Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: According to official guidelines

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Identification: V516690 -- Appearance: Brown to green powder -- Batch: V516690/AY -- Purity/Composition: LC-MSpurity (Area% UV-VIS): 95.8% (Reference: PA064492) -- Test item storage: At room temperature stable under storage conditions until 17 July 2016 (expiry date) -- Molecular formula: C49H49ClN2O6S2 -- MW: 861.52

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
The Sprague Dawley SD rat was used, being a species and strain accepted by many regulatory authorities and since there is ample experience and background data. The oral route of administration is a potential route of exposure during manufacturing, handling or use of the substance.

Procedures and facilities were compliant with the requirements of the Directive 2010/63/EU on the protection of animals used for scientific purposes. The national transposition of the Directive is defined in Decreto Legislativo 26/2014.
RTC test facility is fully accredited by AAALAC. Aspects of the protocol concerning animal welfare have been approved by RTC animal-welfare body.

Age & sex: 6-7 weeks old female rats (nulliparous and non-pregnant)
Supplier: Envigo RMS srl, San Pietro al Natisone (UD), Italy
Weight range at arrival: 157.0 - 170.3 grams
Acclimation period: at least 5 days
Veterinary health check during the accimatisation period

Housing: polisulphone solid bottomed cages measuring 59.5 x 38 x 20 cm with nesting material provided into suitable bedding bags
Animals per cage: 3 during the study; up to 5 during the acclimatisation


Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
The test item was dissolved/suspended in a 0.5% aqueous solution of carboxy- methylcellulose.
Details on oral exposure:
Frequency of treatment: Animals were dosed once only on Day 1.
Fasting procedure: Food was removed from the cages overnight prior to dosing (Day -1) and was made available approximately 4 hours after dosing.
Dose calculation: On the day of dosing (Day 1), the amount of the formulated test item to be administered was calculated for each fasted animal according to body weight.
Dosing method: The formulated test item was administered, by gavage, at a dose volume of 20 mL/kg using a plastic feeding tube attached to a graded syringe.
Doses:
2000 mg/kg
No. of animals per sex per dose:
3 per experiment
Group 1 (step 1): Rat numbers 1, 3, 5
Group 2 (step 2): Rat numbers 7, 9, 11
Details on study design:
Random at arrival. The body weight of each individual was within 20% of the group’s mean.
Animals were unequivocally numbered within the study.
The animal number together with the study number ensured a unique animal numbering for any study employing computerised data collection. The computer system used in this study was Pristima version 6.3.2.

Mortality and morbidity: Throughout the study all animals were checked twice daily.

Clinical signs: Animals were observed for clinical signs as indicated below
Session 2: approximately 0.5 hour after dosing · Session 3: approximately 2 hours after dosing · Session 4: approximately 4 hours after dosing
Daily thereafter for a total of 14 days (Session 1). Body weight
All animals were weighed at allocation to the study (Day -1), on the day of dosing (Day 1) and on Days 2, 8 and 15.
Body weight change calculated for Days 2, 8 and 15 of the dosing phase was relevant to Day 1 of the phase.

Termination: All animals were sacrificed on Day 15.
Euthanasia method: Animals were sacrificed by carbon dioxide narcosis.
Necropsy procedure: Necropsy was carried out on all animals (gross necropsy examination for both external and internal abnormalities, with particular attention to the gastro-intestinal tract).

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD0
Effect level:
>= 2 000 other: mg/kg
Based on:
test mat.
Mortality:
No mortality occurred in the first group of animals initially dosed at 2000 mg/kg (Group 1, Step 1) and in the further group of 3 females dosed at the same dose level (Group 2, Step 2).
Clinical signs:
No clinical signs were observed in the first group of animals initially dosed at 2000 mg/kg (Group 1, Step 1) and in the further group of 3 females dosed at the same dose level (Group 2, Step 2).
Body weight:
Changes in body weight observed during the study were within the expected range for this strain and age of animals.
Gross pathology:
No abnormalities were observed at necropsy examination performed on all animals dosed at 2000 mg/kg (Groups 1 and 2) at the end of the observation period.

Applicant's summary and conclusion

Conclusions:
The acute toxicity of V516690 was investigated following a single oral administration (20 mL/kg in 0.5% aqueous solution of carboxymethylcellulose) to the Sprague Dawley rat followed by a 14-day observation period. No mortality occurred and no signs of toxicity were observed in the 6 animals following dosing at 2000 mg/kg. These results indicate that the test item V516690 did not induce toxic effects in the rat following oral administration of a single dose at a level of 2000 mg/kg. The lack of mortality demonstrates the acute toxicity expected (ATE) to be greater than 2000 mg/kg body weight. European Directives concerning the classification, packaging and labelling of dangerous substances (Council Regulation (EC) No. 1272/2008 and subsequent revisions) would indicate the following: Classification: not required Signal word: none indicated Hazard statement: none indicated