N-{[Benzyl(ethyl)amino]-[(chloro-cyano-nitrophenyl)substituted]phenyl}acetamide

Endpoint summary

Description of key information

The oral LD50 for FAT 41047/A TE is greater than 5000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: In-Vivo Biosciences, Kodigehalli Village Magadi Road, Bangalore, Code-29, Karnataka State.
- Females (if applicable) nulliparous and non-pregnant: yes.
- Age at study initiation: 9 to 10 Weeks.
- Weight at study initiation: 180.8 to 214.5 g
- Fasting period before study: overnight.
- Housing: Rats were housed individually in standard polysulfone cages (size: approximately L 425 x B 266 x H 185 mm), with stainless steel top grill having facilities for pelleted food and drinking water in polycarbonate bottle. Additionally, polycarbonate rat huts were placed inside the cage as an enrichment object and were changed along with the cage once a week.
- Bedding: steam sterilized corn cob was used and changed once a week along with the cage.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: After physical examination for good health and suitability for experiment, the animals were acclimatized six days for G1-FTS, eight days for G1-STS, 10 days for G2-FTS and 12 days for G2- STS before treatment. Animals were observed once daily during acclimatization period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24 °C
- Humidity (%): 65 to 67%
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

Route of administration:

oral: gavage

Vehicle:

other:

Remarks:

0.5% w/v sodium carboxymethyl cellulose (medium viscosity) with 1% v/v Tween 80 in Milli-Q water

Details on oral exposure:

Justification for choice of vehicle: Solubility and Suspend ability test was performed, the test item formed homogenous suspension with 0.5% w/v sodium carboxymethyl cellulose (medium viscosity) with 1% v/v Tween 80 in Milli-Q water. Hence, 0.5% w/v sodium carboxymethyl cellulose (medium viscosity) with 1% v/v Tween 80 in Milli-Q water was used as vehicle to prepare the dose formulations.

Dose selection:
Starting dose
There was no toxicology information available about the test item, hence the study was initiated with the starting dose of 300 mg/kg body weight (G1-FTS). The test was started as per Annex 2c of the OECD 423 test guideline.

Additional treatment groups
As there was no test item-related mortality observed at the starting dose of 300 mg/kg body weight (G1-FTS); the test was continued with same dosewith three additional female rats at second treatment step (G1-STS) and again with next two steps (G2-FTS & G2-STS) at the next higher dose of 2000 mg/kg body weight. The subsequent dosing was done at approximately 48 hours after the previous treatment step.

Doses:

G1 - FTS : 300 mg/kg bw - Group 1, First test step.
G1 - STS : 300 mg/kg bw - Group 1, Second test step.
G2 - FTS : 2000 mg/kg bw - Group 2, First test step.
G2 - STS : 2000 mg/kg bw - Group 2, Second test step.

No. of animals per sex per dose:

Three

Control animals:

no

Details on study design:

Duration of observation - 14 days

Frequency of observation : Animals were observed five times on test day 1 (day of administration) i.e. at 30 minutes and four times at hourly intervals and once daily during days 2 to 15 post administration.
Observations included changes in skin and fur, eyes and mucous membranes, and respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behavior pattern. Attention was directed to the observations of tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma and all observed clinical signs were recorded.

The body weights were recorded on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration).

Necropsy : The rats surviving to the end of the observation period were euthanized by using isoflurane anesthesia and subjected to detailed necropsy. Microscopic examination was not carried out as no gross pathological changes were observed.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality observed at 300 & 2000 mg/kg bw dose levels at any step.

Clinical signs:

No clinical signs were observed at any step.

Body weight:

The body weights of all the rats increased throughout the observation period.

Gross pathology:

There were no gross pathological findings at necropsy.

Dose (mg/kg body weight)	Rat No.	Sex	Body weight (Day 1) (g)	Total volume administered (mL)	Mortality
G1 (FTS) 300	Rm8101	F	187.5	1.88	N
	Rm8102	F	214.5	2.15	N
	Rm8103	F	180.8	1.81	N
G1 (STS) 300	Rm8104	F	196.7	1.97	N
	Rm8105	F	204.9	2.05	N
	Rm8106	F	202	2.02	N
G2 (FTS) 2000	Rm8107	F	201.8	2.02	N
	Rm8108	F	199.6	2	N
	Rm8109	F	185.5	1.86	N
G2 (STS) 2000	Rm8110	F	188	1.88	N
	Rm8111	F	201.6	2.02	N
	Rm8112	F	190.8	1.91	N

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for FAT 41047/A TE was found to be greater than 5000 mg/kg.

Executive summary:

The acute oral toxicity study of FAT 41047/A TE to the Wistar rats was evaluated according to OECD 423 test guideline.

The dose formulation was prepared by using 0.5% w/v sodium carboxymethyl cellulose (medium viscosity) with 1% v/v Tween 80 in Milli-Q water and administered as a single oral gavage to overnight fasted (16 to 18 hours) three female rats (G1-FTS) at the dose of 300 mg/kg body weight (at a dose volume of 10 mL/kg body weight). There were no clinical signs and pre-terminal deaths.

Based on the scheme - Annex 2c, three additional female rats were tested at the same dose of 300 mg/kg body weight (G1-STS). There were no clinical signs and pre-terminal deaths. Additional three female rats were tested at next higher dose of 2000 mg/kg body weight (G2 -FTS).

There were no clinical signs and pre-terminal deaths. Hence, three additional female rats were tested at the same dose of 2000 mg/kg body weight (G2-STS). The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and again on days 8 and 15. Necropsy was performed for all the rats at termination. All survived rats gained weight during experimental period. There were no gross pathological findings at necropsy.

Based on the results of the present study, the LD₅₀for FAT 41047/A TE is greater than 5000 mg/kg as per the LD₅₀cut-off value.

The test item does not meet the classification criteria as per Globally Harmonized Classification system as there were no mortality or clinical signs of toxicity observed at 5000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

High quality GLP study

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The acute oral toxicity of FAT 41047/A was assessed in a study conducted on Wistar rats according to OECD 423 test guideline. As no information regarding toxicity of FAT 41047/A was available, the initial dose was chosen to be 300 mg/kg bw as per the guideline. At each step, 3 female rats were used. No pre-terminal deaths and no clinical steps were observed at 300 mg/kg bw at first and second steps. Hence, additional testing at 2000 mg/kg bw was performed. Again, no pre-terminal deaths and no clinical signs were observed at these steps. Hence, the LD50 of FAT 41047 is estimated to be >5000 mg/kg bw.

 

Justification for classification or non-classification

FAT 41047/A was found to have low acute oral toxicity in the study conducted according to OECD TG 423, hence it does not warrant classification as per the Regulation (EC) No. 1272/2008.