(4aR*, 8aR*)-1-bromo-4a,5,9,10,11,12-hexahydro-3-methoxy-6H-benzofuro[3a,3,2-ef] [2]benzazepin-6-one hydrochloride (1:1)

Endpoint summary

Description of key information

Acute toxicity: Oral
In an acute oral toxicity study in HanBrl:Wist (SPF) rats,  following the acute toxic class method in accordance with the OECD guideline 423 and EU Method B.1 tris, the LD50 was determined to be greater than 2000 mg/kg for both females and males. However, the internal study documents (J&J PRD, 1997) did not include a lot of details (e.g. the vehicle is not known), but there was a clear dose response curve, both in males and females. Based on the results, the expert conclusion is to take 500 mg/kg as the LD50 value.
Acute toxicity: Inhalation
No reliable acute toxicity study via inhalation were available. However, this endpoint is waived as specific data are available for the oral and dermal exposure route.
Acute toxicity: Dermal  
A K1 acute dermal test was performed in male and female HanBrl:WIST (SPF) rats according to OECD Guideline 402 and EC method B3 without deviations. The median lethal dose of the substance after 24-h dermal exposure to rats of both sexes, observed over a period of 14 days is greater than 2000 mg/kg body weight.

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:

weight of evidence

Study period:

no data

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

Although the internal study documents do not include a lot of details (e.g. vehicle is not know), there was a clear dose response curve, both in males and females. Therefore, the information in this study is used in weight of evidence with the available RCC study (Damme, 2001) with this substance.

Principles of method if other than guideline:

Although the internal study documents do not include a lot of details, a clear dose response curve, both in males and females was observed.

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:

250 mg/kg bw, 500 mg/kg bw, 1000 mg/kg bw

No. of animals per sex per dose:

2 animals per sex and per dose

Control animals:

not specified

Details on study design:

No data

Statistics:

No data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

500 mg/kg bw

Based on:

test mat.

95% CL:

309 - 808

Mortality:

250 mg/kg bw: 0/2 mortality for males and females
500 mg/kg bw: 1/2 mortalities for males and females
1000 mg/kg bw: 2/2 mortalities for males and females

Clinical signs:

other: 250 mg/kg bw: hypertonia 1/2 male rats 500 mg/kg bw: tremors and spasms in 2/2 male and 2/2 female rats; clonic convulsions, tonic convulsions of upper limbs, hypotonia, piloerection and hypothermia in 1/2 male rats and clonic convulsions, tonic convulsio

Gross pathology:

No data

Other findings:

No data

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The oral LD50-value of substance T002102 is 500 (309 - 808) mg/kg bw in male and female rats. The test substance is therefore considered as acute tox category 4 according to CLP classification.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2001-07-19 to 2001-08-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted according to OECD guideline 423 (Acute Oral Toxicity-Acute Toxic Class Method) and EU method B.1.tris (Acute Oral Toxicity-Acute Toxic Class Method) without deviation. The endpoint is covered by weight of evidence of the two available studies. The LD50 of the internal study document of 500 mg/kg is considered as a realistic worst-case (J&J PRD, 1997).

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

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: RT002102G4A311
- Expiration date of the lot/batch: 25-Mar-2002
- Purity: 99.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, away from sunlight
- Stability of the test substance in the solvent/vehicle: 2 hours

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: HanBrl:WIST (SPF), RCC Ltd., Biotechnology and Animal Breeding Division, CH-4414 Füllinsdorf, Switzerland
- Age when treated: 8 weeks (males); 10 weeks (females)
- Weight at study initiation: Males: 191.6-204.9 g; Females: 170.4-183.7 g (prior to administration)
- Fasting period before study: 16-20 h (access to water was permitted)
- Housing: In groups of three in Makrolon type 4-cages with wire mesh tops and standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat maintenance diet, batch no. 72/1 ad libitum
- Water (e.g. ad libitum): Community tap water from Itingen, ad libitum
- Acclimation period: Males: 2001-07-23 to 2001-07-29; Females: 2001-07-19 to 2001-07-25 , under laboratory conditions, after health examination

ENVIRONMENTAL CONDITIONS
- Temperature (deg C): 22+-3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): air conditioned, 10-15 air changes per hour
- Photoperiod: 12 hrs dark / 12 hrs fluorescent light, music during the light period

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG 300

Details on oral exposure:

VEHICLE:
- Concentration in vehicle: 0.2 g/mL
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: The vehicle was chosen after a non-GLP solubility trial performed before the study initiation date.
- Lot/batch no. (if required): 412565/1 50501
- Expiration date: 2006-02-1
- Storage: room temperature in the original container away from direct sunlight

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

DOSAGE PREPARATION (if unusual):
- Dose levels were prepared with the test material as delivered. The preparations were made shortly before every dosing. The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight: volume). The mixtures were prepared using a magnetic stirrer in order to maintain homogeneous solutions. Samples of the dose formulation for the analytical investigations (stability for two hours, homogeneity) were taken and analyzed directly with LC-MS.

Doses:

2000 mg/kg tested on two groups (males and females)

No. of animals per sex per dose:

3 animals/treatment group

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations:
- Mortality/viability: Daily during acclimation period and twice daily during days 1-15.
- Body weights: On test days 1 (prior to administration), 8 and 15.
- Clinical signs: Daily during the acclimation period and at least four times on test day 1 after the test substance administration. Once daily during days 2-15. All abnormalities were recorded.
- Necropsy of survivors performed: Yes, all surviving animals were killed at theend of the observation period by intraperitoneal injection of NARCOREN at a dose of at least 2.0 ml/kg bw. Macroscopic examinations were preformed.

Statistics:

No statistical analysis was used.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

One female treated with 2000 mg/kg bw was found dead 1 hour after dosing. No other mortalities were reported during the study.

Clinical signs:

other: Moderate tremor was observed in all females 10 minutes after the test substance application, and one hour after the test substance application in all males. Slightly or moderately ruffled fur, hunched posture, half closed lids and in the two surving fema

Gross pathology:

No macroscopic findings were observed at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose of the test substance after single oral administration to rats of both sexes, observed over a period of 14 days is greater than 2000 mg/kg body weight. Therefore, the substance is not considered to be classified according to the CLP Regulation.
However, the endpoint is covered by weight of evidence of two available studies. Although the LD50 of the internal Jannsen document does not inclue a lot of details, in contrast to this study, there was a clear dose response curve, both in males and females. Based on these results, it was decided to take 500 mg/kg as the LD50 value.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

500 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2001-07-18 to 2001-08-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes

Remarks:

: This study was performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted February 2nd, 2000 [RS 813.016.5].

Test type:

standard acute 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: RT002102G4A311
- Expiration date of the lot/batch: 25-MAR-2002
- Purity:99.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Stability of the test substance in the solvent/vehicle: 2 hours

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: HanBrl:WIST (SPF), RCC Ltd. Biotechnology & Animal Breeding Division, CH-4414 Füllinsdorf, Switzerland
- Age at study treatment: 9 weeks (males); 12 weeks (females)
- Weight at study initiation: males: 229.5-257.7 g; females: 202.3-215.4 g
- Fasting period before study: no data
- Housing: During acclimatization, animals were housed in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. During treatment and observation animals were individually housed in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz).
- Diet: ad libitum, pelleted standard Provimi Kliba 3433, batch no. 72/01 rat maintenance diet
- Water: ad libitum, community tap water from Itingen
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12hrs light/12 hrs dark, music during the light period

Type of coverage:

semiocclusive

Vehicle:

polyethylene glycol

Remarks:

PEG300

Details on dermal exposure:

TEST SITE
- Area of exposure: back of the animals was clipped with an electric clipper. On test day 1, 2000 mg/kg bw evenly on the intact skin with a syringe and covered with a semi-occlusive dressing.
- % coverage: 10 % of the total body surface
- Type of wrap if used: A dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The test sites were washed with lukewarm tap water and dried with disposable paper towels.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 4 mL volume/body weight
- Concentration (if solution): the test substance was diluted in polyethylene glycol (PEG 300) at a concentration of 0.5 g/mL
- Constant volume or concentration used: yes
- For solids, paste formed: no

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 male/female rats per dose

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing:
- mortality/viability: daily during acclimatization and at least 1, 2, 3 and approximately 5 hours after the test item administration. Twice daily during days 2-15.
- body weight: on test days 1(pre-administration), 8, 15
- clinical signs: daily during acclimatization and at least 1, 2, 3 and approximately 5 hours after the test item administration. Once daily during days 2-15
- Necropsy of survivors performed: At the end of the observation period all animals were sacrificed by intraperitoneal injection of Narcoren at a dose of at least 2.0 mL/kg body weight (equivalent to at least 320 mg sodium pentobarbitone/kg body weight). The animals were examined macroscopically and all abnormalities recorded. Thereafter, they were discarded.

Statistics:

No statistical analysis were used.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the study

Clinical signs:

other: No systemic or local signs of toxicity were observed during the study period

Gross pathology:

No macroscopic findings were observed at necropsy

The results of the dose verification analysis were 98.8, 97.7 and 95.8 % of nominal for samples taken from the top, middle and bottom of the mixing container. The individual concentrations varied in the range from -2 % to +1 % of the mean concentrations. Since the concentrations were found to be within the accepted range of 80 - 120 % of the nominal concentration, it was determined that the test substance was homogeneously distributed in the vehicle. The test substance was found to be stable in the vehicle under storage conditions over a period of two hours.

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose after a single dermal administration to rats of both sexes, observed over a period of 14 days is greater than 2000 mg/kg body weight. Therefore, the substance should not be considered to be classified according to CLP regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Acute oral toxicity:

Two studies are available.

An acute oral toxicity with T002102 according to the acute toxic class method in male and female HanBrl:Wist (SPF) rats (OECD guideline 423 and EU Method B.1 tris) was performed (K1, RCC, 2001). The substance was tested at 2000 mg/kg in both males and females. The substance was formulated in polyethylene glycol 300 (PEG300), at the concentration of 0.2 g/mL. Three rats per sex received a single oral dose of test item, and were observed during 14 days following administration. Lethality and viability were observed twice daily, and bodyweight was monitored at day 1 (prior to administration), 8 and 15. Clinical signs were observed at least four times on test day 1 after the test substance administration. Once daily during days 2-15. All abnormalities were recorded. Necropsy of survivors and macroscopic examination were also performed.

One female treated with 2000 mg/kg bw was found dead 1 hour after dosing. No other mortalities were reported during the study. Moderate tremor was observed in all females 10 minutes after the test substance application, and one hour after the test substance application in all males. Slightly or moderately ruffled fur, hunched posture, half closed lids and in the two surviving females, slight sedation, were observed during the first two test days. In the two surviving females, yellow urine was noted on the second test day. Ruffled fur, hunched posture, and half closed lids were noted in all males at the first day, and ruffled fur on the second test day.

The LD50 was determined to be greater than 2000 mg/kg bw for both males and females.

A second acute oral toxicity study is available (K4, J&J PRD, 1997). Although the internal study documents did not include a lot of details, in contrast to the RCC study (LD50>2000 mg/kg), there was a clear dose response curve, both in males and females. The LD50 value was determined to be 500 mg/kg. Based on these results, the expert conclusion was to take 500 mg/kg as the LD50 value for the acute oral toxicity endpoint.

Acute inhalation toxicity:

In addition to the oral route of exposure, for substances other than gases, the information mentioned under REACH section 8.5.2 to 8.5.3 shall be provided for at least one other exposure route (REACH Regulation, column 2 adaptation of Annex VIII). For this substance a key study is available for the dermal route of exposure. Therefore, an acute inhalation toxicity study should not be performed.

 

Acute dermal Toxicity:

The K1 study aimed to assess the acute dermal toxicity of T002102 to the rat using semi-occlusive coverage of test item in male and female Wistar rats (strain HanBrl: WIST (SPF), according to the OECD guideline 402 and EU Method B.3. (RCC, 2001). The dose level tested in this study was 2000 mg/kg for both males and females. The substance was diluted in polyethylene glycol 300 (PEG300), at the concentration of 0.5 g/mL, and then applied on the back of 5 males/females after clipping with an electric clipper. 10% of the total body surface was covered with the test item.

The duration of observation period following 24-h dermal exposure was 14 days. Observations for clinical signs of toxicity were performed daily during acclimatization and at least 1, 2, 3 and approx. 5 hours on test day 1 after test substance administration and once daily during days 2-15. All abnormalities were recorded. Body weights were recorded on test days 1 (pre-administration), 8 and 15. No deaths occurred during the study and no systemic or local signs of toxicity were observed during the study period.

The median lethal dose after a single dermal administration to rats of both sexes, observed over a period of 14 days is greater than 2000 mg/kg body weight.

Justification for classification or non-classification

With an LD50 value of 500 mg/kg bw for the oral route, the substance is classified as acute oral toxic category 4 (H302) according to the criteria of the CLP Regulation.

With an LD50 value greater than 2000 mg/kg bw for the dermal route of exposure, the substance should not be classified as acute toxic via the dermal route according to the criteria of the CLP Regulation.

No data were available to decide on the classification for the inhalation route.