With regard to repeat dose toxicity, twelve males and one female given 250 mg/kg died during the 4th to 6th week of the dosing period. These rats showed progressive development of a paralytic gait and decreased motor activity before death. Body weight gain of the 250 mg/kg males and females, and food consumption of the 250 mg/kg males were significantly reduced. Twelve and 2 pregnant females given 40 and 100 mg/kg, respectively, demonstrated significant decrease in body weight gain during mid and late pregnancy. In the 100 mg/kg males, significantly decreased erythrocyte counts, hemoglobin concentration, hematocrit and A/G ratio, and increased platelet count, percentage of segmented neutrophils, cholinesterase activity, total cholesterol and calcium levels were noted. Similar alteration in hematological and clinical chemistry parameters was observed in one surviving 250 mg/kg male. On histopathological examination, major lesions noted in males and females given 100 mg/kg or more included nephropathy, atrophy of the thymus, liver and testis, and degeneration of nerve fibers in the spinal cord or the peripheral nerves. The incidence and severity of these lesions increased with dose and were greater in males than in females. The NOELs for repeat dose toxicity are considered to be less than 40 mg/kg/day for both sexes. In terms of reproductive/developmental toxicity, the copulation rate of the paired animals was significantly decreased in the 250 mg/kg group and the fertility index and number of implantation sites were decreased significantly in the 100 mg/kg group. In the 40 mg/kg group, intrauterine mortality of embryos was increased significantly and pup weights were significantly greater than those in the control group. The NOELs for reproductive/developmental toxicity are considered to be less than 40 mg/kg/day.

Neither structural chromosomal aberrations nor polyploidy were induced up to the limit concentration of 10 mM, in the absence or presence of an exogenous metabolic activation system in CHL/IU cells.

1 . Repeat Dose and Reproductive/Developmental Toxicity

Purity	:	99.9 wt%
Test species/strain	:	Rat/Crj:CD (Sprague-Dawley)
Test method	:	OECD Combined Repeat Dose and Reproductive/Developmental Toxicity Screening Test
Route	:	Oral (Gavage)
Doses	:	0 (vehicle), 40, 100, 250 mg/kg/day
Number of animals	:	Males, 13; Females, 13/group
Vehicle	:	Distilled Water
Administration period	:	Males, 42 days Females, from 14 days prior to mating to Day 3 of lactation
Terminal killing	:	Males, Day 43 Females, Day 4 of lactation
GLP	:	Yes

　Test results:

<Repeat Dose Toxicity>

Twelve males and one female that received 250 mg/kg died during the 4th to 6th week of the dosing period. These rats showed progressive development of a paralytic gait and decreased motor activity that became evident after the 2nd week. Body weight gain of males and females given 250 mg/kg was significantly lower than in the controls. In addition, 12 and 2 pregnant females given 40 and 100 mg/kg, respectively, demonstrated a significant decrease in body weight gain during mid and late pregnancy. Food consumption of the 250 mg/kg males was significantly lower than that of the controls. In the hematology and clinical chemistry examinations, significantly decreased erythrocyte count, hemoglobin concentration, hematocrit and A/G ratio, and increased platelet count, percentage of segmented neutrophils, cholinesterase activity, total cholesterol and calcium levels were noted in the 100 mg/kg males. Similar alteration in hematological and clinical chemistry parameters was observed in one surviving 250 mg/kg male. At terminal necropsy, compound-related alterations in organ weights included significant increases in the kidney weights of the 40 and 100 mg/kg males and in the thymus weights of the 100 mg/kg males and the 40 mg/kg females, and a significant decrease in the epididymal weights of 100 mg/kg males. On histopathological examination, major lesions noted in males and females given 100 mg/kg or more included nephropathy characterized by tubular and papillary alterations such as increased eosinophilic droplets in tubular epithelium, increased regeneration of tubules and papillary necrosis, atrophy of the thymus, liver and testis, increased atretic follicles in the ovary (250 mg/kg females only), and degeneration of nerve fibers in the spinal cord or the peripheral nerves (e.g., sciatic nerve). The incidence and severity of these lesions increased with dose and were greater in males than in females. The NOELs for repeat dose toxicity are considered to be less than 40 mg/kg/day for both sexes.

<Reproductive and Developmental Toxicity>

The copulation rate of the paired animals was decreased significantly in the 250 mg/kg group and the fertility index and number of implantation sites were decreased significantly in the 100 mg/kg group. Furthermore, intrauterine mortality of embryos was increased significantly in the 40 mg/kg group. However, no significant differences in the pup viability and incidence of morphological abnormalities of pups were found in the groups given 40 mg/kg when compared to the controls. Pup weights in the 40 mg/kg group were significantly greater than those in the control group up until terminal necropsy on day 4 of lactation. The NOELs for reproductive/developmental toxicity are considered to be less than 40 mg/kg/day.

2. Genetic Toxicity

Purity	:	99.9 %
Type of cell used	:	Chinese hamster lung (CHL/IU) cells
Test method	:	Guidelines for Screening Mutagenicity Testing of Chemicals (Japan)
Solvent	:	Acetone
Positive controls	:	-S9, Mitomycin C +S9, Cyclophosphamide
Doses	:	-S9 (continuous treatment): 0, 0.4, 0.7, 1.4 mg/ml -S9 (short-term treatment): 0, 0.4, 0.7, 1.4 mg/ml +S9 (short-term treatment): 0, 0.4, 0.7, 1.4 mg/ml
S-9	:	Rat liver, induced with phenobarbital and 5,6-benzoflavone
Plates/test	:	2
GLP	:	Yes

　Test results:

Cytogenetic effects were not seen under the conditions of this experiment.

　Genotoxic effects:

	clastogenicity				polyploidy
	+	?	-		+	?	-
without metabolic activation:	[ ]	[ ]	[*]		[ ]	[ ]	[*]
with metabolic activation:	[ ]	[ ]	[*]		[ ]	[ ]	[*]

1)	The tests were performed by the Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa, 257, Japan. Tel +81-463-82-4751 Fax +81-463-82-9627