[3-methoxycarbonyl-4-[6-(4-prop-2-enoyloxybutoxycarbonyloxy)naphthalene-2-carbonyl]oxy-phenyl] 6-(4-prop-2-enoyloxybutoxycarbonyloxy)naphthalene-2-carboxylate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Test No. 487: In Vitro Mammalian Cell Micronucleus Test, OECD 26 Sep 2014

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

[3-methoxycarbonyl-4-[6-(4-prop-2-enoyloxybutoxycarbonyloxy)naphthalene-2-carbonyl]oxy-phenyl] 6-(4-prop-2-enoyloxybutoxycarbonyloxy)naphthalene-2-carboxylate

Cas Number:

914918-26-6

Molecular formula:

C46H40O16

IUPAC Name:

[3-methoxycarbonyl-4-[6-(4-prop-2-enoyloxybutoxycarbonyloxy)naphthalene-2-carbonyl]oxy-phenyl] 6-(4-prop-2-enoyloxybutoxycarbonyloxy)naphthalene-2-carboxylate

Method

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

- Type and identity of media: MEM supplemented with 10 % fetal calf serum- Properly maintained: yes- Periodically checked for Mycoplasma contamination: yes- Periodically checked for karyotype stability: yes

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital/ß-naphthoflavone induced rat liver S9 mix

Test concentrations with justification for top dose:

without S9 mix: 3.8 - 243 ug/mlwith S9 mix: 7.6 - 243 ug/ml

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

other: colcemid

Remarks:

Without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

With metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in mediumDURATION- Exposure duration: 4 or 20h (see table 1)- Recovery: 4, 20, 28 or 44h (see table 1)- preparation interval: 24 or 48h (see table 1)For the micronucleus analysis the cultures of two flasks of each treatment group seeded and treated in parallel were trypsinised approx. 4 hours before the end of the recovery period and the cells were counted and then adhered in quadriperm dishes on slides for approx. 3 hours. Afterwards, the slides in the chambers of the quadriperm dishes were treated with a hypotonic solution (1.5 % [w/v] sodium citrate) for 5 min at 37° C. After incubation in the hypotonic solution the cells were fixed twice for < 1 min with a solution containing 3 parts ethanol, 1 part acetic acid and 1.25 % (v/v) formaldehyde. After preparation the cells were stained with May Grünwald and GiemsaNUMBER OF REPLICATIONS: in triplicateNumber of cells evaluated: 2000

Evaluation criteria:

A test substance can be classified as mutagenic if:- the number of micronucleated cells is not in the range of our historical control data (0.0 - 2.0 % micronucleated cells).and- either a concentration-related increase in three test groups or a significant increase of micronucleated cells in at least one test group is observed.A test substance can be classified as non-mutagenic if:- the number of micronucleated cells in all evaluated test groups is in the range of our historical control data (0.0 - 2.0 % micronucleated cells). and/or- no concentration-related increase in the number of micronucleated cells is observed.

Statistics:

Statistical significance can be confirmed by means of the Chi square test. However, both biological and statistical significance should be considered together. lf the criteria above mentioned for the test substance are not clearly met, the classification with regard to the historical data and the biological relevance is discussed and/or a confirmatory experiment is performed.

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Test substance precipitation in culture medium 4 hrs alter start of treatment was observedwith 30.4 μg/mL and above in the absence and the presence of 89 mix in Experiment 1. InExperiment II, precipitation occurred alter treatment with 243 μg/mL in the absence of89 mix and with 121.5 μg/mL and above in the presence of 89 mix.In this study, in the absence and the presence of 89 mix, no clear cytotoxicity indicated byreduced cell numbers below 40 % of control was 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):negativeIn conclusion, it can be stated !hat under the experimental conditions reported, the test substance did not induce micronuclei in V79 cells (Chinese hamster cell line) in vitro in the absence and the presence of metabolic activation. Therefore, the test item has to be considered as non-mutagenic in this in vitro testsystem when tested up to the highest applicable test substance concentrations.