Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 April - 21 May 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well conducted and well described study according to GLP and OECD guideline 423 without any deviation
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
- Name of test material (as cited in study report): PI2080
- Physical state: Limpid amber highly viscous liquid
- Analytical purity: 97 ± 3 %
- Lot/batch No.: 11GP0077
- Date of receipt: 10 February 2012
- Expiration date of the lot/batch: January 2014
- Storage condition of test material: At room temperature and protected from light

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: Approximately 8 weeks
- Weight at study initiation: 200-233 g (mean: 216 g)
- Fasting period before study: Overnight
- Housing: Animals were housed by three from the same group in polycarbonate cages with stainless steel lids.
- Diet (e.g. ad libitum): SSNIFF R/M-H pelleted maintenance diet (SSNIFF Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water (e.g. ad libitum): Tap water (filtered with a 0.22 μm filter), ad libitum
- Acclimation period: 6 or 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 50 ± 20 %
- Air changes: Approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: From 10 April to 09 May 2012

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
DMSO
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 and 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: In a serie of experiments, the solubility assays were performed with drinking water, 0.5 % methylcellulose aqueous solution, corn oil and DMSO. Based on the results, DMSO was selected as the vehicle for the study.
- Lot/batch no. (if required): BCBD5253V

DOSAGE PREPARATION: The flask, containing the test item and the required quantity of vehicle, was put in a water bath at 60 °C for approximately 5 min and then mixed under magnetic agitation for at least 15 min.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Since no relevant toxicity data were available for the estimation of a lethal dose-level, the starting dose-level was 300 mg/kg bw for animal welfare reasons.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
- Groups 1 and 3 (300 mg/kg bw): 3 females/group
- Group 2 (2000 mg/kg bw): 3 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Morbidity and mortality: Each animal was checked for mortality and morbidity, frequently during the hours following administration, then once a day until the end of the observation period, including weekends and public holidays.
Clinical signs: Each animal was observed after treatment as follows: at least once during the first 30 minutes, periodically during the first 4 hours, then once a day, at approximately same time, for the recording of clinical signs.
Body weight: Body weight was recorded on the day of group allocation, then on the day of treatment and on Days 8 and 15.
- Necropsy of survivors performed: Yes; animals were deeply anesthetized by an intraperitoneal injection of pentobarbital sodium, euthanized by exsanguination and then subjected to a macroscopic examination.
Statistics:
None

Results and discussion

Preliminary study:
Not applicable
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
300 - 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- At 2000 mg/kg bw, one female was found dead on Day 6 and another one was sacrificed prematurely on Day 5.
- At 300 mg/kg bw, one female was found dead on Day 13, the cause of death was traumatic (skull fracture).
Clinical signs:
- At 2000 mg/kg bw, piloerection, hypoactivity, hunched posture, half-closed eyes, soiled urogenital and mouth regions, then dyspnea and lateral recumbency were observed in the prematurely sacrificed female. Piloerection was also observed in the surviving female.
- At 300 mg/kg bw, piloerection and ptyalism (3/6 females), hypoactivity and dyspnea (2/6 females), half-closed eyes and chromorhynorrhea (1/6 females) were observed.
Body weight:
When compared to historical control data, a lower body weight gain was noted in 4/5 surviving females treated at 300 mg/kg bw and in the surviving female treated at 2000 mg/kg bw between Day 1 and Day 15 (35-56 g vs. 64 ± 4.7 g in control data base).
Gross pathology:
- No macroscopic abnormalities were observed at necropsy.
Other findings:
None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The oral LD50 for PI2080 is 300-2000 mg/kg bw in rats therefore it is classified as ‘R22 Harmful if swallowed’ according to the Annex VI to the Directive 67/548/EEC and ‘Category 4’ with a signal word ‘warning’ and the hazard statement ‘H302: Harmful if swallowed’ according to the CLP Regulation (EC) N° (1272-2008).
Executive summary:

In an acute oral toxicity study performed according to OECD Guideline 423 and in compliance with GLP, three groups of three female Sprague Dawley rats were given a single oral (gavage) dose of PI2080 at 300 or 2000 mg/kg bw in DMSO. Since no relevant toxicity data were available for the estimation of a lethal dose-level, the starting dose-level was 300 mg/kg bw for animal welfare reasons. After the first assay, the next higher dose-level of 2000 mg/kg bw was tested. Then, in view of toxicity observed at 2000 mg/kg bw, the results previously obtained at 300 mg/kg bw were confirmed in other females. Each animal was then observed for mortality, clinical signs and bodyweights for 14 days and were all sacrificed for macroscopic examination.

At 2000 mg/kg bw, one female was found dead on Day 6 and another one was sacrificed prematurely on Day 5. Piloerection, hypoactivity, hunched posture, half-closed eyes, soiled urogenital and mouth regions, then dyspnea and lateral recumbency were observed in the prematurely sacrificed female. Piloerection was also observed in the surviving female. At 300 mg/kg bw, no unscheduled deaths that could be related to treatment were observed. Piloerection and ptyalism (3/6 females), hypoactivity and dyspnea (2/6 females), half-closed eyes and chromorhynorrhea (1/6 females) were observed. At terminal sacrifice, no gross findings were observed.

The oral LD50 for PI2080 is 300-2000 mg/kg bw in rats therefore it is classified as ‘R22 Harmful if swallowed’ according to the Annex VI to the Directive 67/548/EEC and ‘Category 4’ with a signal word ‘warning’ and the hazard statement ‘H302: Harmful if swallowed’ according to the CLP Regulation (EC) N° (1272-2008).