薬物代謝工学部門 Department
o f Metabolic Engineering
教授 服部 征雄 Professor
Masao H a t t o r i ( P h . D . )
助教授 横津 隆子 AssociateP r o f e s s o r Takako Yokozawa ( P h . D . )
助手 宮代 博継 AssistantP r o f e s s o r H i r o t s u g u M i y a s h i r o ( P h . D . )
技官 中村 憲夫 ResearchA s s o c i a t e N o r i o Nakamura ( P h . D . )
薬物代謝工学部門は和漢薬の薬効，毒性発現に関与する代謝系の分子生物学的研究を発展させる ことを設置目的とし，①和漢薬の薬効発現に関与する腸内細菌の役割，②薬物代謝に関する腸内 細菌遺伝子の解明，③腎毒性物質産生機構の分子生物学的解明とその制御に関する研究を課題とし て取りあげ，和漢薬の薬効発現機構，生体へのレスポンスなどの基礎的研究を通じて，和漢薬の科 学的評価や臨床応用をはかることを目指している。主な研究題目を以下に示す。
1. 天然、物のバイオトランスフォーメイション
2. 和漢薬の薬効発現に関与する腸内細菌遺伝子の解析
3 . AIDS,
C 型肝炎の予防および治療薬の開発 4. 腎疾患における病態の解明と腎臓病治療薬の開発本年度の主な研究を列挙すると：
1 .
ヒト腸内細菌によるゴボウの種子に多量に含まれる arctiin の哨乳動物リグナン（エンテロジ オール，エンテロラクトン）への変換を検討し，この化合物が容易に加水分解，脱メチル化，脱水酸基化反応、を経て種々の代謝物に変換されることを明らかにした。また，これらの代謝物 のエストロジェン様作用，抗エストロジェン様作用を検討した。さらにヒト腸内細菌と化学反 応を併用して簡便なエンテロラクトン，エンテロジオールの合成法を開発した。
2 .
HIVーインテグラーゼ阻害活性を指標にタイ薬用植物を探索し， Coleus pαrvifo l i u s ,
Thevetiα per・uviαnα などに強い酵素阻害活性を見出した。又，これらのエキスから阻害活性成分を単離し，その阻害様式を検討した。
3. 中国およびタイで用いられている薬用植物の抗ヘルペスウイルス活性また， C 型肝炎ウイル ス由来の RNA ポリメラーゼ阻害活性を検討した。
4. 腎疾患増悪因子のラジカルの産生部位を証明するとともに，蛋白・糖修飾を surrogate マーカー として利用し，腎不全合併症における病態把握に着手した。また酸化ストレスを抑制する新た な治療手段を探索するために，地検成分の sanguiin H-6，ソバ成分，温牌湯，桂枝夜苓丸を 中心に＋食言すした。
く〉著書 Books
1) Yokozawa T . , Dong 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 G r e e n Tea Polyphenols. ” Free R a d i c a l s i n F o o d s :
C h e m i s t r y , N u t r i t i o n a n d Health”, edited by M. M o r e l l o , F . S h a h i d i a n d C . T . H o , Quaker O a t s C o . , B a r r i n g t o n , 2 0 0 2 , p p . 2 2 4 ‑ 2 4 0 .
2 ）横津隆子：腎障害を伴う高血圧における緑茶ポリフェノール．“茶の機能一生体機能の新たな可能性”，
村松敬一郎，小園伊太郎，伊勢村護，杉山公男，山本万里，学会出版センター，東京， 2002,
p p . 1 4 5 ‑ 1 5 1 .
3 ）三瀦忠道，横津隆子，二宮裕幸：大黄ならび1こ大黄含有漢方方剤・温牌湯の慢性腎不全に対する効果．“腎とフリーラジカルー第 6 集ーペ横津隆子，湯川 進監修，宗正敏，青柳一正編，東京医学社，東京，
2 0 0 2 , p p . 6 7 ‑ 7 3 .
4 ）家永和治，三上博輝，西端良治，内木充，中村耕，横津隆子，大浦彦吉，青柳一正，遠藤仁： NZ-419
（内因性抗酸化物）の慢性腎不全進展抑制作用．“腎とフリーラジカルー第 6 集一”，横津隆子，湯川 進
監修，宗正敏，青柳一正編，東京医学社，東京， 2002,
p p . 7 4 ‑ 7 6 .
5 ）大久保 勉，レカ・ラジュ・ジュネジャ，横犀隆子：緑茶ポリフェノールの生体内抗酸化と透析患者への 試み“腎とフリーラジカルー第 6 集－”，横津隆子，湯川 進監修，宗正敏，青柳一正編，東京医学社，
東京， 2002,
p p . 1 2 0 ‑ 1 2 3 .
6 ）中川孝子，横津隆子，寺津捷年：糖尿病性腎症における漢方方剤の役割．“腎とフリーラジカルー第 6 集－”，横津隆子，湯川進監修，宗正敏，青柳一正編，東京医学社，東京， 2002,
p p . 1 3 8 ‑ 1 4 1 .
7) Yokozawa T . Kim H . Y . , Cho E . J . , Choi J . S . : Antioxidant A c t i v i t i e s o f Mustard Leaf
(Brαssicαj u
neeα）．“腎とフリーラジカルー第 6 集一”，横津隆子，湯川 進監修，宗正敏，青柳一正編，東京医学 社，東京， 2002,p p . 1 4 2 ‑ 1 4 8 .
8 ）横津隆子，中川孝子：シスプラチン誘発急J性腎不全における緑茶タンニンの関与．“腎とフリーラジカ ルー第 6 集一”，横津隆子，湯川進監修，宗正敏，青柳一正編，東京医学社，東京， 2002,
p p . 2 0 6 ‑ 2 1 0 .
く〉原著 Originalp a p e r s
1)
Abdel・HafezA. ' A . , Nakamura N . , and Hattori M.: Biotransformation of phorbol by human i n t e s t i n a l b a c t e r i a . Chem. Pharm. B u l l . , 5 0 , 160‑164 ( 2 0 0 2 ) .
A n a e r o b i c i n c u b a t i o n o f p h o r b o l ( 1 ) f r o m C r o t o n t i g l i u m w i t h human i n t e s t i n a l b a c t e r i a a f f o r d e d f i v e m e t a b o l i t e s : i s o p h o r b o l ( 2 ) , d e o x y p h o r b o l ( 3 ) , 4 ( 3 ,
9α ， 20-trihydroxy-13,1 5 ‑ s e c o ‑ 1 , 6 , 1 5 ‑ t i g l i a t r i e n e ‑ 3 , 1 3 ‑ d i o n e ( 4 ) ,
4β ， 9α ，2 0 ‑ t r i h y d r o x y ‑ 1 5 , 1 6 , 1 7 ‑ t r i n o r ‑ 1 , 6 ‑ t i g l i a d i e n e ‑ 3 , 1 3 ‑ d i o n e ( 5 ) a n d
4β ， 9α ， 20-trihydroxy-14( 13→ 12)-abeo-12αH-1 , 6 ‑ t i g l i a d i e n e ‑ 3 , 1 3 ‑ d i o n e ( 6 ) . A l l t h e s e m e t a b o l i t e s
(2・6)w e r e i d e n t i f i e d a n d c h a r a c t e r i z e d b y s p e c t r o s c o p i c m e a n s , i n c l u d i n g t w o ‑ d i m e n s i o n a l (2D)‑NMR. N i n e d e f i n e d s t r a i n s f r o m t h e human i n t e s t i n e showed a n a b i l i t y t o t r a n s f o r m 1 t o t h e s e m e t a b o l i t e s .
'CH20H
1
4
OH
。H20H
2
5 Chart 1 .
3
。
2)
El・MekkawyS . , Meselhy M. R . ,
Abdel・HafezA. A . , Nakamura N . , Hattori M., Kawabata T . , and Otake T . : I n h i b i t i o n of cytopathic e f f e c t of human immunodeficiency v i r u s t y p e ‑ 1 by various phorbol ｷ d e r i v a t i v e s . Chem. Pharm. B u l l . , 5 0 ,
523・529( 2 0 0 2 ) .
F o r t y ‑ e i g h t d e r i v a t i v e s o f p h o r b o l ( 9 ) a n d i s o p h o r b o l ( 1 4 ) were e v a l u a t e d f o r t h e i r i n h i b i t i o n o f HIV‑1 i n d u c e d c y t o p a t h i c e f f e c t s (CPE) on MT‑4 c e l l s , a s w e l l a s t h e i r a c t i v a t i o n o f p r o t e i n k i n a s e C ( P K C ) , a s i n d i c e s o f a n t i ‑ H I V ‑ 1 and t u m o r p r o m o t i n g a c t i v i t i e s , r e s p e c t i v e l y . Of t h e s e c o m p o u n d s , t h e most p o t e n t i n h i b i t i o n o f CPE was o b s e r v e d i n 1 2 ‑ 0 ‑ t e t r a d e c a n o y l p h o r b o l 1 3 ‑ a c e t a t e ( 8 ) a n d
12-0-acet）匂horbol1 3 ‑ d e c a n o a t e ( 6 ) . The f o r m e r a l s o showed t h e s t r o n g e s t PKC a c t i v a t i o n a c t i v i t y , w h i l e t h e l a t t e r showed no a c t i v i t y a t 1 0 n g / m l . B o t h a c t i v i t i e s w e r e g e n e r a l l y o b ｭ s e r v e d i n t h o s e p h o r b o l d e r i v a t i v e s w i t h a n AIB
transc o n f i g u r a t i o n , b u t n o t i n t h e i s o p h o r b o l d e r i v a t i v e s w i t h a n AIB
cisc o n f i g u r a t i o n . A c e t y l a t i o n o f 20‑0H i n t h e p h o r b o l d e r i v a t i v e s s i g n i f i c a n t l y r e d u c e d t h e i n h i b i t i o n o f CPE, a s shown i n 1 2 ‑ 0 ‑ ,
20-0・－diacetylphorbol1 3 ‑ d e c a n o a t e ( 6 a ) ( I C 1 0 0 = 1 5 . 6
ｵg / m l )
vscompound 6 ( I C 1 0 0 = 0 . 0 0 7 6 ｵ g / m l ) , a n d 1 2 ‑ 0 ‑ t e t r a d e c a n o y l p h o r b o l 1 3 , 2 0 ‑ d i a c e t a t e ( S a ) ( I C 1 0 0 = 1 5 . 6 ｵ g / m l )
vs1 2 ‑ 0 , . t e t r a d e c a n o y l p h o r b o l 1 3 ‑ a c e t a t e ( 8 ) ( I C 1 0 0 = 0 . 0 0 0 4 8 ｵ g / m l ) , e x c e p t i n t h e c a s e o f
12-0・decanoylphorbol1 3 ‑ ( 2 ‑ m e t h y l b u t y r a t e ) ( 4 ) a n d p h o r b o l ‑ 1 2 ,
13・diacetate( 9 c ) . The r e d u c t i o n of a c a r b o n y l g r o u p a t C
”3 a b r u p t l y r e d u c e d t h e i n h i b i t i o n o f CPE, a s o b s e r v e d i n
3β －hydroxyphorbol1 2 , 1 3 , 2 0 ‑ t r i a c e t a t e
(9め（IC100=500ｵ g / m l )
vsp h o r b o l 1 2 , 1 3 , 2 0 ‑ t r i a c e t a t e ( 9 d ) ( I C 1 0 0 = 6 2 . 5 ｵ g / m l ) .
A l t h o u g h 8 was e q u i p o t e n t i n t h e i n h i b i t i o n o f CPE, a n d a c t i v a t i o n o f PKC, b o t h a c t i v i t i e s w e r e a b r u p t l y d e ｭ c r e a s e d by t h e a c e t y l a t i o n o f 20‑0H a n d m e t h y l a t i o n o f 4‑0H [ a s i n Sa a n d 4 ‑ 0 ‑ m e t h y l ‑ 1 2 ‑ 0 ‑ t e t r a d e c a n o y l p h o r b o l 1 3 , 2 0 ‑ d i a c e t a t e ( S b ) , r e s p e c t i v e l y ] . On t h e o t h e r h a n d , i t s p o s i t i o n a l i s o m e r ( 1 2 ‑ 0 ‑ a c e t y l p h o r b o l 1 3 ‑ t e t r a d e c a n o a t e ( S c ) showed n e i t h e r a c t i v i t i e s . The r e m o v a l o f a l o n g a c y l g r o u p i n 8 l e d t o a s u b s t a n t i a l l o s s o f b o t h a c t i v i t i e s , a s
R4
白L凸LβLPL
H H H H H H H H H M H H H M H H H H H H H M A H H H H H H H R3
00・00700ρ
今31v、3、54’I7・フ－
H H H H
8884HH戸llCCCCCICCCZうJl
c c c H H A H H A A H H A A H C H H H H A A A B H C H C H C Ac
Ac Tig Tig 2‑Me butyryl 2・恥tiebutyryl 2‑Me butyryl C10H190 C10H190 C10H190 C12H230 Ac Ac Ac C14H270 C14H270 H H Ac Ac Ac Ac Ac Bz C6H110 C6H110 C9H170 C9H170 C12H230 C12H230
R1
H Ac H Ac C10H190 C10H190 Tig Ac Ac Ac
2ール1ebutyryl C14H270 C14H270 C14H270 Ac H H Ac H Ac Ac Ac Ac Bz Ac Ac Ac Ac Ac Ac R1
b a a b a b e d a b e d e g a a a 1 1 2 3 4 4 5 6 6 6 7 8 8 8 8 8 9 9 9 9 9 9 9 0 1 1 2 2 3 3
－且’E
・ E
－且咽且EA・E
CH20R3
20
OAc
19
’3
H 勾，
c o c o
句F－Hc
a--
A
CH20Ac
Chart 2 .
9f
Of t h e
12-0-acetyl-13-0・acylphorbold e r i v a t i v e s , t h e h i g h e s t i n h i b i t i o n o f CPE was o b s e r v e d i n 6 , w h i c h h a s a d o d e c a n o y l r e s i d u e a t C ‑ 1 3 . B o t h a n i n c r e a s e a n d d e c r e a s e i n t h e number o f f a t t y a c i d c a r b o n c h a i n s r e s u l t e d i n s i g n i f i c a n t r e d u c t i o n o f t h e i n h i b i t i o n o f CPE.
3) Wang L . , Min B . , Li Y . , Nakamura N . , Qin G . , Li C . , and Hattori M.: Annonaceous acetogenins from t h e l e a v e s of Annona montana. B i o o r g . Med. Chem., 1 0 ,
561・565( 2 0 0 2 ) . A n o v e l Annonaceous a c e t o g e n i n , m o n t a n a c i n F , w i t h a new t y p e o f t e r m i n a l l a c t o n e u n i t , was i s o l a t e d f r o m t h e l e a v e s o f Annonna m o n t a n a . I t s s t r u c t u r e was d e t e r m i n e d on t h e b a s i s o f s p e c t r a l e v i d e n c e s a n d c h e m i c a l m e t h o d s , a n d a p o s s i b l e b i o s y n t h e t i c p a t h w a y was d i s c u s s e d . I n a d d i t i o n , t h e c y t o t o x i c i t y o f m o n t a n a c i n F was e v a l u a t e d i n v i t r o a g a i n s t L e w i s l u n g c a r c i n o m a (LLC) t u m o r c e l l l i n e s . F u r t h e r m o r e , t h e p r e v i o u s l y i s o l a t e d c y t o t o x i c a c e t o g e n i n a n n o n a c i n a g a i n s t LLC was e x a m i n e d f o r i n v i v o a n t i t u m o r a c t i v i t y w i t h LLC t u m o r c e l l s .
4) Tewtrakul S . , Nakamura N . , Hattori M., Fujiwara T . , and Supavita T . : ̲Flavanone and f l a v o n o l g l y c o s i d e s from the l e a v e s of T h e v e t i a p•?ruviana and t h e i r
HIV・1r e v e r s e t r a n s c r i p t a s e and
HIV・1i n t e g r a s e i n h i b i t o r y a c t i v i t i e s . Chem. Pharm. B u l l . , 5 0 , 630‑635 ( 2 0 0 2 ) .
Two new f l a v a n o n e g l u c o s i d e s ,
(2R)‑a n d
(2S)-5-0-/3 ・o-glucopyranosyl-7, 4 二dihydroxy-3 ’， 5 二dimethoxyflavanone[ p e r v i a n o s i d e I ( 3 ) , p e r u v i a n o s i d e I I ( 4 ) ] a n d a new f l a v o n o l g l y c o s i d e , q u e r c e t i n
3-0－｛ β －o-glucopyranosyl (1 →2）ー［ α ーL-rhamnopyranosyl (1 →6）］ー β ーo-galactopyranoside}
( p e r u v i a n o s i d e I I I ,
13)w e r e i s o l a t e d f r o m t h e l e a v e s o f T h e v e t i a
peruνianaS c h u m . , t o g e t h e r w i t h n i n e known f l a v o n o l g l y c o s i d e s a n d two known i r i d o i d g l u c o s i d e s . The s t r u c t u r e s o f a l l compounds w e r e 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 a n d s p e c t r o s c o p i c m e t h o d s . T h e i r i n h i b i t o r y e f f e c t s a g a i n s t HIV‑1 r e v e r s e t r a n s c r i p t a s e a n d HIV‑1 i n t e g r a s e w e r e a l s o i n v e s t i g a t e d .
5) M a C . , Nakamura N . , Hattori M., and Cai S . : I s o l a t i o n of malonyl o l e a n o l i c a c i d hemiester a s anti‑HIV protease substance from t h e stems of
Cynomo，初ms o n g a r i c u m . C h i n . Pharm. J . , 3 7 ,
336・338
( 2 0 0 2 ) .
OBJECTIVE F u r t h e r ̲ i n v e s t i g a t i o n on t h e b i o ‑ a c t i v e c o n s t i t u e n t s o f t h e s t e m s o f C y n o m o r i u m s o n g a r i c u m . METHODS The compounds w e r e i s o l a t e d w i t h c h r o m a t o g r a p h i c t e c h n o l o g y , i n c l u d i n g HPLC. T h e i r s t r u c t u r e s w e r e 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 t h o d s . RESULTS M a l o n y l o l e a n o l i c a c i d h e m i e s t e r
(I)a n d z i n g e r o n e 4 ‑ 0 ‑
β ーo-glucopyranoside
( I I ) w e r e i s o l a t e d from t h e p l a n t . CONCLUSION B o t h compounds w e r e i s o l a t e d f r o m t h i s p l a n t f a m i l y f o r t h e f i r s t t i m e a n d compound I p o t e n t l y i n h i b i t e d t h e a c t i v i t y o f HIV p r o t e a s e .
6) Min B . , Lee H . , Bae K . , Gao J . , Nakamura N . , and Hattori M.: Antitumor a c t i v i t y of cultured mycelia of Ganoderma l u c i d u m . N a t u r a l Product Scienc•?s, 8 , 52‑54 ( 2 0 0 2 ) .
The c u l t u r e d m y c e l i a o f f u n g u s Ganoderma l u c i d u m w e r e i n v e s t i g a t e d f o r t h e i n h i b i t o r y e f f e c t on t h e g r o w t h o f s . c . t r a n s p l a n t e d L e w i s l u n g c a r c i n o m a (LLC) i n BDF‑1 m i c e by i n t r a p e r i t o n e a l ( i . p . ) a d m i n i s t r a t i o n . The c u l t u r e d myｭ c e l i a showed a n t i t u m o r a c t i v i t y w i t h T I C v a l u e s o f 8 9 . 6 a n d 5 0 . 3
%a t d o s e s o f 1 0 0 a n d 500 m g / k g , r e s p e c t i v e l y , compared t o a d r i a m y c i n , which was u s e d a p o s i t i v e c o n t r o l , w i t h T I C v a l u e o f 5 4 . 6
%a t 2 m g / k g .
7) Hur J . , Park J . , Park J . , Hyun K . , l L e e K . , Miyashiro H . , and Hattori M.: I n h i b i t o r y e f f e c t s of n i n e t y nine Korean p l a n t s on human immunodeficiency v i r u s type 1 protease a c t i v i t y . N u t r a c e u t i c a l s and Food, 7 ,
123・ 127( 2 0 0 2 ) .
N i n e t y n i n e e x t r a c t s f r o m K o r e a n p l a n t s w e r e s c r e e n e d f o r t h e i r i n h i b i t o r y a c t i v i t i e s on 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 ) t y p e 1 p r o t e a s e by a n HPLC m e t h o d . The p r o t e a s e i n h i b i t o r y a c t i v i t i e s w e r e d e t e r m i n e d by i n c u b a t i n g
t h e e x t r a c t s i n r e a c t i o n m i x t u r e s c o n t a i n i n g p r o t e a s e a n d s u b s t r a t e
(His-Lys-Ala-Arg-Val-Leu-(p-N02-Phe）ーGlu-Ala-N l e ‑ S e r ‑ N H 2 ) t o p e r f o r m p r o t e o l y t i c c l e a v a g e r e a c t i o n s . Of t h e e x t r a c t s t e s t e d , t h e w a t e r e x t r a c t s o f Viburnum awabuki ( s t e m a n d l e a v e s ) a n d D i s t y l i u m racemosum ( l e a v e s ) h a d t h e h i g h e s t p r o t e a s e i n h i b i t o r y a c t i v i t i e s a t a c o n ｭ c e n t r a t i o n o f 1 0 0 ｵ g l m L . A c t i v i t y ‑ g u i d e d f r a c t i o n a t i o n r e v e a l e d t h a t t h e n ‑ b u t a n o l f r a c t i o n o f t h e V . awabuki e x t r a c t a n d t h e e t h y l a c e t a t e f r a c t i o n from t h e D . racemosum e x t r a c t h a d t h e g r e a t e s t i n h i b i t o r y a c t i v i t y on HIV‑1 p r o t e a s e . 8) M a C . , Nakamura N . , H a t t o r i M., Kawabata T . , and Otake T . : I n h i b i t o r y e f f e c t s of t r i t e r p e n e ｭ azidothymidine conjugates on p r o l i f e r a t i o n of human immunodeficiency v i r u s type 1 and i t s p r o t e a s e . Chem. Pharm. B u l l . , 5 0 ,
877・880( 2 0 0 2 ) .
The c o n j u g a t e s o f some d k 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 t r i t e r p e n e s which show p o t e n t i n h i b i t i o n 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 t y p e 1 p r o t e a s e ( H I V ‑ 1 PR) w i t h a r e v e r s e t r a n s c r i p t a s e i n h i b i t o r a z i d o t h y m i d i n e (AZT) o r a n t i ‑ H I V a l k a l o i d FK 3000 w e r e p r e p a r e d , a n d t h e i r i n h i b i t o r y a c t i v i t i e s w e r e i n v e s t i g a t e d a g a i n s t
HIV寸nducedc y t o p a t h i c e f f e c t s (CPE) a n d HIV‑1 P R . Most o f t h e t r i t e r p e n e ‑ A Z T c o n j u g a t e s showed p o t e n t a n t i ‑ H I V a c t i v i t y a s w e l l a s m o d e r a t e t o p o t e n t PR i n h i b i t o r y a c t i v i t y , t h o u g h AZT i t s e l f showed no PR i n h i b i t o r y a c t i v i t y a t a l l . H o w e v e r , t h e t r i t e r p e n e ‑ F K 3000 c o n j u g a t e s showed n e i t h e r PR i n h i b i t o r y a c t i v i t y n o r a n t i ‑ H I V a c t i v i t y .
9 ) Gao J . , Min B . , Ahn E . , Nakamura N . , Lee H . , and Hattori M.: New t r i t e r p e n e aldehyde, l u c i a l d e h y d e s A‑C, from Ganoderma l u c i d u m and t h e i r c y t o t o x i c i t y a g a i n s t murine and human tumor c e l l s . Chem.
Pharm~B u l l . , 5 0 ,
837・840( 2 0 0 2 ) .
T h r e e new l a n o s t a n t e ‑ t y p e t r i t e r p e n e a l d e h y d e s , named l u c i a l d e h y d e s A‑C ( 1 ‑ 3 ) , w e r e i s o l a t e d from t h e f r u i t i n g b o d ｭ i e s o f Ganoderma l u c i d u m , t o g e t h e r w i t h g a n o d e r m a n o n o l ( 4 ) , g a n o d e r m a d i o l ( 5 ) , g a n o d e r m a n o n d i o l ( 6 ) ,
ganode口nanontriol
( 7 ) , g a n o d e r i c a c i d A ( 8 ) , g a n o d e r i c a c i d B8 ( 9 ) , a n d g a n o d e r i c a c i d C 1 ( 1 0 ) . The s t r u c t u r e s o f t h e new
trite中enesw e r e d e t e r m i n e d a s
(24め－3/3-hydroxy-5α ーlanosta-7 , 9 ( 1 1 ) , 2 4 ‑ t r i e n ‑ 2 6 ‑ a l ( 1 ) , ( 2 4 ｣ ) ‑ 3 , 7 ‑ d i o x o ‑
5α －lanosta-8,24-dien-26-al( 2 ) , a n d
(24£)-3β －hydroxy-7-oxo-5α －lanosta-8,24-dien-26-al( 3 ) , r e s p e c t i v e l y , by s p e c ｭ t r o s c o p i c m e a n s . The c y t o t o x i c i t y o f t h e compounds i s o l a t e d from t h e ganoderma mushroom was t e s t e d i n v i t r o a g a i n s t LLC, T ‑ 4 7 D , Sarcoma 1 8 0 , a n d Meth‑A t u m o r c e l l l i n e s . L u c i a l d e h y d e s B‑C ( 2 ‑ 3 ) , g a n o d e r m a n o n o l ( 4 ) a n d g a n o d e r m a n o n d i o l ( 6 ) showed c y t o t o x i c e f f e c t on t e s t e d t u m o r c e l l s . Of t h e c o m p o u n d s , l u c i a l d e h y d e C ( 3 ) e x h i b ｭ i t e d t h e most p o t e n t c y t o t o x i c i t y a g a i n s t LLC, T ‑ 4 7 D , Sarcoma 1 8 0 , a n d
Meth鈴At u m o r c e l l s w i t h EDso v a l u e s o f 1 0 . 7 , 4 . 7 , 7 . 1 , a n d 3 . 8 ｵ g / m l , r e s p e c t i v e l y .
尽2
R4 R3
R 1 R2 R3 R4 R 1 1
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。4
。 ~24(25)CH20H
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。6
。OH OH CH3 9
。7
。OH OH CH20H 10
。Chart 3 .
R2 R3 R4 Rs
。
H2 H2 H2
。
H2 H2 H2
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Ra R1
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H2 H COOH
1 0 ) Zhao J . , Nakamura N . , Hattori M., Kuboyama T . , Tohda C . , and Komatsu K . : Withanolide d e r i v a t i v e s from the r o o t s of W i t h a n i a somnif e r a and t h e i i r n e u r i t e outgrowth a c t i v i t i e s . Chem.
Pharm. B u l l . , 5 0 ,
760・765( 2 0 0 2 ) .
F i v e new w i t h a n o l i d e d e r i v a t i v e s ( 1 a n d 9 ‑ 1 2 ) w e r e i s o l a t e d f r o m t h e r o o t s o f W i t h a n i a s o m n i f e r a t o g e t h e r w i t h f o u r t e e n known compounds ( 2 ‑ 8 , a n d 1 3 ‑ 1 9 ) . On t h e b a s i s o f s p e c t r o s c o p i c a n d p h y s i o c h e m i c a l e v i d e n c e , comｭ p o u n d s 1 a n d 9 ‑ 1 2 w e r e d e t e r m i n e d t o b e ( 2 0 S ,
22R)-3α ， 6α －epoxy-4β ， 5β ， 27-trihydroxy-1-oxowitha-24-enolide( 1 ) ,
27-0－ β －n-glucopyranosylpubesenolide 3-0－ β －D-glucopyranosyl(
1 → 6）ー β －D-glucopyranoside( w i t h a n o s i d e V I I I , 9 ) ,
27-0－β －D-glucopyranosyl (1 →6）ーβ －n-glucopyranosylpubesenolide 3-0－β ーD-glucopyranosyl (1 →6）－β ーDg l u c o p y r a n o s i d e ( w i t h a n o s i d e I X , 1 0 ) ,
27-0－β ーD-glucopyranosylpubesenolide 3・0－β －D-glucopyranoside( w i t h a n o s i d e X , 1 1 ) , a n d
(20R, 22R）」 α ， 3β ， 20,2 7 ‑ t e t r a h y d r o x y w i t h a ‑ 5 ,
24-dienolide 子0－β ーD-glucopyranoside( w i t h a n o s i d e X I , 1 2 ) . Of t h e i s o l a t e d c o m p o u n d s , 1 , w i t h a n o l i d e A ( 2 ) , ( 2 0 S ,
22R)-4β ， 5β ， 6α ， 27-tetrahydroxy ‑l ‑ o x o w i t h a ‑ 2 , 2 4 ‑ d i e n o l i d e ( 6 ) , w i t h a n o s i d e IV ( 1 4 ) , w i t h a n o s i d e VI ( 1 5 ) a n d c o a g u 1 i n Q ( 1 6 ) showed s i g n i f i c a n t n e u r i t e o u t ｭ g r o w t h a c t i v i t y a t a c o n c e n t r a t i o n o f 1
ｵ Mon a human n e u r o b l a s t o m a SH‑SY5Y c e l l l i n e .
1 1 ) Min B . , Miyashiro H . , and Hattori M.: I n h i b i t o r y e f f e c t s : of quinones on RNase H a c t i v i t y
as・s o c i a t e d with
HIV・ 1r e v e r s e t r a n s c r i p t a s e . P h y t o t h e r .
Rt~s.,1 6 , S57‑62 ( 2 0 0 2 ) .
I n a n e f f o r t t o d e v e l o p new d r u g s p r e v e n t i n g t h e g r o w t h o f 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 ) , we d e v e l o p e d a n
in νitro
a s s a y method o f r i b o n u c l e a s e H (RHase H) a c t i v i t y a s s o c i a t e d w i t h r e v e r s e t r a n s c r i p t a s e ( R T ) from H I V ‑ 1 . Some n a p h t h o q u i n o n e s , s u c h a s 1 , 4 ‑ n a p h t h o q u i n o n e ( 1 ) , v i t a m i n K3 ( 2 ) , j u g l o n e ( 3 ) a n d p l u m b a g i n ( 6 ) , m o d e r a t e l y i n h i b i t e d RNase H a c t i v i t y , a n d o t h e r s , i n c l u d i n g n a p h t h a z a r i n ( 5 ) a n d s h i k o n i n s ( 8 ‑ 9 , 1 8 ‑ 2 3 ) , showed weak i n h i b i ｭ t i o n .
Dite中enoidq u i n o n e s , t a n s h i n o n e s ( 2 4 ‑ 2 8 ) , h a d a l s o m o d e r a t e i n h i l b i t i o n a g a i n s t RNase H a c t i v i t y . Of t h e s e q u i n o n e s , compound 1 showed t h e m o s t p o t e n t i n h i b i t i o n on RNase H a c t i v i t y 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 ( I C s o ) o f 9 . 5
ｵM,t o g e t h e r w i t h m o d e r a t e i n h i b i t i o n a g a i n s t RNA‑dependent a n d DNA‑dependent DNA p o l y m e r a s e (RDDP a n d DDDP) a c t i v i t i e s w i t h I C s o v a l u e s o f 69 a n d 36
ｵM,r e s p e c t i v e l y . Compounds 3 a n d 5 showed s i g n i f i ｭ c a n t i n h i b i t i o n a g a i n s t RDDP ( I C s o
=8 a n d 1 0
ｵM,r e s p e c t i v e l y ) a n d DDDP ( I C s o
=5 a n d 7
ｵM,r e s p e c t i v e l y ) a c ｭ t i v i t i e s . The 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 o f t h e n a p h t h o q u i n o n e s s u g g e s t e d t h a t n o n ‑ h y d r o x y l a t e d n a p h t h o q u i n o n e s ( 1 a n d 2 ) showed s i g n i f i c a n t i n h i b i t i o n o f RNase H a c t i v i t y , w h e r e a s 5 ‑ h y d r o x y l a t e d n a p h t h o q u i n o n e s ( 3 a n d 5 ) showed p o t e n t i n h i b i t i o n a g a i n s t RDDP a n d DDDP a c t i v i t i e s .
1 2 ) M a C . , Nakamura N . , Miyashiro H . , Hattori M., Komatsu K . , Kawahata T . , and Otake T . : Screening of Chinese and Mongolian herbal dmgs f o r anti‑human immunodeficiency v i r u s type 1
(HIV・1)a c t i v i t y . P h y t o t h e r . R e s . , 1 6 ,
-186・ 189( 2 0 0 2 ) .
Water a n d m e t h a n o l e x t r a c t s o f 30 C h i n e s e a n d M o n g o l i a n m e d i c i n a l p l a n t s w e r e t e s t e d f o r t h e i r 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 ‑ I ( H I V ‑ I ) i n h i b i t o r y a c t i v i t y . Of t h e 60 e x t r a c t s , 2 3 showed a n t i ‑ H I V a c t i v i t y . B i o a s s a y ‑ g u i d e d f r a c t i o n a t i o n o f o n e o f t h e most a c t i v e e x t r a c t , t h e m e t h a n o l e x t r a c t o f t h e r o o t t u b e r o f S t e p h a n i a c e p h a r a n t h a , h a s l e d t o t h e i s o l a t i o n o f two a l k a l o i d s , a r o m o l i n e a n d FK‑3000 a s p o t e n t i n h i b i t o r y s u b s t a n c e s . They c o m p l e t e l y i n h i b i t e d t h e c y t o p a t h i c e f f e c t s o f HIV‑1 on MT‑4 c e l l s a t 3 l . 3 a n d 7 . 8
ｵg/mL,r e s p e c t i v e l y .
1 3 ) Ohsaki M., Kurokawa M., Nawawi A . , Nakamura N . , Hattori M., and Shiraki K . : Characterization of a n t i ‑ h e r p e s simplex v i r u s type 1 adivity of an a l k a l o i d FK 3000 from S t e p h a n i a c e p h a r a n t h a . J . T r a d . M e d . , 1 9 ,
129・ 136( 2 0 0 2 ) .
A
mo中hinanea l k a l o i d FK 3000 ( 6 , 7 ‑ d i ‑ 0 ‑ a c e t y l s i n o c o c u l i n e ) f r o m t h e r o o t t u b e r s o f S t e p h a n i a c e p h a r a n t h a
showed a n t i v i r a l a c t i v i t y a g a i n s t a c y c l o v i r ‑a n d p h o s p h o n o a c e t i c a c i d ( P A A ) ‑ r e s i s t a n t h e r p e s s i m p l e x v i r u s t y p e 1
( H S V ‑ 1 ) , i n f l u e n z a v i r u s , m e a s l e s v i r u s , a n d p o l i o v i r u s . The a n t i ‑ H S V a c t i o n o f FK 3000 was a s s e s s e d i n c o m p a r i ｭ
s o n w i t h t h a t o f PAA t h a t i n h i b i t s t h e a c t i v i t y o f HSV DNA p o l y m e r a s e a n d HSV DNA s y n t h e s i s . FK 3000 i n h i b i t e d
t h e g r o w t h o f t h y m i d i n e k i n a s e ‑ d e f i c i e n t a n d ACV a n d P A A ‑ r e s i s t a n t HSV‑1 s t r a i n s , a s w e l l a s w i l d t y p e HSV s t r a i n s i n Vero c e l l s . T h i s compound, a s w e l l a s PAA, i n t e r f e r e d w i t h t h e s y n t h e s i s o f l a t e v i r a l p r o t e i n s b u t n o t e a r l y v i r a l p r o t e i n s . The a n a l y s i s o f HSV DNA s y n t h e s i s by s l o t b l o t h y b r i d i z a t i o n showed t h a t FK 3000 i n h i b i t e d t h e v i r a l DNA s y n t h e s i s i n a d o s e ‑ d e p e n d e n t m a n n e r . However, t h e v i r a l RNA was p a r t i a l l y s y n t h e s i z e d i n t h e p r e s e n c e o f FK 3000 ( e v e n a t a d o s e t h a t HSV DNA s y n t h e s i s was i n h i b i t e d ) and PAA, i n d i c a t i n g t h a t FK 3 0 0 0 , a s w e l l a s PAA, a l l o w e d e a r l y v i r a l RNA s y n t h e s i s b u t n o t v i r a l DNA s y n t h e s i s . S i n c e p a r t i a l l y p u r i f i e d HSV DNA p o l y m e r a s e a c t i v i t y was n o t i n h i b i t e d by FK 3 0 0 0 , t h i s compound was s u g g e s t e d t o i n h i b i t HSV DNA s y n t h e s i s by a mechanism d i f f e r e n t from t h a t o f PAA.
1 4 ) Matsuse I . T . , Nakamura N . , Basnet P . , Hattori M., Kamimura K . , and Funada H . : Amino a c i d s and phosphates s t i m u l a t e hatching of O c h l e r o t a t u s k o r e i c o i d e s ( D i p t e r a : C u l i c i d a e ) e g g s . Med. Entomol. Z o o l . , 5 3 , S47‑54 ( 2 0 0 2 ) .
A t r e e ‑ h o l e m o s q u i t o s p e c i e s , O c h l e r o t a t u s ( F i n l a y a ) k o r e i c o i d e s h a s b e e n r e a r e d i n l a b o r a t o r y by s t i m u l a t i n g t h e i r h a t c h u s i n g d r i e d y e a s t p o w d e r . T h i s method shows t o be more e f f e c t i v e t h a n l o w e r i n g t h e d i s s o l v e d oxygen c o n c e n ｭ t r a t i o n by m e c h a n i c a l , c h e m i c a l o r b i o l o g i c a l m e a n s . From a h a t c h t e s t g u i d e d f r a c t i o n a t i o n o f t h e w a t e r e x t r a c t o f d r i e d y e a s t , g l u t a m i c a c i d was i s o l a t e d a s t h e main compound o f t h e h a t c h s t i m u l a t i n g f r a c t i o n . T h i s amino a c i d a l o n e d i d n o t show h a t c h s t i m u l a t i n g e f f e c t b u t i t was e f f e c t i v e i n c o m b i n a t i o n w i t h t h e p h o s p h a t e s t h a t a r e a l s o c o n ｭ s t i t u e n t s o f d r i e d y e a s t . Among 20 amino a c i d s , o n l y h i s t i d i n e a n d p r o l i n e showed s i m i l a r e f f e c t s a s g l u t a m i c a c i d . A c o n c e n t r a t i o n o f 0 . 1 mg/ml o f e a c h g l u t a m i c a c i d and t r i s o d i u m p h o s p h a t e g a v e h i g h p e r c e n t a g e h a t c h i n 1 8 h .
1 5 ) Zhang C . , Nakamura N . , Tewtrakul S . , Hattori M., Sun Q . , Wang Z . , and Fujiwara T . : Sesquiterpenes and a l k a l o i d s from Lindera c h u n i i and t h e i r 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
HIV・1 i n t e g r a s e . Chem. Pharm. B u l l . , 5 0 , 1195‑1200 ( 2 0 0 2 ) .
T h r e e new eudesmane t y p e s e s q u i t e r p e n o i d l i n d e n a n o l i d e s E ( 1 ) , F ( 2 ) a n d G ( 3 ) , a n d two new a p o r p h i n e a l k a l o i d l i n d e c h u n i n e s A ( 1 8 ) and
B( 2 0 ) were i s o l a t e d from r o o t s o f L i n d e r a c h u n i i
MERR.,t o g e t h e r w i t h s e v e n known
H
亨. 0
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7 R = COCH3
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11 R1 = OCH3 R2 = R3 = H R4 = OH R5 =CH3
12 R1 = R2 =R3 = H R4 = OH R5 = CH3
15 R1 = R3 = H R2 = R5 = CH3 R4 = OH
20 R1 = R2 = H R3 + R4 = OCH20 R5 = H
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19 R1=R2=R5=CH3 R3=R4=H
Chart 4 .
O
sesqui臼rpenes
i n c l u d i n g a new
naturally-occuπingl i n d e n a n o l i d e H ( 4 ) a n d e i g t h known a p o r p h i n e a l k a l o i d s . The s t r u c t u r e s o f t h e s e compounds w e r e 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 . Of t . h e i s o l a t e d c o m p o u n d s , h e r n a n d o n i n e ( 1 4 ) , l a u r o l i s t i n e ( 1 6 ) , 7 ‑ o x o h e r n a n g e r i n e ( 1 7 ) a n d l i n e d e c h u n i n e A ( 1 8 ) showed s i g n i f i c a n t a n t i ‑ h u m a n immunodeｭ f i c i e n c y v i r u s t y p e 1 ( H I V ‑ I ) i n t e g r a s e a c t i v i t y w i t h I C s o v a l u e s o f 1 6 . 3 , 7 . 7 , 1 8 . 2 a n d 2 1 . 1
ｵM,r e s p e c t i v e l y . The m a j o r a l k a l o i d s p r e s e n t e d i n t h e r o o t s o f
L.c h u n i i w e r e q u a n t i t a t i v e l y a n a l y z e d by a n HPLC m e t h o d .
1 6 ) Akao T . , Yoshino T . , Kobashi K . , and
Hattor・iM.: Evaluation of s a l i c i n as an a n t i p y r e t i c prodrug t h a t does not cause g a s t r i c i n j u r y . P l a n t a
Med” 68,714・718( 2 0 0 2 ) .
P h a r m a c o k i n e t i c a n d
ph訂macologicals t u d i e s w e r e
perfo口nedt o compare t h e a n t i p y r e t i c e f f e c t s o f s a l i c i n ( S L ) , s a l i g e n i n ( S G , a n a g l y c o n e o f S L ) a n d s a l i c y l i c a c i d ( S A , a n a c t i v e m e t a b o l i t e o f SL) i n r a t s . When SL was a d m i n ｭ i s t e r e d o r a l l y t o r a t s , SA a p p e a r e d s l o w l y i n t h e p l a s m a a n d l e v e l s i n c r e a s e d g r a d u a l l y , i n c o n t r a s t t o t h e r a p i d a p ｭ p e a r a n c e o b s e r v e d 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 sodium s a l i c y l a t e (SANa) o r SG. O r a l l y a d m i n i s t e r e d SL d i d n o t a f f e c t t h e r e c t a l t e m p e r a t u r e s o f a f e b r i l e r a t s a t a d o s e o f 5 m m o l / k g ; a t t h i s d o s e , SANa a n d SG l o w e r e d body t e m ｭ p e r a t u r e s i g n i f i c a n t l y . H o w e v e r , i t s i g n i f i c a n t l y r e d u c e d
yeast拘inducedf e v e r , p r o d u c i n g a n o r m a l body t e m p e r a t u r e , a n d c o m p l e t e l y p r e v e n t e d f e v e r when a d m i n i s t e r e d s i m u l t a n e o u s l y w i t h y e a s t . SL d i d n o t i n d u c e g a s t r i c l e s i o n s e v e n a t a d o s e o f 5
mmoνkg;c o n v e r s e l y , SANa a n d SG i n d u c e d s e v e r e g a s t r i c l e s i o n s i n a d o s e ‑ d e p e n d e n t manner a t 1 , 2 . 5 a n d 5 m m o l / k g . P o o r
abso中tiono f SL a n d r a p i d a b s o r p t i o n o f SA a n d SG w e r e c o n f i r m e d i n a n i n v i v o s y s t e m , a s w e l l a s i n a n i n v i t r o s y s t e m u s i n g e v e r t e d r a t j e j u n a l s a c s . Only smaH a m o u n t s o f SA a n d SG w e r e d e t e c t e d i n t h e i n t e s t i n a l t r a c t s o f r a t s 1 h a f t e r o r a l a d m i n i s t r a t i o n , w h e r e a s more t h a n 50% o f a n SL d o s e was r e c o v e r e d a n SL a n d SG f r o m t h e i n t e s t i n a l t r a c t s 1 h a f t e r t r e a t m e n t a n d 15.8% o f t h e
do将 wass t i l l p r e s e n t a s SG 4 h a f t e r
admini句s t r a t i o n . When g i v e n t o g e r m ‑ f r e e r a t s , 19.8% o f t h e SL d o s e was r e c o v e r e d i n t a c t , m a i n l y f r o m t h e c e c u m , a n d no SG was d e t e c t e d e v e n a t 4 h a f t e r t r e a t m e n 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 SL i s a p r o d r u g which i s g r a d u a l l y t r a n s ｭ p o r t e d t o t h e l o w e r p a r t o f t h e i n t e s t i n e , h y d r o l y z e d t o SG by i n t e s t i n a l b a c t e r i a , a n d c o n v e r t e d t o SA a f t e r a b s o r p ｭ t i o n . I t t h u s p r o d u c e s a n a n t i p y r e t i c a c t i o n w i t h o u t c a u s i n g g a s t r i c i n j u r y .
1 7 ) Park J . , Hur J . , Park J . , Hatano T . , Yoshida T . , Miyashiro H . , Min B . , and Hattori M.:
I n h i b i t o r y e f f e c t s of Korean medicinal p l a n t s and camelliatannin H from C a m e l l i a j a p o n i c a on human immunodeficiency v i r u s type 1 p r o t e a s e . P h y t o t h e r . R e s . , 1 6 , 422‑426 ( 2 0 0 2 ) . To i d e n t i f y s u b s t a n c e s w i t h a n t i ‑ h u m a n i m m u n o d e f i c i e n c y v i r u s ( H I V ) a c t i v i t y i n t r a d i t i o n a l m e d i c i n e s , 1 0 1 e x t r a c t s o f K o r e a n m e d i c i n a l p l a n t s w e r e s c r e e n 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 HIV t y p e 1 p r o t e a s e ( P R ) . The enzyme a c ｭ t i v i t y was d e t e r m i n e d by HPLC. Of t h e e x t r a c t s t e s t e d , s t r o n g i n h i b i t o r y e f f e c t s w e r e o b s e r v e d i n t h e a c e t o n e e x ｭ t r a c t s o f t h e p e r i c a r p a n d l e a v e s o f C a m e l l i a j a p o n i c a , t h e w a t e r e x t r a c t o f t h e l e a v e s o f S a g e r e t i a t h e e z a n s a n d t h e m e t h a n o l e x t r a c t o f t h e a e r i a l p a r t o f S o p h o r a j l a v e s c e n s . C a m e l l i a t a n n i n H from t h e
perica叩 ofC . j a p o n i c a , showed a p o t e n t i n h i b i t o r y a c t i v i t y on HIV‑1 PR w i t h a n I C s o o f 0 . 9
ｵM.1 8 ) Min B . , Lee H . , Lee S . , Kim Y . , Bae K . , Otake T . , Nakamura N . , and Hattori M.: Anti‑human immunodeficiency v i r u s
・type1 a c t i v i t y of c o n s t i t u e n t s from J u g l a n s m a n d s h u r i c a . A r c h . Pharm. R e s . , 2 5 ,
441・445( 2 0 0 2 ) .
T h r e e n a p h t h a l e n e g l y c o s i d e s ( 1 ‑ 3 ) , f o u r f l a v o n o i d s ( 4 ‑7 ) , a n d two g a l l o y l g l y c o s i d e s ( 8 ‑ 9 ) were i s o l a t e d from t h e
s t e m ‑ b a r k o f J u g l a n s m a n d s h u r i c a ( J u g l a n d a c e a e ) . T h e i r s t r u c t u r e s w e r e d e t e r m i n e d by c h e m i c a l a n d s p e c t r a l
m e a n s , i n c l u d i n g w i t h 2D‑NMR (COSY, HMQC, a n d HMBC) e x p e r i m e n t s . CAmongst t h e i s o l a t e d c o m p o u n d s ,
t a x i f o l i n ( 4 ) showed t h e most p o t e n t H I V ‑ i n d u c e d c y t o p a t h i c a c t i v i t y a g a : i n s t MT‑4 c e l l s w i t h a c o m p l e t e i n h i b i t o r y
c o n c e n t r a t i o n ( I C 1 0 0 ) v a l u e o f 2 5 ｵg/ml a n d a maximum c y t o t o x i c c o n c e n t r a t i o n ( C C 1 0 0 ) v a l u e o f a b o v e 1 0 0 ｵ g l m l .
H o w e v e r , n a p h t h a l e n e g l y c o s i d e s ( 1 ‑ 3 ) , f l a v o n o i d s
(5ヴ），a n d g a l l o y l t a n n i n s ( 8 ‑ 9 ) w e r e i n a c t i v e a g a i n s t a n t i ‑ H I V ‑ I
a c t i v i t y .
1 9 ) Akanitapichat P . , Kurokawa M., Tewtrakul S . , Pramyothin P . , Sripanidkulchal B . , Shiraki K . , and Hattori M.: I n h i b i t o r y a c t i v i t i e s of Thai medicinal p l a n t s a g a i n s t herpes simplex type 1 , p o l i o v i r u s type 1 , and measles v i r u s . J . T r a d . M e d . , 1 9 ,
174・ 180( 2 0 0 2 ) .
F o r t y ‑ e i g h t e t h a n o l ‑a n d 43 w a t e r ‑ e x t r a c t s o f 49 t r a d i t i o n a l T h a i m e d i c i n e s w e r e e v a l u a t e d f o r a n t i v i r a l a c t i v i t i e s by a p l a q u e r e d u c t i o n a s s a y . F o r p r e l i m i n a r y c h a r a c t e r i z a t i o n o f t h e mode o f t h e i r a n t i v i r a l a c t i o n , p o l i o v i r u s t y p e 1 , m e a s l e s v i r u s a n d h e r p e s s i m p l e x v i r u s t y p e 1 (HSV‑1) t h a t a r e d i f f e r e n t i n n u c l e i c a c i d component a n d e n v e l o p e d s t r u c t u r e were u s e d i n t h i s s t u d y . F i f t y ‑ t w o , 28 a n d 29 e x t r a c t s e x h i b i t e d 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 p o l i o v i r u s , meaｭ s l e s v i r u s a n d HSV‑1, r e s p e c t i v e l y . Of 29 e x t r a c t s w i t h a n t i ‑ H S V ‑ 1 a c t i v i t i e s , t h e i n h i b i t o r y a c t i v i t i e s o f R h i n a c a n t h u s n a s u t u s
(le幼， Terminaliac i t r i n a ( f r u i t ) a n d T h e v e t i a p e r u v i a n a ( l e a f ) were o b s e r v e d i n b o t h e t h a n o l a n d w a t e r e x t r a c t s . The e t h a n o l e x t r a c t s o f D e r r i s s c a n d e n s ( l e a f ) a n d Plumbago i n d i c a ( l e a f ) a n d t h e w a t e r e x t r a c t o f Capsicum f r u t e s c e n s
（仕uit)w e r e a c t i v e a g a i n s t o n l y HSV‑1, s u g g e s t i n g t h e mechanism o f t h e i r a n t i v i r a l a c t i o n l i k e l y u n i q u e t o HSV‑1 b u t n e i t h e r p o l i o v i r u s n o r m e a s l e s v i r u s . C o n t r a r i l y , 26 e x t r a c t s d i s p l a y e d i n h i b i t o r y a c t i v i ｭ t i e s a g a i n s t p o l i o v i r u s a n d / o r m e a s l e s v i r u s . T h e s e f i n d i n g s s u g g e s t t h a t t h e 29 e x t r a c t s from t r a d i t i o n a l T h a i m e d i ｭ
cines 訂e
