3-(diethylamino)-7-imino-7H-[1]benzopyrano[3',2':3,4]pyrido[1,2-a]benzimidazole-6-carbonitrile

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Genetic mutation in vitro;

The read across substances share high similarity in structure and functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for test substance. Experimental data from read across chemical have been reviewed to determine the mutagenic nature of3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4] pyrido[1,2-a] benzimidazole-6-carbonitrile (52372-39-1).The studies are as mentioned below

The read across substances share high similarity in functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for test substance. Genetic toxicity in vitro for test chemical was assessed for its possible mutagenic potential .For this purpose Salmonella-microsome assay was performed on Salmonella typhimurium strainTA1535, TA1537 and TA1538,using test substance concentration of 0-1000µg/plate (4.44µmol/plate).The test substance was exposed to the Salmonella typhimurium strainTA1535, TA1537 and TA1538 in the presence and absence of metabolic activation system.S9 mix, Adult male Sprague-Dawley rats (200- 250 g) pre-treated with Aroclor 1254 (500 mg/kg) were used to obtain liver for the metabolic activation system. The strains were checked periodically for relevant genetic markers and for rfa by crystal violet sensitivity. Each experiment included solvent controls as well as known direct-acting mutagens and a mutagen that required metabolic activation. No mutagenic effects were observed during the study. Therefore test chemical was considered to be non mutagenic in Salmonella typhimurium strainTA1535, TA1537 and TA1538 by AMES assay. Hence it cannot be classified as gene mutant in vitro.

The read across substances share high similarity in functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for target substance. The Salmonella/mammalian microsome test was performed to determine the mutagenic nature of test chemical in vitro. Preincubation assay was performed using Salmonella typhimurium TA100, TA1535, TA1537, TA98 with and without S9 metabolic activation system. To select the dose range for the mutagenesis assay, the test chemicals were checked for toxicity to TA100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix. The doses thus selected were 0, 100, 333, 1000, 3333, 5450 µg/plate. Appropriate positive controls were also incorporated in the study. Test chemical is not mutagenic to Salmonella typhimurium TA100, TA1535, TA1537, TA98 in the preincubation assay performed with and without S9 metabolic activation system and hence does not classify as a gene mutant in vitro.

 The data available for the target chemical based on its read across substance and applying weight of evidence 3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4]pyrido [1,2-a]benzimidazole -6-carbonitrile (52372-39-1)does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

WoE derived based on the experimental data from structurally and functionally similar read across chemicals for the target chemical.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Name: 3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4]pyrido[1,2-a]benzimidazole-6-carbonitrile
InChI:1S/C23H19N5O/c1-3-27(4-2)15-10-9-14-11-16-21(29-20(14)12-15)17(13-24)22(25)28-19-8-6-5-7-18(19)26-23(16)28/h5-12,25H,3-4H2,1-2H3
Smiles:c12cc3c4nc5ccccc5n4c(=N)c(C#N)c3oc1cc(N(CC)CC)cc2
Molecular Formula: C23H19N5O
Molecular Weight: 381.437 g/mole

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: TA1535, TA1537 and TA1538

Details on mammalian cell type (if applicable):

Not applicable.

Additional strain / cell type characteristics:

not specified

Species / strain / cell type:

S. typhimurium, other: TA1535, TA1537, TA98, and TA100

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix, Adult male Sprague-Dawley rats (200- 250 g) pretreated with Aroclor 1254 (500 mg/kg) were used to obtain liver for the metabolic activation system

Test concentrations with justification for top dose:

1.0-1000µg/plate (4.44µ/mol)
2,0.0,100.0,333.0,1000.0,3333.0,5450 µg/plate

Vehicle / solvent:

1.Not specified
2,DMSO

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Remarks:

Details not available

Untreated negative controls:

yes

Remarks:

, (choline chloride, glycerol, glycine, mannitol, and sodium phosphate)

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: NOPD: 4-nitro+-phenylenediamine 2-AA :2-aminoanthracene SA: sodium azide 9AAD: 9-aminoacridine

Details on test system and experimental conditions:

1,Details on test system and conditions
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 24 hour


Other: The strains were checked periodically for relevant genetic markers and for rfa by crystal violet sensitivity.
2,Details on test system and conditions
METHOD OF APPLICATION: Preincubation method
DURATION
- Pre incubation period: for 20 min
- Exposure duration: No data available
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS:

NUMBER OF CELLS EVALUATED: 0.05 ml of cells

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available

Rationale for test conditions:

Not specified

Evaluation criteria:

1,The numbers of histidine-independent revertants for each S. typhimurium strain were taken from the linear portion of dose-response curves after the background was subtracted. The range of spontaneous revertants was 25-55 for TA1535 and 7-25 for TA1537. A positive response was defined as a reproducible, dose-related increase in the
2,numbers of his+ revertants in all strains

Statistics:

1,Not specified.
2,The data were evaluated using an analysis based on the models presented by Margolin et al

Species / strain:

S. typhimurium, other: TA1535, TA1537 and TA1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium, other: TA1535, TA1537, TA98, and TA100

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Remarks on result:

other: No mutagenic effect were observed

Conclusions:

Gene mutation toxicity study for 3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4]pyrido[1,2-a]benzimidazole-6-carbonitrile (52372-39-1)as predicted using data from read across chemicals for Salmonella typhimurium bacterial strains in the presence and absence of S9 metabolic activation system is negative and hence the chemical is not likely to classify as a gene mutant in vitro.

Executive summary:

Genetic mutation in vitro;

The read across substances share high similarity in structure and functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for test substance. Experimental data from read across chemical have been reviewed to determine the mutagenic nature of3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4] pyrido[1,2-a] benzimidazole-6-carbonitrile (52372-39-1).The studies are as mentioned below

The read across substances share high similarity in functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for test substance. Genetic toxicity in vitro for test chemical was assessed for its possible mutagenic potential .For this purpose Salmonella-microsome assay was performed on Salmonella typhimurium strainTA1535, TA1537 and TA1538,using test substance concentration of 0-1000µg/plate (4.44µmol/plate).The test substance was exposed to the Salmonella typhimurium strainTA1535, TA1537 and TA1538 in the presence and absence of metabolic activation system.S9 mix, Adult male Sprague-Dawley rats (200- 250 g) pre-treated with Aroclor 1254 (500 mg/kg) were used to obtain liver for the metabolic activation system. The strains were checked periodically for relevant genetic markers and for rfa by crystal violet sensitivity. Each experiment included solvent controls as well as known direct-acting mutagens and a mutagen that required metabolic activation. No mutagenic effects were observed during the study. Therefore test chemical was considered to be non mutagenic in Salmonella typhimurium strainTA1535, TA1537 and TA1538 by AMES assay. Hence it cannot be classified as gene mutant in vitro.

The read across substances share high similarity in functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for target substance. The Salmonella/mammalian microsome test was performed to determine the mutagenic nature of test chemical in vitro. Preincubation assay was performed using Salmonella typhimurium TA100, TA1535, TA1537, TA98 with and without S9 metabolic activation system. To select the dose range for the mutagenesis assay, the test chemicals were checked for toxicity to TA100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix. The doses thus selected were 0, 100, 333, 1000, 3333, 5450 µg/plate. Appropriate positive controls were also incorporated in the study. Test chemical is not mutagenic to Salmonella typhimurium TA100, TA1535, TA1537, TA98 in the preincubation assay performed with and without S9 metabolic activation system and hence does not classify as a gene mutant in vitro.

 The data available for the target chemical based on its read across substance and applying weight of evidence 3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4]pyrido [1,2-a]benzimidazole -6-carbonitrile (52372-39-1)does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Genetic mutation in vitro;

The read across substances share high similarity in structure and functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for test substance. Experimental data from read across chemical have been reviewed to determine the mutagenic nature of3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4] pyrido[1,2-a] benzimidazole-6-carbonitrile (52372-39-1).The studies are as mentioned below

The read across substances share high similarity in functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for test substance. Genetic toxicity in vitro for test chemical was assessed for its possible mutagenic potential .For this purpose Salmonella-microsome assay was performed on Salmonella typhimurium strainTA1535, TA1537 and TA1538,using test substance concentration of 0-1000µg/plate (4.44µmol/plate).The test substance was exposed to the Salmonella typhimurium strainTA1535, TA1537 and TA1538 in the presence and absence of metabolic activation system.S9 mix, Adult male Sprague-Dawley rats (200- 250 g) pre-treated with Aroclor 1254 (500 mg/kg) were used to obtain liver for the metabolic activation system. The strains were checked periodically for relevant genetic markers and for rfa by crystal violet sensitivity. Each experiment included solvent controls as well as known direct-acting mutagens and a mutagen that required metabolic activation. No mutagenic effects were observed during the study. Therefore test chemical was considered to be non mutagenic in Salmonella typhimurium strainTA1535, TA1537 and TA1538 by AMES assay. Hence it cannot be classified as gene mutant in vitro.

The read across substances share high similarity in functional group .Therefore, it is acceptable to derive information on mutation from the analogue substance for target substance. The Salmonella/mammalian microsome test was performed to determine the mutagenic nature of test chemical in vitro. Preincubation assay was performed using Salmonella typhimurium TA100, TA1535, TA1537, TA98 with and without S9 metabolic activation system. To select the dose range for the mutagenesis assay, the test chemicals were checked for toxicity to TA100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix. The doses thus selected were 0, 100, 333, 1000, 3333, 5450 µg/plate. Appropriate positive controls were also incorporated in the study. Test chemical is not mutagenic to Salmonella typhimurium TA100, TA1535, TA1537, TA98 in the preincubation assay performed with and without S9 metabolic activation system and hence does not classify as a gene mutant in vitro.

 The data available for the target chemical based on its read across substance and applying weight of evidence 3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4]pyrido [1,2-a]benzimidazole -6-carbonitrile (52372-39-1)does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

Thus based on the above annotation and CLP criteria for target substance 3-(diethylamino)-7-imino-7H-chromeno[3',2':3,4]pyrido[1,2-a]benzimidazole-6-carbonitrile (52372-39-1)does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.