N*4*-(3-Chloro-4-fluoro-phenyl)-7-[(S)-(tetrahydro-furan-3-yl)oxy]-quinazoline-4,6-diamine

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:

2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N4-(3-chloro-4-fluorophenyl)-7-[(3S)-oxolan-3-yloxy]quinazoline-4,6-diamine

EC Number:

921-874-8

Cas Number:

314771-76-1

Molecular formula:

C18 H16 Cl F N4 O2

IUPAC Name:

N4-(3-chloro-4-fluorophenyl)-7-[(3S)-oxolan-3-yloxy]quinazoline-4,6-diamine

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mixture

Test concentrations with justification for top dose:

A maximum concentration of 5000 μg/plate should be investigated according to relevant
guidelines. CDBA 578 precipitated at concentrations higher than 1250 μg/plate in a non-GLP
solubility test. Therefore, 1000 μg/plate was investigated as the highest concentration.

Vehicle / solvent:

dimethylsulfoxide (DMSO)

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

other: 2-Aminoanthracene

Details on test system and experimental conditions:

The test was performed in the presence and absence of rat liver microsomal enzymes
(S9 mix).

Rationale for test conditions:

The assay was considered valid since the following criteria were met:
All tester strains exhibit a characteristic number of spontaneous revertants per plate. The
addition of the metabolic activation system did not alter significantly the number of
spontaneous revertants per plate and therefore the numbers were combined and given as
ranges (see below)
TA 1535: 5 - 22
TA 1537: 3 - 29
TA 98: 16 - 68
TA 100: 57 - 192
TA 102: 252 – 531
These ranges were taken from about 168 experiments conducted in our
laboratory with Merck plates.
The validation studies have shown that the source of the plates do not influence the outcome
of the study (Maron and Ames [4]). Therefore, the same historical control data range obtained
with Merck plates can be used for the interpretation of the results in this study.
In addition, the reference mutagens induced a distinct increase in the number of revertants,
reflecting also the activity of the metabolizing system.

Evaluation criteria:

A reproducible, concentration-dependent increase in the number of revertants of at least one
tester strain over the vehicle control value and/or outside the historical control range is
indicative of genotoxic activity.

Statistics:

Revertant his+ colonies were counted using the automatic colony counter IPI Analyzer 982 B
after incubation at 37°C for 2 days (TA 102: 3 days) and the data were collected by computer.
For all replicate platings, the mean number of revertants per test concentration was
calculated. The condition of the background bacterial lawn (residual growth on minimum
histidine) was evaluated macroscopically for evidence of bacteriotoxicity induced by the test
substance. If extreme thinning or complete lack of the microcolony lawn compared to the
negative, vehicle control plates was observed, no revertants were counted. Evidence of test
substance precipitates in the agar overlay was recorded.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

CDBA 578 caused neither base-pair substitutions nor frameshift mutations in different strains of S. typhimurium in the presence and absence of
metabolic activation when tested up to bacteriotoxic concentrations. Based on these results it was concluded, that the test substance
is "Ames negative".