2-[[4-[bis(2-hydroxyethyl)amino]-o-tolyl]azo]-5-nitrobenzonitrile

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, gene mutation was predicted for 2-({4-[bis(2-hydroxyethyl)amino]-2- methylphenyl}diazenyl) -5- nitro benzonitrile. The study assumed the use of Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 with S9 metabolic activation system. 2-({4-[bis(2-hydroxyethyl)amino] -2-methylphenyl}diazenyl)-5-nitrobenzonitrile was predicted to not induce gene mutation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence of S9 metabolic activation system and hence, according to the prediction made, it is not likely to classify as a gene mutant in vitro.

 

Based on the predicted result it can be concluded that the substance is considered to not toxic as per the criteria mentioned in CLP regulation.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification

Justification for type of information:

Data is from OECD QSAR Toolbox version 3.4 and the supporting QMRF report has been attached

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

Prediction is done using OECD QSAR Toolbox version 3.4, 2018

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

- Name of the test material: 2-({4-[bis(2-hydroxyethyl)amino]-2-methylphenyl}diazenyl)-5-nitrobenzonitrile
- IUPAC name: 2-({4-[bis(2-hydroxyethyl)amino]-2-methylphenyl}diazenyl)-5-nitrobenzonitrile
- Molecular formula: C18H19N5O4
- Molecular weight: 369.38 g/mole
- Substance type: Organic

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with

Metabolic activation system:

S9 metabolic activation system

Test concentrations with justification for top dose:

No data

Vehicle / solvent:

No data

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

No data

Rationale for test conditions:

No data

Evaluation criteria:

Prediction is done considering a dose dependent increase in the number of revertants/plate

Statistics:

No data

Species / strain:

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

Metabolic activation:

not specified

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

Additional information on results:

No data

Remarks on result:

no mutagenic potential (based on QSAR/QSPR prediction)

The prediction was based on dataset comprised from the following descriptors: "Gene mutation"
Estimation method: Takes highest mode value from the 7 nearest neighbours
Domain  logical expression:Result: In Domain

(((((((((((((("a" or "b" )  and ("c" and ( not "d") )  )  and ("e" and ( not "f") )  )  and ("g" and ( not "h") )  )  and ("i" and ( not "j") )  )  and "k" )  and ("l" and ( not "m") )  )  and ("n" and ( not "o") )  )  and "p" )  and "q" )  and "r" )  and "s" )  and "t" )  and ("u" and "v" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as SN1 OR SN1 >> Nitrenium Ion formation OR SN1 >> Nitrenium Ion formation >> Aromatic azo OR SN1 >> Nitrenium Ion formation >> Aromatic nitro OR SN1 >> Nitrenium Ion formation >> Tertiary aromatic amine by DNA binding by OECD ONLY

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Radical OR Radical >> Radical mechanism via ROS formation (indirect) OR Radical >> Radical mechanism via ROS formation (indirect) >> Nitro Azoarenes OR SN1 OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation >> Nitro Azoarenes by DNA binding by OASIS v.1.4 ONLY

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as Non binder, without OH or NH2 group by Estrogen Receptor Binding

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as Moderate binder, NH2 group OR Moderate binder, OH grooup OR Non binder, impaired OH or NH2 group OR Non binder, MW>500 OR Strong binder, NH2 group OR Strong binder, OH group OR Very strong binder, OH group OR Weak binder, NH2 group OR Weak binder, OH group by Estrogen Receptor Binding

Domain logical expression index: "e"

Referential boundary: The target chemical should be classified as Known precedent reproductive and developmental toxic potential AND NO2-alkyl/NO2-benzene derivatives (8b) AND Toluene and small alkyl toluene derivatives (8a) by DART scheme v.1.0

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as AhR binders.Polycyclic aromatic hydrocarbons (PAHs) (3b-3) OR AhR binders.Tetrachlorodibenzodioxin(TCCD)-like compounds (3b-1) OR Aromatic di-amine derived diazo dyes (12b) OR Aryl triazene derivatives (12d) OR Arylethanamine-like derivatives (11a) OR Bicyclic compounds with aryl fused N containing heterocycle (14 b) OR Bicyclic compounds with aryl fused N containing heterocycle (14 b) >> Benzodiazepines (14b-3) OR Bicyclic compounds with aryl fused N containing heterocycle (14 b) >> Quinolones (14b-2) OR Bicyclic compounds with aryl fused N containing heterocycle (14 b) >> Thalidomide related derivatives (14b-1) OR Not covered by current version of the decision tree OR Not known precedent reproductive and developmental toxic potential OR Organophosphorus compounds (1b) OR Polyhalogenated benzene derivatives (8c) OR Polyhalogenated-, NO2/halogenated-oxydibenzene (8d) OR Triarylmethane dyes (12c) by DART scheme v.1.0

Domain logical expression index: "g"

Referential boundary: The target chemical should be classified as Aromatic diazo AND Nitro-aromatic by in vitro mutagenicity (Ames test) alerts by ISS

Domain logical expression index: "h"

Referential boundary: The target chemical should be classified as 9,10-dihydrophenanthrenes OR Aliphatic halogens OR Alkyl (C<5) or benzyl ester of sulphonic or phosphonic acid OR alpha,beta-unsaturated carbonyls OR Anthrones OR Aromatic mono-and dialkylamine OR Aromatic N-acyl amine OR Azide and triazene groups OR Heterocyclic Polycyclic Aromatic Hydrocarbons OR Hydrazine OR No alert found OR Polycyclic Aromatic Hydrocarbons OR Primary aromatic amine,hydroxyl amine and its derived esters OR Quinones OR Simple aldehyde by in vitro mutagenicity (Ames test) alerts by ISS

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Inclusion rules not met by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as Acid anhydrides OR Aldehydes OR Aromatic amines OR Ketones OR Quaternary organic ammonium compounds by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "k"

Referential boundary: The target chemical should be classified as Bioavailable by Lipinski Rule Oasis ONLY

Domain logical expression index: "l"

Referential boundary: The target chemical should be classified as Non-Metals by Groups of elements

Domain logical expression index: "m"

Referential boundary: The target chemical should be classified as Halogens OR Metalloids by Groups of elements

Domain logical expression index: "n"

Referential boundary: The target chemical should be classified as Group 14 - Carbon C AND Group 15 - Nitrogen N AND Group 16 - Oxygen O by Chemical elements

Domain logical expression index: "o"

Referential boundary: The target chemical should be classified as Group 16 - Sulfur S by Chemical elements

Domain logical expression index: "p"

Similarity boundary:Target: Cc1cc(N(CCO)CCO)ccc1N=Nc1ccc(N(=O)=O)cc1C#N
Threshold=50%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "q"

Similarity boundary:Target: Cc1cc(N(CCO)CCO)ccc1N=Nc1ccc(N(=O)=O)cc1C#N
Threshold=40%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "r"

Similarity boundary:Target: Cc1cc(N(CCO)CCO)ccc1N=Nc1ccc(N(=O)=O)cc1C#N
Threshold=50%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "s"

Similarity boundary:Target: Cc1cc(N(CCO)CCO)ccc1N=Nc1ccc(N(=O)=O)cc1C#N
Threshold=60%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "t"

Similarity boundary:Target: Cc1cc(N(CCO)CCO)ccc1N=Nc1ccc(N(=O)=O)cc1C#N
Threshold=100%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "u"

Parametric boundary:The target chemical should have a value of log Kow which is >= 1.89

Domain logical expression index: "v"

Parametric boundary:The target chemical should have a value of log Kow which is <= 3.35

Conclusions:

2-({4-[bis(2-hydroxyethyl)amino] -2-methylphenyl}diazenyl)-5-nitrobenzonitrile was predicted to not induce gene mutation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence of S9 metabolic activation system and hence, according to the prediction made, it is not likely to classify as a gene mutant in vitro.

Executive summary:

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, gene mutation was predicted for 2-({4-[bis(2-hydroxyethyl)amino]-2- methylphenyl}diazenyl) -5- nitro benzonitrile. The study assumed the use of Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 with S9 metabolic activation system. 2-({4-[bis(2-hydroxyethyl)amino] -2-methylphenyl}diazenyl)-5-nitrobenzonitrile was predicted to not induce gene mutation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence of S9 metabolic activation system and hence, according to the prediction made, it is not likely to classify as a gene mutant in vitro.

 

Based on the predicted result it can be concluded that the substance is considered to not toxic as per the criteria mentioned in CLP regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Gene mutation in vitro:

Prediction model based estimation and data from read across chemicals has been reviewed to determine the mutagenic nature of

2-({4-[bis(2-hydroxyethyl)amino]-2- methylphenyl}diazenyl) -5- nitro benzonitrile. The studies are as mentioned below:

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, gene mutation was predicted for 2-({4-[bis(2-hydroxyethyl)amino]-2- methylphenyl}diazenyl) -5- nitro benzonitrile. The study assumed the use of Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 with and without S9 metabolic activation system. 2-({4-[bis(2-hydroxyethyl)amino] -2-methylphenyl}diazenyl)-5-nitrobenzonitrile was predicted to not induce gene mutation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence and absence of S9 metabolic activation system and hence, according to the prediction made, it is not likely to classify as a gene mutant in vitro.

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, chromosomal aberration was predicted for 2-({4-[bis(2-hydroxyethyl)amino]-2- methylphenyl} diazenyl) -5- nitrobenzonitrile. The study assumed the use of Chinese hamster ovary (CHO) cell line with and without S9 metabolic activation system. 2-({4-[bis(2-hydroxyethyl)amino]-2-methylphenyl} diazenyl)-5-nitrobenzonitrile was predicted to not induce chromosomal aberrations in Chinese hamster ovary (CHO) cell line in the presence and absence of S9 metabolic activation system and hence, according to the prediction made, it is not likely to classify as a gene mutant in vitro.

The predicted data for the target chemical s further supported by the data from read across chemicals as mentoned below:

In a study performed ny Shimizu and Yano (Mutation Research, 1986), gene mutation toxicity study was performed to determine the mutagenic nature of 50 -60% structurally similar read acroos chemical p-Nitrobenzonitrile (RA CAS no 619 -72 -7; IUPAC name: 4-Nitrobenzonitrile). The study was performed using Salmonella typhimurium strains TA98, TA100, TA1535, TA1538 and TA1537 in the presence and absence of S9 metabolic activation system using the preincubation protocol. The chemical was dissolved in DMSO as solvent and used at dose levels 0, 0.01, 0.05, 0.1, 0.5, 1.0 or 5.0 mg/plate by the preincubation for 15 mins. The plates were incubated for 70 hrs in dark and observed for the presence of colonies. Concurrent solvent and positive control chemicals were also included in the study. p-Nitrobenzonitriledid not induce mutation inSalmonella typhimurium strains TA98, TA100, TA1535, TA1538 and TA1537 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

In another study for 50 -60% structurally similar read across chemical by Sayama et al (Mutation Research, 1991), gene mutation toxicity study was conducted to determine the mutagenic nature of 4,4'-Dimethyl-3,3'-dinitroazoxybenzene (RA CAS no 5679 -89 -0). The study was performed as per the method of Ames et al with suspension assay modification using Salmonella typhimurium strains TA98 and TA100 in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 0-0.5µole/plate. The test chemical was considered to be positive if it produced twice the number of spontaneous revertants. 4,4'-Dimethyl-3,3'-dinitroazoxybenzene, however, induced less than twice the number of spontaneous revertants/plate. Based on these considerations, 4,4'-Dimethyl-3,3'-dinitroazoxybenzene did not induce gene mutation toxicity in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Based on the data available for the target chemical and its read across, 2-({4-[bis(2-hydroxyethyl)amino]-2- methylphenyl} diazenyl) -5- nitrobenzonitrile does not exhibit gene mutation in vitro. Hence the test chemical is not likely to be a mutant as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available for the target chemical and its read across, 2-({4-[bis(2-hydroxyethyl)amino]-2- methylphenyl} diazenyl) -5- nitrobenzonitrile (CAS no 12236 -25 -8) does not exhibit gene mutation in vitro. Hence the test chemical is not likely to be a mutant as per the criteria mentioned in CLP regulation.