N,N’-(ジスルファンー1,2-ジイルジー2,1-フェニレン)

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1 4-(3-phenylpropyl) pyridine [CAS No ] 1-Phenyl-3-(4-Pyridyl) Propane Molecular formula: C14H15N Molecular weight: N ABSTRACT The acute oral median lethal dose (LD 50 ) of 4-(3-phenylpropyl) pyridine in the female Sprague-Dawley CD strain rat was approximately 500 mg/kg bodyweight (Globally Harmonised Classification System - Category 4 > mg/kg bodyweight). The oral administration of 4-(3-phenylpropyl) pyridine to rats for a period of up to forty seven days (including two weeks pre-mating, gestation and early lactation period for females) at dose levels of 30, 100 and 250 mg/kg/day (reduced to 150 mg/kg/day on Day 6), resulted in treatment-related effects in animals of either sex from all treatment groups. For male animals, the No Observed Adverse Effect Level (NOAEL) for systemic toxicity was considered to be 30 mg/kg/day because of the reduced bodyweight gain and spermatid retention in testes at 100 mg/kg/day. The No Observed Effect Level (NOEL) was considered to be less than 30 mg/kg/day because of increase in albumin/globulin ratio, total protein and albumin, increase of liver weight, centrilobular hepatocyte enlargement, periportal lipid-type vacuolation in liver, thyroid follicular cell hypertrophy and globular accumulations of eosinophilic material in kidney at 30 mg/kg/day. For female animals, the No Observed Adverse Effect Level (NOAEL) for systemic toxicity was considered to be 30 mg/kg/day because of mortality (killed in extremis),

2 reduction in bodyweight gain and statistically significant blood chemical changes (increases in total protein, albumin, cholesterol, calcium, inorganic phosphorus and reduction in chloride) at 100 mg/kg/day. The No Observed Effect Level (NOEL) was considered to be less than 30 mg/kg/day because of microscopic change (mononuclear cell foci in liver) at 30 mg/kg/day. The No Observed Adverse Effect Level (NOAEL) for reproductive toxicity in female was considered to be 30 mg/kg/day because of reduction in delivery index, reduced litter size with live birth index and viability index and total litter losses in 6 animals out of 8 animals delivered the offspring at 100 mg/kg/day. The No Observed Effect Level (NOEL) was considered to be less than 30 mg/kg/day because of longer gestation length at 30 mg/kg/day. For F1 offspring, the No Observed Adverse Effect Level (NOAEL) and No Observed Effect Level (NOEL) was 30 mg/kg/day under these test conditions because of reduced bodyweight gains at 100 mg/kg/day. 4-(3-phenylpropyl) pyridine was considered to be non-mutagenic in Salmonella typhimurium, TA100, TA1535, TA98, TA1537 and Escherichia coli WP2 uvra regardless of the presence or absence of a liver enzyme metabolic activation system. 4-(3-phenylpropyl) pyridine did not induce any statistically significant, dose-related increases in the frequency of cells with structural or numerical chromosome aberrations either in the absence or presence of a liver enzyme metabolic activation system. The test material was therefore considered to be non-clastogenic to CHL cells in vitro.

3 SUMMARIZED DATA FROM THE STUDIES 1. Single Dose Oral Toxicity 1) Purity : 98.31% Test species/strain : Rat/Crj:CD (SD) Test method : OECD Test Guideline 423 Route : Oral (gavage) Doses : 0 (Vehicle), 300, 2000, 300 mg/kg/day Number of animals/group : Females, 3 Vehicle : Arachis oil BP GLP : Yes Test results: All animals treated at a dose level of 2000mg/kg were humanely killed one day after dosing. There were no deaths noted at a dose level of 300 mg/kg. Signs of systemic toxicity noted in animals treated at a dose level of 2000 mg/kg were prostration, lethargy, decreased respiratory rate, laboured respiration, tonic convulsions, hunched posture, ataxia, pilo-erection, hypothermia, pallor of the extremities and dehydration. All animals treated at a dose level of 2000 mg/kg were comatose one day after dosing. There were no signs of systemic toxicity noted in animals treated at a dose level of 300 mg/kg. Surviving animals showed expected gains in bodyweight except for one animal which showed no gain in bodyweight during the first week but expected gain in bodyweight during the second week. No abnormalities were noted at necropsy. The acute oral median lethal dose (LD50) of the test material in the female Sprague- Dawley CD strain rat was approximately 500 mg/kg bodyweight (Globally Harmonised Classification System Category 4 > mg/kg bodyweight).

4 4-(3- フェニルプロピル ) ピリジンのラットを用いる単回経口投与毒性試験 Single Dose Oral Toxicity Test of 4-(3-phenylpropyl)pyridine in Rats 要約 4-(3-フェニルプロピル ) ピリジンの急性毒性を OECD 試験ガイドライン 急性毒性等級法 に従って検討した. 動物は,8~12 週の Sprague-Dawley 系ラットに,1 投与段階 (1 群 ) につき雌 3 匹ずつ用いて強制経口投与を行った mg/kg を投与した全動物を投与 1 日後に安楽死させた.300 mg/kg 群では死亡は認められなかった.2000 mg/kg を投与した動物では全身毒性の徴候として, 腹臥位, 嗜眠, 呼吸数減少, 努力性呼吸, 強直性痙攣, 円背位, 運動失調, 立毛, 体温低下, 四肢の蒼白および脱水が認められ, 投与 1 日後には全動物が昏睡状態になった. 体重については,300mg/kg 投与時の 1 匹で第 1 週に体重増加が認められなかったことを除き, 順調な推移を示した. 剖検では異常は認められなかった. 方法 1. 被験物質および投与液の調製 4-(3-フェニルプロピル ) ピリジン ( ロット番号 :F9369A, 純度 :98.31%) は, 黄色液体である. 本物質については,IR スペクトラムと純度を測定して, 同一性を確認した mg/kg の用量には, 被験物質を提供された状態のまま使用した. 比重を測定し, その値を用いて, 投与用量となるように投与容量を算出した.300 mg/kg の用量には, 被験物質のラッカセイ油 ( 英国局方 ) 溶液を用時調製した. 2. 試験動物および飼育条件 Sprague-Dawley CD(Hsd:Sprague Dawley (CD )) 系雌ラットを Harlan Laboratories UK Limited(Bicester,Oxon, 英国 ) から入手した.5 日間以上の馴化期間後, 動物を無作為に選んで識別番号を付した後試験に供した. 試験開始時, 動物は 8~12 週齢であった. 体重は, 最初の投与群の投与時における平均体重の ±20% 以内であった. 動物は, 温度 :19~25, 相対湿度 :30~70%, 換気回数 : 毎時 15 回以上,12 時間連続照明 (6 時 ~18 時 ) および 12 時間連続消灯になるように制御された飼育室で, 針葉樹フレークを敷いた吊り下げ型ポリプロピレン製平底ケージで 3 匹ずつ飼育した. 投与直前の一晩と投与後約 3~4 時間の絶食期間を除き, 水道水および飼料 (2014 Teklad Global げっ歯類用飼料,Harlan Teklad,Blackthorn,Bicester,Oxon, 英国 ) を試験を通じて自由に摂取させた. 1

5 3. 投与量および投与方法 絶食させた雌ラットに被験物質を単回強制経口投与した. 被験物質の毒性に関する既存データから, 開始投与用量として 300 mg/kg を選択した. 第 1 段 階の投与において死亡動物がみられなかったことから, 第 2 段階の投与量は 2000mg/kg を選択 した. 以下に群構成 ( 投与用量, 濃度および投与容量 ) を示す. 投与用量 (mg/kg) 濃度 (mg/ml) 比重 投与容量 (ml/kg) 動物数雌 検査項目生死および明らかな毒性徴候の有無を投与後 30 分,1,2,4 時間に, またそれ以降は 1 日 1 回, 14 日間観察した. 各個体の体重を投与前 ( 投与容量の算出に用いた ) および投与後 7,14 日, または死亡時に測定した. 観察期間終了時に生存動物を頚椎脱臼により死亡させ, 肉眼的観察を実施し, 外表を調べ, 腹腔および胸腔を開けて主要器官を観察した. 結果 1. 死亡状況 (Table 1) 2000 mg/kg を投与した全動物を投与 1 日後に安楽死させた.300 mg/kg を投与した動物では死 亡は認められなかった. 2. 一般状態 2000 mg/kg を投与した動物では全身毒性の徴候として, 腹臥位, 嗜眠, 呼吸数減少, 努力性呼吸, 強直性痙攣, 円背位, 運動失調, 立毛, 体温低下, 四肢の蒼白および脱水が認められ, 投与 1 日後には全動物が昏睡状態になった.300 mg/kg を投与した動物では全身毒性の徴候はみられなかった. 3. 体重 300mg/kg 投与した動物の 1 匹で第 1 週に体重増加が認められなかったことを除き, 順調な推移 を示した. 4. 剖検 剖検では異常は認められなかった. 2

6

7 Table 1 Mortality of rats treated orally with 4-(3-phenylpropyl)pyridine Dose Level mg/kg Sex Number of Animals Treated Deaths During Day Deaths During Period After Dosing of Dosing (Days) (Hours) ½ Deaths Female /3 300 Female / Female /3 = Humanely killed 4

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