of the high dose group showed irregular respiration as well as respiratory sounds at day 11 and 12 of the study. The following clinical signs were observed in female animals of the high dose group sporadically between day 11 and 24 of the study: decreased spontaneous activity, stilted gait, swollen abdomen, respiratory sounds, gasping and panting. The clinical signs were mainly seen in those females which died intercurrently. Behavior and state of health remained unaffected by the administration of the test compound in all other dose groups.
BODY WEIGHT AND WEIGHT GAIN
Body weight development was not impaired by the administration of the test compound and was comparable in all groups.

FOOD CONSUMPTION AND COMPOUND INTAKE
Absolute and relative food consumption remained unaffected by the administration of the test compound throughout the study.

WATER CONSUMPTION AND COMPOUND INTAKE
Likewise, the administration of the test compound did not alter the water consumption.

OPHTHALMOSCOPIC EXAMINATION
No abnormalities were observed

HAEMATOLOGY
Hematological examinations revealed statistically significant decreases in erythrocyte counts as well as in hemoglobine and hematocrit values in males of the intermediate dose group. Haematocrit values were also decreased in males of the high dose group. Leukocyte counts were statistically significantly decreased in males of the intermediate and females of the low dose group. In all cases there was no dose dependency or the values were within the physiological range of rats. Therefore, a compound-related effect is not evident.

CLINICAL CHEMISTRY
Statistical evaluation revealed increases in aspartate aminotransferase and decreases in total protein values in females from the high dose group. lnorganic phosphorus values were decreased in females of the low dose group. Males of the intermediate dose group showed statistically significant decreases in total bilirubin values. In all dose groups of the females the uric acid values were statistically significantly decreased. In all cases there was no dose dependency or the values were within the physiological range of rats. Therefore, a compound-related effect is not evident.

URINALYSIS
Examination of the urine did not reveal any abnormalities.

ORGAN WEIGHTS
In males of the intermediate dose group statistical evaluation of the organ weights revealed increases in relative liver weights. As there was no dose dependency a compound-related effect is not evident.

GROSS PATHOLOGY
In terminally killed animals, no macroscopically visible organ alterations attributable to the compound administration were observed. Dilatation of renal pelvices was found in same male or female animals of all treatment groups and therefore was not considered to be compound-related but to be a strain specific alteration. In the four high dose females which died intercurrently gross findings included discoloration of Iungs with multiple red spots an its surfaces and foamy content. One female also showed a small spleen.

HISTOPATHOLOGY: NON-NEOPLASTIC
Histopathological examinations revealed lesions in the four females of the high dose group dying intercurrently. These lesions included congestion of organs, pulmanary hemorrhage, and edema, consistent with cardiorespiratory failure as cause of death. In addition, one of these females exhibited marked loss of lymphatic follicles of the spleen with massive marginal zone and periarteriolar lymphoid sheath atrophy, probably reflecting chronic stress due to treatment. In the one high dose male rat which had shown clinical signs, focal ballooning degeneration of the stratum corneum of the squamous epithellum of the forestomach with granulocytic infiltration of the submucosa was found, most likely due to the locally irritating effect of the compound.

Dose descriptor:
NOAEL
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no treatment-related adverse effects were seen.
Dose descriptor:
LOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: 3-Dimethylaminopropylamin caused clinical symptoms and mortality in male and female Wistar rats when administered 28 times at the dose level of 250 mg/kg body weight. The female rats seem to be more sensitive.
Critical effects observed:
not specified
Conclusions:
The „No-Observed-Adverse-Effect-Level“ (NOAEL) was 50 mg DMAPA/kg body weight/day, by gavage for 28 days.
Executive summary:

Groups of 5 male and 5 female Wistar rats received DMAPA by oral gavage at dose levels of 0, 10, 50 or 250 mg/kg body weight per day for 28 days and were necropsied on day 29. Because 3 females of the 250 mg/kg bw group died during the study period, 5 females received DMAPA at the dose level of 250 mg/kg bw per day for 28 days again and were necropsied on day 29. Toxicological examinations were carried out on mortalities, clinical signs, body weight, food consumption, examination of eyes, haematology, clinical biochemistry, urinalysis, organ weights, macroscopic post mortem findings and histopathology.

Intercurrent mortality occurred in four of ten females of the high dose group. One male animal of the high dose group showed irregular respiration as well as respiratory sounds at day 11 and 12 of the study. The following clinical signs were observed in female animals of the high dose group sporadically between day 11 and 24 of the study: decreased spontaneous activity, stilted gait, swollen abdomen as well as impaired respiration. The clinical signs were mainly seen in those females which died intercurrently. Behavior and state of health remained unaffected by the administration of the test compound in all other dose groups. Body weight development, food and water consumption remained unaffected by the administration of the test compound in all groups.

No treatment-related changes were detected by urine analysis. Clinicochemical and haematological examination also failed to reveal any significant changes compared with the untreated control animals. Organ weights, as well as their gross and histological examination, in those animals killed following termination of the study, also failed to reveal any substance-related effects.

The four high dose female rats which died intercurrently showed macroscopically visible changes such as discoloration of lungs with multiple red spots on its surfaces and foamy content. One of them also showed a small spleen. Histopathological examinations revealed lesions in the four females of the high dose group dying intercurrently. These lesions included congestion of organs, pulmonary hemorrhage, and edema, consistent with cardiorespiratory failure as cause of death. In addition, one of these females exhibited marked loss of lymphatic follicles of the spleen with massive marginal zone and periarteriolar lymphooid sheath atrophy. In the one high dose male rat which had shown clinical signs, focal ballooning degeneration of the squamous epithelium of the forestomach was found. In particular, no pathological findings were reported for the genitals. Neither were any neurological

disturbances observed. In conclusion DMAPA caused clinical symptoms and mortality in male and female Wistar rats when administered 28 times during 29 days at the dose level of 250 mg/kg body weight. The female rats seem to be more sensitive.

No compound-related adverse effects were observed after repeated administration of DMAPA at the dose levels of 50 or 10 mg/kg body weight per day. Based on the findings the „No-Observed-Adverse-Effect-Level“ (NOAEL) was 50 mg DMAPA/kg body weight/day, by gavage for 28 days.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
50 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Read across to an analogue substance.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

No repeated dose toxicity is available for the N, N, N', N'-tetramethyltrimethylenediamine. However, a 28-day oral toxicity study performed following the OECD TG 407 is available on a structural analogue, 3-aminopropyldimethylamine (DMAPA).

Groups of 5 male and 5 female Wistar rats received DMAPA by oral gavage at dose levels of 0, 10, 50 or 250 mg/kg body weight per day for 28 days and were necropsied on day 29. Because 3 females of the 250 mg/kg bw group died during the study period, 5 females received DMAPA at the dose level of 250 mg/kg bw per day for 28 days again and were necropsied on day 29. Toxicological examinations were carried out on mortalities, clinical signs, body weight, food consumption, examination of eyes, haematology, clinical biochemistry, urinalysis, organ weights, macroscopic post mortem findings and histopathology. Intercurrent mortality occurred in four of ten females of the high dose group. One male animal of the high dose group showed irregular respiration as well as respiratory sounds at day 11 and 12 of the study. The following clinical signs were observed in female animals of the high dose group sporadically between day 11 and 24 of the study: decreased spontaneous activity, stilted gait, swollen abdomen as well as impaired respiration. The clinical signs were mainly seen in those females which died intercurrently. Behavior and state of health remained unaffected by the administration of the test compound in all other dose groups. Body weight development, food and water consumption remained unaffected by the administration of the test compound in all groups. No treatment-related changes were detected by urine analysis. Clinicochemical and haematological examination also failed to reveal any significant changes compared with the untreated control animals. Organ weights, as well as their gross and histological examination, in those animals killed following termination of the study, also failed to reveal any substance-related effects. The four high dose female rats which died intercurrently showed macroscopically visible changes such as discoloration of lungs with multiple red spots on its surfaces and foamy content. One of them also showed a small spleen. Histopathological examinations revealed lesions in the four females of the high dose group dying intercurrently. These lesions included congestion of organs, pulmonary hemorrhage, and edema, consistent with cardiorespiratory failure as cause of death. In addition, one of these females exhibited marked loss of lymphatic follicles of the spleen with massive marginal zone and periarteriolar lymphoid sheath atrophy. In the one high dose male rat which had shown clinical signs, focal ballooning degeneration of the squamous epithelium of the forestomach was found. In particular, no pathological findings were reported for the genitals. Neither were any neurological disturbances observed. In conclusion DMAPA caused clinical symptoms and mortality in male and female Wistar rats when administered 28 times during 29 days at the dose level of 250 mg/kg body weight. The female rats seem to be more sensitive. No compound-related adverse effects were observed after repeated administration of DMAPA at the dose levels of 50 or 10 mg/kg body weight per day. Based on the findings the „No-Observed-Adverse-Effect-Level“ (NOAEL) was 50 mg DMAPA/kg body weight/day, by gavage for 28 days.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Key study

Repeated dose toxicity: via oral route - systemic effects (target organ) digestive: stomach

Justification for classification or non-classification

According to DSD and CLP criteria, no classification is warranted for the specific target organ toxicity after repeated exposure.