1,4,7,10-Tetraazacyclododecane, 1,4,7,10-tetrakis(phenylmethyl)-

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

449-410-5

EC Name:

-

Cas Number:

18084-64-5

Molecular formula:

C36H44N4

IUPAC Name:

1,4,7,10-Tetrabenzyl-1,4,7,10-tetraazacyclododecane

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Sulzfeld, Germany
- Weight at study initiation: males 169-212 g, females 144-176 g
- Acclimatization time: 14 days

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 9 mg physiological saline + 0.85 mg Myrj 53 (polyoxyl-50-stearate) ad 1 ml aqua p.i.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Recovery of test substance in formulations varied between 79.5 and 99.4% of the nominal (analytically verified). This is deemed an acceptable level for the toxicological evaluation considering that ZK 118759 is an intermediate in a production process.

Duration of treatment / exposure:

4 weeks

Frequency of treatment:

once daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 8, 50, and 200 mg/kg bw; from day 8 onwards the high dose was reduced to 100 mg/kg bw due to severe intolerance in all animals
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: once daily

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean weekly diet consumption: Yes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: in week 1 and 4 of the study in 5 males/females per group

HAEMATOLOGY: Yes

CLINICAL CHEMISTRY: Yes

URINALYSIS: Yes

NEUROBEHAVIOURAL EXAMINATION: Yes

OTHER:
- bone marrow examination on days 29 to 32

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

DUNNETT test

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

200/100 mg/kg bw: 4 male and 4 female animals died or were killed in moribund condition between days 8 and 15. Clinical findings manly observed in agonizing animals consisted of e.g. apathy, irregular respiration, gait anomalies, ruffled fur, diarrhea

Mortality:

mortality observed, treatment-related

Description (incidence):

200/100 mg/kg bw: 4 male and 4 female animals died or were killed in moribund condition between days 8 and 15. Clinical findings manly observed in agonizing animals consisted of e.g. apathy, irregular respiration, gait anomalies, ruffled fur, diarrhea

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

200/100 mg/kg bw: body weight loss between days 1 and 8 after administration of 200 mg/kg bw; after reduction to 100 mg/kg bw animals gained weight during weeks 2 and 3; during week 4 body weight again decreased in males

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

200/100 mg/kg bw: reduced

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

effects observed, treatment-related

Description (incidence and severity):

200/100 mg/kg bw: reduced

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

200/100 mg/kg bw: increase in neutrophil counts in males on day 28; slight increase in platelet count (n.s.) in males and females on day 28

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

200/100 mg/kg bw: increase in AST in males (mainly due to animal no. 65), and decrease in total protein in males and females and gamma globuline in females on day 28

Urinalysis findings:

no effects observed

Behaviour (functional findings):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

200/100 mg/kg bw: reduced thymus (males) and spleen (males and females) weights

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

200/100 mg/kg bw: erosions and hyperkeratosis in the forestomach of one male; atrophy in prostate, seminal vesicles, thymus as well as lymphoid depletion in spleen and hypocellularity in the bone marrow was observed in single animals

Details on results:

200/100 mg/kg bw: it is likely that the effects in the forestomach are due to local irritation; with exeption of the effects in the gastro-intestinal tract all other clinical, macroscopic and microscopic observations noted were considered to be secondary to emaciation, moribundity and stress:

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

8 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

2

Dose descriptor:

LOAEL

Effect level:

50 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: see 'Remark'

3

Dose descriptor:

dose level:

Effect level:

>= 100 - <= 200 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: The daily dose of 200 mg/kg bw, reduced on day 8 to 100 mg/kg bw, led to severe organ damage and deaths in males and females.

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Executive summary:

In a 4 week oral toxicity study on 10 male and 10 female rats per group the daily oral administration of the test substance in doses of 8 and 50 mg/kg bw was well tolerated. The findings (increased sialorrhea; increased incidences of porphyrin deposits in the Harderian gland of male and female animals) in the 50 mg/kg bw dose group were judged as of minor toxicological relevance.

Signs of severe intolerance including deaths were found after 7 days of treatment with 200 mg/kg bw, whereafter the dose was reduced to 100 mg/kg bw. The signs of intolerance were reduced after reduction of dose, however, the animals still showed signs of toxicity such as reduced body weight, changes in blood parameters, atrophy of male sexual organs and changes in the lymphatic system. Microscopic lesions were noted in the stomach, thymus, spleen, lymph nodes and bone marrow of both sexes and in the male sexual organs. Severe forestomach lesions were observed in some animals which died or were sacrificed during administration of the high dose which may be caused by local irritation. It is suggested by the author of the study that the other substance related effects are secondary to these lesions and the subsequently degenerating status of the animals.

The NOAEL was established in this study with 8 mg/kg bw/day, while the findings at 50 mg/kg bw (LOAEL) were considered as of minor toxicological relevance. The daily dose of 200 mg/kg bw, reduced on day 8 to 100 mg/kg bw, led to severe organ damage and deaths in males and females.