Name: COMPLETE / SUBSTANCE : benzenesulfonamide / benzenesulfonamide / 98-10-2
Fri, 16 Dec 2022, 16:24:13+0900 /Legal entity owner: National Institute of Health Sciences
Printing date: 2022-12-16T16:24:13.265+09:00

iTable of Contents
0/0 ............................................................................................................................. 1
National Institute of Health Science ..................................................................... 2
benzenesulfonamide ........................................................................................... 3
CORE .......................................................................................................... 3
1 General information .......................................................................... 3
1.10 Assessment approach (assessment entities) ......................... 3
Assessment approach (assessment entities) ........................ 3
OECD .......................................................................................................... 4
D Health Effects .................................................................................. 4
67 Repeated dose toxicity: oral .................................................... 4
Repeated dose toxicity: oral.001 ........................................... 470 Genetic toxicity in vitro ......................................................... 12
Genetic toxicity in vitro.001 ................................................ 12
Genetic toxicity in vitro.002 ................................................ 18
73 Toxicity to reproduction ........................................................ 22
Toxicity to reproduction.001 ............................................... 22
DOMAIN .................................................................................................... 30
Substance ......................................................................................... 30
Substance ................................................................................ 30
References ................................................................................................................ 31
Reference Substances ...................................................................................... 31
benzenesulphonamide ............................................................................... 31
Test Materials .................................................................................................. 33
benzenesulfonamide .................................................................................. 33
Literatures ....................................................................................................... 34
A reproduction/developmental toxicity screening test in rats treated orallywith benzensulphonamide .......................................................................... 34
In Vitro Chromosomal Aberration Test of benzenesulfonamide on Cultured
Chinese Hamster Cells. ............................................................................. 35
Reverse mutation test of benzenesulfonamide in Bacteria ........................... 36
Twenty-eight-day Repeat Dose Oral Toxicity Test of benzenesulfonamide
in Rats ...................................................................................................... 37
Legal Entities ................................................................................................... 38
National Institute of Health Sciences .......................................................... 38ii

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benzenesulfonamide / benzenesulfonamide / 98-10-2
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LEGAL_ENTITY: National Institute of Health
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General information
Legal entity nameNational Institute of Health Science2

benzenesulfonamide
CORE
General information
Assessment approach (assessment entities)
FIXED_RECORD: Assessment approach

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OECD
Health Effects
Repeated dose toxicity: oral
ENDPOINT_STUDY_RECORD: Repeated dose toxicity: oral.001

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Administrative data
Endpoint
short-term repeated dose toxicity: oral
Type of information
experimental study
Adequacy of study
key study
Robust study summary
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
Twenty-eight-day Repeat Dose Oral Toxicity Test of benzenesulfonamide in Rats / MHLW, Japan / study
reportData accessdata published

Materials and methods
4Test guideline
Qualifierequivalent or similar to guideline
Guideline
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Qualifieraccording to guideline
Guideline
other: Guideline for 28-Day Repeated Dose Toxicity Test in Mammalian Species (Chemical
Substances Control Law of Japan)
GLP compliance
yes

Test material
Test material information
benzenesulfonamide
Specific details on test material used for the study- Name of test material (as cited in study report): benzenesulfonamide- Analytical purity: 99.6%- Lot No.: GF01
- Storage condition of test material: at a cold (temperature 2-6 ℃) and dark place, with airtightstopper.
- Stability under test conditions: The stability of test material was identified by analysis of theremainder.

Test animals
Speciesratcommon rodent species
Strainother: Crl:CD(SD)Sexmale/female
Details on test animals or test system and environmental conditions
TEST ANIMALS- Source: Charles River Japan, Inc.
- Age at study initiation: 5 weeks old- Weight at study initiation: male 168 g (150-177 g), female 143 g (135-152 g)- Housing: Animals were individually housed in a metallic cage with wire mesh bottoms- Diet: Solid feed (MR stock: Nosan Corporation) was given ad libitum.
- Water: Tap water was given ad libitum.
- Acclimation and quarantine period:7-8 days
ENVIRONMENTAL CONDITIONS- Temperature (°C): 22.0-22.8 ℃- Humidity (%): 53-62%- Air changes (per hr): 10 or more
- Photoperiod (hrs dark / hrs light): 12 hr dark/12 hr light (light: 7:00~19:00)
5Administration / exposure
Route of administration
oral: gavage
Vehiclemethylcellulose
Analytical verification of doses or concentrations
yes
Duration of treatment / exposure
28 daysFrequency of treatment
once a day
Doses / concentrations
Dose / conc.
0mg/kg bw/day (actual dose received)

Dose / conc.
6

mg/kg bw/day (actual dose received)

Dose / conc.
30

mg/kg bw/day (actual dose received)

Dose / conc.
150

mg/kg bw/day (actual dose received)

No. of animals per sex per dose
10/sex (0, 150 mg/kg bw/day)
5/sex (16, 30 mg/kg bw/day)Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale: Doses in this test were set based on the results of the following study: 14day repeated dose oral toxicity test (Crl:CD(SD) rats, doses: 0 (olive oil), 2, 6, 20, 60, or 200 mg/kg bw/day). At 60 mg/kg/day and higher, increased relative liver and kidney weights and changes in blood
chemistry were observed. At 200 mg/kg bw/day, changes in body weight were observed. On the basisof these effects, a dose level of 150 mg/kg was selected as the maximum dose expecting to induce
the toxic changes, while a dose level of 6 mg/kg bw/day was selected as the lowest dose expecting
to induce no toxicological effects. A dose level of 30 mg/kg bw/day was selected as a middle dose.

6

- Rationale for animal assignment (if not random): Body weight-balanced randomization- Post-exposure recovery period in satellite groups: 14 days

Examinations
Observations and examinations performed and frequency
LINICAL OBSERVATIONS: Yes- Time schedule: every day during the administration period (4 times a day) and the recovery period(at least once a day)
DETAILED CLINICAL OBSERVATIONS: Yes
The functional observational battery testing (FOB) was performed on all animals. Among the mea
sures in the FOB, detailed clinical observations were made before the initiation of dosing. Thereafter,
detailed clinical observations were made once a week in dosing and recovery periods.
Sensory motor reflexes, forelimb and hindlimb grip strengths, and motor activity were measured
on week 4 of administration period (main/recovery group animals) and week 2 of recovery period (recovery group animals).
BODY WEIGHT: Yes- Time schedule for examinations: Before administration (on days 1, 7, 14, 21 and 28 of the admin
istration period, days 7 and 14 of the recovery period) and the necropsy days after completion of every
period.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):- Food consumption: Yes. Once a week for 24-h (males: on days 5, 12, 19 and 26 of the administration
period and days 5 and 12 of the recovery period. females: on days 4, 11, 18 and 25 of the administr
ation period and days 4 and 11 of the recovery period)
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes- Time schedule for collection of blood: the after completion of the administration and recovery
periods- Anaesthetic used for blood collection: ether- Animals fasted: Yes (overnight)- How many animals: all animals
CLINICAL CHEMISTRY: Yes- Time schedule for collection of blood: the day after completion of the administration and recovery
periods- Anaesthetic used for blood collection: ether- Animals fasted: Yes (overnight)- How many animals: all animals
URINALYSIS: Yes- Time schedule for collection of urine: on weeks 4 of the administration period and weeks 2 of the recovery period.
- Metabolism cages used for collection of urine: Yes
NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology
GROSS PATHOLOGY: Yes
ORGAN WEIBHT: Yes [brain, pituitary gland, thyroid, adrenal, spleen, heart, liver, kidney, thymus, testis,
epididymis, ovary]

7

HISTOPATHOLOGY: Yes [brain (cerebrum, cerebellum and medulla oblongata), pituitary gland, spinal
cord (cervical, thoracical, lumber), thymus, thyroid (including parathyroid), adrenal glands, spleen, heart, stomach, liver, duodenum, jejunum, ileum (including Peyer's patches), cecum, colon, rectal, mes
enteric lymph nodes, submandibular lymph nodes, trachea, lung, kidney, bladder, testis, epididymis,
prostate, seminal vesicles, ovary, uterus, vagina, eye, bone marrow (femur) and the sciatic nerve.
Statistics
As for parametric data (grip strength, locomotor activity, body weight, body weight gain, food consu
mption, hematology and clinical chemistry data, organ weights), the values of means and standard
deviations were calculated per group. When more than three groups exist in the test group, Bartlett
test for variance was done, and if the variance was homogenous, ANOVA was applied. If the variance
was not homogenous or data was non-parametric (differential WBC percentage, urinalysis data),
Kruskal-Wallis rank sum test was used. Consequently, if the result was significant, Dunnett multiple
comparison or Dunnett-typed method was used for detection of statistical significance against control group. When the number of the test group was two, F-test was used as for parametric data. Then, student's t-test or Aspin-Welch's t-test was applied depending on the result of homogeneity of variance.
While, as for non-parametric data, Man-Whitney's U-test was applied. Furthermore, as for categorized
data (incidence of abnormal findings in clinical observation, detailed observation, sensory functional
examination, necropsy and histopathology), Fischer's exact test was used. In any tests, level of signi
ficance was set at 5%.

Results and discussion
Results of examinations
Clinical signs
effects observed, treatment-related
Description (incidence and severity)
At 150 mg/kg bw/day, one male died on day 21. This animal showed decrease in locomotor activity,
pale skin, prone position, and convulsion. Decreased locomotor activity was also observed in 4 out of
9 males and one out of 10 females at 150 mg/kg bw/day. Transient salivation was also observed at
this dose.
Mortality
mortality observed, treatment-related
Description (incidence)
At 150 mg/kg bw/day, one male died on day 21.
Body weight and weight changes
effects observed, treatment-related
Description (incidence and severity)
Body weight and body weight gain were significantly decreased at 30 mg/kg bw/day and higher inboth sexes. After the recovery period, body weight was still significantly low, but body weight gain
was significantly increased.
Food consumption and compound intake (if feeding study)
effects observed, treatment-related
Description (incidence and severity)
Food consumption was significantly decreased at 6 mg/kg bw/day and higher in males and 30 mg/kg
bw/day and higher in females.
Haematological findings
effects observed, treatment-related8Description (incidence and severity)
At the end of the administration period, no effects were observed. Decreased Hb and Ht wereobserved at the end of the recovery period. These effects were considered to be due to low food consumption and low body weight.
Clinical biochemistry findings
effects observed, treatment-related
Description (incidence and severity)
At 150 mg/kg bw/day, significantly high values of ALT, T-Cho, and T-Bil in males, and significantly
high values of BUN and T-Cho in females were observed. At the end of the recovery period, there weresignificantly decreased Alb and Crea in both sexes, and significantly increased ALT and decreased
Glu and BUN in males. These changes observed at the end of the administration period dented torecover.
Urinalysis findings
no effects observed
Organ weight findings including organ / body weight ratios
effects observed, treatment-related
Description (incidence and severity)
Absolute weights of the heart and pituitary gland in both sexes, spleen and epididymis in males, andbrain in females were significantly decreased due to low body weights at 150 mg/kg bw/day. Relative
weights of the brain were increased in males at 30 and 150 mg/kg bw/day and in females at 150 mg/
kg bw/day. Relative weights of the heart, liver, and kidney in both sexes and adrenal gland and testis
in males were also significantly increased at 150 mg/kg bw/day. These changes observed at the endof the administration period dented to recover.
Gross pathological findings
no effects observed
Histopathological findings: non-neoplastic
effects observed, treatment-related
Description (incidence and severity)
Mineralization of the lung alvenolar septa and kidney papilla/pelvis was observed in both sexes at 150 mg/kg bw/day. Hypertrophy of the centrilobular hepatocytes was observed in both sexes at 150
mg/kg bw/day. Hyaline droplet of the proximal tubular epithelium in the kidney observed in maleanimals at 30 mg/kg bw/day and higher were considered to be male rat-specific α-2u globulin nephro
pathy. Hyperplasia of transitional cells in the urinary bladder was observed at 30 mg/kg bw/day andhigher in both sexes. Sulfonamides are known to produce urinary bladder hyperplasia, but the effectis specific to rats due to urinary composition. At the end of the recovery period, all of the changes
except mineralization in the lung had resolved or showed a tendency to resolve.

Effect levels
Key result
false
Dose descriptor
NOAEL
Effect level6

mg/kg bw/day (actual dose received)

Based onact. ingr.
9

Sex
male/female
Basis for effect level
body weight and weight gain
Male and female rats administered 30 and 150 mg/kg bw/day showed significantly decreased b
ody weight.
food consumption and compound intake
Male and female rats administered 30 and 150 mg/kg bw/day showed significantly decreased foodconsumption.
histopathology: non-neoplasticchanges in the urinary bladder were observed at 30 mg/kg bw/day

Target system / organ toxicity
Key result
false
Critical effects observed
yes
Lowest effective dose / conc.
30

mg/kg bw/day (nominal)

Organ
bladder
Treatment related
yes
Dose response relationshipyes

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF98-10-2b.pdf

Applicant's summary and conclusion
Executive summary
A repeated-dose 28-day oral toxicity study was performed in accordance with the Japanese guidelines(similar to OECD TG 407). Male and female rats (5 or 10 animals/sex/dose) were administeredbenzenesulfonamide for 28 days at 0 (vehicle:1 w/v% methyl cellulose solution), 6, 30, and 150 mg/
kg bw/day. Five out of 10 males with this administration at 0 and 150 mg/kg bw/day were used as arecovery assessment group and examined after a 14-day recovery period. At 150 mg/kg bw/day, one
male died and decrease in locomotor activity was observed in both sexes during the administration
period. Male and female rats administered 30 and 150 mg/kg bw/day showed significantly decreasedfood consumption and body weight. Upon the administration of benzenesulfonamide at 150 mg/kg bw/
day, the following effects on the liver were also observed in both sexes: significantly increased relative
10

organ weight with centrilobular hypertrophy and changes in blood chemical parameters. Mineralization
was observed in the kidney and lung at 150 mg/kg bw/day in both sexes. Moreover, hyperplasia oftransitional cells in the urinary bladder was observed at 30 mg/kg bw/day and higher in both sexes.
Sulfonamides are known to produce urinary bladder hyperplasia, but the effect is specific to rats due
to urinary composition. At the end of the recovery period, all of the changes except mineralization in
the lung had resolved or showed a tendency to resolve. Based on decreases in food consumption andbody weight and histopathological changes in the urinary bladder at 30 mg/kg bw/day, the NOAEL of
the repeated-dose toxicity was determined to be 6 mg/kg bw/day for male and female rats.

11

Genetic toxicity in vitro
ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.001

UUID: e8c3ace4-eb02-488a-a13f-f7add8d8aca7
Dossier UUID:
Author:Date: 2022-12-16T16:21:23.677+09:00
Remarks:

Administrative data
Endpoint
in vitro gene mutation study in bacteria
Type of information
experimental study
Adequacy of study
key study
Robust study summary
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
Reverse mutation test of benzenesulfonamide in Bacteria / MHLW, Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
in vitro gene mutation study in bacteria

12Qualifieraccording to guideline
Guideline
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
genetic toxicity in vitro, other
GLP compliance
yesType of assay
bacterial reverse mutation assay
in vitro gene mutation study in bacteria

Test material
Test material information
benzenesulfonamide
Specific details on test material used for the study- Name of test material (as cited in study report): benzenesulfonamide- Analytical purity: 99.2%- Lot/batch No.:EWG6537 (Wako)- Stability under test conditions: Stable- Storage condition of test material: dark place at room temperature (21.1-25.3 C degree )MethodSpecies / strain
Species / strain / cell typeS. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2bacteria
Metabolic activation
with and without
Metabolic activation system
rat liver, induced by phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose
Dose–range finding test (–S9 mix and +S9 mix): 0 (vehicle), 1.22, 4.88, 19.5, 78.1, 313, 1250, and 5000
μg/plate
In range-finding studies, growth inhibition was not observed on all plates with concentration of up to
5000 μg/plate with/without metabolic activation.
Main bacterial reverse mutation test (–S9 mix and +S9 mix): 0 (vehicle), 313, 625, 1250, 2500, and5000 μg/plate
Vehicle / solvent- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test substance was soluble in DMSO, but not inwater.

13

Controls
Untreated negative controls
yes
Negative solvent / vehicle controls
yes
Positive controls
yes
Positive control substance
9-aminoacridine(9-aminoacridine acid), -S9 mix, TA1537
sodium azide-S9 mix, TA1535
other: 2-aminoanthracene (+S9 mix, all strains), 2-methoxy-6-chloro-9-[3-(2-chloroethyl)-aminopro
pylamino]acridine-2HCl, -S9mix TA100, WP2, TA98
Details on test system and experimental conditions
METHOD OF APPLICATION: Preincubation
The mixtures were incubated at 37 C degree with shaking (90 times/min).
DURATION- Preincubation period: 20 min- Exposure duration: 48 hours
NUMBER OF REPLICATIONS: 3
DETERMINATION OF CYTOTOXICITY- Method: Cell growth
Evaluation criteria
Criteria for determining a positive result were as follows; A 2–fold or more increase in the number ofrevertant colonies compared with the solvent control, a concentration–related increase in the numberof revertant colonies, and a reproducible increase in the number of revertant colonies.
Statistics
No statistic method was used for judging of results.

Results and discussion
Test results
Key result
false
Species / strain
S. typhimurium TA 1535
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
14

Vehicle controls validity
valid
Untreated negative controls validity
valid
Positive controls validity
valid
Key result
false
Species / strain
S. typhimurium TA 1537
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity
valid
Untreated negative controls validity
valid
Positive controls validity
valid
Key result
false
Species / strain
S. typhimurium TA 98
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity
valid
Untreated negative controls validity
valid
Positive controls validity
valid
Key result
false
15

Species / strain
S. typhimurium TA 100
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity
valid
Untreated negative controls validity
valid
Positive controls validity
valid
Key result
false
Species / strain
E. coli WP2 uvr A pKM 101
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity
valid
Untreated negative controls validity
valid
Positive controls validity
valid
Additional information on results
TEST-SPECIFIC CONFOUNDING FACTORS- Precipitation: Precipitation was not observed on any plates with/without metabolic activation.
COMPARISON WITH HISTORICAL CONTROL DATA:
In all test conditions and in all tested strains, the number of revertant colonies of solvent controls andpositive controls were within the range of historical control data.

Any other information on results incl. tables
Figures and Tables (in Japanese) are available in the following full report of the study.http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF98-10-2e.pdf
16

Tables (in English) are attachted to this document. Please download the export file to see the Tables.

Overall remarks, attachments
Attachments
Attached (sanitised) documents for publication
98-10-2_Ames.xlsx / 26.894 KB (application/vnd.openxmlformatsofficedocument.spreadsheetml.sheet)

Applicant's summary and conclusion
Conclusions
Interpretation of results: Negative
Executive summary
In a bacterial reverse mutation assay using S. typhimurium TA100, TA1535, TA98, and TA1537 and E.
coli WP2uvrA (OECD TG 471), benzenesulfonamidewas negative with or without metabolic activation.

17

ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.002

UUID: 2fde2394-2aba-4eeb-ab5a-acaf0d8feaee
Dossier UUID:
Author:Date: 2022-12-16T16:22:01.200+09:00
Remarks:

Administrative data
Endpoint
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information
experimental study
Adequacy of study
key study
Robust study summary
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
In Vitro Chromosomal Aberration Test of benzenesulfonamide on Cultured Chinese Hamster Cells. /MHLW, Japan / study report

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)in vitro cytogenicity / chromosome aberration study in mammalian cells
Qualifieraccording to guideline
18

Guideline
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
genetic toxicity in vitro, other
GLP compliance
yesType of assay
in vitro mammalian chromosome aberration testin vitro cytogenicity / chromosome aberration study in mammalian cells

Test material
Test material information
benzenesulfonamide
Specific details on test material used for the study- Name of test material (as cited in study report): benzenesulfonamide- Analytical purity: 99.2%- Lot/batch No.:EWG6537 (Wako)- Stability under test conditions: Stable- Storage condition of test material: dark place at room temperature (21.1-25.3 C degree)

MethodTarget gene
Chromosome
Species / strain
Species / strain / cell typeother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
with and without
Metabolic activation system
rat liver, induced by phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose+/-S9 mix (short-term treatment): 0, 100, 200, 400, 800, 1600 μg/mL-S9 mix (continuous treatment): 0, 100, 200, 400, 800, 1600 μg/mL
Test concentrations were set based on the following results of a preliminary study:
short term treatment (-S9): growth inhibition was not observed at 5000 μg/mL
short term treatment (+S9): growth inhibition was not observed at 5000 μg/mL
continuous treatment (-S9): growth inhibition was not observed at 500 μg/mL, but 90% growth inhib
ition was observed at 5000 μg/mL
According to the Guideline, the maximum concentration was set at 10 mmol/L (1600 ug/mL).

Vehicle / solvent- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test substance was soluble in DMSO, but not inwater.

19

Controls
Negative solvent / vehicle controls
yes
Positive controls
yes
Positive control substancebenzo(a)pyrene+S9 mix
mitomycin C-S9 mix
Details on test system and experimental conditions
METHOD OF APPLICATION: Exposure duration: [continuous treatment]: 24 hrs[short-term treatment]:6 hrs + 18 hr
SPINDLE INHIBITOR: Colcemid
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 500 cells / plate (1000 cells/concentration)
DETERMINATION OF CYTOTOXICITY- Method: relative total growth
Evaluation criteria
For the evaluation of the frequencies of structural aberrations and of polyploidy induced, the following
criteria were employed.
Appearance incidence of cells with chromosomal aberrations: Negative (-): < 5%; equivocal (±): 5-10%;
positive (+): > 10%.
Finally, the substance is positive when the incidence is considered to be dose-related and repro
ducible.
Statistics
not used

Results and discussion
Test results
Key result
false
Species / strainother: Chinese hamster lung (CHL/IU) cells
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity
valid
Positive controls validityvalid

Any other information on results incl. tables
20

Figures and Tables (in Japanese) are available in the following full report of the study.
http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF98-10-2f.pdf
Tables (in English) are attachted to this document. Please download the export file to see the Tables.

Overall remarks, attachments
Attachments
Attached (sanitised) documents for publication
98-10-2_CA Tables.xlsx / 25.046 KB (application/vnd.openxmlformatsofficedocument.spreadsheetml.sheet)

Applicant's summary and conclusion
Conclusions
Interpretation of results: Negative
Executive summary
An in vitro chromosomal aberration test using CHL/IU cells (OECD TG 473) showed positive result withmetabolic activation.

21

Toxicity to reproduction
ENDPOINT_STUDY_RECORD: Toxicity to reproduction.001

UUID: 05f7bda6-7b60-4623-befb-3860a7656a92
Dossier UUID:
Author:Date: 2022-12-16T16:23:53.886+09:00
Remarks:

Administrative data
Endpoint
screening for reproductive / developmental toxicity
Type of information
experimental study
Adequacy of study
key study
Robust study summary
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
A reproduction/developmental toxicity screening test in rats treated orally with benzensulphonamide /MHLW, Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
GLP complianceyes
22Test material
Test material information
benzenesulfonamide
Specific details on test material used for the study- Name of test material (as cited in study report): benzenesulfonamide- Purity: 99.9%- Lot/batch No.: FQ2GG- Stability under test conditions: Stable- Storage condition of test material: a cool (3-8 °C) and dark place (in a refrigerator), with an airtight s
topper- Dosing solution storage condition: under a cool (3-6 °C) place (in a refrigerator), in a brown glass
bottleTest animals
Speciesrat
Strainother: Crl:CD(SD)Sexmale/female
Details on test animals or test system and environmental conditions
TEST ANIMALS- Source: Charles River Laboratories Japan, Inc. Atsugi- Age at study initiation: 10 weeks old- Weight at study initiation: Males: 374-509 (average 442) g; Females: 228-286 (average 260) g- Housing: Steel wire-mesh cage (254 mm x 350 mm x 170 mm), pregnant rats were housed in a pla
stic Econ cage with bedding (340 mm x 400 mm x 185 mm) on GD17 and after.
- Diet: ad libitum- Water: ad libitum- Acclimation period: 20 days
ENVIRONMENTAL CONDITIONS- Temperature (°C): 22-24- Humidity (%): 49-61- Air changes: 10-15 times / hr
- Photoperiod: 12 hrs dark / 12 hrs light (07:00-19:00)

Administration / exposure
Route of administration
oral: gavage
Vehicleother: 1 w/v% methyl cellulose solution
Details on mating procedure- M/F ratio per cage: 1:1- Length of cohabitation: up to 14 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0] of pregnancy
23

Analytical verification of doses or concentrations
yes
Duration of treatment / exposure
Males were dosed for 28 days, including a 14-day pre-mating period and subsequent mating period.
Females were dosed for 40–53 days, including 14-day pre-mating, mating, and gestation periods, anduntil lactation day 3.
Frequency of treatment
once a day
Doses / concentrations
Dose / conc.
0mg/kg bw/day (actual dose received)

Dose / conc.
3

mg/kg bw/day (actual dose received)

Dose / conc.
10

mg/kg bw/day (actual dose received)

Dose / conc.
30

mg/kg bw/day (actual dose received)

No. of animals per sex per dose
12/sex/doseControl animals
yes, concurrent vehicle
Details on study design- Dose selection rationale: Doses in this test were set based on the 28-day repeated dose oral toxicity
test (Repeated dose toxicity: oral.001 ). A dose level of 30 mg/kg was selected as the maximum dose
expecting to induce the toxic changes, and then dose levels of 10 and 3 mg/kg bw/day were selected
as a middle dose and a minimum dose levels, respectively, in accordance with a common ratio of 3.
- Rationale for animal assignment (if not random): Body weight-balanced randomization

Examinations
Parental animals: Observations and examinations
AGE SIDE OBSERVATIONS: Yes- Time schedule:
Males and females: 3 times/day

24

BODY WEIGHT: Yes- Time schedule for examinations:
Males: Days 1, 8, 15, and 22 and the day of necropsy
Females: Days 1, 8, 15, and 22 during the precopulation period; gestation days 0, 7, 14, and 20; lactation days 0 and 4; and the day of necropsy.
FOOD CONSUMPTION: Yes
Males: Days 2, 8, 15 in dosing period
Females: Days 2, 8, 15; gestation days 1, 7, 14, and 20; lactation days 2 and 4
HAEMATOLOGY: No
CLINICAL CHEMISTRY: No
URINALYSIS: No
Oestrous cyclicity (parental animals)
Vaginal smears were collected from all females in the main groups and microscopically examined
every day from the day after the start of administration until the day copulation was confirmed.
During the pre-mating administration period, vaginal smear pictures were classified as proestrus,
estrus, metestrus or diestrus and examined for the frequency of estrus and interval between estruses(estrous cycle). During the mating period, vaginal smears were examined for the presence of sperm.
Litter observations
PARAMETERS EXAMINED:The following parameters were examined in F1 offspring [number and sexof pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, and weight].
GROSS EXAMINATION OF DEAD PUPS: Yes, for external abnormalities.
Postmortem examinations (parental animals)
SACRIFICE:
Male animals: Rats were euthanized by exsanguination under ether anesthesia on the day after the last administration.
Maternal animals: Rats were euthanized by exsanguination under ether anesthesia on day 4 of lactati
on.
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes (epididymis, prostate, seminal vesicle, testis, ovary, uterus, vagina, and grossabnormal sites)
ORGAN WEIGHTS, Yes: Testes and epididymis
Statistics
The data were analyzed for homogeneity of variance by the Bartlett test. If variances were
homogeneous, data was analyzed by the Dunnett test, whereas heterogeneous data was analyzed by
the Steel test (p<0.05, two-sided).
2 groups: The data were analyzed for homogeneity of variance by the F test. If variances were homogeneous, data was analyzed by the Student t test, whereas heterogeneous data was analyzed by the
Aspin-Welch t test (p<0.05, two-sided).
Especially,
Implantation index, Stillborn index, Liveborn index, External abnormalities, Viability index: the Steeltest (p<0.05 and <0.01, two-sided)
Copulation index, Fertility index, Insemination index, Delivery index: Fisher's exact test (p<0.05 and<0.01, two-sided)
Reproductive indices
Each parameter was determined by the following equations:
Copulation index (%) = (No. of copulated animals/No. of co-housed animals) × 100
25

Fertility index (%) = (No. of pregnant females/No. of copulated females) × 100
Insemination index (%) = (No. of pregnant females/No. of copulated males) × 100
Duration of gestation (days) = day 0 of lactation – day 0 of gestation
Delivery index (%) = (No. of females delivered liveborn pups/No. of pregnant females) × 100
Implantation index (%) = (No. of implantation sites/No. of corpora lutea) × 100
Stillborn index (%) = (No. of stillborn pups/Total No. of pups born) × 100
Liveborn index (%) = (No. of liveborn pups/Total No. of pups born) × 100
External abnormalities (%) = (No. of pups with external abnormalities/No. of liveborn pups) × 100
Sex ratio = No. of liveborn male pups/(No. of liveborn male pups + No. of liveborn female pups)
Offspring viability indices
Viability index (%) = (No. of surviving pus on day 4 after birth/No. of liveborn pups on day 0 after birth)
× 100

Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
Clinical signs
no effects observed
Mortality
no mortality observed
Body weight and weight changes
effects observed, treatment-related
Description (incidence and severity)
Body weights were significantly decreased at 30 mg/kg bw/day and body weight gain was sign
ificantly decreased at 100 mg/kg bw/day in males.
Food consumption and compound intake (if feeding study)
effects observed, treatment-related
Description (incidence and severity)
Food consumption was significantly low at 30 mg/kg bw/day in males.
Organ weight findings including organ / body weight ratios
effects observed, treatment-related
Description (incidence and severity)
Absolute and relative weights of the ovary were also significantly increased at 30 mg/kg bw/day.
Gross pathological findings
effects observed, non-treatment-related
Description (incidence and severity)
White focus in the epididymis was found in one males at 10 mg/kg bw/day.
Histopathological findings: non-neoplastic
effects observed, treatment-related
Description (incidence and severity)
Hyperplasia/hypertrophy of the transitional cells in the urinary bladder was observed at 10 and 30 mg/
kg bw/day in males. In females, hyperplasia/hypertrophy of the transitional cells in the urinary bladder
was observed upon administration of benzenesulfonamide at doses ≥ 3 mg/kg bw/day.
26

Reproductive function / performance (P0)
Reproductive function: oestrous cycle
no effects observed
Reproductive performanceno effects observed

Effect levels (P0)
Key result
false
Dose descriptor
NOAEL
Effect level30

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect level
reproductive performanceNo effects on fertility
Key result
false
Dose descriptor
LOAEL
Effect level3

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect level
histopathology: non-neoplastic
histopathological change in the urinary bladder

Results: F1 generation
General toxicity (F1)Mortality / viability
no mortality observed
27

Body weight and weight changes
effects observed, treatment-related
Description (incidence and severity)
The body weights of pups on postnatal day (PND) 0 and/or 4 were found to be significantly
decreased at 30 mg/kg bw/day in both sexes.

Effect levels (F1)
Key result
false
Dose descriptor
NOAEL
Generation
F1
Effect level10

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect level
body weight and weight gain
the body weights of pups on postnatal day (PND) 0 and/or 4 were found to be significantly
decreased at 30 mg/kg bw/day in both sexes.

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study. http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF98-10-2c.pdf

Applicant's summary and conclusion
Conclusions
The NOAEL for the rat reproductive/developmental toxicity of benzenesulfonamide was determined tobe 10 mg/kg bw/day based on decreased pup body weights at 30 mg/kg bw/day
Executive summary
A reproduction/developmental toxicity screening test (OECD TG 421) was also performed using rats.
In this study, benzenesulfonamide was administered via gavage to 12 animals/sex/dose at 0 (vehicle:1
w/v% methyl cellulose solution), 3, 10, and 30 mg/kg bw/day. Males were dosed for 28 days, including a
14-day pre-mating period and subsequent mating period. Females were dosed for 40–53 days, including
14-day pre-mating, mating, and gestation periods, and until lactation day 3. Significantly decreased bodyweight and hyperplasia/hypertrophy of the transitional cells in the urinary bladder were observed at
10 and 30 mg/kg bw/day in males. In females, hyperplasia/hypertrophy of the transitional cells in theurinary bladder was observed upon administration of benzenesulfonamide at doses ≥ 3 mg/kg bw/
day. Absolute and relative weights of the ovary were also significantly increased at 30 mg/kg bw/day.
28

There were no effects on fertility, but the body weights of pups on postnatal day (PND) 0 and/or 4
were found to be significantly decreased at 30 mg/kg bw/day in both sexes. The NOAEL for the rat
reproductive/developmental toxicity of benzenesulfonamide was determined to be 10 mg/kg bw/day
based on decreased pup body weights at 30 mg/kg bw/day while the lowest-observed-adverse-effectlevel for parental general toxicity was 3 mg/kg bw/day, based on the histopathological change in theurinary bladder.

29

DOMAIN
Substance
SUBSTANCE: benzenesulfonamide
UUID: f596d537-c3e7-44bf-b585-e989d5d06385
Dossier UUID:
Author:Date: 2022-12-16T16:23:53.886+09:00
Remarks:
Substance namebenzenesulfonamide
Legal entity
National Institute of Health Sciences / Kawasaki / Japan

Identification of substance
Reference substancebenzenesulphonamide / benzenesulfonamide / 98-10-2 / 202-637-1
EC number

EC name202-637-1

EC Inventory

CAS number

CAS name

98-10-2
IUPAC namebenzenesulfonamide

Role in the supply chain
Manufacturer
false
Importer
false
Only representativefalse
Downstream user
false

30

References
Reference Substances
REFERENCE_SUBSTANCE: benzenesulphonamide
UUID: ECB5-7ab84e8b-3c5d-4bed-bae6-4b4361df8447
Dossier UUID:
Author:Date: 2007-05-10T18:00:00.000+09:00
Remarks:
Reference substance namebenzenesulphonamide
IUPAC namebenzenesulfonamide

Inventory
Inventory number
Inventory namebenzenesulphonamide
Inventory
EC Inventory
Inventory number
202-637-1
CAS number
98-10-2
Molecular formula
C6H7NO2S
Description
CAS number
98-10-2

Synonyms
Synonyms
Identity
Benzenesulfonamide
Identity
Benzenesulfonamide

Molecular and structural information
31

Molecular formula
C6H7NO2S
Molecular weight157.1903
SMILES notation
NS(=O)(=O)c1ccccc1
InChl
InChI=1/C6H7NO2S/c7-10(8,9)6-4-2-1-3-5-6/h1-5H,(H2,7,8,9)
Structural formula

Related substancesGroup / category information
DSL Category: Organics

32

Test Materials
TEST_MATERIAL_INFORMATION: benzenesulfonamide
UUID: 5ef05df4-03fa-4af0-9669-2bfb6f6f6e3b
Dossier UUID:
Author:Date: 2018-03-06T15:00:30.000+09:00
Remarks:Namebenzenesulfonamide

Composition
Composition
Reference substancebenzenesulphonamide / benzenesulfonamide / 98-10-2 / 202-637-1
EC number

EC name202-637-1

EC Inventory

CAS number

CAS name

98-10-2
IUPAC namebenzenesulfonamide

33

Literatures
LITERATURE: A reproduction/developmental
toxicity screening test in rats treated orally withbenzensulphonamide
UUID: 344feefd-0a81-43ec-a80b-54f7a38428a1
Dossier UUID:
Author:Date: 2018-03-07T11:24:27.000+09:00
Remarks:

General information
Reference Typestudy report
Title
A reproduction/developmental toxicity screening test in rats treated orally with benzensulphonamide
Author
MHLW, Japan
Year
2014
Bibliographic source
available in the web of Japan Existing Chemical Data Base (JECDB) at http://dra4.nihs.go.jp/
mhlw_data/jsp/SearchPageENG.jsp
Testing facility
BoZo Research Center
Report number
R-1131

34

LITERATURE: In Vitro Chromosomal Aberration Test ofbenzenesulfonamide on Cultured Chinese Hamster Cells.
UUID: 68ce0a97-383f-4a44-b004-96474746b2c3
Dossier UUID:
Author:Date: 2018-03-07T10:16:56.000+09:00
Remarks:

General information
Reference Typestudy report
Title
In Vitro Chromosomal Aberration Test of benzenesulfonamide on Cultured Chinese Hamster Cells.
Author
MHLW, Japan
Year
2007
Bibliographic source
available in the web of Japan Existing Chemical Data Base (JECDB) at http://dra4.nihs.go.jp/
mhlw_data/jsp/SearchPageENG.jsp
Testing facility
BoZo Research Center
Report number
B06031735

LITERATURE: Reverse mutation test ofbenzenesulfonamide in Bacteria
UUID: 99eda9cb-6b68-4c85-9db1-5708603fd6a8
Dossier UUID:
Author:Date: 2018-03-07T08:55:20.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Reverse mutation test of benzenesulfonamide in Bacteria
Author
MHLW, Japan
Year
2007
Bibliographic source
available in the web of Japan Existing Chemical Data Base (JECDB) at http://dra4.nihs.go.jp/
mhlw_data/jsp/SearchPageENG.jsp
Testing facility
Bozo Research Center Inc.

36

LITERATURE: Twenty-eight-day Repeat Dose Oral Toxicity
Test of benzenesulfonamide in RatsUUID: ebf5708e-b097-4a1b-81f5-621e08563109
Dossier UUID:
Author:Date: 2018-03-06T14:53:53.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Twenty-eight-day Repeat Dose Oral Toxicity Test of benzenesulfonamide in Rats
Author
MHLW, Japan
Year
2011
Bibliographic source
available in the web of Japan Existing Chemical Data Base (JECDB) at http://dra4.nihs.go.jp/
mhlw_data/jsp/SearchPageENG.jsp
Testing facility
Research institute for animal science in biochemistry and toxicology (RIAS)
Report number
06-08837

Legal Entities
LEGAL_ENTITY: National Institute of Health Sciences
UUID: IUC4-b036ff75-0f3c-323b-b200-ed5f46cf5101
Dossier UUID:
Author:Date: 2022-11-07T15:49:29.000+09:00
Remarks:

General information
Legal entity nameNational Institute of Health Sciences
Remarks
Disclaimer: The contents in this document were created based on the MHLW (Ministry of Health,Labour and Welfare) peer reviewed study reports (in Japanese) in JECDB (Japan Existing Chemical
Database) at http://dra4.nihs.go.jp/mhlw_data/jsp/SearchPageENG.jsp. Authorship is in the Divisionof Risk Assessment, the National Institute of Health Sciences, and the contents do not reflect any official MHLW opinions or any other regulatory policies.

Address
Address 1
Tonomachi 3-25-26
Address 2
Kawasaki-ku
Postal code
210-9501
Town
KawasakiRegion / State
Kanagawa
CountryJapan
JP

Identifiers
Other IT system identifiers
IT system
LEO
ID
10767
IT systemIUCLID438

ID
16558402024DIV750

39