6-fluoro-3-(piperidin-4-yl)-1,2-benzoxazole hydrochloride

Endpoint summary

Description of key information

Acute toxicity: Oral

In an acute oral toxicity study in female Wistar rats, following the acute toxic class method in accordance with the OECD Guideline 423 and EU Method B.1 tris (Ott M; 2006), the LD50 of T001492 was established to be within the range of 300 - 2000 mg/kg bw. T001492 was found to be slightly toxic by the oral route and should be classified as Category 4 according to CLP regulation (EC) No 1272/2008.

 

In an acute oral toxicity study in rats, the LD50 of T001492 was found to be 77.10 (58.99 - 100.8) mg/kg (Janssen; 1988).

 

In a 5-day range-finding oral toxicity study in male and female Wistar rats, dose levels of 10, 30 or 100 mg/kg bw are proposed for the subsequent 28-day study with T001492 (RCC; 2006).

 

The studies  were used in a weight-of-evidence approach. 

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

no data

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

No data

GLP compliance:

not specified

Test type:

other: No data

Species:

rat

Strain:

not specified

Sex:

not specified

Details on test animals or test system and environmental conditions:

No data

Route of administration:

oral: unspecified

Vehicle:

not specified

Details on oral exposure:

No data

Doses:

640 mg/kg bw
160 mg/kg bw
80 mg/kg bw
40 mg/kg bw

No. of animals per sex per dose:

5 rats per dose

Control animals:

not specified

Details on study design:

No data

Statistics:

No data

Preliminary study:

No data

Dose descriptor:

LD50

Effect level:

77.1 mg/kg bw

95% CL:

58.99 - 100.8

Mortality:

640 mg/kg bw: 5/5 mortality after 30 minutes
160 mg/kg bw: 5/5 mortality after 30 minutes
80 mg/kg bw: 3/5 mortality within 3 hours after administration
40 mg/kg bw: 0/5 mortality

Clinical signs:

other: 80 mg/kg bw: convulsions or tremors. The 2 surviving rats behaved normal 24 hours afterwards.

Body weight:

other body weight observations

Remarks:

No data

Gross pathology:

No data

Other findings:

No data

Not applicable

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The oral LD50-value of substance T001492 is 77.10 (58.99 - 100.8) mg/kg bw.

Reference 2

Endpoint:

acute toxicity: oral

Remarks:

5-day range-finding oral toxicity study

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2006

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

dose range-finding toxicity study, daily administration by oral gavage

GLP compliance:

not specified

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
No information was provided in the report on the test item.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
No information was provided in the report on the test item.

FORM AS APPLIED IN THE TEST/
No information was provided in the report on the test item.

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: male and female rats, SPF-bred; no data available on source
- Acclimation: yes, no data available on acclimation period
No further information was provided in the report on the test animals.

ENVIRONMENTAL CONDITIONS
No information was provided in the report on the environmental conditions.

IN-LIFE DATES: No information was provided in the report on the in-life dates.

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG 300

Details on oral exposure:

VEHICLE
- Concentration in vehicle: not indicated
- Amount of vehicle: 5 mL/kg

MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg

Doses:

0, 50, 150 and 500 mg/kg bw (daily administration for a period of 5 days)

No. of animals per sex per dose:

4 animals per dose (two animals of each sex per group), 4 groups in total

Control animals:

yes

Remarks:

4 animals, 2 males and 2 females

Details on study design:

- Duration of observation period following administration: animals were observed during acclimatization and treatment period only
- Frequency of observations and weighing: clinical signs, food consumption and body weights were recorded periodically. No further data available on frequency of observations.
- Necropsy of survivors performed: yes, at the end of the dosing period, the surviving animals were killed, necropsied and examined post mortem.

Statistics:

No statistical analysis was performed.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 150 - < 500 mg/kg bw

Based on:

test mat.

Mortality:

Both females treated with 500 mg/kg bw of T1492 died on treatment day 2, the males treated with 500 mg/kg bw of T1492 died on day 3 and 4, respectively.

Clinical signs:

bodyweight loss

convulsions

irregular respiration

other:

Body weight:

lower than 10% body weight loss

Remarks:

Both males treated with 500 mg/kg bw of T1492 showed obvious weight loss (-9%) between treatment days 1 and 3. Males and females treated with 150 mg/kg bw of T1492 showed lower body weight gains than controls between days 1 and 5, although this was more pronounced in males.

Gross pathology:

No test item related microscopical findings were present at any dose level. Both males treated with 500 mg/kg bw of T1492 showed foci in the thymus. One male treated with 500 mg/kg bw showed unilateral pelvic dilatation of the kidney.

Other findings:

Food consumption:
Food consumption during treatment was clearly reduced in all animals treated with 500 mg/kg bw and slightly reduced in males treated with 150 mg/kg bw of T1492.

Organ weight findings including organ / body weight ratios:
Absolute and relative thymus weights were reduced in males and females treated with 150 mg/kg bw of T1492 and absolute and relative spleen weights were reduced in males treated with 150 mg/kg bw of T1492. These changes are considered to be test-item related.

Interpretation of results:

study cannot be used for classification

Conclusions:

Based on the results of this five-day dose range-finding study, dose levels of 10, 30 or 100 mg/kg bw/day are proposed for the subsequent 28-day study (RCC Study Number A46620) with T1492.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006-04-04 (date of report to study director and test facility management)

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: IRA091
- Expiration date of the lot/batch: Unknown
- Purity: 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature (20 ± 5 °C), light protected
- Stability under test conditions: Unknown in PEG 300

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: female rats (nulliparous and non-pregnant), HanRCC:WIST (SPF-Quality); RCC Ltd., Laboratory Animal Services
- Age at study initiation: 12 weeks
- Weight at study initiation: 0.177 - 0.197 kg (range of three treatment groups)
- Fasting period before study: animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test item. Water was available ad libitum.
- Housing: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).
- Diet (e.g. ad libitum): ad libitum, free access to pelleted rodent diet
- Water (e.g. ad libitum): ad libitum, free access to tap water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 30-70% (values above 70 % during cleaning process possible)
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: exact dates not reported

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG 300

Details on oral exposure:

VEHICLE
- Concentration in vehicle: The test item was diluted in vehicle (PEG 300) at a concentration of 0.03 g/mL and 0.2 g/mL
- Justification for choice of vehicle: The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. PEG 300 was found to be a suitable vehicle.

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

DOSAGE PREPARATION (if unusual):
- Dose levels are in terms of the test item as supplied by the sponsor.
- The dose formulations were made shortly before each dosing occasion using a magnetic stirrer, a spatula and an Ultra-Turrax (Janke & Kunkel, D-79219 Staufen) as homogenizers.
The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume). Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: not indicated

TREATMENT
The animals received a single dose of the test item by oral gavage administration at 300 mg/kg body weight or 2000 mg/kg body weight after being fasted for approximately 17 to 18 hours (access to water was permitted). Food was provided again approximately 3 hours after dosing.

Doses:

2000 mg/kg, 300 mg/kg (single dosage each)

No. of animals per sex per dose:

3 females per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (until day 15)
- Frequency of observations and weighing:
mortality/viability: Daily during the acclimatization period, approximately 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15.
body weights: days 1 (pre-administration), 8 and 15.
clinical signs: Daily during the acclimatization period, approximately 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15. All abnormalities were recorded.
- Necropsy of survivors performed: The two 2000 mg/kg treated animals and one 300 mg/kg treated animal which died spontaneously approximately 30 minutes or 3 hours after test item administration and the third animal of the 2000 mg/kg treated group which was killed in extremis approximately one hour after treatment were necropsied as soon as they were found dead or killed. The animal sacrificed for ethical reasons was killed by an intraperitoneal injection of Vetanarcol.
All surviving animals were killed at the end of the observation period by Carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. No organs or tissues were retained.

Statistics:

No statistical analysis was performed.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Two 2000 mg/kg treated animals were found dead approximately 30 minutes after treatment and the remaining animal treated at the same dose was killed in extremis for ethical reasons approximately 1 hour after treatment. Two out of six 300 mg/kg treated animals were found dead approximately 3 and 6 hour after treatment, respectively.

Clinical signs:

convulsions

observations of tremors

other:

Body weight:

other body weight observations

Remarks:

The body weight of the animals was within the range commonly recorded for this strain and age.

Gross pathology:

Liquid contents in the stomach, duodenum, jejunum, and ileum was noted in the two animals treated at 2000 mg/kg at the unscheduled necropsy. Lungs not collapsed were also noted in one of them. Liquid contents in the stomach and ileum and an empty jejunum was also noted in one 300 mg/kg treated animals at the unscheduled necropsy. Due to a technical error of the animal technician, no macroscopic findings were recorded for one 300 mg/kg treated animal.

Number of dead animals in each test group with 3 animals each:

Females 2000 mg/kg: two out of three

Females 300 mg/kg: two out of six (1 out of three in first group, 1 out of three in second group)

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The median lethal dose of T001492 after single oral administration to female rats, observed over a period of 14 days is: 300 mg/kg body weight < LD50 (female rat) < 2000 mg/kg body weight.

T001492 is to be classified as Category 4, harmful if swallowed (H302) following this study conclusion.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

> 50 - < 300 mg/kg bw

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

Acute oral toxicity:

An acute oral toxicity study with T001492 according to the acute toxic class method 5 female Wistar rats was performed by Ott (2006). Single dosages of 300 mg/kg bw (2 groups of 3 animals) and 2000 mg/kg bw (1 group of 3 animals) were applied. The test material was formulated in propylene glycol (PEG 300) at concentrations of 0.03 g/mL and 0.2 g/mL respectively. The animals were observed for 14 days following administration. Mortality/viability was observed approximately 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 and twice daily during days 2-15. Body weight was observed at days 1, 8 and 15 and clinical signs were observed approximately 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 and once daily during days 2-15. Necrospy of survivors and macroscopic examination were also performed.

At 2000 mg/kg, two animals were found dead approximately 30 minutes after treatment and the remaining animal treated at the same dose was killed in extremis for ethical reasons approximately 1 hour after treatment. Two out of six 300 mg/kg treated animals were found dead approximately 3 and 6 hour after treatment, respectively.

At 2000 mg/kg, hunched posture, slight to moderate ruffled fur with slight to moderate sedation, slight to moderate tremor, half closed to closed eyes as well as ventral recumbency were noted. At 300 mg/kg, half closed to closed eyes, hunched posture, slightly to moderately ruffled fur and slight to moderate sedation, slight poor coordination, convulsions and tachypnea were noted. Furthermore, each animal was observed sitting individually in a corner of the cage one hour after treatment.

Liquid contents in the stomach, duodenum, jejunum, and ileum was noted in the two animals treated at 2000 mg/kg at the unscheduled necropsy. Lungs not collapsed were also noted in one of them. Liquid contents in the stomach and ileum and an empty jejunum was also noted in one 300 mg/kg treated animal at the unscheduled necropsy. Due to a technical error of the animal technician, no macroscopic findings were recorded for one 300 mg/kg treated animal.

The oral LD50 value of T001492 in Wistar rats was established to be within the range of 300-2000 mg/kg body weight, which classifies T001492 as slightly toxic by the oral route and should be classified as Category 4 according to the CLP regulation.

 

In addition, in an acute oral toxicty study performed by Janssen (1988), T001492 was orally administered at 40, 80, 160 and 460 mg/kg in rats (5 animals per dose). All animals died at 640 and 160 mg/kg and 3 out of 5 animals died at 80 mg/kg. The oral LD50-value of T001492 was considered to be 77.10 (58.99 - 100.8) mg/kg.

 

Also, a dose range-finding study was performed by Broich and Braun (2006). T001492 was daily administered at 50, 150 and 500 mg/kg bw as 5mL/kg in PEG300 by oral gavage in Wistar rats for a period of 5 days. The animals were observed during acclimatization and dosing period. Mortality, clinical signs, body weight and food consumption were observed and necropsy of survivors and macroscopic examination were also performed. 

Both females treated with 500 mg/kg bw of T1492 died on treatment day 2, the males treated with 500 mg/kg bw of T1492 died on day 3 and 4, respectively.

Clinical signs included convulsions, half closed eyes, hunched posture, ruffled fur, body weight loss and labored breathing. 

Necropsy of the decendents revealed no test item related macroscopical findings were present at any dose level. Absolute and relative thymus weights were reduced in males and females treated with 150 mg/kg bw of T1492 and absolute and relative spleen weights were reduced in males treated with 150 mg/kg bw of T1492. These changes are considered to be test-item related.

Dose levels of 10, 30 or 100 mg/kg bw/d are proposed for the subsequent 28-day study with T001492.

 

In a Janssen Expert Report performed by De Smedt, Van Gompel and Lammens (Expert Report JNJ-1782820-AAC, 2014-04-28) it was concluded that based on a (1) mortality percentage of 33% at 300 mg/kg in the study of 1998, (2) the high number of mortalities (2/2 F on day 2 and 2/2 M on day 3 and 4) in the 5-day range findings and (3) the internal communication (1988) with an LD50 value of 77.1 mg/kg, to consider an LD50 value between 50 and 300 mg/kg instead of the range of 300 -2000 mg/kg.

Studies referred to in the expert judgement:

Experiment Number	Study Type	Report conclusion
RCC A28967	Acute oral toxicity study in rats	300 mg/kg < LD50 < 2000 mg/kg
Internal communication	Acute oral toxicity study in rats	LD50 = 77.1 mg/kg
RCC A71447	5 Day Range finding oral Toxicity study in rats	Selected dose levels for 28 days repeated dose tox study are 10, 30 and 100 mg/kg

T001492 is to be classified as toxic if swallowed (Category 3) based on the Janssen expert statement.

Justification for classification or non-classification

Acute oral toxicity

Based on the results of the acute oral toxicity studies, the 5-day range finding study and the expert statement, an LD50 between 50 - 300 mg/kg bw is considered. According to the criteria laid down in Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, T001492 should be classified as Category 3.