3,7-Diazabicyclo[3.3.1]nonane-1,5-dicarboxylic acid, 3-methyl-9-oxo-2,4-di-2-pyridinyl-7-(2-pyridinylmethyl)-, 1,5-dimethyl ester, (1R,2S,4R,5S)-rel-

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 9 December 2003 to 5 December 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

Adopted : 17th December 2001

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

618-281-9

EC Number:

618-281-9

Cas Number:

895520-71-5

Molecular formula:

C28 H29 N5 O5

IUPAC Name:

618-281-9

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: E-699-2003
- Expiration date of the lot/batch: 1 July 2004
- Purity test date: 27 and 28 October 2003

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: white to light beige powder stored at approximately 20°C in a fume cupboard
- Stability under test conditions: is guaranteed for 4 hours in dionized water by the sponsor (dated 04 December 2003)
- Solubility and stability of the test substance in the solvent/vehicle: is guaranteed for 4 hours in dionized water by the sponsor (dated 04 December 2003)

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: DP-2 was suspended in the stated concentrations in deionozed water and distributed homogenously by means of magnetic stirrer
- Final dilution of a dissolved solid, stock liquid or gel: 10% w/v in vehicle for the high dose and 3% for the low dose

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

SD

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann GmbH
- Females (if applicable) nulliparous and non-pregnant: not specified
- Age at study initiation: 6 to 10 weeks
- Weight at study initiation: mean : 179g±10.9g (168 to 201 g)
- Fasting period before study: Fasted, no more details
- Housing: In transparent macrolon cages (type IV) on soft wood granulate in air conditionel room, 3 animals per cage
- Diet (e.g. ad libitum): ssniff R/M-H (V 1534) ad libitum
- Water (e.g. ad libitum): tap water in plastic bottles ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 50±20%
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12 hours of light / 12 hours of dark

IN-LIFE DATES: From: 9 December 2003 To: 5 December 2004

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

deionized water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 10% w/v for 2000 mg/kg dose and 3% for 300 mg/kg dose level
- Amount of vehicle (if gavage): 10 ml/kg for 300 mg/kg and 2 x 10 ml/kg for high dose 2000 mg/kg
- Justification for choice of vehicle: not specified
- Purity: not specified

DURATION OF TREATMENT :
Single dose (300 mg/kg) and two applications in 30 minutes interval (2000 mg/kg)

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: not specified

Doses:

300 mg/kg and 2000 mg/kg

No. of animals per sex per dose:

6 females for 300 mg/kg group and 3 for 2000 mg/kg dose group

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Symptoms and lethality were recorded twice everyday. During this time, animals were weighed weekly.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology (macroscopic examinations of organs)

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

2000 mg/kg : 3/3 animals died
300 mg/kg : 0/6 animals died

Clinical signs:

2000 mg/kg :
The following clinical signs were observed in female animals starting between 0 and 10 minutes after administration of DP-2 : stupor, moderate ataxia, hypoactivity, prone or lateral postion, forward crawling, straddled hind limbs, squatting posture, stilted gait, drawn in flanks, irregular respiration, bristling coat, narrowed palpebral fissures and dilated pupils, absent paw reflex to pinching, negative righing reflex and clonic convulsions.

300 mg/kg
The following clinical signs were observed in female animals starting 10 to 30 minutes after the treatment : stupor, hypoactivity, squatting posture, moderate ataxia, stilted or uncoordinated gait, drawn in flanks, irregular respiration, bristling coat, narrowed palpebral fissures and trembling. From day 3 until the end of the study no symptoms were observed

Body weight:

Development of the body weight was not impaired in the surviving animals.

Gross pathology:

The decedent animals and the animals killed at the end of the observation period showed no macroscopically visible changes.