3-Quinolinecarboxylic acid, 7-[6-(benzoylmethylamino)-5-methyl-3-pyridinyl]-1-cyclopropyl-1,4-dihydro-8-methyl-4-oxo-, ethyl ester

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 August 2020 - 19 August 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

yes

Remarks:

A relative humidity higher than 70% was registered on 15 and 16 August 2020. The maximum value measured was 72%. As no effect was noted on the health of the animals, this deviation is considered as without impact on the conclusion of the study.

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3-Quinolinecarboxylic acid, 7-[6-(benzoylmethylamino)-5-methyl-3-pyridinyl]-1-cyclopropyl-1,4-dihydro-8-methyl-4-oxo-, ethyl ester

EC Number:

610-204-7

Cas Number:

446299-90-7

Molecular formula:

C30H29N3O4

IUPAC Name:

3-Quinolinecarboxylic acid, 7-[6-(benzoylmethylamino)-5-methyl-3-pyridinyl]-1-cyclopropyl-1,4-dihydro-8-methyl-4-oxo-, ethyl ester

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER LABS (53940 Le Genest St Isle – France)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 weeks
- Weight at study initiation: 212.0 g (SD = 7.4 g)
- Fasting period before study: Food was removed on D-1 and then redistributed 4 hours after the test item administration.
- Housing: Rats were housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains sawdust bedding which was changed at least 2 times a week. Each cage was installed in conventional air conditioned animal husbandry.
- Diet (e.g. ad libitum): Ad libitum. Teklad Global 16% Protein Rodent Diet (ENVIGO 2016).
- Water (e.g. ad libitum): Ad libitum. Drinking water (tap-water from public distribution system). Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas – Eurofins (FRANCE).
- Acclimation period: the animals were acclimatized for at least 5 days before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19ºC to 25ºC
- Humidity (%): 30 to 70%
- Air changes (per hr): ≥ 10 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light (07.00 to 19.00) and 12 hours dark cycle.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: Olive oil was chosen as it produced the most suitable formulation at the requested concentration.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION (if unusual): The preparations were stirred by vortex to obtain yellow homogeneous suspensions just before the administration.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Step 1 (2000 mg/kg bw): 1 animal
Step 2 (2000 mg/kg bw): 4 animals.

Control animals:

yes

Remarks:

(study performed on three animals receiving olive oil under requirements of OECD Guideline 423)

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At each step, the animals were observed four times on test day 0 (day of administration), i.e. at T0+30 min, T0+1h, T0+3h and T0+4h, and once daily during days 1 to 14 post administration. The body weights were recorded on test day 0 (just before administration) then on D2, D7, and D14.
- Necropsy of survivors performed: yes.
- Clinical signs including body weight: spontaneous activity, Preyer’s reflex (noise), respiratory rate, convulsions, tremors, body temperature, muscle tone, palpebral opening, pupil appearance, salivation, lachrymation, righting reflex, back hair appearance. The body weight evolution of animals treated with the test item was compared with the body weight evolution of the control group.
- Other examinations performed: Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. No organ was removed and preserved in view to microscopic examinations.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: No clinical signs related to the administration of the test item were observed during the study.

Gross pathology:

The macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

Any other information on results incl. tables

Table 1: Body weight and weight gain in grams. Steps 1 & 2 (2000 mg/kg bw)

FEMALES	D0	D2	D2-D0	D7	D7-D0	D14	D14-D0
Rf5082	208	222	14	248	40	267	59
Rf5093	223	234	11	250	27	265	42
Rf5094	216	220	4	231	15	245	29
Rf5095	208	217	9	233	25	235	27
Rf5096	205	213	8	220	15	239	34
MEAN	212.0	221.2	9.2	236.4	24.4	250.2	38.2
Standard deviation	7.4	7.9	3.7	12.5	10.3	14.9	13.0

Applicant's summary and conclusion

Interpretation of results:

other: No category (CLP Regulation EC no. 1272/2008)

Conclusions:

The LD50 of the test item is higher than 2000 mg/ kg body weight by oral route in the rat.

Executive summary:

The acute oral toxicity of the test item has been tested in accordance with OECD Test Guideline 420, following GLP. 5 female Sprague-Dawley rats were administered by oral gavage in 2 steps with test item diluted in vehicle olive oil. The sighting study was performed at the dose of 2000 mg/kg bw in one female rat. As no effects were observed in the preliminary test, the subsequent main study was performed with 4 females administered with test item at the same dose of 2000 mg/kg bw. No mortality occurred during the study. No clinical signs related to the administration of the test item were observed during the study. The body weight evolution of the animals remained normal throughout the study. The macroscopic examination of the animals at the end of the study did not reveal treatment related changes. Based on these results, the LD50 of the test item is determined to be higher than 2000 mg/ kg bw by oral route in the rat.