5-Amino-N,N'-Bis(2,3-Dihydroxypropyl)-2,4,6-Triiodoisophtalamide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20.1. 2020 – 5.2. 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

474-080-4

EC Name:

-

Cas Number:

76801-93-9

Molecular formula:

C14H18I3N3O6

IUPAC Name:

5-amino-N1,N3-bis(2,3-dihydroxypropyl)-2,4,6-triiodobenzene-1,3-dicarboxamide

impurity 1

Reference substance name:

Unknown impurities

Molecular formula:

Not available

IUPAC Name:

Unknown impurities

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 00104016
- Expiration date of the lot/batch: 12/2020

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in cool place. Keep container tightly closed in a dry and well-ventilated place.
- Stability under test conditions: Stable

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SPF breeding, VELAZ s.r.o., Lysolajské údolí 15/53, 165 00 Prague 6, Czech Republic, RČH CZ 11760500
- Females (if applicable) nulliparous and non-pregnant: yes
- Weight at study initiation: 160-224 g
- Fasting period before study: 20 hrs
- Housing: three animals per cage
- Diet (e.g. ad libitum): Pelleted standard diet for experimental animals ad libitum (Brand - Altromin International)
- Water (e.g. ad libitum): Drinking tap water
- Acclimation period: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Room temperature 22 ± 3 C, permanently monitored
- Humidity (%): Relative humidity 30 – 70 %, permanently monitored
- Photoperiod (hrs dark / hrs light): Light: 12 hours light/dark cycle: 6am-6pm/6pm-6am

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
- Lot/batch no. (if required): Olivae oleum raffinatum, Batch No.: 8003998001, Expiration date: 01/2020, Supplier: Dr. Kulich Pharma, s.r.o. Hradec Králové, Czech Republic

MAXIMUM DOSE VOLUME APPLIED: 1 mL/100 g of animal body weight

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The dose level of 300 mg/kg of body weight was used as the starting dose, according to the test guideline, because there is no information about toxicity.

Doses:

300 mg/kg bw (group 1) and 2000 mg/kg bw (group 2 and 3)

No. of animals per sex per dose:

3 females per group

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Body weight: before application, 8th day and before euthanasia of animals
- Mortality: daily
- Clinical examination: daily
After application the animals were observed individually:
- the first day: twice (30 minutes and 3 hours after application)
- the second day: twice (in the morning and in the afternoon) and daily thereafter for 14 days.
Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, and presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system.
- Pathological examination: 15th day
- Necropsy of survivors performed: yes
- Testing schedule: START: 300 mg/kg bw - 3 females (Step No. 1) - no deaths ► 2000 mg/kg bw – 3 females (Step No. 2) – no deaths ►2000 mg/kg bw - 3 females (Step No. 3) - no deaths ► END of study.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Clinical signs:

No clinical signs of intoxication were observed in all 3 females at the dose 300 mg/kg (group No. 1) during the clinical observation after the application.
No clinical signs of intoxication were observed in all 6 females at the dose 2000 mg/kg (group No. 2 and No. 3) during the clinical observation after the application.

Body weight:

Weight increments in groups No. 1, 2 and 3 were adequate to species, sex and age of animals in experiment.

Gross pathology:

No pathological macroscopic changes were diagnosed during pathological examination of animals at the dose 300 mg/kg in group No. 1.
No pathological macroscopic changes were diagnosed during pathological examination of animals at the dose 2000 mg/kg in group No. 2 and No. 3.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item toxicity was evaluated on the basis of mortality, clinical signs of intoxication, body weight increments during the observation period and necropsy findings at the end of study.
The test item administered at the doses of 300 and 2000 mg/kg of body weight caused no death of any animal. No serious clinical signs of intoxication were detected at these doses during the whole study. Weight increments were adequate to species, sex and age of animals in experiment. No pathological macroscopic changes were diagnosed during the pathological examination.
According to the study results, the LD50 value of the test item, ATIBA-A, for female rats is higher than 2000 mg/kg of body weight.

Executive summary:

The aim of the study was to investigate acute toxic effects of the test item ATIBA-A, after a single oral administration to Wistar Han rats.

The testing was performed according Method B.1 tris: Acute Oral Toxicity - Acute Toxic Class Method, Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

The test item was administered in a single dose as a suspension in olive oil, given orally via gavage to female Wistar rats. The volume of administered suspension was 1 ml/100 g body weight of animals.

The dosing was performed sequentially in three groups of three females: Group No. 1 (the first step) using the starting dose of 300 mg/kg of body weight, this dose caused no death of animals. Therefore the dose of 2000 mg/kg of body weight was applied to another three animals (Group No. 2) in the second step. Afterwards the dose of 2000 mg/kg of body weight was applied for confirmation to another three animals (Group No. 3) in the third step.

The test item administered at the dose of 2000 mg/kg of body weight caused no death of all 6 females in group No. 2 and No. 3. No clinical signs of intoxication were detected during the whole study in 6 animals. No pathological macroscopic changes were diagnosed during pathological examination.

According to the study results, the LD50 value of the test item, ATIBA-A, for female rats is higher than 2000 mg/ kg of body weight.