Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
31 May 2018 - 21 Jun 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study was conducted in accordance with international guidelines and in accordance with GLP. All guideline validity criteria were met.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Version / remarks:
Commission Regulation (EC) No. 440/2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, No. 2-1-1 "Acute oral toxicity studies", 12 Nousan No. 8147
Version / remarks:
24 Nov 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature in the dark
- Stability under test conditions: Not reported
- Solubility and stability of the test substance in the solvent/vehicle: Not reported
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not reported

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item formulated in arachis oil BP within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level.
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: Not reported
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material): Light yellow liquid

OTHER SPECIFICS: Homogeneity of formulation confirmed by visual inspection.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Oxon, UK
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: body weight variation did not exceed ±20% of the mean body weight at the start of treatment
- Fasting period before study: yes overnight and for approximately 3 - 4 hours post dosing
- Housing: housed in groups of up to 4 individuals in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet (e.g. ad libitum): free access to food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK)
- Water (e.g. ad libitum): free access to mains drinking water
- Acclimation period: minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 ºC
- Humidity (%): 30 - 70 %
- Air changes (per hr): minimum of 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 : 12

IN-LIFE DATES: Not clarified in the study report

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
arachis oil
Remarks:
Arachis oil BP used for 300 mg/kg bw dose only
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg total
- Justification for choice of vehicle: Arachic oil was used because the test item did not dissolve/suspend in distilled water
- Lot/batch no. (if required): n/d
- Purity: n/d

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw (300 mg/kg bw) and 2.24 mL/kg bw (2000 mg/kg bw)

DOSAGE PREPARATION (if unusual): applied unchanged

CLASS METHOD (if applicable) - a fixed dose procedure was used
- Rationale for the selection of the starting dose: In the absence of data regarding toxicity of the test item, 300 mg/kg bw was chosen as the start dose for one female. In the absence of effects at this dose level a further female was tested at 2000 mg/kg bw. As not effects were found here a further 4 females were tested at the maximum guideline required concentration of 2000 mg/kg bw. This is in accordance with the OECD 420 (2001).
Doses:
300, 2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Each animal was given single dose of 10 mL/kg dose volume of 2000 mg/kg of the test item of 30 mg/mL concentration.
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 0.5, 1, 2 and 4 hours after dosing then daily thereafter until day 14 post-dose.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs and body weight
Statistics:
not required

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No deaths reported
Clinical signs:
There were no signs of systemic toxicity noted during the course of the study with excepttion of hunched posture was noted within four hours post dosing in animals treated at a dose level of 300 mg/kg and four animal treated at 2000 mg/kg.
Body weight:
All animals showed expected weight gain. The animal dosed at 300 mg/kg bw gained 16 g in the first week and 11 g in the second week (27 g total). The animals dosed at 200 mg/kg bw gained 16 g in the first week and 11 g in the second week (27 g total).
Gross pathology:
No abnormality noted at necropsy

Any other information on results incl. tables

Table 1:   Number of animals dead (and with evident toxicity)

Dose

(mg/kg bw)

Mortality

(# dead / total)

Time range of deaths

(hours)

Number with evident toxicity

(# / total)

Male

Female

Combined

Male

Female

Combined

300

-

0 / 5

0 / 5

n/a

 

0 / 5

0 / 5

2000

-

0 / 5

0 / 5

n/a

-

0 / 5

0 / 5

Table 2:   Number of animals hunched at observations time x

Dose

(mg/kg bw)

Hunched

(# hunched / total)

Time (h)

Male

Female

Combined

300

0.5

-

0 / 1

0 / 1

1

 

1 / 1

1 / 1

2

 

1 / 1

1 / 1

3

 

1 / 1

1 / 1

4

 

1 / 1

1 / 1

1-14 (days)

 

0 / 1

0 / 1

2000

0.5

 

4 / 5

4 /5

1

 

4 / 5

4 /5

2

 

4 / 5

4 /5

3

 

4 / 5

4 /5

4

 

4 / 5

4 /5

1-14

-

0 / 5

0 / 5

Table 3:   Body weight gain (g)

Body weight gain (g)

Animal #

Week 1

Week 2

Female 1-0

16

11

Female 2-0

19

16

Female 3-0

23

16

Female 3-1

23

27

Female 3-2

27

9

Female 3-3

27

14

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System - Unclassified).
Executive summary:

OECD 420 (2018) - In an acute oral toxicity study, a group of fasted, 8-12 week old female Wistar rats were given a single oral dose of AD-464 at a single dose rate of 2000 mg/kg bw (limit test) and observed for 14 days. A sighting study in line with the guideline with 1 animal dosed at 300 mg/kg bw was also conducted.

In the absence of mortality during the observation period, the oral LD50 was estimated to be greater than 2000 mg/kg bw.

In addition, there were no treatment related clinical signs, necropsy findings or changes in body weight observed in any of the individuals.

In conclusion, the test item, AD-464 did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixture.