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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
Data is from publication

Data source

Reference
Reference Type:
publication
Title:
Microbial mutagenic effects of the DNA minor groove binder pibenzimol (Hoechst 33258) and a series of mustard analogues
Author:
Lynnette R. Ferguson *, William A. Denny
Year:
1995
Bibliographic source:
Mutation Research 329 (1995) 19-27

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: As mentiuon below
Principles of method if other than guideline:
To evaluate the mutagenic potential of N,N-BIS(2-CHLOROETHYL)-4-(5-((5-(4-METHYL-1-PIPERAZINYL)-1H-BENZIMIDAZOL-2-YL)METHYL)-1H-BENZIMIDAZO L-2-YL)-BENZENAMINE in Salmonella strains TA98, TA100 and TA102 by bacterial reverse mutation assay.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
N,N-bis(2-chloroethyl)-4-[6-[[6-(4-methylpiperazin-1-yl)-1H-benzimidazol-2-yl]methyl]-1H-benzimidazol-2-yl]aniline
Cas Number:
166546-20-9
Molecular formula:
C30H33Cl2N7
IUPAC Name:
N,N-bis(2-chloroethyl)-4-[6-[[6-(4-methylpiperazin-1-yl)-1H-benzimidazol-2-yl]methyl]-1H-benzimidazol-2-yl]aniline
Details on test material:
Name of test material (as cited in study report): N,N-BIS(2-CHLOROETHYL)-4-(5-((5-(4-METHYL-1-PIPERAZINYL)-1H-BENZIMIDAZOL-2-YL)METHYL)-1H-BENZIMIDAZO L-2-YL)-BENZENAMINE
- Molecular weight : 562.546g/mol
- Substance type: Organic

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA98, TA100 and TA102
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
other: Other: All are deep rough derivatives of the LT2 subline of Salmonella typhimurium.
Cytokinesis block (if used):
not specified
Metabolic activation:
without
Metabolic activation system:
not specified
Test concentrations with justification for top dose:
0,4,10,20,40,80 and 120 µg/plate
Vehicle / solvent:
Vehicle
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: TA98: daunomycin and cl-nitro-o-phenylene diamine TA100: sodium azide TA102: bleomycin
Details on test system and experimental conditions:
Details on test system and conditions
METHOD OF APPLICATION: plate incorporation assay

DURATION
- Preincubation period:
- Exposure duration: 3days

NUMBER OF REPLICATIONS: All assays were performed in triplicate,
and repeated at least once
Rationale for test conditions:
Characteristics of these strains and the rationale for their selection
Evaluation criteria:
Evaluation was done considering a dose dependent increase in the number of revertants/plate.
Statistics:
Mean± standard deviation was observed.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA98, TA100 and TA102
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: No mutagenic effect were observed

Any other information on results incl. tables

Bacterial mutagenicity data for compounds 2

Compound

Concentration

(µg/plate)

 

Bacterial strainsa

 

       2

0

60±9

187±4

296±41

4

70±3

196±23

406±20

10

62±7

162±21

417±2

20

67±9

172±13

442±62

40

52±7

226±23

442±43

80

65±3

207±12

(194±60)

120

(53±39)

(155±39)

(82±48)

 

a Strains of S. typhimurium (Maron and Ames, 1983).

b Results are mean values (two experiments, plated in triplicate) f standard deviation.

c Square brackets denote toxicity was becoming apparent on visual inspection of the plates.

Applicant's summary and conclusion

Conclusions:
(N-(2-CHLOROETHYL)-N-ETHYL-4-(5-(4-METHYL-1-PIPERAZINYL)(2,5'-BI-1H-BENZIMIDAZOL)-2'-YL)-BENZENAMINE(166546-18-5) was tested for its mutagenic potential in Salmonella strains TA98, TA100 and TA102.The test result was considered to be negative both in the presence and absence of metabolic activation.
Executive summary:

In Vitro Genetoxicity study for (N-(2-CHLOROETHYL)-N-ETHYL-4-(5-(4-METHYL-1-PIPERAZINYL)(2,5'-BI-1H-BENZIMIDAZOL)-2'-YL)-BENZENAMINE was assessed for its mutagenic potential. For this purpose are Bacterial Reverse mutation assay was performed in Salmonella strains TA98, TA100 and TA102. The test material was exposed at the concentration of 0,4,10,20,40,80 and 120µg/plate in the presence and absence of metabolic activation. No mutagenic effects were observed. Therefore (N-(2-CHLOROETHYL) -N-ETHYL -4-(5- (4-METHYL-1-PIPERAZINYL)(2,5'-BI-1H-BENZIMIDAZOL)-2'-YL)-BENZENAMINE was considered to be non mutagenic in Salmonella strains TA98, TA100 and TA102 by bacterial reverse mutation assay. Hence the substance cannot be classified as gene mutant in vitro.