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Administrative data

Endpoint:
repeated dose toxicity: oral, other
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 day dosing period. Study dates from May 09, 2017 to September 26, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction products of 2,7-Naphthalenedisulfonic acid, 5-[[4-chloro-6-(ethylphenylamino)-1,3,5-triazin-2-yl]amino]-4-hydroxy-3-[(2-sulfophenyl)azo]-, trisodium salt and lithium chloride
EC Number:
943-066-4
Molecular formula:
Not applicable; this UVCB substance contains: C27H19ClN7O10S3.xLi.yNa, (x + y) = 3; 0 < (x,y) < 3 with 753.9 < MW < 802.0 g/mol (UVCB substance) and traces of NaCl.
IUPAC Name:
Reaction products of 2,7-Naphthalenedisulfonic acid, 5-[[4-chloro-6-(ethylphenylamino)-1,3,5-triazin-2-yl]amino]-4-hydroxy-3-[(2-sulfophenyl)azo]-, trisodium salt and lithium chloride
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
Batch No.: 5503
CAS No.: 1809145-88-7
Purity: 95%
Appearance: Brown powder

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl:WI
Sex:
male/female

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distilled water
Details on oral exposure:
A constant dose volume of 5 mL/kg bw was administered to all animals in all groups. The actual volume to be administered was calculated and adjusted based on each animal’s most recent body weight.
Details on analytical verification of doses or concentrations:
Test item concentration and homogeneity of the dosing formulations was determined on two occasions during the treatment period
All formulations were found to be in the range of 100 to 104 % of nominal concentrations.
Duration of treatment / exposure:
28 days
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
for low dose
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
for mid dose
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
for high dose
No. of animals per sex per dose:
Five
Control animals:
yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:
Animals were inspected for signs of morbidity and mortality twice daily (at the beginning and end of each day).
Sacrifice and pathology:
Gross necropsy was performed on all animals at the end of treatment period on Day 28 (after the sample collection for clinical pathology evaluation). The animals were euthanized by exsanguination under pentobarbital anaesthesia
Other examinations:
The animals were monitored for changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern), or changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behaviour (e.g. self-mutilation, walking backwards); special attention were directed towards the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

During the last week of treatment, each animal was subjected to the functional observation battery, including qualitative assessment of the grip strength, and to measurements of the landing foot splay and fore/hind grip strength.
Statistics:
Data was collected using the software PROVANTIS v.9 or recorded on data collection sheets taken from the relevant SOPs, then tabulated using PROVANTIS v.9, Microsoft Office Word and/or Excel, as appropriate.
Group mean and standard deviation were calculated for numerical data. Statistical analysis was performed using SAS 9.2 (built in Provantis System).

Results and discussion

Results of examinations

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
For the male and female cages in the high doses, red coloured faeces were observed from Day 1 until the end of the study. For the male and female cages in the mid doses, red coloured faeces were observed from Day 15 until the end of the study. (Although the test item was a brown powder, the prepared formulations were red in colour.)
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
For male rats:
Significantly higher (p<0.05, p<0.01) mean platelet volume (MPV) in all dose groups and significantly higher (p<0.05) relative reticulocyte percentage in the mid and high dose groups were recorded. The PTT (Prothrombin Time (sec)) was statistically lower in the mid (p<0.01) and high dose (p<0.05) groups compared to the control.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
For male rats:
Significantly higher (p<0.01) potassium concentration in the Low dose, significantly higher (p<0.01) total bilirubin, significantly higher (p<0.05) cholesterol, significantly higher (p<0.05) albumin/globulin ratio in the High dose and significantly higher (p<0.01, p<0.05) phosphorus concentration in the Mid and High dose groups.

For female rats:
Significantly higher (p<0.05) urea in the Low dose, significantly lower (p<0.05) potassium concentration in the Mid dose, significantly higher phosphorus (p<0.05) and calcium (p<0.01) concentration in the High dose, significantly higher total bilirubin and bile acid concentration (p<0.01, p<0.05) in the Mid and High dose groups.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No evidence of discolouration of urine; ano-genital areas were stained red in higher dose animals, but this was considered as a result of containation from red faeces / grooming
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
For male rats:
Significantly higher (p<0.05) absolute and relative epididymides weights in the mid dose group were recorded.

For female rats:
Significantly higher (p<0.05, p<0.01) absolute and relative adrenal gland weight in the low and high dose groups and significantly higher (p<0.01) absolute kidney weight in the High dose group were recorded.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
For male:
Enlarged spleen in one Control (#1003) and enlarged thyroid glands in one High dose (#4003) was seen at necropsy.
Neuropathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Two animals (#1002, 1004) were observed that focal or multifocal, minimal tubular basophilia in the kidney. Four animals (#1004, 4004, 4005, 4502) were recorded that focal or multifocal, minimal congestion or haemorrhage in the thymus. One animal (#1003) was recorded that diffuse, moderate congestion or haemorrhage in the spleen.
Other effects:
no effects observed
Description (incidence and severity):
No aderse effects obsrved that were considered to be treatment related.

Effect levels

Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food efficiency
gross pathology
haematology
histopathology: non-neoplastic
mortality
neuropathology
organ weights and organ / body weight ratios

Target system / organ toxicity

Critical effects observed:
no

Applicant's summary and conclusion

Conclusions:
According to OECD 407 test method, the NOAEL of CJ306 was greater than 1000 mg/kg bw/day.
Executive summary:

This test using the procedures outlined in the CiToxLAB Study Plan for 16/378-100P and OECD 407 (OECD, 2008). The test article was administered to the three treatment groups by oral gavage in a dose of 100, 300 and 1000 mg/kg bw for 28 consecutive days. There were five male and five female Wistar rats in each group.The examinations included clinical signs, mortality, body weights, food consumption, neurological assessment, clinical pathology, vaginal smears, gross pathology, organ weights, histopathology. The observations of clinical signs, body weights, food consumption, clinical pathology, gross pathology and organ weights were not associated with any adverse effects that could be ascribed to treatment.On the basis of the test results given above, the NOAEL of CJ306 for the rats was greater than 1000 mg/kg bw/day.

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