Benzeneacetic acid, 4-[4-[4-(hydroxydiphenylmethyl)-1-piperidinyl]-1-oxobutyl]-.alpha.,.alpha.-dimethyl-, ethyl ester

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:

Start of study: 17-09-1997 / End of study: 17-10-1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to international guideline

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-[4-[4-(Hydroxydiphenylmethyl)-1-piperidinyl]-1-oxobutyl]-α,α-dimethyl-ethyl ester benzeneacetic acid

EC Number:

616-391-1

Cas Number:

76812-02-7

Molecular formula:

C34 H41 N O4

IUPAC Name:

4-[4-[4-(Hydroxydiphenylmethyl)-1-piperidinyl]-1-oxobutyl]-α,α-dimethyl-ethyl ester benzeneacetic acid

Constituent 2

Reference substance name:

4-(4-(4-Hydroxydiphenylmethyl)-1-piperidinyl)-1-oxobutyl- alpha,alphadimethylbenzeneacetic acid ethyl ester

IUPAC Name:

4-(4-(4-Hydroxydiphenylmethyl)-1-piperidinyl)-1-oxobutyl- alpha,alphadimethylbenzeneacetic acid ethyl ester

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Product number/Code ((I.N.N. or U.S.A.N): HOE CG 0529 OY ZD.. 0001
- Name or number of compound: Fexofenadonethylester
- Synonyms: Terfenadonethylester
- Chemical name: 4-[4-[4-hydroxydiphenylmethyl]-1-piperidinyl]-1- oxobutyl]- a.,a.-dimethylbenzeneacetic acid ethyl ester
- Retention Sample: L00446/001
- Batch number: TAM 40-96-13
- Certificate of analysis: dated Augusf 19th, 1997, CR&T Analytisches Laboratorium Dr. D. Andert
- Content: 98.65 % (w/w)
- Appearance: white powder
- Date of production: August 27th, 1996
- Date of receipt: August 28th, 1997
- Date of expiry: August 27th, 1998
- Storage conditions: darkness at approximately 5°C in a refrigerator
- Solubility: solution in DMSO
- Stability in the solvent: confirmed over 4 hours in DMSO
- Concentration of stock solution: 5% (w/v)

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: rfa uvrB for all strains. All 4 Salmonella strains are deficient in the structure of their LPS layer and in DNA excision repair system. TA 98 and TA 100 also possess a modified postreplication DNA repair system.

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

Test groups
First experiment:
a: without metabolic activation: 4, 20, 100, 500, 2500, 5000 µg/plate
b: with metabolic activation: 4, 20, 100, 500, 2500, 5000 µg/plate

Second experiment:
a: without metabolic activation: 0.032, 0.16, 0.8, 4, 20 and 100 µg/plate
b: with metabolic activation: 0.16, 0.8, 4, 20, 100 and 500 µg/plate

Vehicle / solvent:

Vehicle(s)/solvent(s) used: DMSO

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

untreated control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

without metabolic activation for strains TA 100 and TA 1535

Controls 2

Untreated negative controls:

yes

Remarks:

untreated control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without metabolic activation for strains TA 1537

Controls 3

Untreated negative controls:

yes

Remarks:

untreated control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

without metabolic activation for strains TA 98

Controls 4

Untreated negative controls:

yes

Remarks:

untreated control

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

with metabolic activation for all strains

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
Incubation period: 48h at approx. 37°C
NUMBER OF REPLICATIONS: 3
DETERMINATION OF CYTOTOXICITY: reduced rate of spontaneously occurring colonies and visible thinning of the bacterial lawns

OTHER: A toxicity test using histidine-enriched agar plates and a dilution of the tester strain TA 100 (designated TA 100 D) was performed in parallel with the second mutation experiment.

Evaluation criteria:

Criteria for a valid assay
The assay is considered valid if the following criteria are met:
- the solvent control data are within the laboratory's normal control range for the spontaneous mutant frequency
- the positive controls induced increases in the mutation frequency which were both statistically significant and within the laboratory's normal range

Criteria for a positive response
A test compound is classified as mutagenic if it has either of the following effects:
a) it produces at least a 2-fold increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control at complete bacterial background lawn
b) it induces a dose-related increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control in at least two to three concentrations of the test compound at complete bacterial background lawn.

The test results must be reproducible.

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

The test compound proved to be toxic to most of the bacterial strains at concentrations of 100 µg/plate and above in the presence of S9-mix

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

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

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

The test compound proved to be toxic to most of the bacterial strains at concentrations of 20 µg/plate and above in the absence of S9-mix.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The results lead to the conclusion that Fexofenadonethylester is not mutagenic in these bacterial test systems either in the absence or in the presence of an exogenous metabolizing system.