Propanamide, N,N',N''-1,3,5-benzenetriyltris[2,2-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:

2004-11-08 to 2004-12-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant; Guideline Study (OECD 471)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

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:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: ICH Guideline "Genotoxicity: Guidance on Specific Aspects of Regulatory Genotoxicity Tests for Pharmaceuticals"

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: EMEA Guideline"ICH Topic S2A - Genotoxicity: Guidance on Specific Aspects of Regulatory Genotoxicity Tests for Pharmaceuticals" (CPMP/ICH/141/95)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Method

Target gene:

his +/- (S. typhimurium); trp+/- (E. coli)

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:

S9 fraction derived from liver of phenobarbitone/B-naphtoflavone treated rats

Test concentrations with justification for top dose:

1, 0.316, 0.1, 0.0316, and 0.01 mg/plate (main study)
0.2, 0.1, 0.05, 0.025, and 0.0125 mg/plate (repeat study)

Vehicle / solvent:

- Vehicle: DMSO
- Justification for choice of solvent/vehicle: DMSO was found to be suitable for dissolving the test item

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

without S9

Migrated to IUCLID6: S. typh. TA 100, TA 1535

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

without S9

Migrated to IUCLID6: S.typh. TA 98

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without S9

Migrated to IUCLID6: S.typh. TA 1537

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

without S9

Migrated to IUCLID6: E.coli WP2 uvrA

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminofluorene; S.typh. TA 98, TA 100, TA 1537

Remarks:

with S9

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene; S.typh. TA 1535, E.coli WP2 uvrA

Remarks:

with S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 60 - 70 minutes in an agitated water bath at 37 ± 0.5°C
- Incubation of plates: main study: 72 h at 37 ± 2°C; repeat study: 65 h at 37 ± 2°C

NUMBER OF STUDIES: 2 (main and repeat study)

NUMBER OF REPLICATIONS: triplicates

DETERMINATION OF CYTOTOXICITY AND PRECIPITATION:
- Cytotoxicity was determined by microscopic analysis (100 - 200 fold magnification) of the plates tested with the highest test item concentration.
- Precipitation was determined by visual control after pre-incubation and microscopically after incubation of plates tested.

Evaluation criteria:

A mutagenic potential of a test item was assumed if one quotient of the mean value of the respective revertant colonies over the mean value of the respective negative control is 2.0 or higher. A dose effect relationship could underline this conclusion.
A possible mutagenic potential of a test item was assumed if one quotient of the mean value of the respective revertant colonies over the mean value of the respective negative control ranges between 1.7 to 1.9 in combination with a dose effect relationship.
No mutagenic potential of a test item was assumed if all quotients of the mean value of the respective revertant colonies over the mean value of the respective negative control range between 1.0 (or lower) to 1.6. The absence of a dose effect relationship could underline this conclusion.

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:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

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:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

PRECIPITATION:
- Main study: after the pre-incubation period precipitation at test item concentrations of 1 and 0.316 mg/plate, after the incubation phase precipitation at a concentration of 0.1 mg/plate.
- Repeat study: after the pre-incubation period precipitation at test item concentrations of 0.2 and 0.1 mg/plate, after the incubation phase precipitation at a concentration of 0.05 mg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: The number of revertants in control and test item plates were within the historical control data range.

ADDITIONAL INFORMATION ON CYTOTOXICITY: Neither in the main nor in the repeat studies any signs of cytotoxicity were observed.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative