Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental starting date: 29 November 2018 and experimental completion date: 28 December 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
Adopted 17 December 2001
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
Name: XTJ-785, experimental
Lot No.: 9570-2-6738
CAS No.: Not listed
Purity: >92% C1214 alcohol, propoxylated, aminated, ethyoxylated
Expiry Date: No date established

Test animals

Species:
rat
Strain:
other: WIST albino
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: ENVIGO RMS (UK) Ltd.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 12 weeks of age prior to dosing (Day 1).
- Weight at study initiation: 156 to 169 grams (the body weight variation did not exceed +/- 20% of the mean body weight of any previously treated animal.
- Fasting period before study: Yes, for overnight prior to and approximately four hours after dosing.
- Housing: the cages were solid bottomed polycarbonate cages with a stainless steel mesh lid.
- Diet (e.g. ad libitum): free access to standard rodent diet.
- Water (e.g. ad libitum): free access of potable water taken from public supply via polycarbonate bottles fitted with sipper tubes.
- Acclimation period: five days before treatment
Each animal was identified uniquely within the study by tail marking.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 40% to 70 %
- Air changes (per hr): The animal room was kept at positive pressure with respect to the outside by its own supply of filtered fresh air, which was passed to atmosphere and not re-circulated.
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours continuous dark per 24 hours.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
The test item was formulated at concentrations of 30 and 200 mg/mL in the vehicle and administered at a volume of 10 mL/kg body weight.
The test item formulations were prepared on the day of dosing

Doses:
300 and 2000 mg/kg
No. of animals per sex per dose:
- First step: 2 single animals were treaated at 300 and 2000 mg/kg.

- Second step: additonnal group of 4 rats treated at 3000 mg/kg.

A total of 5 animals were therefore treated at a dose level of 300 mg/kg in the study and one rat at 2000 mg/kg.
Control animals:
no
Details on study design:
STUDY DESIGN:
In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
Two single animals were treated as follows:
Dose level (mg/kg) Concentration (mg/mL) Dose volume (mL/kg) Number of rats Female
300 30 10 1
2000 200 10 1


Due to mortality and signs of systemic toxicity at a dose level of 2000 mg/kg, an additional group of animals was treated as follows:
Dose level (mg/kg) Concentration (mg/mL) Dose volume (mL/kg) Number of rats Female
300 30 10 4

A total of five animals were therefore treated at a dose level of 300 mg/kg in the study.

- Duration of observation period following administration: 14 days


SERIAL OBSERVATIONS:
- Mortality: Cages of rats were checked at least twice daily for any mortalities.

- Clinical observations: Animals were observed soon after dosing and at frequent intervals (at least 0.5, 1, 2 and 4 hours after dosing) on Day 1. On subsequent days, surviving animals were observed at least once in the morning and again at the end of the experimental day (with the exception of Day 15 - morning only). The nature and severity, where appropriate, of the clinical signs and the time were recorded at each observation.

- Body Weight: The weight of each rat was recorded on Days 1 (prior to dosing), 8 and 15 or at death. Individual weekly body weight changes and group mean body weights were calculated.

- Method of Kill: All surviving animals were humanely killed on Day 15 by carbon dioxide asphyxiation.

- Macroscopic Pathology: All animals were subject to a macroscopic examination which consisted of opening the cranial, thoracic and abdominal cavities. The macroscopic appearance of the brain, caecum, duodenum, heart, kidneys, small and large intestine, liver, lungs and bronchi, spleen, stomach, subcutaneous tissue and urinary bladder was recorded.
Statistics:
The computer systems that were used on this study to acquire and quantify data include: XYBION Pristima: used for pharmacy test item managment.

Results and discussion

Preliminary study:
not available
Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 300 - <= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Clinical signs prior to death comprised piloerection, elevated and unsteady gait, reduced body temperature, blue extremities and underactivity. These signs were seen on Day 2 of treatment. A loss in body weight was noted. Macroscopic examination of the animal revealed congestion (characterized by darkened tissues/organs) of the heart, liver, kidneys, stomach, duodenum and small intestines, enlarged, swollen, or thickened tissues of the stomach, duodenum, caecum and small and large intestines. A red fluid was seen in the caecum and small and large intestines. Gaseous distension of the caecum was also observed.
Clinical signs:
There were no clinical signs of reaction to treatment for animals dosed at 300 mg/kg in the sighting study or main study.
Body weight:
All animals dosed at 300 mg/kg in the sighting study or main study were considered to have achieved satisfactory body weight gains throughout the study.
Gross pathology:
No abnormalities were noted in any animal dosed at 300 mg/kg in the sighting study or main study at the macroscopic examination at study termination on Day 15.

Any other information on results incl. tables

Mortality data:

Dose (mg/kg)

No. of deaths

Day*

1

2 to 3

4

5 to 15

 

a

b

a

b

a

b

Sighting study

300

0/1F

0

0

0

0

0

0

0

2000

1/1F

0

0

0

1

-

-

-

Main study

300

0/4F

0

0

0

0

0

0

0

*            The day indicated is the time that the animal was found dead

F            Female

a            First observation

b            Second observation

-         Not applicable

Signs associated with dosing:

Dose (mg/kg)

Animal number

Day 1

Pre dose

IAD

~30 minutes after exposure

~1 hour after exposure

~2 hours after exposure

~4 hours after exposure

~5 hours after exposure

Sighting investigations

2000

82

/

-

/

/

/

/

/

Only animals with observations are presented

/ : No abnormalities detected

Signs associated with dosing - continued

Dose (mg/kg)

Animal number

Day 2

Day 3

Day 4

Days 5 to 15

~21.5 hours after dosing

~25.5 hours after dosing

~26.5 hours after dosing

~28.5 hours after dosing

~31 hours after dosing

 

Initial observation

 

Sighting investigations

2000

82

PE, EL, UA

PE, EL, RT

PE, EL, RT, UNS, BE

PE, EL, UNS, BE

PE, EL, UNS

/

Dead

-

Only animals with observations are presented

/           No abnormalities detected

-            Not applicable

PE         Piloerection

EL         Elevated gait

UA        Underactive

RT         Reduced body temperature

UNS      Unsteady gait

BE         Blue extremities

Individual body weight change (g)

Dose (mg/kg)

Sex

Animal Number

Body weight change (g) at Day

1-8

8-15

Sighting investigations

300

Female

81

21

13

2000

Female

82

-

-

Main study

300

Female

83

23

8

84

21

12

85

17

8

86

18

7

Macroscopic findings

 

Dose (mg/kg)

Sighting investigations

Main study

300

2000

300

Female

Female

Animal Number

81

82

83

84

85

86

Fate of animal

K

D

K

K

K

K

Day of death

15

4

15

15

15

15

Tissues examined

Findings

Subcutaneous tissue

ü

ü

ü

ü

ü

ü

Brain

ü

ü

ü

ü

ü

ü

Heart

ü

2

ü

ü

ü

ü

Lungs

ü

ü

ü

ü

ü

ü

Liver

ü

2

ü

ü

ü

ü

Spleen

ü

ü

ü

ü

ü

ü

Kidneys

ü

2

ü

ü

ü

ü

Stomach

ü

2, 5

ü

ü

ü

ü

Duodenum

ü

2, 5

ü

ü

ü

ü

Small Intestines

ü

2, 5, 9

ü

ü

ü

ü

Large Intestines

ü

5, 9

ü

ü

ü

ü

Caecum

ü

5, 8, 9

ü

ü

ü

ü

Urinary Bladder

ü

ü

ü

ü

ü

ü

K           Killed at study termination

D           Sporadic death

ü           No abnormalities detected

2            Congestion (darkened tissue/organs)

5            Enlarged, swollen, or thickened tissues

8            Gaseous distension

9            Fluid contents (red liquid)

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute median lethal oral dose (LD50) to rats of XTJ-785 was demonstrated to be between 300 and 2000 mg/kg body weight.
XTJ-785 is included in Category 4, according to the Globally Harmonised System (GHS).
Executive summary:

The study was performed to assess the acute oral toxicity of XTJ-785 to the rat.

Fasted female rats received a single oral gavage dose of the test item, formulated in corn oil,

The study was performed to assess the acute oral toxicity of XTJ-785 to the rat.

Fasted female rats received a single oral gavage dose of the test item, formulated in corn oil, at the following dose levels:

Sighting investigations: 300 and 2000 mg/kg body weight

Main study: Based on the results of the sighting investigations a further four fasted females were similarly dosed at 300 mg/kg body weight.

During the study, clinical condition, body weight and macropathology investigations were undertaken.

Results

One female (number 82) dosed at 2000 mg/kg in the sighting study was found dead on Day 4.

Clinical signs prior to death comprised piloerection, elevated and unsteady gait, reduced body temperature, blue extremities and underactivity. These signs were seen on Day 2 of treatment. A loss in body weight was noted. Macroscopic examination of the animal revealed congestion (characterized by darkened tissues/organs) of the heart, liver, kidneys, stomach, duodenum and small intestines, enlarged, swollen, or thickened tissues of the stomach, duodenum, caecum and small and large intestines. A red fluid was seen in the caecum and small and large intestines. Gaseous distension of the caecum was also observed.

There were no clinical signs of reaction to treatment for animals dosed at 300 mg/kg in the sighting study or main study.

All animals dosed at 300 mg/kg in the sighting study or main study were considered to have achieved satisfactory body weight gains throughout the study.

No abnormalities were noted in any animal dosed at 300 mg/kg at the macroscopic examination at study termination on Day 15.

The acute median lethal oral dose (LD50) to rats of XTJ-785 was demonstrated to be between 300 and 2000 mg/kg body weight.

XTJ-785 is included in Category 4, according to the Globally Harmonised System (GHS).