4-[(4,6-bis{[3-(diethylamino)propyl]amino}-heteromonocycle-2-yl)amino]-N-[2-(4,5,6,7-tetrachloro-1,3-dioxo-2,3-dihydro-1H-inden-2-yl)quinolin-8-yl]benzenesulfonamide

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26-Oct-2005 to 30-Nov-2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to the guideline stipulated by the Japanese Ministry of health, labour and welfare.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

No. 476 Yellow

IUPAC Name:

No. 476 Yellow

Details on test material:

- Substance type: yellow powder
- Lot/batch No.: 50162

Method

Target gene:

- S. typhimurium: Histidine gene
- E. coli: Tryptophan gene

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone

Test concentrations with justification for top dose:

Experiment 1:
Without and with S9-mix: 20, 78, 313, 1250 and 5000 µg/plate
Experiment 2:
TA100
Without S9-mix: 78, 156, 313, 625 and 1250 µg/plate
With S9-mix: 313, 625, 1250, 2500 and 5000 µg/plate
TA1535, TA1537, TA98 and WP2uvrA
Without S9-mix: 313, 625, 1250, 2500 and 5000 µg/plate
With S9-mix: 313, 625, 1250, 2500 and 5000 µg/plate
Experiment 2:
TA100
Without S9-mix: 78, 156, 313, 625 and 1250 µg/plate
With S9-mix: 313, 625, 1250, 2500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Test compound was soluble in DMSO and DMSO has been accepted and approved by authorities and international guidelines

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-(2-Furyl)-3-(5-Nitro-2-Furyl)acrylamide for TA98, TA100 and WP2uvrA

Remarks:

without S9

Controls 2

Positive control substance:

9-aminoacridine

Remarks:

without S9

Migrated to IUCLID6: for TA1537

Controls 3

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

without S9

Migrated to IUCLID6: for TA1535

Controls 4

Positive control substance:

other: 2-aminoanthracene in DMSO for all tester strains

Remarks:

with S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (preincubation, 20 min 37 °C)

DURATION
- Exposure duration: 48 hour

NUMBER OF REPLICATIONS:
- Doses of the test substance were tested in duplo in each strain. Two independent experiments were conducted. An additional experiment with TA100 was performed.

NUMBER OF CELLS EVALUATED: 10E8 per plate

DETERMINATION OF CYTOTOXICITY
- Method: The reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies.

OTHER EXAMINATIONS:
- The presence of precipitation of the test compound on the plates was determined.

Evaluation criteria:

A compound is deemed to provide evidence of mutagenic potential if:
(1) a statistically significant dose-related increase in the number of revertant colonies is obtained in two seperate experiments and
(2) the increase in the number of revertant colonies is at least twice the concurrent solvent control value and
(3) the reappearance of (1) and (2) is certificated

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

But tested beyond toxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

But tested beyond toxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation was observed at dose levels of 78 µg/plate

RANGE-FINDING/SCREENING STUDIES:
- In tester strain TA100, toxicity was observed at dose levels of 1250 μg/plate and above in the absence of S9-mix. In the tester strains TA98 and TA1537, toxicity was observed at 5000 µg/plate in the absence of S9-mix.
EXPERIMENT 2
- In the tester strains WP2uvrA and TA1537, toxicity was observed at 5000 µg/plate in the absence of S9-mix. In tester strain TA1537, toxicity was observed at 5000 µg/plate in the presence of S9-mix.
EXPERIMENT 3
- In tester strain TA100, toxicity was observed at 1250 and 5000 µg/plate in the absence and presence of S9-mix, respectively.

COMPARISON WITH HISTORICAL CONTROL DATA:
- The negative and strain-specific positive control values were within our laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

It is concluded that the test substance shows no evidence of mutagenic activity when tested in this bacterial system.