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Toxicological information

Acute Toxicity: other routes

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Administrative data

Endpoint:
acute toxicity: other routes
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
from 21 August, 2007 to 4 September 2007
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: in compliance with GLP, OECD guideline No. 423 (as the data is used in a read-across approach, a maximal reliability score of 2 was attributed).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD Guideline 423 (Acute toxicity - Acute toxic Class Method)
Deviations:
yes
Remarks:
Analysis of results, Subsection 1.10.2, page 23: No statistical analysis was performed on body weight for animals dosed with LAB 3822 at 2 g/kg of DEI. Reason: Most of animals were found dead on D1.
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: LAB 3822.
Supplier: ROQUETTE FRERES.
Batch Number: FAL 07/25.
Galenic form: slightly yellowish liquid.
Molecular weight (salt form): not applicable.
Molecular weight (base form): about 426 g/mol.
Salt/Base ratio: not applicable.
Expiry date: March 2009.
On 17.Jul.2007, 26.2 kg samples of test item were received, in vial labelled ”LAB 3822, batch No.FAL 07/25”. The Sponsor confirmed that ”LAB 3822 - Di-ester-isosorbide” in the certificate of analysis corresponds to ”LAB 3822”, name used throughout the study plan.
Storage condition: Immediately upon receipt, LAB 3822 was registered, then stored at ambient temperature in accordance with the Sponsor's instructions.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories - Domaines des Oncins, BP 0109, 69592 L'Arbresle Cedex, France.

- Age at study initiation: 8-9 weeks on the day of randomisation.

- Weight at study initiation:
• Between 210.2 g and 241.5 g on the day of randomisation for females.
• Between 271.8 g and 299.8 g on the day of randomisation for males.

- Fasting period before study: Food and water consumption was not measured.

- Housing:Observations were performed at the time of delivery of the animals and daily during the period of acclimatisation. Animals were housed in cages of standard dimensions with sawdust bedding (SAFE, Reference B8/20). Cages were cleaned according to CERB internal SOPs.

- Diet (e.g. ad libitum): RM1 (E)-SQC SDS/DIETEX feed (quality controlled/radiation sterilized) was available ad libitum except during the fasting experimental period. The criteria for acceptable levels of contaminants in the feed supplied were within the limits of the analytical specifications established by the diet manufacturer.

- Water (e.g. ad libitum):Drinking water was available ad libitum in polycarbonate feeder bottles with a stainless steel nipple. A specimen of water is obtained every 6 months and sent to the Laboratoire Départemental d0Analyse du Cher - 216, Rue Louis Mallet - 18014 Bourges Cedex, France, for analysis.
The criteria for acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.

- Acclimation period:Minimum of five days before treatment in the laboratory animal house where the experiment took place. Only animals without any visible sign of illness were used for the study.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): in an air-conditioned 20-24°C
- Humidity (%): between 45% and 65%
- Air changes (per hr): 10 times per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours darkness

Administration / exposure

Route of administration:
intravenous
Vehicle:
water
Doses:
0;0.25 ; 0.5; 1; 2 g/kg of DEI
No. of animals per sex per dose:
10 SPF Sprague-Dawley rats (each of 5 males and 5 females)
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:at least once a day
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs

Results and discussion

Mortality:
On D1, 1 hour after treatment, 3 males dosed with LAB 3822 at 1 g/kg of DEI, all males and 4 females dosed with LAB 3822 at 2 g/kg of DEI were found dead
Clinical signs:
On D1, 1 hour post-dosing:
- males dosed with LAB 3822 at 0.5 g/kg of DEI had mainly an absence or reduced spontanous locomotor activity, absence of reactivity and a decrease in body, abdominal and limb tone
- females dosed with LAB 3822 at 1 g/kg of DEI had mainly absence or reduced spontaneous locomotor activity, recumbent position, absence of reactivity and decreases in body, abdominal and limb tone.
On D1, 3-4 hours post-dosing, all females dosed with LAB 3822 at 1 g/kg of DEI showed lack of spontaneous locomotor activity and polypnoea.

Body weight:
No effect on body weight gain was seen in animals dosed with LAB 3822 whatever the dose when compared with the control group
Gross pathology:
No gross lesion was seen in organs examined at necropsy.

Applicant's summary and conclusion

Conclusions:
Under the experimental conditions adopted, LAB 3822 (batch FAL 07/25) induced mortality in male Sprague-Dawley rats when administered once by the intravenous route at 1 g/kg and 2 g/kg of DEI and in female Sprague-Dawley rats when administered once by the intravenous route at 2 g/kg of DEI. Moreover, signs of toxicity began to appear in the male Sprague-Dawley rats dosed with LAB 3822 at 0.5 g/kg of DEI and in the female Sprague-Dawley rats dosed with LAB 3822 at 1 g/kg of DEI.
As a result, the Maximum Non-Lethal Dose is 0.5 g/kg of DEI (0.575 g/kg of LAB 3822) in male Sprague-Dawley rats, and 1 g/kg of DEI (1.15 g/kg of LAB 3822) in female Sprague-Dawley rats.
Executive summary:

Toxicity of the test item LAB 3822 (batch FAL 07/25) was evaluated after a single administration by the intravenous route in the rat.

The study involved 5 groups of 10 SPF Sprague-Dawley rats (each of 5 males and 5 females), 8 to 9 weeks old and weighing between 271.8 g and 299.8 g for males and between 210.2 g and 241.5 g for females on the day of randomisation. Animals were purchased from Charles River Laboratories France (Domaine des Oncins - 69592 L0Arbresle Cedex, France).

Groups were as follows:

• Group 1: control group dosed with sterile water.

• Group 2: dosed with LAB 3822 at the dose of 0.25 g/kg of DEI (0.29 g/kg of LAB 3822).

• Group 3: dosed with LAB 3822 at the dose of 0.5 g/kg of DEI (0.575 g/kg of LAB 3822).

• Group 4: dosed with LAB 3822 at the dose of 1 g/kg of DEI (1.15 g/kg of LAB 3822).

• Group 5: dosed with LAB 3822 at the dose of 2 g/kg of DEI (2.3 g/kg of LAB 3822).

LAB 3822 or sterile water were administered by the intravenous route at the volume below:

• Group 1: 2.35 mL/kg.

• Group 2: 0.30 mL/kg.

• Group 3: 0.59 mL/kg.

• Group 4: 1.17 mL/kg.

• Group 5: 2.35 mL/kg.

Experimental procedure:

Animals were weighed on the day of randomisation (D-1), on D7, D14 and D15. Mortality was recorded twice a day for 14 days. General observations were performed on D1, 60 minutes ± 30 minutes after dosing, again between 3 and 4 hours post-dose and then once a day for 14 days. Functional and neurobehavioural tests were also assessed on D1 60 minutes ±30 minutes after dosing and then on D7. All animals surviving at the end of the 14-day period were submitted to gross necropsy.

Results:

No mortality was seen in animals dosed with sterile water.

No effect on body weight gain, no clinical sign and no abnormality in organs examined at the necropsy was seen in animals dosed with sterile water.

On D1, 1 hour after treatment, 3 males dosed with LAB 3822 at 1 g/kg of DEI, all males and 4 females dosed with LAB 3822 at 2 g/kg of DEI were found dead.

No effect on body weight gain was seen in animals dosed with LAB 3822 whatever the dose when compared with the control group. On D1, 1 hour post-dosing

• males dosed with LAB 3822 at 0.5 g/kg of DEI had mainly an absence or reduced spontaneous locomotor activity, absence of reactivity and a decrease in body, abdominal and limb tone.

• females dosed with LAB 3822 at 1 g/kg of DEI had mainly absence or reduced spontaneous locomotor activity, recumbent position, absence of reactivity and decreases in body, abdominal and limb tone.

On D1, 3-4 hours post-dosing, all females dosed with LAB 3822 at 1 g/kg of DEI showed lack of spontaneous locomotor activity and polypnoea.

No gross lesion was seen in organs examined at necropsy.

Under the experimental conditions adopted, LAB 3822 (batch FAL 07/25) induced mortality in male Sprague-Dawley rats when administered once by the intravenous route at 1 g/kg and 2 g/kg of DEI and in female Sprague-Dawley rats when administered once by the intravenous route at 2 g/kg of DEI. Moreover, signs of toxicity began to appear in the male Sprague-Dawley rats dosed with LAB 3822 at 0.5 g/kg of DEI and in the female Sprague-Dawley rats dosed with LAB 3822 at 1 g/kg of DEI.

As a result, the Maximum Non-Lethal Dose is 0.5 g/kg of DEI (0.575 g/kg of LAB 3822) in male Sprague-Dawley rats, and 1 g/kg of DEI (1.15 g/kg of LAB 3822) in female Sprague-Dawley rats.