2-{[(2-{3-[(4-{Ethyl[3-(vinylsulfonyl)phenyl]amino}-6-fluoro-heteromonocycle-2-yl)amino]-2- (hydroxy-O)phenyl}hydrazono-N2)(phenyl)methyl]diazenyl-N1}benzoato(6-)-O]cuprate(4-) sodium salt, polysulfo

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-12-08 to 2012 -1.-6

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliance

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

Test guideline 3

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:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Species/strain: healthy rats, WISTAR rats Crl: WI(Han) (full barrier)
Source: Charles River, 97633 Sulzfeld, Germany
Sex: female, non-pregnant, nulliparous
Number of animals: 3 per step
Age
at the beginning of the study: 8 - 11 weeks old
Body weight
on the day of administration: Step 1 / animals no. 1 - 3: 174 – 184 g;
Step 2 / animals no. 4 – 6: 143 – 153 g;
The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to Art. 9.2, No. 7 of the German Act on Animal Welfare the animals were bred for experimental purposes.

Housing and Feeding Conditions:
- Full barrier in an air-conditioned room
- Temperature: 22  3 °C
- Relative humidity: 55  10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 1956)
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 190711)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

The test item was administered at a single dose by gavage using a feeding tube.
The test item was administered at a dose volume of 10 mL/kg body weight.

Doses:

The starting dose was selected to be 2000 mg/kg body weight. No compound-related mortality was recorded for any animal of step 1 or 2. Based on these results and according to the acute toxic class method regime no further testing was required.

No. of animals per sex per dose:

Number of animals: 3 per step

Control animals:

no

Details on study design:

Preparation of the Animals:
The animals were marked for individual identification by tail painting.
Prior to the administration a detailed clinical observation was made of all animals.

Observation Period:
All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.

Weight Assessment:
The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.

Clinical Examination:
A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Pathology:
At the end of the observation period the animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial; lot no.: 212041; expiry date: 04/2014) at a dosage of approximately 8 mL/kg bw.
All animals were subjected to gross necropsy. All gross pathological changes were recorded and in case of findings the tissues were preserved for a possible histopathological evaluation. The preserved tissues of which no histopathological evaluation was made will be discarded 3 months after the release of the final report unless otherwise agreed upon with the sponsor.

Prior to the administration food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All animals survived until the end of the study without showing any signs of toxicity.

Clinical signs:

other: no clinical signs were observed during the observation period

Gross pathology:

With the exception of acute injection of blood vessels in the abdominal region, which is due to the euthanasia injection, no specific gross pathological changes were recorded for any animal

Other findings:

All animals survived until the end of the study without showing any signs of toxicity.

Any other information on results incl. tables

Clinical Signs - Individual Data:

Animal No. / Sex	Time of Observation Post-Dose	Observations
Step 1 (2000 mg/kg Body Weight)
1 / female	during the whole observation period	no signs of toxicity
2 / female	during the whole observation period	no signs of toxicity
3 / female	during the whole observation period	no signs of toxicity
Step 2 (2000 mg/kg Body Weight)
4 / female	during the whole observation period	no signs of toxicity
5 / female	during the whole observation period	no signs of toxicity
6 / female	during the whole observation period	no signs of toxicity

d = day; h = hour/s, min = minute/s

Based on these results and according to the acute toxic class method regime no further testing was required.Therefore, according to OECD Guideline 423, a sufficient estimation of the acute oral toxicity of the test item is provided.

Absolute Body Weights in g and Body Weight Gain in %:

Animal No. / Sex	g Day 1	g Day 8	g Day 15	% Day 1-15
Step 1 (2000 mg/kg Body Weight)
1 / female	176	202	199	13
2 / female	174	202	207	19
3 / female	184	213	220	20
Step 2 (2000 mg/kg Body Weight)
4 / female	153	189	207	35
5 / female	143	165	171	20
6 / female	144	168	180	25

Findings of the Necropsy - Individual Data:

Animal No./ Sex	Organ	Macroscopic Findings
Step 1 (2000 mg/kg Body Weight)
1 / female	-	nsf
2 / female	-	nsf
3 / female	-	nsf
Step 2 (2000 mg/kg Body Weight)
4 / female	-	nsf
5 / female	-	nsf
6 /female	-	nsf

nsf = no specific findings

Table5:  LD50 Cut-Off:

Dose (unit)	Number of Animals Investigated	Number of Intercurrent Deaths	LD50 Cut-Off
2000 mg/kg bw	6	0	unclassified

bw = body weight

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

Under the conditions of the present study, a single oral application of the test item FAT 40853/A TE to rats at a dose of 2000 mg/kg body weight was associated with no signs of toxicity or mortality.
The median lethal dose of FAT 40853/A TE after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat): unclassified
In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC, the test item FAT 40853/A TE has no obligatory labelling requirement for toxicity.
According to Annex I of Regulation (EC) 1272/2008, the test item FAT 40853/A TE has no obligatory labelling requirement for toxicity and is not classified.
According to GHS (Globally Harmonized Classification System), the test item FAT 40853/A TE has no obligatory labelling requirement for toxicity and is not classified

Executive summary:

Two groups, each of three female WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was dissolved in the vehicle aqua ad injectionem (sterile water) at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

All animals used in the study after their entrance at BSL were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15.All animals were necropsied and examined macroscopically.

Results per Step:

Step	Sex/No.	Dose (mg/kg)	Number of Animals	Number of Intercurrent Deaths
1	female/1-3	2000	3	0
2	female/4-6	2000	3	0

All animals survived until the end of the study without showing any signs of toxicity.

Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain.

At necropsy, no macroscopic findings were observed in any animal of any step.

On the basis of the test results given below and in conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC, the substance should be:

classified asvery toxic
classified astoxic
classified as harmful
not classified	X
limit test	X

On the basis of the test results given below and in conformity with the criteria given in inAnnex I of Regulation (EC) 1272/2008, the substance should be:

classified into category 1
classified into category 2
classifiedinto category 3
classifiedinto category 4
not classified	X

On the basis of the test results given below and in conformity with the criteria given in inGHS (Globally Harmonized System of Classification and Labelling of Chemicals), the substance should be:

classified into category 1
classified into category 2
classifiedinto category 3
classifiedinto category 4
classifiedinto category 5
not classified	X

LD₅₀cut-off:                                                     unclassified

Species/strain:                                                   WISTAR Crl: WI(Han) rats

Number of animals:                                           3 per step / 2 steps performed

Vehicle:                                                            aqua ad injectionem

Method:  OECD 423
    EC 440/2008, Method B.1 tris
    OPPTS 870.1000
    OPPTS 870.1100