4,4'-Oxybis(benzenesulfonylhydrazide)

4,4'-オキシビス(ベンゼンスルホニルヒドラジド)


[CAS No. 80-51-3]

Molecular formula: C12H14N4O5S2　Molecular weight: 358.40

ABSTRACT

4,4'-Oxybis(benzenesulfonylhydrazide) was studied for oral toxicity in rats in a single dose toxicity test at doses of 0, 1000, 1500 and 2000 mg/kg, in a repeat dose toxicity test at doses of 0, 10, 30, 100 and 200 mg/kg/day, and in a preliminary reproduction toxicity screening test at doses of 0, 3, 10 and 30 mg/kg/day.

With regard to the single dose toxicity, one male dosed with 1500 mg/kg and 2 animals of each sex dosed with 2000 mg/kg died after showing paralytic or tiptoe gait and/or soiling of fur.

The LD50 value was estimated to be more than 2000 mg/kg for both sexes.

With regard to repeat dose toxicity, decrease of grip strength, a paralytic gait, degeneration of peripheral nerve fibers, increase of kidney weights, fatty degeneration of proximal tubular epithelium, and shortening of prothrombin time were observed in animals receiving 30 mg/kg/day or more. An anemic condition, decrease of body weight and food consumption, increase of total cholesterol and liver weight, and hypertrophy of centrilobular hepatocytes were observed in animals receiving 100 mg/kg or more per day. Death following paralytic gait, diarrhea, decrease of body weight and food consumption was observed in some animals receiving 200 mg/kg/day. Others receiving this dose were killed in a moribund condition.

The NOEL is considered to be 10 mg/kg/day for both sexes.

With regard to the preliminary reproduction toxicity, increase of liver and/or kidney weights was observed in parent animals receiving 10 mg/kg/day or more, but no effects were observed on the reproductive performance of the parents or on the developmental performance in the pups.

The NOELs for systemic toxicity in the parents and reproductive and developmental toxicity are considered to be 3 mg kg/day, and 30 mg/kg/day respectively.

Reverse mutation tests using microorganisms (Salmonella typhimurium, Escherichia coli) were conducted to assess the potential of 4,4'-oxybis(benzenesulfonylhydrazide) to induce gene mutations. 4,4'-Oxybis(benzenesulfonylhydrazide) induced gene mutations in TA100, TA1535 and WP2 uvrA under the conditions of this study.

In vitro chromosomal aberration tests using cultured Chinese hamster cells (CHL/IU) were conducted to assess the potential of 4,4'-oxybis(benzenesulfonylhydrazide) to induce chromosomal aberrations. 4,4'-Oxybis(benzenesulfonylhydrazide) induced chromosomal aberrations in cultured cells under the conditions of this study.

SUMMARIZED DATA FROM THE STUDIES

1. Single Dose Oral Toxicity 1)

Purity:99.3 %
Test species/strain:Rat/Crj:CD(SD)IGS
Test method:OECD Test Guideline 401
　Route:Oral (Gavage)
　Dosage:0 (Vehicle), 1000, 1500, 2000 mg/kg
　Number of animals/group:Males, 5; females, 5
　Vehicle:Corn oil
GLP:Yes

　Test results:

One male dosed with 1500 mg/kg and 2 animals of each sex dosed with 2000 mg/kg died 2 or 3 days after the administration after showing paralytic gait, tiptoe gait and/or soiling of fur.

The LD50 value was estimated to be more than 2000 mg/kg for both sexes.

2. Repeated Dose Oral Toxicity 1)

Purity:99.3 %
Test species/strain:Rat/Crj:CD(SD)IGS
Test method:OECD Test Guideline 407
　Route:Oral (Gavage)
　Dosage:0 (Vehicle), 1000, 1500, 2000 mg/kg
　Number of animals/group:Males, 5; females, 5
　Vehicle:Corn oil
　Administration period:Males and females, 28 days
　Terminal killing:Males and females, days 29 or 43
GLP:Yes

　Test results:

Decrease of grip strength, a paralytic gait, degeneration of peripheral nerve fiber, increase of kidney weight, fatty degeneration of proximal tubular epithelium, and shortening of prothrombin time were observed in animals given 30 mg/kg/day or more. An anemic condition, decrease of body weight and food consumption, increase of total cholesterol and liver weight, and hypertrophy of centrilobular hepatocytes were observed in animals given 100 mg/kg or more per day. Death following paralytic gait, diarrhea, decrease of body weight and food consumption was observed in animals given 200 mg/kg/day. Others receiving this dosage were killed in a moribund condition.

The NOEL is considered to be 10 mg/kg/day for both sexes.

3. Preliminary Reproductive Toxicity 1)

Purity:99.3 %
Test species/strain:Rat/Crj:CD(SD)IGS
Test method:OECD Test Guideline 421
　Route:Oral (Gavage)
　Dosage:0 (Vehicle), 3, 10, 30 mg/kg
　Number of animals/group:Males, 10; females, 10
　Vehicle:Corn oil
　Administration period:Males, 46 days
Females, from 14 days before mating to day 4 of lactation
　Terminal killing:Males, day 47 of the administration
Females, day 5 of lactation
GLP:Yes

　Test results:

Increase of liver and/or kidney weights was observed in parent animals given 10 mg/kg/day or more, but no effects were observed on reproductive performance of parents or on developmental performance in the pups.

The NOEL for systemic toxicity in parents is considered to be 3 mg/kg/day, and that for reproductive and developmental toxicity is considered to be 30 mg/kg/day.

4. Genetic Toxicity

4-1. Bacterial test 2)

Purity:99.3 %
Test species/strains:Salmonella typhimurium TA100, TA1535, TA98, TA1537, Escherichia coli WP2 uvrA
Test method:Guidelines for Screening Mutagenicity Testing of Chemicals (Chemical Substances Control Law of Japan) and OECD Test Guideline 471
　Procedures:Pre-incubation method
　Solvent:Dimethyl sulfoxide
　Positive controls:-S9 mix; 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (TA100, TA98, WP2 uvrA), Sodium azide (TA1535) and 9-Amino-acridine hydrochloride (TA1537)
+S9 mix; 2-Aminoanthracene (all strains)
　Dosage:-S9 mix; 0, 2.29, 6.86, 20.6, 61.7, 185, 556, 1667, 5000 μg/plate (all strains for the dose-finding study)
+S9 mix; 0, 2.29, 6.86, 20.6, 61.7, 185, 556, 1667, 5000 μg/plate (all strains for the dose-finding study)
-S9 mix; 0, 4.88, 9.77, 19.5, 39.1, 78.1, 156 μg/plate (TA100 for the main study)
-S9 mix; 0, 2.44, 4.88, 9.77, 19.5, 39.1, 78.1 μg/plate (TA1535 for the main study)
-S9 mix; 0, 39.1, 78.1, 156, 313, 625, 1250 μg/plate (WP2 uvrA for the main study)
-S9 mix; 0, 313, 625, 1250, 2500, 5000 μg/plate (TA98, TA1537 for the main study)
+S9 mix; 0, 4.88, 9.77, 19.5, 39.1, 78.1, 156 μg/plate (TA100 for the main study)
+S9 mix; 0, 0.610, 1.22, 2.44, 4.88, 9.77, 19.5 μg/plate (TA1535 for the main study)
+S9 mix; 0, 39.1, 78.1, 156, 313, 625, 1250 μg/plate (WP2 uvrA for the main study)
+S9 mix; 0, 313, 625, 1250, 2500, 5000 μg/plate (TA98, TA1537 for the main study)
-S9 mix; 0, 439, 658, 988, 1481, 2222, 3333, 5000 μg/plate (TA1537 for the confirmation study)
　S9:Rat liver, induced with phenobarbital and 5,6-benzoflavone
　Plates/test:3
　Number of replicates:3
GLP:Yes

　Test results:

Increase in revertant colonies was observed for TA100, TA1535 and WP2 uvrA with both the non-activation method (-S9 mix) and the activation method (+S9 mix).

Genetic effects:
Salmonella typhimurium TA100, TA1535
+?-
Without metabolic activation:[*][ ][ ]
With metabolic activation:[*][ ][ ]

Salmonella typhimurium TA98, TA1537
+?-
Without metabolic activation:[ ][ ][*]
With metabolic activation:[ ][ ][*]

Escherichia coli WP2 uvrA
+?-
Without metabolic activation:[*][ ][ ]
With metabolic activation:[*][ ][ ]

4-2. Non-bacterial in vitro test (chromosomal aberration test) 2)

Purity:99.3 %
Type of cell used:Chinese hamster lung (CHL/IU) cells
Test method:Guidelines for Screening Mutagenicity Testing of Chemicals (Chemical Substances Control Law of Japan) and OECD Test Guideline 473
　Solvent:Dimethyl sulfoxide
　Positive controls:-S9 mix; Mitomycin C
+S9 mix; Cyclophosphamide
　Dosage:-S9 mix (short-term treatment); 0, 557, 696, 870, 1088 μg/mL
+S9 mix (short-term treatment); 0, 357, 446, 557 μg/mL
　S9:Rat liver, induced with phenobarbital and 5,6-benzoflavone
　Plates/test:2
GLP:Yes

　Test results:

A dose-dependent increase in structural aberrations was observed in the test with the short-term treatment(-S9 mix and +S9 mix).

Lowest concentration producing cytogenetic effects in vitro:
Without metabolic activation(short-term treatment):557 μg/mL (clastogenicity)
With metabolic activation (short-term treatment):446 μg/mL (clastogenicity)

Genetic effects:
clastogenicitypolyploidy
+?-+?-
Without metabolic activation:[*][ ][ ][ ][ ][*]
With metabolic activation:[*][ ][ ][ ][ ][*]

1)The tests were performed by the Safety Research Institute for Chemical Compounds Co., Ltd., 363-24 Shin-ei, Kiyota-ku, Sapporo, Hokkaido, 004-0839, Japan. Tel +81-11-885-5031 Fax +81-11-885-5313
2)The tests were performed by the Biosafety Research Center, Foods, Drugs and Pesticides(An-pyo Center), 582-2 Shioshinden, Fukude-cho, Iwata-gun, Shizuoka, 437-1213, Japan. Tel +81-538-58-1266 Fax +81-538-58-1393