Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2017-09-05 to 2017-11-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl:CD(SD), SPF
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: ORIENTBIO INC., Republic of Korea
- Age at study initiation: 8 − 9 weeks
- Weight at study initiation: 180.7 − 201.4 g
- Fasting period before study: Animals were fasted overnight, approximately 16 hours prior to dosing.
- Housing: One animal/cage in Stainless wire mesh cage
- Diet: Pelleted rodent chow (Teklad Certified Irradiated Global 18 % Protein Rodent Diet 2918C); Envigo RMS, Inc., U.S.A., ad libitum
- Water: Public tap water, ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5 − 23.0
- Humidity (%): 46.0 − 58.3
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 60 and 400 mg/mL
- Amount of vehicle: 5 mL/kg bw
- Justification for choice of vehicle: As a result of the vehicle review, the test substance was not dissolved in water for injection, and it was not uniformly suspended. Therefore, corn oil was selected as the vehicle since the test substance was soluble in it.
- Lot no: MKCC0462

MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg bw

DOSAGE PREPARATION: The required amount of the test substance was weighed using an electronic balance (CP423S, Sartorius, Germany) and placed in a bottle. A small amount of vehicle, corn oil, was added and mixed using a vortex mixer until dissolved. The vehicle was gradually added to yield the desired concentrations (60 and 400 mg/mL). All preparations were conducted just prior to use.

CLASS METHOD
- Rationale for the selection of the starting dose: The starting dose level for this study was selected at 300 mg/kg because there were no available toxicity information on the test substance.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
2 x 3 females per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed for mortality, general condition and clinical signs (type, severity, time of onset and recovery) at 30 minutes after dosing and at 1, 2, 4 and 6 hours after dosing on Day 0 and once daily thereafter for 14 days (Day 1−Day 14). The body weight was recorded prior to dosing on Day 0 and on Days 1, 3, 7 and on the day of necropsy (Day 14).
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology
Statistics:
Statistical analysis was not performed. Mean scores and values were determined.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
2 500 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths of animals at 300 mg/kg bw throughout the study. Two animals at 2000 mg/kg bw were found dead on Day 1 and 4. The death was considered to be a test substance-related effect
Clinical signs:
At 300 mg/kg bw, abnormal gait was observed in all animals at 0.5, 1, 2, 4, and 6 hours after dosing, and lacrimation was observed in 4 − 5 animals at 1 and 2 hours after dosing, and they disappeared on Day 1.
At 2000 mg/kg bw, abnormal gait was observed in all animals at 0.5, 1, 2, 4, and 6 hours after dosing, and salivation and/or lacrimation were observed in 1 − 6 animals at 0.5, 1, 2, and 4 hours after dosing. Then, one animal was found dead in a state of prone position on Day 1. A decrease of fecal volume, chromaturia (red), decrease in locomotor activity and/or lacrimation were observed in the other animal on Days 1 − 3, and the animal was found dead in a state of prone position on Day 4. In other surviving animals, no stool, decrease of fecal volume, abnormal gait and/or decrease in locomotor activity were observed in 1 − 4 animals on Days 1 − 3 after dosing, and they disappeared on Day 4.
These clinical signs at 300 and 2000 mg/kg bw were considered to be test substance-related effects.
Body weight:
At 300 mg/kg bw, a tendency for suppression of body weight gain was observed three animals on Day 1.
In surviving animals at 2000 mg/kg bw, a tendency for suppression of body weight gain was observed in two animals on Day 1 and a decrease in body weight was observed in two animals each on Day 1 and Day 3. Then, normal body weight gain was observed in these animals on Day 7. In one of dead animals at 2000 mg/kg bw, a decrease in body weight was observed on Days 1 and 3.
These body weights changes at 300 and 2000 mg/kg bw were considered to be test substance-related effects.
Gross pathology:
No grossly visible findings were observed in any animal at 300 mg/kg bw.
Enlarged adrenal, small spleen, small thymus and multiple black spots in the glandular stomach were noted in one dead animal at 2000 mg/kg bw, and these changes were considered to be stress-related changes.

Any other information on results incl. tables

Table 1: Summary of Mortality

Step /Dose (mg/kg bw)

No of Animals

Days after Dosing

Mortality

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Step 1

300

3

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/3

Step 2

300

3

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/3

Step 1

2000

3

0

0

0

0

1

0

0

0

0

0

0

0

0

0

0

1/3

Step 2

2000

3

0

1

0

0

0

0

0

0

0

0

0

0

0

0

0

1/3

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
In an acute oral toxicity study according to OECD Guideline 423 in Sprague-Dawley rats, the test item was classified as Category 5 according to the GHS classification and the median lethal dose derived was LD50cut off = 2,500 mg/kg bw
Executive summary:

In an acute oral toxicity study according to OECD Guideline 423 the potential toxicity of the test item following a single oral dose administration (gavage) to female Sprague-Dawley rats was assessed. All animals were monitored for clinical signs and body weight changes during the 14-day observation period after administration. They were subjected to a gross necropsy at the end of the observation period. A dose of 300 mg/kg bw was sequentially administered to groups of 3 femals. There were no deaths of animals at 300 mg/kg. Abnormal gait and/or lacrimation were observed in animals at 300 mg/kg on the day of dosing, and the observations disappeared on Day 1. A tendency for suppression of body weight gain was observed in three animals on Day 1. No test substance-related effects were observed in necropsy findings in any animal at 300 mg/kg. Following the guideline, a dose of 2000 mg/kg bw was sequentially administered to groups of 3 femals. Two animals were found dead at 2000 mg/kg bw on Days 1 and 4. Abnormal gait, salivation and/or lacrimation were observed in animals on the day of dosing. Then, one animal was found dead on Day 1. A decrease of fecal volume, chromaturia (red), decrease in locomotor activity and/or lacrimation were observed in the other animal on Days 1 − 3, and the animal was found dead on Day 4. In other surviving animals, no stool, decrease of fecal volume, abnormal gait and/or decrease in locomotor activity were observed in animals on Days 1 − 3 after dosing, and the observations disappeared on Day 4. In the surviving animals at 2000 mg/kg bw, a tendency for suppression of body weight gain was observed in two animals on Day 1 and a decrease in body weight was observed in two animals each on Day 1 and Day 3. Then, normal body weight gain was observed in these animals on Day 7. In one of dead animals at 2000 mg/kg bw, a decrease in body weight was observed on Days 1 and 3. Macroscopic examination revealed enlarged adrenal, small spleen, small thymus and multiple black spots in the glandular stomach in one dead animal at 2000 mg/kg bw.

Based on the result of the acute oral toxicity study in Sprague-Dawley rats, the test item, was classified as Category 5 according to the GHS classification and the median lethal dose derived was LD50 cut off = 2,500 mg/kg bw.