1H-Benz[e]indolium, 2-[2-[2-chloro-1-[2-[1,3-dihydro-1,1-dimethyl-3-(4-sulfobutyl)-2H-benz[e]indol-2-ylidene]ethylidene]-1H-inden-3-yl]ethenyl]-1,1-dimethyl-3-(4-sulfobutyl)-, inner salt

Acute Toxicity: oral

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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-[(1E)-2-[(1E)-2-chloro-1-{2-[(2E)-1,1-dimethyl-3-(4-sulfobutyl)-1H,2H,3H-benzo[e]indol-2-ylidene]ethylidene}-1H-inden-3-yl]ethenyl]-1,1-dimethyl-3-(4-sulfonatobutyl)-1H-benzo[e]indol-3-ium

EC Number:

942-705-4

Cas Number:

2151868-08-3

Molecular formula:

C49 H49 Cl N2 O6 S2

IUPAC Name:

2-[(1E)-2-[(1E)-2-chloro-1-{2-[(2E)-1,1-dimethyl-3-(4-sulfobutyl)-1H,2H,3H-benzo[e]indol-2-ylidene]ethylidene}-1H-inden-3-yl]ethenyl]-1,1-dimethyl-3-(4-sulfonatobutyl)-1H-benzo[e]indol-3-ium

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 or test system 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:

other: 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).

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