細 胞 資 源 工 学
教 授 服 部 征 雄 （ 薬 学 博 士 ）
助教授 横 津 隆 子 （ 学 術 博 士 ） 助 手 宮 代 博 継 （ 薬 学 博 士 ） 技 官 中 村 憲 夫 （ 学 術 博 士 ）
本部門では，和漢薬資源の恒久的維持，育成を図るため，薬用生物に関する細胞工学的研究を 行なうと同時に，動植物細胞の持つ遺伝 情報を解析して，その薬用資源開発への応用，あるいは 微生物および動物細胞を用いて生理活性物質の探索，和漢薬の薬効発現機構の解明を行なうこと を目指している。本年度の主な研究テーマと成果は下記の通りである。
I.腸内嫌気性菌によるバイオトランスフォーメーション
腸内細菌による
s h i k o n i n
の変換反応を検討し，生成物と各種フラピン型補酵素との関係を 調べた。また，Eub α c t e r i u ms p . A‑44
感染ノトバイオートラットを作製し，経口投与後の血 中サイコサポニンおよび代謝物の濃度を通常ラット，無菌ラットの場合と比較検討した。II.腸内細菌による薬物の代謝活性化に関与する遺伝子
Senn a s i d e
の加水分解に関与する，B i j i d o bαc t e r i u ms p .
由来の酵素遺伝子を大腸菌に導入，発現させ大量の加水分解酵素を精製した。
皿．抗ウイルス薬の開発
巴豆から種々のホルボール系の化合物を単離し，その抗
HIV
作用を検討した。また，各種 韓国産生薬のHIV‑1
プロテアーゼ阻害作用を検討した。抗へルペスウイルス作用を有する蒙 古産生薬を検索し，S t e p h α n i ac e p hα r αn t h α
に強い活性を見いだした。活性を指標に成分の 単離を試み，FK‑3000
を得た。この化合物は1 2 5 ‑ 2 5 0mg/kg
で，ヘルペスウイルスを皮膚に 感染させたラットの帯状庖疹の進展，生存率を有意に抑制した。しかし，大量では毒性が発現した。
町．腎疾患における病態の解明と治療薬の開発
増悪因子のフリーラジカルの関与について，
g i n s e n o s i d e ‑ R d ,magnesium l i t h o s p e r m a t e B ,
t a n n i n , f l a v o n o i d
，茶，大黄，黄嘗，紅花，温牌湯を用い検討した。。 著 書
1
）横津隆子，董而博，河合悦子，玄番宗一：Magnesiuml i t h o s p e r m a t e B
はc i s p l a t i n
による腎細胞障害を軽減する． 腎とフリーラジカルー第4
集一 遠藤 仁，下条文武監修，武田理夫，長谷川俊雄編，東京医学社，東京，
1 9 9 8 ,1 5 3 ‑ 1 5 7 .
2）横津隆子，劉 中武，董而博：腎障害における大黄牡丹皮湯の関与について 腎とフ リーラジカルー第
4
集一 ，遠藤 仁，下条文武監修，武田理夫，長谷川俊雄編，東京医学 社，東京，1 9 9 8 ,1 8 8 ‑ 1 9 1 .
3
）横津隆子：生化学的アプローチ． 改訂第3
版生薬学概論 難波恒雄，津田喜典編，南江 堂，東京，1 9 9 8 ,1 9 4 ‑ 2 0 1 .
4 ) Y okoza wa T . : The r o l e o f g r e e n t e a t a n n i n s i n r e n a l d i s e a s e . R e c e n t R e s e a r c h Developments i n A g r i c u l t u r a l & B i o l o g i c a l Chemistry
，e d i t e d by S . G . P a n d a l a i , R e s e a r c h S i g n p o s t , I n d i a , 1 9 9 8 , 4 6 7 ‑ 4 8 3 .
。 原 著
1 ) E l
・MekkawyS . , Meselhy M. R . , Nakamura N . , Tezuka Y . , H a t t o r i M . , Kakiuchi N . , Shimotohno K . , Kawahata T . and Otake T . : Anti‑HIV‑1 and A n t i ‑ H I V ‑ I ‑ P r o t e a s e Substances from Ganoderm α l u c i d u m . P h y t o c h e m i s t r y , 4 9 : 1 6 5 1 ‑ 1 6 5 7 ; 1 9 9 8 .
A new h i g h l y oxygenated t r i t e r p e n e named g a n o d e r i c acidαhas b e e n i s o l a t e d from a methanol e x t r a c t o f t h e f r u i t i n g b o d i e s o f G α noderm α l u c i d u m t o g e t h e r w i t h t w e l v e known compounds. The s t r u c t u r e s o f t h e i s o l a t e d compounds were d e t e r m i n e d by s p e c t r o s c o p i c means i n c l u d i n g 2D‑NMR. G a n o d e r i o l F and g a n o d e r m a n o n t r i o l were found a c t i v e a s a n t i ‑ HIV w i t h an i n h i b i t o r y c o n c e n t r a t i o n o f 7 . 8 μg/ml f o r b o t h , and g a n o d e r i c a c i d B , g a n o d e r i o l B , g a n o d e r i c a c i d C l , 3 β ， 5 α − d i h y d r o x y ‑ 6 β − m e t h o x y e r g o s t a ‑ 7 , 2 2 ‑ d i e n e , g a n o d e r i c a c i d α ， g a n o d e r i c a c i d Hand g a n o d e r i o l A were m o d e r a t e l y a c t i v e i n h i b i t o r s a g a i n s t HIV‑1 PR w i t h a 50% i n h i b i t o r y c o n c e n t r a t i o n o f 0 . 1 7 ‑ 0 . 2 3 mM.
2 ) Ma C . , Nakamura N. and H a t t o r i M . : Saponins and C ‑ G l u c o s y l Flavones from t h e S e e d s o f Abrus p r e c a t o r i u s . Chem. Pharm. B u l l . , 4 6 : 9 8 2 ‑ 9 8 7 , 1 9 9 8 .
Two new s a p o n i n s , 3‑0
［−β
幽D ‑ g l u c u r o n o p y r a n o s y l ‑ ( 1
→2
−）β
−D ‑ g l u c o p y r a n o s y l ]h e d e r a g e n i n (named a b r u s ‑ s a p o n i n I ) and 3‑0
［−β
ーD ‑ g l u c u r o n o p y r a n o s y l ‑ ( 1
→2
−）β
−D‑g 1 u c o p y r a n o s y 1 J
o l e a n o l i c a c i d 2 8 − β − D ‑ g l u c o p y r a n o s y l e s t e r ( a b r u s ‑ s a p o n i n I I ) , and t h r e e new f l a v o n e s , 6 ‑ C ‑ β
−D ‑ g l u c o p y r a n o s y l ‑ 4 ' , 5 ‑ d i h y d r o x y ‑ 7 , 8
・d i m e t h o x y f l a v o n e( p r e c a t o r i n ) , 6‑C
［−β
−D ‑ a p i o f u r a ‑ n o s y l ‑ C l
→2
ー）β − D ‑ g lu c o p y r a n o s y l ] ‑ 4
，5 ‑ d i h y d r o x y ‑ 7 , 8 ‑ d i m e t h o x y f l a v o n e ( p r e c a t o r i n I I ) , 6 ‑ C
［−β
−D ‑ a p i o f u r a n o s y 1 ‑( 1
→2
−）β D ‑ g l u c o p y r a n o s y l ] ‑ 4 ' , 5 ‑ d i h y d r o x y ‑7
・m e t h o x y f l a v o n e ( p r e c a t o r i n I I I ) , were i s o l a t e d from t h e s e e d s o f Abrus p r e c a t o r i u s L . t o g e t h e r w i t h t w e l v e known com‑
pounds i n c l u d i n g a n a t u r a l l y new s a p o n i n , 3‑0
〔−β
−D ‑ g l u c u r o n o p y r a n o s y l ‑ C l
→2
ー）β
−D‑
g l u c o p y r a n o s y l ] o l e a n o l i c a c i d . T h e i r s t r u c t u r e s were d e t e r m i n e d on t h e b a s i s o f c h e m i c a l
and s p e c t r o s c o p i c m e t h o d s . I n a d d i t i o n , t h e u n u s u a l NMR s p e c t r a l b e h a v i o r o f t h e f l a v o n e
C ‑ g l y c o s i d e s i s a l s o d i s c u s s e d .
3 ) Kakuda H . , H e l a l A. M . , Nakamura N . , H a t t o r i M . : 8 α
−Hydroxy‑11α ， 1 3d i h y d r o ‑ z a l u z a n i n C .
Actα
Cry stα
llogrα
：phfoα ， C54 : 1 1 3 ‑ 1 1 4 , 1 9 9 8 .
I n t h e s t r u c t u r e o f t h e t i t l e compound [ 3 α
，4 , 5 , 6 , 6 α
，7 , 8 , 9 , 9 α
，9 β − d e c a h y d r o ‑ 4 , 8 ‑ d i h y d r o x y ‑ 3 ‑ m e t h y l ‑ 6 , 9 ‑ b i s ( m e t h y l e n e ) a z u l e n o [ 4 , 5 β
］−f u r a n ‑ 2 ( 3 H ) ‑ o n e , C 1 s H 2 0 0 4 ] , t h e s e c o n d a r y h y ‑ d r o x y l groups p a r t i c i p a t e i n i n t e r m o l e c u l a r hydrogen bonds b o t h a s d o n o r s and a c c e p t o r s
( F i g . l ) .
H30 03
F i g . 1
H15A
H14B
Hll
4 ) Meselhy M. R . , H e i k a l 0 . , Akao T . , H a t t o r i M . , Ono H. and Sadakane C . : D i s p o s i t i o n o f P a e o n i f l o r i n and Paeonimetabolin I i n Rats a f t e r Oral A dm i n i st r at i o n o f Toki‑
Shakuyaku‑San ( T S ) and Shakuyaku
聞Kanzo‑To ( S K ) . Nα t . M e d . , 5 2 : 2 6 5 ‑ 2 6 8 , 1 9 9 8 .
A f t e r o r a l a d m i n i s t r a t i o n o f a t r a d i t i o n a l C h i n e s e p r e s c r i p t i o n , Toki‑Shakuyaku‑San ( T S ) a t d o s e s o f 1 0 0 and 5 0 0 mg t o r a t s , t h e Cmax v a l u e s o f p a e o n i f l o r i n ( P F ) i n t h e plasma were 1 4 6 . 3 and 1 6 5 . 1 ng/ml a t 6 0 and c α . 4 5 min ( w i t h AUC v a l u e s o f 1 4 3 0 5 and 1 9 3 8 5 ng ・ min/
m l ) , w h e r e a s t h e Cmax v a l u e s o f p a e o n i m e t a b o l i n I ( P M ‑ I , a major m e t a b o l i t e o f PF) were 1 8 4 and 4 0 0 . 3 ng/ml a t 1 2 0 and 1 8 0 min ( w i t h AUC v a l u e s o f 9 8 4 9 7 and 1 8 2 1 8 8 ng•min/ml).
When a n o t h e r p r e s c r i p t i o n , Shakuyaku‑Kanzo‑To (SK) was o r a l l y g i v e n a t t h e same d o s e s ,
t h e Cmax v a l u e s o f PF and PM‑I were 1 2 8 . 5 and 1 5 3 . 5 n g / m l , and 1 4 1 . 7 and 7 2 6 . 5 n g / m l , r e ‑
s p e c t i v e l y a t 5 m i n , and a t 3 6 0 and 4 8 0 m i n , w i t h AUC v a l u e s o f PF and PM
幽I( 4 8 8 5 7 and
3 2 5 1 8 ng•min/ml, and 1 0 2 1 3 6 and 4 6 9 3 0 5 ng ・ min/ml),r e s p e c t i v e l y . C o n s i d e r a b l e amounts o f
PM‑I were s t i l l d e t e c t e d i n t h e r a t plasma 2 4 hr a f t e r p . o . a d m i n i s t r a t i o n o f SK.
Iti s c o n ‑
e l u d e d t h a t f o l l o w i n g p . o . a d m i n i s t r a t i o n o f TS o r SK, P F , o n e o f t h e main c o n s t i t u e n t s o f
t h e s e p r e s c r i p t i o n s , was s c a r c e l y a b s o r b e d b u t P M ‑ I , t h e i n t e s t i n a l b a c t e r i a l m e t a b o l i t e , l a r g e l y a b s o r b e d from t h e i n t e s t i n a l t r a c t . M o r e o v e r , i t seems l i k e l y t h a t SK, which p o t e n ‑ t i a l l y r e d u c e s t h e g a s t r i c emptying r a t e o r p e r i s t a l t i c movement o f t h e i n t e s t i n e , t a k e s l o n g e r t i m e t o r e a c h t h e l a r g e b o w e l where most o f PF i s t r a n s f o r m e d t o PM‑I by i n t e s t i n a l b a c t e r i a .
5 ) Yang X . , Gu Z . , Ma C . , H a t t o r i M. and Namba T . : A New I n d o l e D e r i v a t i v e I s o l a t e d from t h e Root o f Tuber F l e e c e f l o w e r (Polygonum m u l t i f l o r u m
）.中草薬，2 9 :5 ‑ 1 1 , 1 9 9 8 .
From t h e r o o t o f Polygonum m u l t i j l o r u m T h u n b . , a new i n d o l e d e r i v a t i v e , named i n d o l e ‑ 3(L
ールamino − α − h y d r o x y p r o p i o n i ca c i d ) methyl e s t e r ( X I ) , was i s o l a t e d t o g e t h e r w i t h t h e t e n known compounds, c h r y s o p h a n o l ( I ) , p h y s c i o n e ( I I ) , emodin ( I I I ) , c i t r e o r o s e i n ( I V ) , c h r y s o p h a n o l 8‑0 − β − D ‑ g l u c o p y r a n o s i d e ( V ) , p h y s c i o n e 8‑0 − β − D ‑ g l u c o p y r a n o s i d e ( V I ) , emodin 8‑0 − β − D ‑ g l u c o p y r a n o s i d e ( V I I ) , t o r a c h r y s o n e 8‑0 − β − D ‑ g l u c o p y r a n o s i d e ( V I I I ) , 2 , 3 , 5 , 4 ' ‑ t e t r a h y d r o x y s t i l b e n e 2‑0 − β ー D ‑ g l u c o p y r a n o s i d e( I X ) , and m e t h y l g a l l a t e ( X ) . T h e i r s t r u c t u r e s were d e t e r m i n e d by s p e c t r o s c o p i c m e a n s . These a n t h r a q u i n o n e compounds and a l o e ‑ e m o d i n ( X I I ) , r h e i n ( X I I I ) , a l o e
司emodin8 ‑ 0 ‑ , 8 ‑ D ‑ g l u c o p y r a n o s i d e ( X I V ) , c h r y s o p h a n o l 8‑0 − β − D ‑ ( 6 ' ‑ 0 ‑ m a l o n y l ) g l u c o p y r a n o i s i d e (XV), s e n n o s i d e A (XVI) and s e n n o s i d e B ( X V I I ) had no i n h i b i t o r y e f f e c t a g a i n s t recombinant HIV‑1 p r o t e a s e a t a c o n c e n t r a t i o n o f 1 0 0 mmol/L i n v i t r o .
6 ) Kida H . , Akao T . , Meselhy M. R . and H a t t o r i M.: Metabolism and P h a r m a c o k i n e t i c s o f O r a l l y A d m i n i s t e r e d S a i k o s a p o n i n b l i n C o n v e n t i o n a l , Germ‑Free and Eub
αc t e r i u m s p . A‑44 I n f e c t e d Gnotobiote R a t s . B i o l . Ph α r m . B u l l . , 2 1 : 5 8 8 ‑ 5 9 3 , 1 9 9 8 .
The m e t a b o l i c f a t e o f s a i k o s a p o n i n b l ( 1 ) was i n v e s t i g a t e d u s i n g c o n v e n t i o n a l , g e r m ‑ f r e e and Eub α c t e r i u m s p . A ‑ 4 4 ‑ i n f e c t e d g n o t o b i o t e r a t s . A f t e r t h e o r a l a d m i n i s t r a t i o n o f 1 t o g e r m ‑ f r e e r a t s a t a d o s e o f 5 0 mg/kg, no m e t a b o l i t e was d e t e c t e d i n t h e p l a s m a , t h e c e c a l c o n ‑ t e n t s o r t h e c u m u l a t i v e f e c e s through t h e e x p e r i m e n t . On t h e o t h e r h a n d , when 1 was o r a l l y g i v e n t o t h e Eub α c t e r i u m s p . A ‑ 4 4 ‑ i n f e c t e d g n o t o b i o t e r a t s , c o n s i d e r a b l e amounts o f i t s me‑
t a b o l i t e s , p r o s a i k o g e n i n A ( 2 ) and s a i k o g e n i n A ( 3 ) , were d e t e c t e d i n t h e r a t plasma w i t h t h e r e s p e c t i v e A U C 0 ‑ 1 0 h v a l u e s o f 1 7 4 2 4 and 2 2 2 6 0 pmol•min/ml, s i m i l a r t o t h e c a s e o f i t s o r a l a d m i n i s t r a t i o n t o c o n v e n t i o n a l r a t s ( A U C o ‑ l O h v a l u e s o f 9 9 3 6 and 1 2 4 1 4 pmol •min/ml f o r 2 and 3 , r e s p e c t i v e l y ) . F u r t h e r m o r e , s i g n i f i c a n t amounts o f b o t h m e t a b o l i t e s were d e t e c t e d i n t h e c e c a l c o n t e n t s and t h e c u m u l a t i v e f e c e s o f t h e g n o t o b i o t e and c o n v e n t i o n a l r a t s , b u t n o t i n t h o s e o f t h e germ
目f r e er a t s , w i t h i n 1 0 h a f t e r t h e a d m i n i s t r a t i o n .
F e c a l and c e c a l a c t i v i t i e s o f h y d r o l y z i n g 1 and 2 were found i n t h e g n o t o b i o t e and c o n v e n ‑
t i o n a l r a t s , though t h e r e were no d e t e c t a b l e a c t i v i t i e s i n t h e g e r m ‑ f r e e r a t s . A c c o r d i n g l y ,
b o t h h y d r o l y z i n g a c t i v i t i e s i n t h e i n t e s t i n a l b a c t e r i a , s u c h a s Eub α c t e r i u m s p . A ‑ 4 4 , a r e e s ‑
s e n t i a l f o r t h e a p p e a r a n c e o f 2 and 3 i n t h e r a t plasma and c u m u l a t i v e f e c e s , s i n c e o r a l l y a d ‑
m i n i s t e r e d 1 was p o o r l y a b s o r b e d from t h e g a s t r o i n t e s t i n a l t r a c t .
7 ) Ma C . , Nakamura N . , Miyashiro H . , H a t t o r i M. and Shimothono K . : I n h i b i t o r y E f f e c t s o f U r s o l i c Acid D e r i v a t i v e s from Cynomorium s o n g α r i c u m , and R e l a t e d T r i t e r p e n e s on Human Immunodeficiency V i r a l P r o t e a s e . P h y t o t h e r . R e s . , 1 2 : 1 3 8 ‑ 1 4 2 , 1 9 9 8 .
From t h e stems o f Cynomorium s o n g α r i c u m Rupr. ( C y n o m o r i a c e a e ) , u r s o l i c a c i d and i t s hydrogen malonate were i s o l a t e d a s i n h i b i t o r y s u b s t a n c e s a g a i n s t human i m m u n o d e f i c i e n c y v i r u s ( H I V ‑ 1 ) p r o t e a s e . T h e i r 50% i n h i b i t o r y c o n c e n t r a t i o n s C I C s o ) were 8 μMand 6 μM, r e ‑ s p e c t i v e l y . Of v a r i o u s d i c a r b o x y l i c a c i d h e m i e s t e r s o f r e l a t e d t r i t e r p e n e s s y n t h e s i z e d , t h e i n ‑ h i b i t o r y a c t i v i t i e s t e n d e d t o i n c r e a s e i n t h e o r d e r o f o x a l i c , m a l o n y l , s u c c i n y l and g l u t a r y l h e m i e s t e r s o f t r i t e r p e n e s s u c h a s u r s o l i c a c i d , o l e a n o l i c a c i d and b e t u l i n i c a c i d . The most p o ‑ t e n t i n h i b i t i o n was o b s e r v e d f o r t h e g l u t a r y l h e m i e s t e r s w i t h I C s o o f 4 μM.
8 ) Abdel‑Hafez A. A . , Meselhy M. R . , Nakamura N . , H a t t o r i M . , Watanabe H . , Murakami Y . , E l
・G e n t l yM. A . , Mahfouz N. M. and Mohamed T . A . : E f f e c t s o f P a e o n i f l o r i n D e r i v a t i v e s on t h e Scopolamine‑lnduced Amnesia Using a P a s s i v e Avoidance Task i n M i c e ; S t r u c t u r e ‑ A c t i v i t y R e l a t i o n s h i p . Bwl. Ph α r m . B u l l . , 2 1 , 1 1 7 4 ‑ 1 1 7 9 , 1 9 9 8 .
P a e o n i f l o r i n ( 1 ) and i t s d e r i v a t i v e s having a common c a g e ‑ l i k e p i n a n e s k e l e t o n w i t h h e m i k e t a l ‑ a c e t a l s y s t e m , were e v a l u a t e d f o r t h e i r e f f e c t s on memory impairment i n d u c e d by s c o p o l a m i n e i n m i c e u s i n g a s t e p ‑ d o w n t y p e p a s s i v e a v o i d a n c e t a s k . I n t h e t e s t s e s s i o n , 1 and i t s d e r i v a t i v e s ( F i g . 2 ) were i n t r a p e r i t o n e a l l y C i . p . ) a d m i n i s t e r e d a t d o s e s o f 0 . 0 0 2 , 0 . 0 1 , 0 . 0 2 and 0 . 2 mmol/kg, and 3 0 min l a t e r ( 1 5 min b e f o r e t h e e x p e r i m e n t ) , s c o p o l a m i n e ( 1 mg/kg, i . p . ) was g i v e n . These compounds showed d o s e ‑ d e p e n d e n t a t t e n u a t i o n i n a d o s e range o f 0 . 0 0 2 ‑ 0 . 0 2 mmol/kg and a l s o enhancement o f s c o p o l a m i n e ‑ i n d u c e d d e c r e a s e i n s t e p ‑ d o w n l a t e n c y . The e f f e c t s o f t h e s e compounds, e x c e p t t h a t o f 2 ' , 3 ' , 4 ' , 5 ' ‑ 0 ‑ t e t r a a c e t y l ‑ 3 ‑ 0 ‑ m e t h y l p a e o n i f l o r i n ( 8 ) , f o l l o w e d a b e l l
司s h a p e d d o s e r e s p o n s e p r o f i l e . 8 ‑ D e b e n z o y l − ι
d e g l u c o s y l ‑ 3 ‑ 0 ‑ m e t h y l p a e o n i f l o r i n ( 6 ) showed no s i g n i f i c a n t i n c r e a s e i n t h e s t e p ‑ d o w n
l a t e n c y a t a l l t e s t e d d o s e s . Maximum s t e p ‑ d o w n l a t e n c y was o b t a i n e d by 3 ‑ 0 ‑ m e t h y l p a e o n i f l o r i n
( 3 ) and 2 ' , 3 , 3 ' , 4
，5 ' ‑ p e n t a
叩0 ‑ m e t h y l p a e o n i f l o r i n ( 7 ) ( t h e minimal e f f e c t i v e d o s e was 0 . 0 0 2
mmol/kg). R e l a t i v e t o 3 , d e b e n z o y l a t i o n , a s i n 8 ‑ d e b e n z o y l ‑ 3 ‑ 0 ‑ m e t h y l p a e o n i f l o r i n ( 4 ) ,
s l i g h t l y i n c r e a s e d t h e l a t e n c y , w h i l e d e g l u c o s y l a t i o n , a s i n 6 ‑ d e g l u c o s y l ‑3 ‑ 0 ‑ m e t h y l p a e o n i f l o r i n
( 5 ) , s i g n i f i c a n t l y r e d u c e d t h e p r o l o n g a t i o n o f l a t e n c y . Removal o f b o t h g l u c o s e and b e n z o y l
m o i e t i e s r e s u l t e d i n t h e l o s s o f a c t i v i t y a s s e e n i n 6 . T h e s e r e s u l t s r e v e a l e d t h a t , i n a d d i t i o n
t o t h e c a g e ‑ l i k e p i n a n e s k e l e t o n , t h e b e n z o y l and t h e g l u c o s y l m o i e t i e s a r e i m p o r t a n t s t r u c ‑
t u r a l e l e m e n t s o f t h e p a e o n i f l o r i n s k e l e t o n a s i t s e f f e c t s on s c o p o l a m i n e ‑ i n d u c e d a m n e s i a .
CH3
H
R20
8 4
OR1Compound No R I R z R 3
1 H Bz Gk
2
H H Gk
3 Me Bz Gk
4 Me H Gk
5 Me Bz H
6 Me H H
7 Me Bz G l c ( 0 M e ) 4 8 Me Bz G l c ( 0 A c ) 4
9
H p‑HO‑Bz Gk
10 H Bz 6 ' ‑ B z Gk
F i g . 2
9 ) Abdel‑Hafez A. A . , Meselhy M. R . , Nakamura N . , H a t t o r i M . , Watanabe H . , Mohamed T . A . , Mahfouz N.
島' 1 . ,and E l
・G e n t l yM. A . : P o t e n t A n t i c o n v u l s a n t Paeonimetabolin ・ ID e r i v a t i v e s Obtained by I n c u b a t i o n o f P a e o n i f l o r i n and T h i o l Compounds with L α c t ob α c i l l u s b r e v i s . Chem. Ph α r m . B u l l . , 4 6 : 1 4 8 6 ‑ 1 4 8 7 , 1 9 9 8 . S e v e n t e e n t h i o p a e o n i m e t a b o l i n ‑ I a d d u c t s were o b t a i n e d a s m i x t u r e s o f d i a s t e r e o i s o m e r s a f t e r i n c u b a t i o n o f p a e o n i f l o r i n w i t h L α c t o b α c i l l u s b r e u i s i n t h e p r e s e n c e o f v a r i o u s t h i o l s ( F i g . 3 ) . Four compounds, 8 ‑ ( n ‑ h e x y l t h i o
）ー（8 ) ,8 ‑ c y c l o p e n t y l t h i o ‑ , 8
（・p ‑ t o l y l ) t h i o
・and8 ‑ b e n z o y l t h i o
ベ1 8 ) p a e o n i m e t a b o l i n s , showed 1 0 0 % p r o t e c t i o n a g a i n s t p e n t y l e n e t e t r a z o l e ‑ i n d u c e d c o n v u l s i o n s a t d o s e s o f 0 . 1 2 5 , 0 . 2 5 , o r 0 . 5 0 mmoljkg, r e l a t i v e t o v a l p r o i c a c i d ( 1 0 0 % p r o t e c t i o n a t 1 . 5 mmol/kg). For 8 and 1 8 , t h e p r i n c i p l e a n t i c o n v u l s a n t a c t i v i t y r e s i d e d i n t h e
( 7 S ) ‑ i s o m e r s , w h i l e ( 7 R ) ‑ i s o m e r s showed m u s c l e r e l a x a t i o n e f f e c t s .
1 0 ) Min B . , Nakamura N . , Miyashiro H . , Bae K. and H a t t o r i M . : T r i t e r p e n e s from t h e Spores o f G α noderm α l u c i d u m and T h e i r I n h i b i t o r y A c t i v i t y a g a i n s t HIV‑1 P r o t e a s e . Chem. Ph α r m . B u l l . , 4 6 : 1 6 0 7 ‑ 1 6 1 2 , 1 9 9 8 .
Two new l a n o s t a n e ‑ t y p e t r i t e r p e n e s , l u c i d u m o l A and g a n o d e r i c a c i d β
，were i s o l a t e d from
t h e s p o r e s o f G α n o d e rm α
（G . ) l u c i d u m , t o g e t h e r w i t h a new n a t u r a l o n e and s e v e n t h a t were
known. The s t r u c t u r e s o f t h e new t r i t e r p e n e s w e r e d e t e r m i n e d a s ( 2 4 8 ) ‑ 2 4 , 2 5 ‑ d i h y d r o x y l a n o s t
1
2
F i g . 3
3 ,
R~n-propyl
4 ,
R= i s o ‑ p r o p y l 5 ,
R= n ‑ b u t y l 6 ,
R= s e c ‑ b u t y l 7 ,
R= i s o ・ b u t y l 8 ,
R= n ‑ h e x y l 9 ,
R= a l l y ! 1 0 ,
R= c y c l o p e n t y l 1 1 ,
R= c y c l o h e x y l
1 2 ,
R= p h e n y l 1 3 ,
R= o ‑ t o l y l
1 4 ,
R= m ‑ t o l y l 1 5 ,
R= p ‑ t o l y l 1 6 ,
R= 2 ‑ n a p h t h y l 1 7 ,
R= b e n z y l 1 8 ,
R= b e n z o y l 1 9 ,
R= a c e t y l
8 ‑ e n e ‑ 3 , 7 ‑ d i o n e and 3 β
，7 β
司d i h y d r o x y ‑ 1 1 , 1 5d i o x o l a n o s t a ‑ 8 , 2 4 ( E ) ‑ d i e n ‑ 2 6 ‑ o i c a c i d , r e s p e c ‑ t i v e l y , by c h e m i c a l and s p e c t r o s c o p i c m e a n s . The q u a n t i t a t i v e a n a l y s e s o f 5 f r u i t i n g b o d i e s , a n t l e r e d form and s p o r e s o f G . l u c i d u m were p e r f o r m e d by h i g h p e r f o r m a n c e l i q u i d chroma‑
tography and d e m o n s t r a t e d t h a t g a n o d e r i c a l c o h o l and a c i d c o n t e n t s were q u i t e h i g h i n t h e s p o r e . Of t h e compound i s o l a t e d , g a n o d e r i c a c i d β
，（248
ー）l a n o s t a ‑ 7 , 9 ( 1 1 ) ‑ d i e n e ‑ 3 β
，2 4 , 2 5 ‑ t r i o l ( c a l l e d l u c i d u m o l B ) , g a n o d e r m a n o n d i o l , g a n o d e r m a n o n t r i o l and g a n o l u c i d i c a c i d A showed s i g n i f i c a n t anti‑human i m m u n o d e f i c i e n c y v i r u s ( a n t i ‑ H I V ) ‑ 1 p r o t e a s e a c t i v i t y w i t h I C 5 o v a l ‑ u e s o f 20‑9 0
μM.1 1 ) Akao T . , Kida H . , H a t t o r i M . , Kobashi K . : I n t e s t i n a l B a c t e r i a l H y d r o l y s i s i s R e q u i r e d f o r t h e Appearance o f Compound K i n Rat Plasma a f t e r O r a l A d m i n i s t r a t i o n o f G i n s e n o s i d e Rbl from Pαnαx g i n s e n g .
J.Ph α rm.Ph α r m a c o l . , 5 0 : 1 1 5 5 ‑ 1 1 6 0 , 1 9 9 8 . G i n s e n o s i d e Rbl from P α n αx g i n s e n g r o o t i s t r a n s f o r m e d i n t o compound K v i α g i n s e n o s i d e s Rd and F2 by i n t e s t i n a l b a c t e r i a l f l o r a . Among 3 1 d e f i n e d i n t e s t i n a l s t r a i n s from man, o n l y Eub α c t e r i u m s p . A‑44 t r a n s f o r m e d g i n s e n o s i d e Rbl i n t o compound K v i α g i n s e n o s i d e R d . The g i n s e n o s i d e R b l ‑ h y d r o l y s i n g enzyme i s o l a t e d from Eub α c t e r i u m s p . A‑
4 4 was i d e n t i c a l t o a p r e v i o u s l y p u r i f i e d g e n i p o s i d e ‑ h y d r o l y s i n g β − D ‑ g l u c o s i d a s e .
When g i n s e n o s i d e Rbl ( 2 0 0 mg kg 1 ) was a d m i n i s t e r e d o r a l l y t o g e r m ‑ f r e e r a t s , n e i t h e r compound K nor any o t h e r m e t a b o l i t e was d e t e c t e d i n t h e p l a s m a , i n t e s t i n a l t r a c t o r cumu‑
l a t i v e f a e c e s 7 o r 1 5 h a f t e r a d m i n i s t r a t i o n . Most o f t h e g i n s e n o s i d e Rbl a d m i n i s t e r e d was r e
『c o v e r e d from t h e i n t e s t i n a l t r a c t , e s p e c i a l l y t h e c a e c a , and c u m u l a t i v e f a e c e s i n d i c a t i n g p o o r a b s o r p t i o n o f g i n s e n o s i d e R b l . When g i n s e n o s i d e Rbl was a d m i n i s t e r e d o r a l l y t o g n o t o b i o t e r a t s m o n o ‑ a s s o c i a t e d w i t h Eub α c t e r i u m s p . A ‑ 4 4 , a s i g n i f i c a n t amount o f compound K was d e t e c t e d i n t h e plasma and c o n s i d e r a b l e amounts were found i n t h e c a e c a l c o n t e n t s and c u ‑ m u l a t i v e f a e c e s 7 and 1 5 h a f t e r a d m i n i s t r a t i o n . A s m a l l amount o f g i n s e n o s i d e Rbl was d e ‑ t e c t e d i n t h e c a e c a l c o n t e n t s o n l y 7 h a f t e r a d m i n i s t r a t i o n .
T h e s e r e s u l t s i n d i c a t e t h a t o r a l l y a d m i n i s t e r e d g i n s e n o s i d e Rbl i s p o o r l y a b s o r b e d from
t h e gut b u t t h a t i t s m e t a b o l i t e compound K , p r o d u c e d by g i n s e n o s i d e R b l ‑ h y d r o l y s i n g b a c ‑ t e r i a s u c h a s Eub α c t e r i u m s p . A ‑ 4 4 i n t h e l o w e r p a r t o f i n t e s t i n e , i s a b s o r b e d .
1 2 ) Min B . S . , Bae K . , Kim Y. H . , Shimotohno K . , Miyashiro H . , H a t t o r i M . , I n h i b i t o r y A c t i v i t i e s o f Korean P l a n t s on HIV
幽1P r o t e a s e . N α t u r a l P r o d u c t S c i e n c e s , 4 : 2 4 1 ‑ 2 4 4 , 1 9 9 8 .
For t h e d e v e l o p m e n t o f a n t i ‑ A I D S a g e n t s , t h i r t y ‑ s e v e n methanol e x t r a c t s o f Korean p l a n t m a t e r i a l s were t e s t e d f o r t h e i r i n h i b i t o r y e f f e c t s on human i m m u n o d e f i c i e n c y v i r u s t y p e ‑ 1 ( H I V ‑ 1 ) p r o t e a s e . E x t r a c t s o f s e v e n p l a n t s showed more t h a n 30% i n h i b i t o r y a c t i v i t i e s on HIV‑1 p r o t e a s e a t a c o n c e n t r a t i o n o f 1 0 0 μg/mL The bark o f B e r c h e m i a b e r c h e m i a e f o l i α
，t h e l e a f o f L i n d e r α e r y t h r o c α r p αand t h e whole p l a n t o f S i e g e s b e c k i α p u b e s c e n s e x h i b i t e d s i g n i f i ‑ c a n t i n h i b i t i t o r y a c t i v i t i e s on HIV‑1 p r o t e a s e w i t h 5 6 . 2 , 5 0 . 8 , and 4 6 . 6 % , r e s p e c t i v e l y .
1 3 ) Kim D . , Yokozawa T . , H a t t o r i M . , Kadota S . , Namba T . : E f f e c t s o f Aqueous E x t r a c t s o f Apocynum venetum Leaves on H y p e r c h o l e s t e r o l a e m i c " R a t s . P h y t o t h e r . R e s . , 1 2 : 4 6 ‑ 4 8 , 1 9 9 8 .
The e x t r a c t s o f d r i e d l e a v e s o f Apocynum v e n e t u m c o l l e c t e d i n d i f f e r e n t p l a c e s i n China (Luobuma I and I I ) , and two p r o c e s s e d l e a v e s (Luobuma I I I and I V ) , were i n v e s t i g a t e d on serum l i p i d and h e p a t i c t o t a l c h o l e s t e r o l l e v e l s i n c h o l e s t e r o l ‑ f e d r a t s .
The serum t o t a l c h o l e s t e r o l l e v e l s were s i g n i f i c a n t l y r e d u c e d on day 4 0 i n r a t s g i v e n t h e r o a s t e d l e a f e x t r a c t s (Luobuma I I I and I V ) w i t h p
<0 . 0 5 and p
<0 . 0 1 , r e s p e c t i v e l y , when compared w i t h a c o n t r o l v a l u e o f c h o l e s t e r o l ‑ f e d r a t s . I n t h e serum l i p o p r o t e i n s , t h e LDL‑
c h o l e s t e r o l l e v e l s were s i g n i f i c a n t l y l o w e r i n r a t s g i v e n e a c h s a m p l e e x t r a c t on day 4 0 , com‑
p a r e d w i t h t h e c o n t r o l v a l u e , and t h e H D L ‑ c h o l e s t e r o l l e v e l s were s i g n i f i c a n t l y h i g h e r than t h a t o f a c o n t r o l g r o u p , e x c e p t f o r a group g i v e n Luobuma
I.F u r t h e r m o r e , t h e r e s p e c t i v e a t h e r o g e n i c i n d i c e s were s i g n i f i c a n t l y l o w e r i n a l l groups g i v e n Luobuma I t o I V , compared w i t h t h e c o n t r o l v a l u e . The h e p a t i c t o t a l c h o l e s t e r o l l e v e l s were a l s o s i g n i f i c a n t l y l o w e r i n a l l o f t h e g r o u p s g i v e n t h e Luobuma e x t r a c t s than t h a t o f c o n t r o l r a t s on day 4 0 . T h e s e r e s u l t s s u g g e s t t h a t t h e e x t r a c t o f A . v e n e t u m l e a v e s i s e f f e c t i v e f o r h y p e r c h o l e s t e r a e m i a and t h e p r e v e n t i o n o f a t h e r o s c l e r o s i s .
1 4 ) Yokozawa T . , Liu Z.W. and Dong E . : A Study o f Ginsenoside‑Rd i n a Renal I s c h e m i a ‑ R e p e r f u s i o n Model. N e p h r o n , 7 8 : 2 0 1 ‑ 2 0 6 , 1 9 9 8 .
The e f f e c t o f g i n s e n o s i d e ‑ R d i n i s c h e m i c ‑ r e p e r f u s e d r a t s was e x a m i n e d . I n c o n t r o l r a t s ,
b l o o d and r e n a l p a r a m e t e r s and t h e a c t i v i t i e s o f a n t i o x i d a t i v e enzymes i n r e n a l t i s s u e d e v i ‑
a t e d from t h e normal r a n g e , i n d i c a t i n g d y s f u n c t i o n o f t h e k i d n e y s .
In co n t r a s t , when
g i n s e n o s i d e ‑ R d was g i v e n o r a l l y f o r 3 0 c o n s e c u t i v e days p r i o r t o i s c h e m i a and r e p e r f u s i o n ,
t h e a c t i v i t i e s o f t h e a n t i o x i d a t i o n enzymes s u p e r o x i d e d i s m u t a s e , c a t a l a s e and g l u t a t h i o n e
p e r o x i d a s e were h i g h e r , w h i l e m a l o n d i a l d e h y d e l e v e l s i n serum and r e n a l t i s s u e were l o w e r
i n t h e t r e a t e d r a t s t h a n i n t h e c o n t r o l s . D e c r e a s e d l e v e l s o f u r e a n i t r o g e n and c r e a t i n i n e i n
serum d e m o n s t r a t e d a p r o t e c t i v e a c t i o n a g a i n s t t h e r e n a l d y s f u n c t i o n c a u s e d by i s c h e m i a and r e c i r c u l a t i o n . On t h e o t h e r h a n d , i t was d e m o n s t r a t e d t h a t g i n s e n o s i d e ‑ R d a f f e c t e d c u l ‑ t u r e d p r o x i m a l t u b u l e c e l l s s u b j e c t e d t o h y p o x i a ‑ r e o x y g e n a t i o n , p r o b a b l y by p r e v e n t i n g oxygen f r e e r a d i c a l s from a t t a c k i n g t h e c e l l membranes.
1 5 ) Yokozawa T . , Dong E . , Nakagawa T . , Kim D . W . , H a t t o r i M. and Nakagawa H . : E f f e c t s o f Japanese Black Tea on A t h e r o s c l e r o t i c D i s o r d e r s . B i o s c i . B i o t e c h n o l . B i o c h e m . , 6 2 : 4 4 ‑ 4 8 , 1 9 9 8 .
The a t h e r o g e n i c i n d e x was found t o b e s i g n i f i c a n t l y b e t t e r i n r a t s f e d a h i g h ‑ c h o l e s t e r o l d i e t s u p p l e m e n t e d w i t h b l a c k t e a e x t r a c t than i n t h e o n e s n o t g i v e n t h e e x t r a c t .
Itwas a l s o e v i d e n t t h a t b l a c k t e a i n h i b i t e d t h e p r o l i f e r a t i o n o f smooth m u s c l e c e l l s i n v o l v e d i n t h e d e v e l ‑ opment and p r o g r e s s i o n o f a t h e r o s c l e r o s i s , and s u p p r e s s e d t h e p r o d u c t i o n o f o x i d i z e d l o w ‑ d e n s i t y l i p o p r o t e i n , a c a u s e o f l i p i d a c c u m u l a t i o n .
Itt h u s seems l i k e l y t h a t b l a c k t e a h a s an a n t i a t h e r o s c l e r o t i c a c t i o n .
1 6 ) Yokozawa T . , Chen C . P . and Liu Z . W . : E f f e c t o f T r a d i t i o n a l Chinese P r e s c r i p t i o n s and Their Main Crude Drugs on l , l ‑ D i p h e n y l ‑ 2 ‑ p i c r y l h y d r a z y l R a d i c a l . P h y t o t h e r . R e s . , 1 2 : 9 4 ‑ 9 7 , 1 9 9 8 .
The i n h i b i t o r y e f f e c t s o f 7 9 t r a d i t i o n a l C h i n e s e p r e s c r i p t i o n s and 2 8 c r u d e drugs on t h e 1 , 1 ‑ d i p h e n y l ‑ 2 ‑ p i c r y l h y d r a z y l (DPPH) r a d i c a l were e x a m i n e d , and many demonstrated s i g n i f i ‑ c a n t i n h i b i t i o n . The most e f f e c t i v e c r u d e drugs were G a l l a e Rhois and Rhei Rhizoma, f o l l o w e d i n o r d e r by Cinnamomi C o r t e x , Ephedrae H e r b a , S c u t e l l a r i a e R a d i x , P e r i l l a e H e r b a , P a e o n i a e Ruber Radix and Moutan C o r t e x , w h i l e p r e s c r i p t i o n s composed o f one o r more o f t h e s e c r u d e drugs a l s o showed s t r o n g f r e e r a d i c a l s c a v e n g i n g a c t i v i t y . These r e s u l t s p r e d i c t t h a t t r a d i t i o n a l C h i n e s e m e d i c i n e s would b e p r o m i s i n g a g e n t s f o r s c a v e n g i n g f r e e r a d i c a l s , and f o r c u r i n g d i s e a s e s r e l a t e d t o f r e e r a d i c a l r e a c t i o n s .
1 7 ) Yokozawa T . , Dong E . , Yasui T . and Muraguchi A . : P r o t e c t i v e E f f e c t o f Wen‑Pi‑
Tang a g a i n s t Apoptosis o f Cultured Renal E p i t h e l i a l C e l l s . P h y t o t h e r . R e s . , 1 2 : 1 3 5 ‑ 1 3 7 ' 1 9 9 8 .
I n terms o f t h e r a t e o f DNA f r a g m e n t a t i o n and e l e c t r o p h o r e s i s p a t t e r n s , we demonstrated t h a t a p o p t o s i s was i n d u c e d i n LLC‑PK1 c e l l s s u b j e c t e d t o t h e Fenton r e a c t i o n w i t h H202 and F e 2 + . I n a d d i t i o n , Wen‑Pi‑Tang, which i s known t o i n h i b i t t h e p r o g r e s s i o n o f r e n a l f a i l u r e i n b o t h an e x p e r i m e n t a l and a c l i n i c a l s e t t i n g , was found t o s u p p r e s s t h i s a p o p t o s i s .
1 8 ) Kim D . W . , Yokozawa T . , H a t t o r i M . , Kadota S . and Namba T . : Luobuma Leaf I n h i b i t s Oxidation o f Low‑density L i po p r o t e i n i n C h o l e s t e r o l ‑ f e d R a t s .
よ Trα
d.M e d . , 1 5 : 4 0 ‑ 4 4 , 1 9 9 8 .
P e r o x i d a t i o n was found t o b e s i g n i f i c a n t l y r e d u c e d when l o w ‑ d e n s i t y l i p o p r o t e i n (LDL)
i s o l a t e d from t h e plasma o f r a t s t r e a t e d w i t h Luobuma l e a f e x t r a c t o r a l l y was p r e s e n t i n t h e
i n c u b a t i o n medium i n c o m p a r i s o n w i t h plasma from u n t r e a t e d r a t s . Of t h e two Luobuma e x
四t r a c t s t e s t e d , Luobuma B e x t r a c t showed c o n s i d e r a b l y s t r o n g s u p p r e s s i o n o f t h e p e r o x i d a ‑ t i o n i n d u c e d by c o p p e r .
Itwas a l s o e v i d e n t t h a t Luobuma e x t r a c t d e c r e a s e d t h e l e v e l s o f f r e e c h o l e s t e r o l and L D L ‑ c h o l e s t e r o l i n s e r u m , and t h i o b a r b i t u r i c a c i d ‑ r e a c t i v e s u b s t a n c e i n serum and l i v e r . From t h e s e r e s u l t s , i t s e e m s l i k e l y t h a t Luobuma e x t r a c t h a s an a n t i a t h e ‑ r o s c l e r o t i c a c t i o n .
1 9 ) Mitsuma T . , Yokozawa T . , Non
叫王aG . , l t o h T . , Shimada Y. and Terasawa K . : D i f f e r e n c e s i n C a t h a r t i c A c t i o n among D i f f e r e n t Types o f t h e O r i e n t a l Drug Rhubarb.
J.T r a d . M e d . , 1 5 : 4 5 ‑ 5 1 , 1 9 9 8 .
T h r e e d i f f e r e n t t y p e s o f r h u b a r b : r h u b a r b A ( p r o d u c e d i n t h e P r o v i n c e o f S i ‑ C h u a n
（四 Jll),C h i n a ) , B ( c u l t i v a t e d and p r o c e s s e d i n J a p a n ) and C ( t a b l e t s m a n u f a c t u r e d w i t h t h e p r o c e s s e d r h i z o m a o f Rheum palm α tum L . from t h e P r o v i n c e o f Q i n g ‑ H a i
（青海），C h i n a ) were a d m i n i s t e r e d t o 1 2 h e a l t h y male v o l u n t e e r s f o r t h r e e d a y s e a c h , and t h e i r q u a l i t y was e v a l u a t e d i n t e r m s o f t h e number o f b o w e l m o v e m e n t s , b o w e l s o u n d s , u r i n a r y volume and v a r i o u s b l o o d c h e m i c a l p a r a m e t e r s . Upon o v e r a l l a s s e s s m e n t o f t h e s e p a r a m e t e r s , p r o c e s s e d r h u b a r b , w i t h a weaker c a t h a r t i c a c t i o n , i s s u i t a b l e f o r t h e r a p e u t i c u s e i n p a t i e n t s w i t h c h r o n i c r e n a l f a i l u r e .
2 0 ) Yokozawa T . , Dong E . , Nakagawa T . , Kashiwagi H . , Nakagawa H . , Takeuchi S . and Chung H . Y . : I n V i t r o and i n Vivo S t u d i e s on t h e R a d i c a l ‑ S c a v e n g i n g A c t i v i t y o f T e a .
J.A g r i c . Food C h e m . , 4 6 : 2 1 4 3 ‑ 2 1 5 0 , 1 9 9 8 .
The e f f e c t s o f t e a ( C αm e l l i α s i n e n s i s L . ) o f t h r e e t y p e s on e x c e s s i v e f r e e r a d i c a l s w e r e e x ‑ amined u t i l i z i n g s p i n t r a p p i n g , 1 , 1 ‑ d i p h e n y l ‑ 2 ‑ p i c r y l h y d r a z y l r a d i c a l , l i p i d p e r o x i d a t i o n , and l a c t a t e d e h y d r o g e n a s e l e a k a g e from c u l t u r e d c e l l s . Green t e a e x t r a c t p r e s e n t e d s i g n i f i c a n t a n t i r a d i c a l e f f e c t s i n t h e s e f o u r a s s a y s y s t e m s , w h e r e a s o o l o n g t e a and b l a c k t e a e x t r a c t s showed a r a t h e r weak p r o t e c t i v e e f f e c t a g a i n s t f r e e r a d i c a l s . A more p o t e n t s c a v e n g e r e f f e c t u s i n g c u l t u r e d c e l l s was f o u n d w i t h a g r e e n t e a t a n n i n m i x t u r e . S i m i l a r l y t o t h e e f f e c t s o f t h e g r e e n t e a t a n n i n m i x t u r e ,
（ー）ーe p i g a l l o c a t e c h i n
与0 ‑ g a l l a t e ,i t s main i n g r e d i e n t , had an i n h i b i t o r y e f f e c t on o x i d a t i v e s t r e s s ‑ i n d u c e d a p o p t o s i s . The a c t i v i t i e s o f t h e a n t i o x i d a t i o n enzymes i n r a t s a f t e r s u b t o t a l nephrectomy w e r e i n c r e a s e d , s u g g e s t i n g a p r o t e c t i v e a c t i o n a g a i n s t o x i d a t i v e s t r e s s . The i n c r e a s e d l e v e l s o f u r e m i c t o x i n s i n t h e b l o o d w e r e a l s o r e d u c e d i n r a t s g i v e n
（ー）−e p i g a l l o c a t e c h i n3‑0
晴g a l l a t e .T h e s e f i n d i n g s i n d i c a t e t h a t
（ー）−e p i g a l l o c a t e c h i n 3‑0 ・ g a l l a t eh e l p s t o i n h i b i t t h e p r o g r e s s i o n o f r e n a l f a i l u r e by s c a v e n g i n g r a d i c a l s .
2 1 ) Yokozawa T . , Oura H . and N i s h i o k a I . : C o n f i r m a t i o n t h a t Magnesium L i t h o s p e r m a t e B A m e l i o r a t e s Paraquat
・Induced I n j u r y i n C u l t u r e d Renal E p i t h e l i a l C e l l s . N e p h r o n , 7 9 : 3 7 3 ‑ 3 7 4 , 1 9 9 8 .
F o c u s i n g a t t e n t i o n on t h e c l i n i c a l f i n d i n g t h a t d e c r e a s e d u r i n a r y e x c r e t i o n o f p a r a q u a t i s
a s s o c i a t e d w i t h a g g r a v a t i o n o f t h e p a t h o l o g i c a l c o n d i t i o n , we i n v e s t i g a t e d t h e e f f e c t o f
magnesium l i t h o s p e r m a t e B , a s u b s t a n c e known t o i n t e r f e r e w i t h t h e p r o g r e s s i o n o f r e n a l f a i l u r e , u s i n g a c u l t u r e d c e l l l i n e . When magnesium l i t h o s p e r m a t e B was added t o t h e me‑
dium a t g r a d e d c o n c e n t r a t i o n s , LDH l e a k a g e was s u p p r e s s e d i n p r o p o r t i o n t o an i n c r e a s e i n t h e magnesium l i t h o s p e r m a t e B c o n c e n t r a t i o n . Leakage o f MDA i n t o t h e medium o c c u r r e d i n p a r a l l e l w i t h v a r i a t i o n i n t h e LDH c o n c e n t r a t i o n when magnesium l i t h o s p e r m a t e B was added t o t h e medium. T h i s f i n d i n g i n d i c a t e s t h a t magnesium l i t h o s p e r m a t e B may b e a p r o m i s i n g a g e n t f o r u s e a g a i n s t p a r a q u a t ‑ i n d u c e d c e l l i n j u r y .
2 2 ) Yokozawa T . , Chen C . P . , Dong E . , Tanaka T . , Nonaka G . and Nishioka
I.: Study ont h e I n h i b i t o r y E f f e c t o f Tannins and F l a v o n o i d s a g a i n s t t h e 1 , 1 ・ D i p h e n y l ‑ 2 ‑ p i c r y l h y d r a z y l R a d i c a l . B i o c h e m . Ph α rm α c o l . , 5 6 : 2 1 3 ‑ 2 2 2 , 1 9 9 8 .
F i f t y
時o n et a n n i n s and f o r t y ‑ o n e f l a v o n o i d s i s o l a t e d from O r i e n t a l m e d i c i n a l h e r b s were e v a l u a t e d f o r t h e i r a n t i o x i d a n t a b i l i t y w i t h a 1 , 1 ‑ d i p h e n y l ‑ 2 ‑ p i c r y l h y d r a z y l (DPPH) r a d i c a l ‑ g e n e r a t i n g s y s t e m . The r e s u l t s showed t h a t t a n n i n s and c e r t a i n f l a v o n o i d s a r e p o t e n t i a l f r e e ‑ r a d i c a l s c a v e n g e r s , and t h a t t h e i r a c t i v i t y a g a i n s t t h e DPPH r a d i c a l i s c l o s e l y a s s o c i a t e d w i t h t h e i r c h e m i c a l s t r u c t u r e . A comparison o f t h e two c l a s s e s o f compounds showed t h a t t a n n i n s h a v e more p o t e n t i a l than f l a v o n o i d s b e c a u s e a l m o s t a l l t h e t a n n i n s d e m o n s t r a t e d s i g n i f i c a n t s c a v e n g i n g a c t i o n w i t h i n a low c o n c e n t r a t i o n r a n g e , whereas t h e a c t i v i t y o f f l a v o n o i d s v a r i e d d i s t i n c t i v e l y among t h e d i f f e r e n t compounds. An i n c r e a s e o f g a l l o y l g r o u p s , m o l e c u l a r w e i g h t , and o r t h o ‑ h y d r o x y l s t r u c t u r e e n h a n c e d t h e a c t i v i t y o f t a n n i n s , whereas t h e number and p o s i t i o n o f hydroxyl groups were i m p o r t a n t f e a t u r e s f o r t h e s c a v ‑ e n g i n g o f f r e e r a d i c a l s by f l a v o n o i d s . M o r e o v e r , i t a p p e a r e d t h a t when t h e f r e e hydroxyl group was m e t h o x y l a t e d o r g l y c o s y l a t e d , t h e i n h i b i t o r y a c t i v i t y was o b v i o u s l y d e c r e a s e d o r e v e n a b o l i s h e d .
2 3 ) Ninomiya H . , Mitsuma T . , Takara M . , Yokozawa T . , Terasawa K. and Okuda H . : E f f e c t s o f t h e O r i e n t a l Medical P r e s c r i p t i o n Wen‑Pi‑Tang i n P a t i e n t s R e c e i v i n g D i a l y s i s . P h y t o m e d i c i n e , 5 : 2 4 5 ‑ 2 5 2 , 1 9 9 8 .
H e m o d i a l y s i s p a t i e n t s were t r e a t e d w i t h Wen‑Pi‑Tang C a t y p e o f t r a d i t i o n a l C h i n e s e (K
，α mpo
）明p r e s c r i p t i o n ) f o r 8 w e e k s , and t h e c h a n g e s i n a c t i v e oxygen p r o d u c t i o n by n e u t r o p h i l s , s u p e r o x i d e d i s m u t a s e (SOD) a c t i v i t y , m e t h y l g u a n i d i n e (MG) / c r e a t i n i n e ( C r ) r a t i o , b l o o d c h e m i s t r y and s u b j e c t i v e symptoms were e x a m i n e d . A d e c r e a s e i n a c t i v e oxygen p r o d u c t i o n by n e u t r o p h i l s was o b s e r v e d i n p a t i e n t s w i t h and w i t h o u t p h o r b o l m y r i s t a t e a c e ‑ t a t e s t i m u l a t i o n . SOD a c t i v i t y and MG/Cr r a t i o were a l s o r e d u c e d by t h e t r e a t m e n t . I n a d ‑ d i t i o n , c o l d n e s s o f t h e l i m b s , c o n s t i p a t i o n and e a s y f a t i g a b i l i t y were improved by W e n ‑ P i ‑ Tang a d m i n i s t r a t i o n .
2 4 ) Yokozawa T . , Chen C . P . , Tanaka T . and Kouno
I.:I s o l a t i o n from Wen‑Pi‑Tang o f
t h e A c t i v e P r i n c i p l e P o s s e s s i n g A n t i o x i d a t i o n and R a d i c a l ‑ s c a v e n g i n g A c t i v i t i e s .
P h y t o m e d i c i n e , 5 : 3 6 7 ‑ 3 7 3 , 1 9 9 8 .
The a c t i v e p r i n c i p l e s o f Wen‑Pi‑Tang p o s s e s s i n g a n t i o x i d a t i o n and f r e e r a d i c a l ‑ s c a v e n g i n g a c t i v i t i e s were i s o l a t e d , and i t s p r o p e r t i e s were d e t e r m i n e d u s i n g t h e t h i o b a r b i t u r i c a c i d ‑ r e a c t i v e s u b s t a n c e (TEARS) p r o c e d u r e and a 1 , 1
・d i p h e n y l ‑ 2 ‑ p i c r y l h y d r a z y l(DPPH) r a d i c a l g e n e r a t i n g s y s t e m . S i g n i f i c a n t i n h i b i t o r y a c t i v i t i e s a g a i n s t b o t h t h e f o r m a t i o n o f TEARS and g e n e r a t i o n o f f r e e r a d i c a l s were shown by f r a c t i o n s 5 , 6 and 7 , which c o n t a i n e d mainly p o l y p h e n o l i c compounds and p h e n o l i c g l y c o s i d e s a s w e l l a s a n t h r a q u i n o n e g l y c o s i d e s . The most a c t i v e compounds were p r o c y a n i d i n B ‑ 2 3 , 3 ' ‑ d i ‑ 0 ‑ g a l l a t e ,
（ー）−e p i c a t e c h i n3‑0 ・ g a l l a t e , p r o c y a n i d i n B ‑ 2 and 1 , 2 , 6 ‑ t r i
・0
司g a l l o y l − β − g l u c o s e ,t h e former two h a v i n g b e e n c o n f i r m e d t o b e t h e most a c t i v e p r i n c i p l e s o f Wen‑Pi
t h a t t a n n i n s make a prominent c o n t r i b u t i o n t o b o t h t h e b i o l o g i c a l a c t i v i t y and p h a r m a c o l ‑ o g i c a l e f f e c t s o f Wen‑Pi‑Tang.
2 5 ) Chung H . Y . , Yokozawa T . , Soung D . Y . , Kye I
.S . , No J . K . and Baek B . S . : P e r o x y n i t r i t e ‑ s c a v e n g i n g A c t i v i t y o f Green Tea T a n n i n .
J.A g r i c . Food C h e m . , 4 6 : 4 4 8 4 ‑ 4 4 8 6 , 1 9 9 8 .
P e r o x y n i t r i t e formed from s u p e r o x i d e and n i t r i c o x i d e a c t s a s a s t r o n g r e a c t i v e o x i d a n t . H o w e v e r , among g r e e n t e a c o m p o n e n t s , c a t e c h i n s w i t h a g a l l o y l group i n h i b i t e d p e r o x y n i t r i t e f o r m a t i o n by 3 ‑ m o r p h o l i n o s y d n o n i m i n e and s c a v e n g e d p e r o x y n i t r i t e i t s e l f . E s p e c i a l l y when compared w i t h p e n i c i l l a m i n e a s a p o s i t i v e c o n t r o l , t h e g r e e n t e a components ( ‑ ) ‑ e p i g a l l o c a t e c h i n 3 ‑ 0 ‑ g a l l a t e and ( ‑ ) ‑ g a l l o c a t e c h i n 3 ‑ 0g a l l a t e , which h a v e two g a l l o y l g r o u p s , showed t h e most p o t e n t p e r o x y n i t r i t e ‑ s c a v e n g i n g a c t i v i t y , i n d i c a t i n g t h a t t h e g a l l o y l group may c o n t r i b u t e t o t h i s a c t i v i t y . A s t r u c t u r e o f f l a v a n ‑ 3 ‑ o l l i n k e d t o g a l l i c a c i d may b e e s s e n ‑ t i a l f o r t h e p e r o x y n i t r i t e ‑ s c a v e n g i n g a c t i v i t y .
2 6 ) Yokozawa T . , Liu Z.W. and Chen C . P . : E v a l u a t i o n o f O r i e n t a l M e d i c i n e s Using a C u l t u r e d Renal E p i t h e l i a l C e l l L i n e , LLC‑PK1: E f f e c t s o f Carthami F l o s , Rhei Rhizoma and A s t r a g a l i R a d i x .
よTr α d .M e d . , 1 5 : 2 0 2 ‑ 2 0 7 , 1 9 9 8 .
The e f f e c t s o f Carthami F l o s , R h e i Rhizoma and A s t r a g a l i Radix e x t r a c t , which h a v e b e e n p r o v e d t o a m e l i o r a t e r e n a l f a i l u r e i n r a t s w i t h g l y c e r o l ‑ i n d u c e d r e n a l f a i l u r e , were examined i n c e l l c u l t u r e under t h r e e d i f f e r e n t c u l t u r e c o n d i t i o n s . R h e i Rhizoma e x t r a c t showed t h e most p o t e n t e f f e c t among t h e t h r e e c r u d e drugs under any o f t h e r o u t i n e , h y p o x i a ‑ r e o x y g e n a t i o n and c i s p l a t i n e x p o s u r e c o n d i t i o n s e m p l o y e d , s u g g e s t i n g t h a t i t s f a v o r a b l e e f ‑ f e c t on p r o x i m a l t u b u l e f u n c t i o n i s d u e t o s u p p r e s s i o n o f l i p i d p e r o x i d a t i o n v i a f r e e r a d i c a l s . I n c o n t r a s t , t h i s a n t i ‑ c y t o t o x i c a c t i v i t y was low i n A s t r a g a l i Radix e x t r a c t , and a l m o s t n i l i n Carthami F l o s e x t r a c t . T h e s e r e s u l t s i n d i c a t e t h a t R h e i Rhizoma, A s t r a g a l i Radix and Carthami F l o s e x e r t t h e i r a c t i o n s on d i f f e r e n t s i t e s i n t h e k i d n e y .
2 7 ) Soung D . Y . , Chung H.Y. and Yokozawa T . : A Novel A c t i o n o f 5‑Hydroxytryptamine
a s a P e r o x y n i t r i t e S c a v e n g e r . Ph α rm.Ph α r m a c o l . Commun., 4 : 5 8 3 ‑ 5 8 6 , 1 9 9 8 .
P e r o x y n i t r i t e i s a s t r o n g o x i d a n t formed by t h e r e a c t i o n o f a s u p e r o x i d e a n i o n r a d i c a l and
n i t r i c o x i d e .
Itp l a y s an i m p o r t a n t r o l e i n i n d u c i n g a p o p t o s i s , damaging t h e m i t o c h o n d r i a l r e s p i r a t o r y c h a i n and c a u s i n g t h e o c c u r r e n c e o f n i t r o t y r o s i n e i n c o r t i c a l n e u r o n s .
We h a v e d e m o n s t r a t e d t h a t 5 ‑ h y d r o x y t r y p t a m i n e ( 5 ‑ H T ) d o e s n o t i n h i b i t p e r o x y n i t r i t e f o r m a t i o n by 3 ‑ m o r p h o l i n o s y d n o n i m i n e w i t h t h e r e l e a s e o f a s u p e r o x i d e a n i o n r a d i c a l and n i t r i c o x i d e , h o w e v e r , i t d o e s markedly s c a v e n g e p e r o x y n i t r i t e i t s e l f .
5‑HT i s d i r e c t l y i n v o l v e d i n t h e s c a v e n g i n g o f p e r o x y n i t r i t e , and might p l a y a r o l e i n r e ‑ t a r d i n g t h e d e v e l o p m e n t o f b r a i n d i s e a s e .
。学会報告
1 ) El‑Mekkawy S . , Meselhy M.
R.,Nakamura N . , H a t t o r i , M . , Kawahata T . and Otake T . : A n t i ‑ H I V ‑ 1 A c t i v i t y o f P l a n t s Used i n t h e E g y p t i a n F o l k M e d i c i n e .
日本薬学会第1 1 8
年会，1 9 9 8 ,3
，京都．2）佐藤利江，小松かっ子，中村憲夫，服部征雄，門田重利，難波恒雄，周 光春：「丹参」類 の生薬学的研究（
I I
）一基源と水溶性成分による品質評価一．日本薬学会第1 1 8
年会，1 9 9 8 , 3
，京都3) Min B . S . , Nakamura N . , M i y a s h i r o H . , Bae K . H . , H a t t o r i M . : Anti‑HIV P r o t e a s e T r i t e r p e n e s from t h e S p o r e s o f G α noderm α l u c i d um.
日本薬学会第1 1 8
年会，1 9 9 8 .3 .
京都．4
）西本栄里，MeselhyM.
R.，来田浩明，中村憲夫，服部征雄，赤尾光昭：腸内細菌によるシ コニン代謝反応の検討．日本薬学会第1 1 8
年会，1 9 9 8 ,3
，京都．5) H u s s e i n G .
，中村憲夫，宮代博継，服部征雄：G l y c o s i d eand P r o a n t h o c y a n i d i n s from Mα . y t e n u s s e n e g α l e n s i s .
日本薬学会第1 1 8
年会，1 9 9 8 ,3
，京都．6
）林 隆幸，宮代博継，服部征雄，赤尾光昭：ヒト腸内細菌由来s e n n o s i d e
水解β − g l u c o s i ‑ d a s e
遺伝子の同定．日本薬学会第1 1 8
年会，1 9 9 8 ,3
，京都．7
）横津隆子，劉 中武，董而博：腎虚血一再濯流モデルを用いたG i n s e n o s i d e ‑ R d
の検討．日本薬学会第
1 1 8
年会，1 9 9 8 ,3
，京都．8
）横津隆子，中川孝子，服部征雄，中川 昨：動脈硬化性障害における黒茶の役割について．日本薬学会第
1 1 8
年会，1 9 9 8 ,3
，京都．9
）横津隆子，劉中武，董而博：C e p h a l o r i d i n e
による腎障害に対するG i n s e n o s i d e ‑ R d
の 関与．第1 3
回薬用人参研究会，1 9 9 8 ,3
，神戸．1 0 ) H a t t o r i M . : M e d i c i n a l P l a n t s a s a S o u r c e o f Anti‑HIV A g e n t s . Vth Symposium on P l a n t L i f e i n South West and C e n t r a l A s i a . 1 9 9 8 , 5 , T a s h k e n t , U z b e k i s t a n .
1 1 ) H a t t o r i M . : B i o t r a n s f o r m a t i o n o f P a e o n i f l o r i n by Human I n t e s t i n a l B a c t e r i a . JSPS
・KOSEF J o i n t Seminar on T r a d i t i o n a l O r i e n t a l M e d i c i n e s : Modern S c i e n t i f i c Approach and H a r m o n i z a t i o n , 1 9 9 8 , 7
，富山．1 2 ) P a b l o N . S o l i s , N o r i o Nakamura, D i o n i s i o O l m e d o , Masao H a t t o r i , Mahabir P . G u p t a : P a r a t h e s i l a c t o n e ‑ A N o v e l Compound from P a r a t h e s i s A m p l i f o l i a L u n d . ASP m e e t i n g , 1 9 9 8 , 7 . USA.
1 3
）来田浩明，赤尾光昭，MeselhyRagab Meselhy
，服部征雄：C o n v e n t i o n a l ,G e r m ‑ f r e e ,
G n o t o b i o t e
ラットでのサイコサポニンの体内動態．第1 5
回和漢医薬学会大会，1 9 9 8 ,8 ,
富山．
1 4
）横津隆子，劉中武，大和田滋：C e p h a l o r i d i n e
による腎障害に対するG i n s e n o s i d e ‑ d
の関与．第1 5
回和漢医薬学会大会，1 9 9 8 ,8
，富山．1 5
）王天山，金東郁，横津隆子，服部征雄：イボツヅラフジの高コレステロール食投与ラッ トに及ぼす効果．第1 5
回和漢医薬学会大会，1 9 9 8 ,8
，富山．1 6
）陳翠捧，王天山，中川孝子，横津隆子，服部征雄：NO
におけるイボツヅラフジの影響．第
1 5
回和漢医薬学会大会，1 9 9 8 ,8
，富山．1 7
）中川孝子，横津隆子，李 慶責任，竹内茂禰，服部征雄：P e r o x y n i t r i t e
による細胞障害に対 する緑茶タンニンの役割．第1 5
回和漢医薬学会大会，1 9 9 8 ,8
，富山．1 8
）金東部，横津隆子，服部征雄，門田重利，難波恒雄：LDL
の酸化による内皮細胞傷害と マクロファージ泡沫化における羅布麻の関与．第1 5
回和漢医薬学会大会，1 9 9 8 ,8
，富山．1 9
）田中 隆，横津隆子，河野功：温牌湯の抗酸化及びラジカル消去活性成分．第1 5
回和漢 医薬学会大会，1 9 9 8 ,8
，富山．2 0
） 鄭 海 泳 ， 横 津 隆 子 ， 桂 仁 淑 ， 成 道 裕 ， 直 載 鏡 ， 白 奉 淑 ：P e r o x y n i t r i t e
消去活性 を有する緑茶タンニン．第1 5
回和漢医薬学会大会，1 9 9 8 ,8
，富山．2 1 ) I e n a g a
K.,Mikami H . , T a k e u c h i S . , Nakamura
K.,Aoyagi
K.,Yokozawa T . , Oura H . , Nakano
K.,Endou H . : NZ
『4 1 9 ,a n o v e l i n t r i n s i c a n t i o x i d a n t , a s a t h e r a p e u t i c a g e n t a g a i n s t p r o g r e s s i v e c h r o n i c r e n a l f a i l u r e and g u a n i d i n o c o m p o u n d s . The 5 t h I n t e r ‑ n a t i o n a l Symposium on G u a n i d i n o Compounds i n B i o l o g y and M e d i c i n e , 1 9 9 8 , 9 , Yokohama.
2 2 ) Meselhy Ragab M e s e l h y , H i r o a k i K i d a , T e r u a k i Akao, Masao H a t t o r i : I s o l a t i o n o f Human I n t e s t i n a l B a c t e r i a C a p a b l e o f Transforming Rhein t o Rhein A n t h r o n e .
第1 5
回和漢医薬学会大会，1 9 9 8 ,8
，富山．2 3
）馬 超美，中村憲夫，服部征雄，喬俊厘，子海藍：蒙薬文冠木のHIV‑1
プロテアーゼ阻 害物質について．日本生薬学会第4 5
回年会，1 9 9 8 ,9
，仙台．2 4 ) Yokozawa T . , L i u Z . W . , Dong E . : P r o t e c t i v e e f f e c t s o f g i n s e n o s i d e ‑ R d i n a r e n a l i s c h e ‑ m i a ‑ r e p e r f u s i o n m o d e l . The 7 t h I n t e r n a t i o n a l Symposium on G i n s e n g , 1 9 9 8 , 9 , S e o u l . 2 5 ) Yokozawa T . , Chung H . Y . : E f f e c t o f g i n s e n o s i d e ‑ R d i n c e p h a l o r i d i n e ‑n d u c e d r e n a l
d i s o r d e r . The 7 t h I n t e r n a t i o n a l Symposium on G i n s e n g , 1 9 9 8 , 9 , S e o u l .
26）横津隆子，田中 隆，河野功：腎不全に有効な温牌湯の活性成分の探索．第
