Benzeneacetic acid, 4-[4-[4-(hydroxydiphenylmethyl)-1-piperidinyl]-1-butyn-1-yl]-.alpha.,.alpha.-dimethyl-

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:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP, Guideline study. Two deviations occured (quoted in the full test report) but they were judged as minor deviations and did not influence the quality and integrity so the reliability of the study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

FEXO-07

IUPAC Name:

FEXO-07

Constituent 2

Reference substance name:

4-( 4-( 4-(1-hydroxydiphenylmethyl)-1- piperinidyl)-1 butinyl)-a,a-dimethylbenzeneacetic acid

IUPAC Name:

4-( 4-( 4-(1-hydroxydiphenylmethyl)-1- piperinidyl)-1 butinyl)-a,a-dimethylbenzeneacetic acid

Test material form:

other: liquid

Details on test material:

Name: FEX0-07
Chemical Name: 4-( 4-( 4-(1-hydroxydiphenylmethyl)-1-piperinidyl)-1 butinyl)-a,a-dimethylbenzeneaceticacid
Colour: colourless
Purity: >99%
Storage: at room temperature, protected from light

Method

Target gene:

uvrB gene, nitrate reductase (chl) and biotin (bio) genes

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

The test item was tested in the pre-experiment at the following concentrations:
3.16, 10, 31.6, 100,316, 1000,2500 and 5000 µg/plate.

The test item concentrations to be applied in the main experiments were chosen according to the results of the pre-experiment; two independent
experiments were performed at the following concentrations:
Experiment 1:
10, 31.6, 100, 316, 1000,2500 and 5000 µg/plate.
Experiment II:
50, 100, 190, 375, 750, 1500 and 3000 µg/plate (TA 98, TA 1535, TA 1537, TA 102)
250, 500, 700, l 000, 2000, 3000, 4000 and 5000µg/plate (only for TA 100)

Controls

Untreated negative controls:

yes

Remarks:

Solvent controls, consisting of solvent alone and treated in the same way as the treatment groups were included.

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Remarks:

Strain specific positive controls were inclnded in the assay, which demonstrated the effective performance of the test.

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine, 2-aminoanthracene

Details on test system and experimental conditions:

Pre-incubation method and the plate incorporation method were applyed.

Evaluation criteria:

A test item is considered as mutagenic if:
- a clear and dose-related increase in the number of revertants occurs
and/or
- a biologically relevant positive response for at least one of the dose groups occurs in at least one tester strain with or without metabolic activation.

A biologically relevant increase is described as follows:
- if in tester strains T A 100 and TA 102 the number of reversions is at least twice as high
- if in tester strains TA 98, TA 1535 and TA 1537 the number of reversions is at least three times higher as compared to the spontaneous reversion rate

A test item producing neither a dose related increase in the number of revertants nor a reproducible biologically relevant positive response at any ofthe dose groups is considered to be non-mutagenic in this system

Statistics:

According to the OECD guidelines, the biological relevance ofthe results was the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA98 and TA102

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic effects were observed at a dose of 1000 µg/plate and higher

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium, other: TA 98 and TA 102

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic effects were observed at a dose of 2500 µg/plate and higher.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

Toxic effects were observed at a dose of 1000 µg/plate and higher.

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium, other: TA 1535 and TA 1537

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

Toxic effects of the test item were found at a dose of 1000 µg/plate and higher. (No biologically relevant increases in revertant colony numbers were noted for these two strains).

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic effects of the test item were found at a dose of 1000 µg/plate and higher

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 98

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic effects of the test item were found at a dose of 1500 µg/plate and higher

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 6

Species / strain:

S. typhimurium TA 100

Metabolic activation:

without

Genotoxicity:

positive

Remarks:

Toxic effects of the test item were found at a dose of 750 µg/plate and higher

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 7

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

Toxic effects of the test item were found at a dose of 2000 µg/plate and higher

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 8

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic effects of the test item were found at a dose of 750 µg/plate and higher

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 9

Species / strain:

S. typhimurium, other: TA 1537 and TA 102

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic effects of the test item were found at a dose of 750 µg/plate and higher

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 10

Species / strain:

S. typhimurium, other: TA 1537 and TA 102

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic effects of the test item were found at a dose of 1500 µg/plate and higher

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

For the results relative to (i) pre-experiment, (ii) experiment I and (iii) experiment II see the file attached below.

Remarks on result:

other: strain/cell type: S. typhimurium TA98 and TA102 specifically

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

No biologically relevant increases in revertant colony numbers were noted in tester strains TA 98, TA 1535, TA 1537 and TA 102. Biologically relevant increases of revertant colony numbers were observed in tester strain TA 100 at a dose of 1000 µg/plate (without metabolic activation) and at a dose of l000 µg/plate and higher (with metabolic activation) in experiment I.

In experiment II biologically relevant increases of revertant colony numbers were observed in the same tester strain at a dose of 1000 µg/plate (without metabolic activation) and at a dose of 750 µg/plate and higher (with metabolic activation). The threshold value of 2.0 was exceeded and a maximum mutation factor of 4.2 was reached at a dose of 1000 µg/plate (with metabolic activation) in experiment I.

The reference mutagens induced a distinct increase of revertant colonies indicating the validity of the experiments.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive

FEX0-07 caused gene mutations by base pair changes in the genome of the tester strain TA 100.
Therefore, FEX0-07 is considered to be mutagenic in this bacterial reverse mutation assay.

Executive summary:

In order to investigate the potential of FEX0 -07 for its ability to induce gene mutations the plate incorporation test (experiment I and II) was performed with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102. In two independent experiments several concentrations of the test item were used. Each assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate. The following concentrations of the test item were prepared and used in the experiments:

* Experiment I:

- 10, 31.6, l 00, 316, l 000, 2500 and 5000 µg/plate

* Experimcnt II:

-50, 100, 190, 375,750, 1500 and 3000 µg/plate (TA 98, TA 1535, TA 1537, TA 102)

-250, 500, 700, l 000, 2000, 3000, 4000 and 5000 µg/plate (only for TA 100) In experiment I toxic effects of the test item were observed at concentrations of 1000 µg/plate and higher (with and without metabolic activation), depending on the particular tester strain. In experiment II toxic effects of the test item were noted at concentrations of 750 µg/plate and higher (with and without metabolic activation), depending on the particular tester strain. No biologically relevant increases in revertant colony numbers were noted in tester strains TA 98, TA 1535, TA 1537 and TA 102. Biologically relevant increases of revertant colony numbers were observed in tester strain TA 100 at a dose of 1000 µg/plate (without metabolic activation) and at a dose of l000 µg/plate and higher (with metabolic activation) in experiment I. In experiment II biologically relevant increases of revertant colony numbers were observed in the same tester strain at a dose of 1000 µg/plate (without metabolic activation) and at a dose of 750 µg/platea and higher (with metabolic activation). The threshold value of 2.0 was exceeded and a maximum mutation factor of 4.2 was reached at a dose of l000 µg/plate (with metabolic activation) in experiment I. The reference mutagens induced a distinct increase of revertant colonies indicating the validity of the experiments. In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, FEX0-07 caused gene mutations by base pair changes in the genome of the tester strain TA 100. Therefore, FEX0-07 is considered to be mutagenic in this bacterial reverse mutation assay.