Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Acute toxicity: Oral: In an acute oral toxicity study in female Crl:WI (Han) (SPF) rats, following the acute toxic class method in accordance with the OECD Guideline 423 and EU Method B.1 tris, the LD50 was established to be within the range of 50 - 300 mg/kg. According to the OECD 423 guideline, the LD50 cut-off value was considered to be 300 mg/kg body weight (Latour, 2016). Toxicity was observed on central nervous system after single exposure (hunched posture, uncoordinated movements and/or piloerection).

Acute toxicity: Inhalation: 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 T000750, a key study is available for the dermal route of exposure. Therefore, an acute inhalation toxicity study should not be performed.

Acute toxicity: Dermal: In an acute dermal toxicity study in male and female Crl:WI (Han) (SPF) rats, following the standard acute method according to OECD Guideline 402 and EC method B.3, the LD50 was established to exceed 2000 mg/kg body weight (Latour, 2016).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2016-01-19 to 2016-02-23
Reliability:
1 (reliable without restriction)
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
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
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: Janssen Pharmaceutica N.V., M15B1451
- Expiration date of the lot/batch: 30 January 2001
- Purity correction factor: 1

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Analysis of stability, homogeneity and concentration of the test item under test
conditions was not performed as part of this study.
- Solubility and stability of the test substance in the solvent/vehicle:


Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Wistar strain Crl:WI (Han) (outbred, SPF-Quality); Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approx. 8-10 weeks
- Weight at study initiation: 138 - 182 grams
- 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: group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- 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): 18-24 °C
- Humidity (%): 40-70%
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
1% aqueous
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL, 300 mg/mL, 5 mg/mL
- Amount of vehicle (if gavage): no data
- Justification for choice of vehicle: The vehicle was selected based on trial preparations performed at WIL Research Europe and on test item data supplied by the Sponsor. The vehicle was chosen from (in order of preference): water (Elix) (test item did not dissolve), 1% aq. carboxymethyl cellulose (turbid solution), propylene glycol (spec.gravity 1.036), polyethylene glycol 400 (spec. gravity 1.125) and corn oil (spec. gravity 0.92). There was no information available regarding the solubility or stability in vehicle.

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

DOSAGE PREPARATION (if unusual):
- The preparations (w/w) were kept at room temperature and were dosed within 4 hours after adding the vehicle to the test item. Homogeneity was obtained to visually acceptable levels and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies.
- No correction was made for purity of the test item as correction factor is 1

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
The toxicity of the test item was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg.
Doses:
2000 mg/kg body weight
300 mg/kg body weight (2 groups)
50 mg/kg body weight (2 groups)
No. of animals per sex per dose:
3 females per group
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: twice daily.
- body weights: days 1 (pre-administration), 8 and 15 and at death (if found dead or sacrificed after day 1);
- clinical signs: at periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The signs were graded according to fixed scales and the time of onset, degree and duration were recorded: maximum grade 4: grading slight (1) to very severe (4); maximum grade 3: grading slight (1) to severe (3); maximum grade 1: presence is scored (1).
- Necropsy of survivors performed: yes, the moribund animals and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.
Statistics:
No statistical analysis was performed
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 50 - <= 300 mg/kg bw
Based on:
test mat.
Mortality:
At 2000 mg/kg, all animals were found dead or were sacrificed for humane reasons on Day 1.(3/3)
At 300 mg/kg, two animals were found dead on Day 1 and one animal was found dead on Day 2. No mortality occurred in the remaining three animals. (3/6)
At 50 mg/kg, no mortality occurred. (0/6)
Clinical signs:
At 2000 mg/kg, hunched posture and piloerection were noted for all animals on Day 1. Additionally, the animal sacrificed for humane reasons showed clonic spasms, flat posture, abnormal gait, shallow respiration and hypersensitivity to touch and sound.
At 300 mg/kg, tremors, hunched posture, uncoordinated movements and/or piloerection were noted for all animals between Days 1 and 5.
At 50 mg/kg, tremors, hunched posture, uncoordinated movements and/or piloerection were noted for all animals between Days 1 and 3.
Body weight:
The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.
Gross pathology:
At 2000 mg/kg, gray-white discolouration of the stomach contents was noted for all animals at macroscopic examination.
At 300 mg/kg, gray-white discolouration of the stomach contents was noted for one animal at macroscopic examination. No abnormalities were noted for the other animals at this dosage.
At 50 mg/kg, macroscopic post mortem examination did not reveal any abnormalities.
Beginning of autolysis was noted for one of the animals found dead at 300 mg/kg. This was considered not toxicologically relevant.
Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
The oral LD50 value of T000750 in Wistar rats was established to be within the range of 50-300 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 300 mg/kg body weight.

Based on these results:
- according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), T000750 should be classified as: Toxic if swallowed (Category 3) for acute toxicity by the oral route;
- according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments), T000750 should be classified as Category 3 and should be labeled as H301: Toxic if swallowed.

In addition, given the toxicity observed on central nervous system after single exposure (hunched posture, uncoordinated movements and/or piloerection), T000750 should also be classified as STOT SE 1 Category 1 and should be labeled as H370 - Causes damage to organs (Central Nervous System).
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
300 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
other justification
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:
From 2016-02-18 to 2016-03-03
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
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute 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: Janssen Pharmaceutica N.V., M15BB1451
- Expiration date of the lot/batch: 2021-01-31 (retest date)
- Purity correction factor: 1

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: No data
- Solubility and stability of the test substance in the solvent/vehicle: No data

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Wistar strain Crl:WI (Han) (outbred, SPF-Quality); Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 10 weeks old females and 11 weeks old males)
- Weight at study initiation: 300-333 g (males), 191-208 g (females)
- Housing: Individually housed in labeled Makrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- 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. During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24 °C
- Humidity (%): 40-70%
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
1% aqueous
Details on dermal exposure:
TEST SITE
- Area of exposure: on the back of the animal
- % coverage: approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females
- Type of wrap if used: surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): after removal of dressing, the skin was cleaned of residual test item using tap water
- Time after start of exposure: 24 h
Duration of exposure:
24 h
Doses:
2000 mg/kg (single dosage)
No. of animals per sex per dose:
5
Control animals:
not specified
Details on study design:
Preparation of test item:
The preparation (w/w) was dosed within 4 hours after adding the vehicle to the test item. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficiently for these kinds of studies.
No correction was made for purity of the test item as the correction factor is 1.

Treatment of animals and application of test item:
Method: Dermal application. Test preparation was stirred on a magnetic stirrer during application.
Clipping: One day before exposure (Day -1) an area of approximately 5x7 cm on the back of each animal was clipped.

Observations:
Observation period: until day 15 after treatment
- Mortality/Viability: Twice daily.
- Body weights: Days 1 (pre-administration), 8 and 15.
- Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
- Necropsy At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.
Statistics:
No statistical analysis was performed.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred
Clinical signs:
Chromodacryorrhoea (nose) was noted for two male and three female animals on Days 1 and/or 2.
General erythema, scars, scales, scabs and/or white staining were seen in the treated skin-area of all animals during the observation period. These local effects were considered not to have affected the conclusion of the study.
Body weight:
The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
Abnormalities of the epididymides (right side, tail: yellowish, soft nodule) or liver (left lateral lobe: irregular surface) were found in two male animals at macroscopic post mortem examination. Macroscopic post mortem examination of the other animals did not reveal any abnormalities.
Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD50 value of T000750 in Wistar rats was established to exceed 2000 mg/kg body weight. Based on these results, T000750 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1271/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Acute oral toxicity:

An acute oral toxicity study with T000750 according to the acute toxic class method in female Crl:WI (Han) (SPF) rats (OECD guideline 423 and EU Method B.1 tris) was performed (Latour, 2016). Initially, the substance was administered to three female Wistar rats at 300 mg/kg body weight. In a stepwise procedure two additional groups of three females were dosed at 2000 and 300 mg/kg body weight. The substance was formulated in 1% aqueous CMC (carboxymethyl cellulose) at concentrations of 5 mg/ml, 30 mg/mL and 200 mg/mL, respectively. The rats received a single oral dose of test item, and were observed during 14 days following administration. Mortality/viability was observed twice daily, and bodyweight was monitored at day 1 (prior to administration), 8 and 15. Clinical signs were observed at periodic intervals on the day of dosing (day 1) and once daily thereafter. Necropsy of survivors and macroscopic examination were also performed. Internal macroscopic abnormalities were recorded.

Mortality: At 2000 mg/kg, all animals were found dead or were sacrificed for humane reasons on Day 1.(3/3) At 300 mg/kg, two animals were found dead on Day 1 and one animal was found dead on Day 2. No mortality occurred in the remaining three animals. (3/6). At 50 mg/kg, no mortality occurred. (0/6)

Clinical signs observed were: at 2000 mg/kg, hunched posture and piloerection were noted for all animals on Day 1. Additionally, the animal sacrificed for humane reasons showed clonic spasms, flat posture, abnormal gait, shallow respiration and hypersensitivity to touch and sound. At 300 mg/kg, tremors, hunched posture, uncoordinated movements and/or piloerection were noted for all animals between Days 1 and 5. At 50 mg/kg, tremors, hunched posture, uncoordinated movements and/or piloerection were noted for all animals between Days 1 and 3.

The body weight gain shown by most of the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

Additionally, a gray-white discoloration of the stomach contents was noted for all animals at macroscopic examination at 2000 mg/kg.

Beginning of autolysis was noted for one of the animals found dead at 300 mg/kg. This was considered not toxicologically relevant.

The oral LD50 value of T000750 in Wistar rats was established to be within the range of 50-300 mg/ kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 300 mg/kg body weight. Toxicity was observed on central nervous system after single exposure (hunched posture, uncoordinated movements and/or piloerection).

In addition, a K4 study was available in which the LD50 was determined to be 113.6 mg/kg bw (J&J PRD, 1984). As the no sufficient documentation was available to allow a good assessment of reliability and as the LD50 in this study is situated in the range of LD50 values report in the K1 study, this K4 study is added as a supporting study.

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:

An acute dermal toxicity study with T000750 according to OECD guideline 402 and EU Method B.3 in male and female Crl:WI (Han) (SPF) rats was performed (Latour, 2016). The substance was dissolved in 1% aqueous CMC (carboxymethyl cellulose) and applied on a clipped area on the back at 2000 mg/ kg bw. The test item formulation was held in contact with the skin with a dressing consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only. After 24 hours of exposure remainings of the test item were washed-off using tap water. The animals were observed during 14 days. Mortality/viability was observed twice daily, and bodyweight was monitored at day 1 (prior to administration), 8 and 15. Clinical signs were observed at periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. Necropsy of survivors and macroscopic examination were also performed. Internal macroscopic abnormalities were recorded.

No mortality occurred. Clinical signs observed were: Chromodacryorrhoea (nose) was noted for two male and three female animals on Days 1 and/or 2. General erythema, scars, scales, scabs and/or white staining were seen in the treated skin-area of all animals during the observation period. These local effects were considered not to have affected the conclusion of the study.

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

Abnormalities of the epididymides (right side, tail: yellowish, soft nodule) or liver (left lateral lobe: irregular surface) were found in two male animals at macroscopic post mortem examination. Macroscopic post mortem examination of the other animals did not reveal any abnormalities.

The dermal LD50 value of T000750 in Wistar rats was established to exceed 2000 mg/kg body weight.

Justification for classification or non-classification

Acute Oral Classification:

Based on the results showing an acute oral LD50 between 50300 mg/kg bw and according to the criteria laid down in Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, T000750 should be classified as acute oral toxic Category 3 and should be labelled as H301: Toxic if swallowed.

Acute Inhalation Classification:

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

Acute Dermal Classification:

Based on the results showing an acute dermal LD50 >2000 mg/kg bw and according to the criteria laid down in Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, T000750 should not be classified for acute dermal toxicity.

STOT SE Classification:

Based on the results showing toxicity on central nervous system after single exposure (hunched posture, uncoordinated movements and/or piloerection

, T000750 should also be classified as STOT SE 1 Category 1 and should be labeled as H370 - Causes damage to organs (Central Nervous System).