m-(4,5-dihydro-5-imino-3-methyl-1H-pyrazol-1-yl)benzenesulphonic acid

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

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 is from J-check

Data source

Reference

Reference Type:

other: J check

Title:

28 days repeated oral toxicity study for 7-Amino-4-hydroxy-2-naphthalenesulfonic acid

Author:

National Institute of Technology and Evaluation

Year:

2016

Bibliographic source:

J check, 2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

28 days repeated oral toxicity study was performed for the test compound 7-Amino-4-hydroxy-2-naphthalenesulfonic acid using rats

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

7-amino-4-hydroxynaphthalene-2-sulphonic acid

EC Number:

201-718-9

EC Name:

7-amino-4-hydroxynaphthalene-2-sulphonic acid

Cas Number:

87-02-5

Molecular formula:

C10H9NO4S

IUPAC Name:

7-Amino-4-hydroxy-2-naphthalenesulfonic acid

Details on test material:

- Name of test material : 7-Amino-4-hydroxy-2-naphthalenesulfonic acid
- Molecular formula: C10H9NO4S
- Molecular weight: 239.25 g/mol
- Substance type: Organic

Specific details on test material used for the study:

- Name of test material : 7-Amino-4-hydroxy-2-naphthalenesulfonic acid
- Molecular formula: C10H9NO4S
- Molecular weight: 239.25 g/mol
- Substance type: Organic
- Physical state: Grey white powder
- Impurities: 91.8% pure

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

No data

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan, Inc
- Age at study initiation: 6 weeks old
- Weight at study initiation:
Males: 208.4 to 228.6 g
Females: 137.6 to 165.0 g
- Fasting period before study:
- Housing: Animals were placed in a stainless steel hanger cages. 3 rats were housed together during the quarantine period and then housed individually during the study period.
- Diet (e.g. ad libitum): High pressure steam sterilized solid feed (MF, Oriental Yeast Industry
Co., Ltd.)
- Water (e.g. ad libitum): water added with sodium hypochlorite added (about 2 ppm)
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 2 ° C
- Humidity (%): 55 ± 10%
- Air changes (per hr): ventilation times of 13 / hour
- Photoperiod (hrs dark / hrs light): lighting time of 7 to 19 h

IN-LIFE DATES: From: To: No data

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

No data

Vehicle:

other: 5% aqueous gum arabic solution

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test substance was suspended at concentrations of 2.5, 5 and 10 w / v% in the vehicle to prepare a solution.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): 5% aqueous gum arabic solution
- Concentration in vehicle: 0, 250, 500 or 1000 mg/Kg bw
- Amount of vehicle (if gavage): 10mL/kg
- Lot/batch no. (if required): PTG 0424
- Purity: No data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

28 days

Frequency of treatment:

Once daily

Doses / concentrations

Remarks:

0, 250, 500 or 1000 mg/Kg bw

No. of animals per sex per dose:

Total: 96
0 mg/Kg bw: 12 male and 12 female
250 mg/Kg bw: 6 male and 6 female
500 mg/Kg bw: 6 male and 6 female
1000 mg/Kg bw: 12 male and 12 female

Recovery group:
0 mg/Kg bw: 6 male and 6 female
1000 mg/Kg bw: 6 male and 6 female

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose was set from the results of preliminary test (dose: 0, 250, 500 and 1000 mg / kg) by repeated administration for 2 weeks. That is, since no toxicity was expressed by the test substance even at the dose of 1000 mg / kg in the test, the dose in this study was 1000 mg / kg as a high dose according to the guidelines of the Chemical Substitution Law, and 500 and 250 Mg/ kg in total 3 doses were set.
- Rationale for animal assignment (if not random): The grouping was carried out by stratified continuous randomization method based on the body weight on the day before starting the administration.
- Rationale for selecting satellite groups: No data
- Post-exposure recovery period in satellite groups: Yes
- Section schedule rationale (if not random): No data

Positive control:

No data

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During the administration period, the general condition and the presence or absence of death were observed twice in total, twice a day before and after administration and twice in the morning and afternoon every day during the recovery period.
- Cage side observations checked in table [No.?] were included. General condition, mortality

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data

BODY WEIGHT: Yes
- Time schedule for examinations: Twice weekly during the administration period and recovery period.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, Twice weekly during the administration period and recovery period.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the end of the administration period and at the end of the recovery period
- Anaesthetic used for blood collection: Yes, pentobarbital sodium anesthesia
- Animals fasted: No data
- How many animals: 6/dose
- Parameters checked in table [No.?] were examined. the number of white blood cells, the number of red blood cells, The amount of hemoglobin, the hematocrit value and the number of platelets were measured, and the average red blood cell volume (MCV), the average red blood cell hemoglobin amount (MCH ) and average red blood cell hemoglobin concentration (MCHC) were calculated. Furthermore, 3.8% sodium citrate added blood was centrifuged at 3000 rpm for 15
minutes, and the plasma obtained was used to perform a total automatic blood coagulation measurement time and activated partial thromboplastin time were measured.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the end of the administration period and at the end of the recovery period
- Animals fasted: No data
- How many animals: 6/dose
- Parameters checked in table [No.?] were examined. Total protein, albumin, A / G ratio, total bilirubin, GOT, GPT, glutamyl transpeptidase, alkaline phosphatase, total cholesterol, triglyceride, phospholipids, glucose , urea nitrogen, creatinine, inorganic phosphate And calcium were measured. Sodium, potassium, and crawl were measured.

URINALYSIS: Yes
- Time schedule for collection of urine: On the 4 th week of administration and 2 weeks of recovery
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table [No.?] were examined. PH, protein, glucose, ketone bodies, bilirubin, occult blood, and urobilinogen were examined. Further, fresh urine was centrifuged at 1,500 rpm for 5 minutes and the obtained urine sediment was microscopically examined. In addition, urine volume, color tone, osmotic pressure and specific gravity was also measured.

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

OTHER: No data

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, organ weight (absolute weight) was measured by removing brain, heart, lung (including bronchus), thymus, liver, spleen, kidney, adrenal glands, testis and ovaries, and the body weight ratio on the day of necropsy. Relative organ weight was also calculated.

HISTOPATHOLOGY: Yes, in addition to the organs tested for gross pathology, hypophysis, spinal cord, eyeball, thyroid (including parathyroid gland), pancreas, stomach, bladder, femur (including bone marrow) and macroscopic abnormal sites were collected and resuspended in 10% neutral buffered formalin The solution was fixed with a solution (glutaraldehyde solution for eyeballs, prefixed with Bouin solution for testis).

Other examinations:

No data

Statistics:

The mean value and standard deviation were determined for each group for body weight, food intake, urinalysis (excluding
qualitative reaction), hematology examination, blood biochemical examination, organ weight and organ weight ratio, for each group, Homogeneity was tested. If the variance is uniform, a one-way analysis of variance was performed, and when a significant difference was observed between the groups in this analysis, a pairwise comparison test of each group was performed by the Dunnett method. When the variance is not uniform, a rank test is performed by the Kruskal-Wallis method. In the case where a significant difference is observed between the groups in this test, a pairwise comparison test of the Dunnett type was performed. When there was no significant difference between the groups by the variance analysis described above or the rank test by the Kruskal-Wallis method, multiple comparison between each group was not carried out. In both cases, the significance level was set at 5%.

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

At 4 weeks of dosing, loss of the maxillary incisor tip was observed in one male in the 1000 mg / kg group, but it was considered to be an accidental change. In addition to this there was no change in general condition.

Mortality:

no mortality observed

Description (incidence):

No mortality was noted

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No significant difference between the control group and each administration group throughout the administration period and recovery period was noted

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

A transient increase in food intake was observed in males of the 1000 mg / kg group at the 4th week of administration, but it was thought that there was minor fluctuation and was of no toxicological significance

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Description (incidence and severity):

No significant alterations were noted

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

There was no change at the end of the administration period, and as a minor change only at the end of the recovery period, GOT increased in male at 1000 mg / kg group and calcium was decreased in female of the same group.

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

A slight rise in urine specific gravity in males at 500 mg / kg group at 4 weeks of dosing was noted.

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

A decrease in the relative weight of the heart in females in all treatment groups was noted. In addition, in the 250 and 1000 mg / kg group, the absolute weight of the heart was decreased, and the same tendency was observed in the females of the 500 mg / kg group. However, these were considered to be minor fluctuations, and because there was no constant trend in the dose and the degree of change, it was considered to be an accidental fluctuation unrelated to administration.

At the end of the recovery period, there was an increase in the absolute weight of the lungs and a decrease in the relative weight of the heart in males in the 1000 mg / kg group.

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

Test group: A single grayish white spot and mild pyeloid dilatation in the right kidney was noted in one male in the 1000 mg / kg group. The same extent of renal pelvic dilation in the right kidney was also seen in 2 males in the 250 mg / kg group, and in one of them, mild atrophy of the left testis was also observed. In these histopathological examinations, renal pelvic dilation was observed in the kidneys, and 1 of them in the 1000 mg / kg group had basophilization of localized proximal renal tubular epithelium, dilation of distal tubule, and Lymphocyte infiltration into the interstitium as well as calcification of the papilla were observed. Atrophy was also confirmed in the testis by histopathological examination.

These changes were frequently observed changes in normal rats, and since their frequency of appearance was not related to the dose, it was considered that these changes were spontaneous changes. No other macroscopic and histopathological changes were observed at the end of the administration period.

Recovery group: No changes were noted

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Details on results:

No data

Effect levels

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No significant alteration were noted

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The No Observed Adverse Effect Level (NOAEL) for the test compound 7-Amino-4-hydroxy-2-naphthalenesulfonic acid is found to be 1000 mg/Kg bw.

Executive summary:

28 days repeated oral toxicity study was performed for the test compound 7-Amino-4-hydroxy-2-naphthalenesulfonic acid using rats. The animals were dosed at dose levels of 0, 250, 500 or 1000 mg/Kg bw. In addition, recovery group was also included in the study at dose levels of 0 or 1000 mg/Kg bw for 14 days.There was no change due to administration in any of the general condition, body weight, food intake, urinalysis, hematology examination, blood biochemical examination, organ weight, necropsy, and histopathological examination. At 4 weeks of dosing, loss of the maxillary incisor tip was observed in one male in the 1000 mg / kg group, but it was considered to be an accidental change. In addition to this there was no change in general condition. No mortality was noted in the treated animals. No significant difference between the control group and each administration group throughout the administration period and recovery period was noted. A transient increase in food intake was observed in males of the 1000 mg / kg group at the 4th week of administration, but it was thought that there was minor fluctuation and was of no toxicological significance. There was no change at the end of the administration period, and as a minor change only at the end of the recovery period, GOT increased in male at 1000 mg / kg group and calcium was decreased in female of the same group. A slight rise in urine specific gravity in males at 500 mg / kg group at 4 weeks of dosing was noted. A decrease in the relative weight of the heart in females in all treatment groups was noted. In addition, in the 250 and 1000 mg / kg group, the absolute weight of the heart was decreased, and the same tendency was observed in the females of the 500 mg / kg group. However, these were considered to be minor fluctuations, and because there was no constant trend in the dose and the degree of change, it was considered to be an accidental fluctuation unrelated to administration. At the end of the recovery period, there was an increase in the absolute weight of the lungs and a decrease in the relative weight of the heart in males in the 1000 mg / kg group. A single grayish white spot and mild pyeloid dilatation in the right kidney was noted in one male in the 1000 mg / kg test group. The same extent of renal pelvic dilation in the right kidney was also seen in 2 males in the 250 mg / kg group, and in one of them, mild atrophy of the left testis was also observed. In these histopathological examinations, renal pelvic dilation was observed in the kidneys, and 1 of them in the 1000 mg / kg group had basophilization of localized proximal renal tubular epithelium, dilation of distal tubule, and Lymphocyte infiltration into the interstitium as well as calcification of the papilla were observed. Atrophy was also confirmed in the testis by histopathological examination. These changes were frequently observed changes in normal rats, and since their frequency of appearance was not related to the dose, it was considered that these changes were spontaneous changes. No other macroscopic and histopathological changes were observed at the end of the administration period. No changes were noted in the recovery group. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for the test compound 7-Amino-4-hydroxy-2-naphthalenesulfonic acid is found to be 1000 mg/Kg bw.