3-hydroxy-4-[(2-methoxy-5-nitrophenyl)azo]-N-(3-nitrophenyl)naphthalene-2-carboxamide

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

NOAEL was estimated to be 1015 mg/kg bw when rats were orally exposed with 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide. 

Thus, as per criteria of CLP regulation, 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl) diazen-1-yl]-N-(3-nitrophenyl) naphthalene-2-carboxamide can be not classified for reproductive toxicity.  

Link to relevant study records

Reference

Endpoint:

toxicity to reproduction

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 predicted using OECD QSAR toolbox version 3.3 and the supporting QMRF report has been attached

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Principles of method if other than guideline:

Prediction is done using QSAR Toolbox version 3.3

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

Name: 3-hydroxy-4-[(2-methoxy-5-nitrophenyl)azo]-N-(3-nitrophenyl)naphthalene-2-carboxamide
SMILES:COc1ccc(N(=O)=O)cc1N=Nc1c2ccccc2cc(C(=O)Nc2cccc(N(=O)=O)c2)c1O
InChI:1S/C24H17N5O7/c1-36-21-10-9-17(29(34)35)13-20(21)26-27-22-18-8-3-2-5-14(18)11-19(23(22)30)24(31)25-15-6-4-7-16(12-15)28(32)33/h2-13,30H,1H3,(H,25,31)/b27-26+
Molecular Formula: C24H17N5O7
Molecular Weight: 487.4263 g/mole

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

polyethylene glycol

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

2 weeks prior to mating, throughout mating, and pregnancy and at least up to, and including the day before sacrifice

Frequency of treatment:

once daily

Details on study schedule:

not specified

Dose / conc.:

1 015 mg/kg bw/day

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

not specified

Positive control:

not specified

Parental animals: Observations and examinations:

not specified

Oestrous cyclicity (parental animals):

not specified

Sperm parameters (parental animals):

not specified

Litter observations:

not specified

Postmortem examinations (parental animals):

not specified

Postmortem examinations (offspring):

not specified

Statistics:

not specified

Reproductive indices:

not specified

Offspring viability indices:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Dose descriptor:

NOAEL

Effect level:

1 015 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect observed

Remarks on result:

other: No effect observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Dose descriptor:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

other: not specified

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not specified

Histopathological findings:

not examined

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

other: not specified

Generation:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

other: not specified

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

other: not specified

Generation:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

other: not specified

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Reproductive effects observed:

not specified

Treatment related:

not specified

Relation to other toxic effects:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

The prediction was based on dataset comprised from the following descriptors: NOAEL
Estimation method: Takes average value from the 8 nearest neighbours
Domain  logical expression:Result: In Domain

(((((("a" or "b" or "c" or "d" or "e" )  and ("f" and ( not "g") )  )  and ("h" and ( not "i") )  )  and "j" )  and ("k" and ( not "l") )  )  and ("m" and "n" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Non-covalent interaction AND Non-covalent interaction >> DNA intercalation AND Non-covalent interaction >> DNA intercalation >> DNA Intercalators with Carboxamide Side Chain by DNA binding by OASIS v.1.3

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as SN1 AND SN1 >> Nitrenium Ion formation AND SN1 >> Nitrenium Ion formation >> Aromatic azo AND SN1 >> Nitrenium Ion formation >> Aromatic nitro by DNA binding by OECD

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as Acylation AND Acylation >> Ester aminolysis AND Acylation >> Ester aminolysis >> Amides by Protein binding by OASIS v1.3

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as Acylation AND Acylation >> Direct Acylation Involving a Leaving group AND Acylation >> Direct Acylation Involving a Leaving group >> Acetates by Protein binding by OECD

Domain logical expression index: "e"

Referential boundary: The target chemical should be classified as Amides AND Phenol Amines AND Phenols by Aquatic toxicity classification by ECOSAR

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as SN1 AND SN1 >> Nitrenium Ion formation AND SN1 >> Nitrenium Ion formation >> Aromatic azo AND SN1 >> Nitrenium Ion formation >> Aromatic nitro by DNA binding by OECD

Domain logical expression index: "g"

Referential boundary: The target chemical should be classified as Michael addition OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals >> Alkyl phenols OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals >> Hydroquinones OR Michael addition >> Polarised Alkenes-Michael addition OR Michael addition >> Polarised Alkenes-Michael addition >> Alpha, beta- unsaturated amides OR No alert found OR SN1 >> Iminium Ion Formation OR SN1 >> Iminium Ion Formation >> Aliphatic tertiary amines OR SN1 >> Nitrenium Ion formation >> Unsaturated heterocyclic azo OR SN2 OR SN2 >> SN2 at an sp3 Carbon atom OR SN2 >> SN2 at an sp3 Carbon atom >> Aliphatic halides by DNA binding by OECD

Domain logical expression index: "h"

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

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Not known precedent reproductive and developmental toxic potential by DART scheme v.1.0

Domain logical expression index: "j"

Similarity boundary:Target: COc1ccc(N(=O)=O)cc1N=Nc1c2ccccc2cc(C(=O)Nc2cccc(N(=O)=O)c2)c1O
Threshold=60%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "k"

Referential boundary: The target chemical should be classified as Alkylarylether AND Aromatic compound AND Azo compound AND Carbonic acid derivative AND Carboxylic acid derivative AND Carboxylic acid sec. amide AND Ether AND Hydroxy compound AND Nitro compound AND Phenol by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "l"

Referential boundary: The target chemical should be classified as Heterocyclic compound by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "m"

Parametric boundary:The target chemical should have a value of log Kow which is >= 3.93

Domain logical expression index: "n"

Parametric boundary:The target chemical should have a value of log Kow which is <= 9.41

Conclusions:

NOAEL was estimated to be 1015 mg/kg bw when rats were orally exposed with 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide.

Executive summary:

In a prediction done by SSS (2017) using the OECD QSAR toolbox with log kow as the primary descriptor, the reproductive toxicity was estimated for 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide. The NOAEL was estimated to be 1015 mg/kg bw when rats were orally exposed with 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide. 

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 015 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Data is Klimisch 2 and from OECD QSAR toolbox

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive toxicity:

In different studies, 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl) diazen-1-yl]-N-(3-nitrophenyl) naphthalene-2-carboxamide has been investigated for reproductive oral toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments and estimated data in rodents, i.e. most commonly in rats and mice for 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide along with the study available on structurally similar read across substance CI Pigment Red 22 (2-Hydroxy-1-[(2-methyl-5-nitrophenyl)azo]-N-phenylnaphthalene-3-carboxamide) (CAS no 6448-95-9). The predicted data using the OECD QSAR toolbox has also been compared with the experimental studies.

In a prediction done by SSS (2017) using the OECD QSAR toolbox with log kow as the primary descriptor, the reproductive toxicity was estimated for 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide. The NOAEL was estimated to be 1015 mg/kg bw when rats were orally exposed with 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide. 

In another experimental study conducted by National Toxicology Program (National Toxicology Program. Technical Report Series. No.411, U.S. Department of commerce, National Technical Information Service 1992),F344male and female rat were treated withC.1. Pigment Red (3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide)in the concentration of 0,425, 1100 and 2100 mg/kg bw for male and 0, 500, 1300 and 2600 mg/kg/day for female orally in dietfor 2 years. The average daily ingestion of C.1. Pigment Red 23 was approximately 425, 1,100, or 2,100 mg/kg body weight per day for male rats and 500, 1,300, or 2,600 mg/kg for females. Significant increase in survival of treated male and female rats were observed at 2100 and 2600 mg/kg bw as compared to control. Significant increase in survival of treated male rats were observed at 1100 mg/kg bw as compared to control. The greater survival of the exposed groups was due principally to the chemically related decreased incidence of mononuclear cell leukemia. No clinical sign were considered to be treatment related. Significant decrease in body weights of female rats were observed at 1300 and 2600 mg/kg bw as compared to control. No effect on food consumption of treated male and female rats was observed as compared to control. In female, significant decrease in Hematocrit values, hemoglobin concentration, and erythrocyte counts at 2600 mg/kg bw as compared to control. Which indicate mild anemia. No effect in male rats was observed as compared to control. Significant increase in Serum total bilirubin were observed in female rats at 2600 mg/kg bw as compared to control. This finding coupled with the mild anemia suggests a mild hemolytic process. No effect in male rats was observed as compared to control. Similarly, at 1300 and 2600 mg/kg bw in female rats, significant increase in relative kidney and brain weight were observed. In addition, at 2100 mg/kg bw in male rats, Four renal tubule cell adenomas or carcinomas, renal tubule focal hyperplasia and renal tubule adenomas were observed. At 1100 and 2100 mg/kg bw in male rats, Four renal tubule cell adenomas or carcinomas were observed. At 2500 mg/kg bw in female rats, one renal tubule adenoma was observed. Although the trend for these renal neoplasms is significant, the incidences are low and do not exceed the historical control range of 0% to 6% in male rats. At 2100 and 2600 mg/kg bw in male and 1300 and 2600 mg/kg bw in female,significant dose-related decrease in the incidence of mononuclear cell leukemia was observed as compared to control.No significant histopathological changes were observed in reproductive organ such as testes and mammary gland of treated male and female rats as compared to control.Therefore, NOAEL was considered to be 500 mg/kg/day for P and F1 generation whenWistarmale and female rats treated withIndigo Carmineorally in dietfor 2 years. Therefore,NOAEL was considered to be 2100 mg/kg/day for male and 2600 mg/kg bw for female P generation whenF344male and female rats treated withC.1. Pigment Red (3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide) orally in dietfor 2 years.

Further supported by experimental study conducted by National Toxicology Program (National Toxicology Program. Technical Report Series. No.411, U.S. Department of commerce, National Technical Information Service 1992),B6C3F1male and female mice were treated withC.1. Pigment Red (3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide)in the concentration of 0,1,900, 4,500 and 9,500 mg/kg body/day for males and 0, 2,100, 5,240 and 10,800 mg/ kg for femalesorally in dietfor 2 years. The average daily ingestion of C.I. Pigment Red 23 was approximately 1,900, 4,500, or 9,500 mg/kg body weight per day for males and 2,100, 5,240, or 10,800 mg/kg for females. Significant decrease in survival of treated male mice were observed at 1900 mg/kg bw as compared to control. This decrease in survival was associated with evidence of trauma and secondary septicemia caused by fighting. No effect on female mice was observed as compared to control. Red stained fur, extremities, and feces were observed in treated mice. Similarly, No effect on body weight, food consumption,Hematology andClinical chemistry parameters of treated male and female rats were observed as compared to control. In addition, Increase in forestomach epithelial hyperplasia with dose were observed in male and female mice as compared to control.The presence of red pigment, presumed to be C.I. Pigment Red 23 or a metabolite, was observed within intestinal lymphoid tissue (Peyer's patches), and to a lesser extent withinmandibular and inguinal lymph nodes in mice at the 15-month interim evaluation and at the end of the 2-year study. The pigment was bright red and was observed in small intracellular granules and in largeextracellular clumps. No significant histopathological changes were observed in reproductive organ such as testes and mammary gland of treated male and female mice as compared to control. Therefore,NOAEL was considered to be 9500 mg/kg/day for male and 10800 mg/kg bw for female P generation whenB6C3F1male and femalemicetreated withC.1. Pigment Red (3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl)diazen-1-yl]-N-(3-nitrophenyl)naphthalene-2-carboxamide) orally in dietfor 2 years.

Above studies are supported by experimental study conducted by J-check (Ministry of Health, Labour and Welfare", "Ministry of the Environment" and "National Institute of Technology and Evaluation, J-check, 2010) on structurally similar read across substanceCI Pigment Red 22 (2-Hydroxy-1-[(2-methyl-5-nitrophenyl)azo]-N-phenylnaphthalene-3-carboxamide) (CAS no 6448-95-9), Crj:CD(SD) male and female rats treated with CI Pigment Red 22 (2-Hydroxy-1-[(2-methyl-5-nitrophenyl)azo]-N-phenylnaphthalene-3-carboxamide) in the concentration of 0 (vehicle), 100, 300, 1000 mg/kg orally by gavage for 42 to 47 days. No effect on survival of treated male and female rat was observed as compared to control. Red feces exhibiting the same color tone as the test substance administered in all sexes were observed daily from the day following the administration start date. At 1000 mg/kg bw, in 2 male loose stools were observed transiently on 22 days after the start of administration, but abnormalities in general conditions considered to be toxic changes. No significant effect on body weight and body weight gain and food consumption of treated male and female rats were observed as compared to control. Significant decrease in prothrombin time were observed in male rats at 100 mg / kg/day, but no change was observed in the 300 and 1000 mg / kg group, so it was judged to be an accidental change. In females, there was no significant difference between the test substances administered group and the control group. Significant increase in A / G ratio were observed in male rats. At 1000 mg / kg/day However, since the total protein and albumin were not changed and were slightly high, it was considered to be fluctuating within the physiological range and it was judged that it was not caused by administration of the test substance. Significant increase in ASAT (GOT) in female rats at 100 mg / kg/day, but no change was observed in the 300 and 1000 mg / kg group, so it was judged to be an accidental change. Similarly, No effect on reproductive parameters such as sexual cycle, mating index, the fertility index, numbers of corpora lutea or implantations, the implantation index, the delivery index, the gestation index, gestation length, parturition or maternal behavior, numbers of offspring or live offspring of treated rats as compared to control. Significant increase in relative liver weight in male and absolute and relative weight in female were observed at 1000 mg / kg bw as compared to control. Significant decrease in absolute spleen weight and significant increase in relative adrenal gland weight were observed at 300 mg / kg bw in male rats. Significant increase in relative adrenal gland weight were observed in male rats at 100 mg / kg bw. But, since there was no change in the 1000 mg / kg group, it was judged to be an accidental change. Colored stomach at 1000 mg / kg and red, colored contents were observed in small intestine and large intestine at 100 mg / kg. Focal bleeding of the lung, contraction failure and white spots, stomach tar-like contents, white spots of the liver, dilation of the renal pelvic cavity, uterine bloating, vaginal distension and ventricular dilation were observed. However, since there was no certain tendency in its expression status, it was judged to be an accidental change unrelated to administration of the test substance. No histopathological changes were observed in treated male and female rats as compared to control. In addition, No effect on numbers of live offspring, clinical signs and body weight of offspring were observed as compared to control. No gross pathological changes were observed in offspring. Therefore, NOAEL was considered to be 1000 mg/kg/day for P and F1 generation when Crj:CD (SD) male and female rats treated with CI Pigment Red 22 (2-Hydroxy-1-[(2-methyl-5-nitrophenyl)azo]-N-phenylnaphthalene-3-carboxamide) orally by gavage for 42 to 47 days.

Thus, based on the above study and predictions on sodium 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl) diazen-1-yl]-N-(3-nitrophenyl) naphthalene-2-carboxamide and its read across substances, it can be concluded that NOAEL value is 1015 mg/kg bw with no effect on reproduction. Thus, as per criteria of CLP regulation, 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl) diazen-1-yl]-N-(3-nitrophenyl) naphthalene-2-carboxamide can be not classified for reproductive toxicity.  

Effects on developmental toxicity

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the above study and predictions on sodium 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl) diazen-1-yl]-N-(3-nitrophenyl) naphthalene-2-carboxamide and its read across substances, it can be concluded that NOAEL value is 1015 mg/kg bw with no effect on reproduction. Thus, as per criteria of CLP regulation, 3-hydroxy-4-[(E)-2-(2-methoxy-5-nitrophenyl) diazen-1-yl]-N-(3-nitrophenyl) naphthalene-2-carboxamide can be not classified for reproductive toxicity.