Chemistry and Pharmacology of Diterpenoids of Siegesbeckia pubescens

Journal of the Pharmaceutical Society of Korea 19. 129-165 C1975)

Chemistry and Pharmacology of Diterpenoids of

Siegesheckia pubescens*

K o o D o n g H a n , Ja e H o o n K i m a n d S e a J o n g O h

N a t u r a l P r o d u c t s R e s e a r c h I n s t i t u t e , S e o u l N a t i o n a l U n i v e r s i t y , S e o u l 1 1 0

**Hi-cheom", Korean name of S i e g e s h e c k i a p u b e s c e n s Ma k in g, w hich is grow n a b u n d a n tly not only in Korea, but also in other A sian countries, has long been used quite m uch i n Korea from olden times, as folk medicine for the treatment of hypertension and in fla m m a tio n . Its medicinal values were described in the famous **Pen-Tsao-Kang-mu*" and in the other several description도그 indicating us that **Hi-chem** possesses antihypertensive and a n tip h lo ­ gistic effect.

In recent, there appeared several reports on the ingredients of S ie g e s h e c k i a species. T h e first one was reported in 1931 by Wehmer^^ who separated a bitter principle from S i e g e s - b e c k i a o r ie n t a lis e nam ing it darutin, but it^ chemical structure was not know n. In 1960, A n n ie D iara^그 separated a glucoside w ith pim arane skeleton from S i e g e s h e c k i a o r i e n t a l i s , n a m ed it darutoside, and elucidated its chemical structure.

In our laboratory, extraction of the plant, *"Hi-cheom**, w ith methanol and then extensive fractionation has yielded five diterpenes (compound A , B, C, D and F ) w ich are show n in the Table L

As shown in Table I, among these five kinds of diterpenoids, two components ( A a n d C ) were diterpene (A ) or diterpene glucoside (C ) w ith pimarane skeleton and other three components (B, D and F ) w ith kaurane skeleton.

W e found that compound B has remarkable antihypertensive activity by the a nim al test.

In addition to this activity, we also found that the diterpene B has remarkable a n tiin fla m ­ matory action by the prelim inary album in coagulation test and rat-paw edema test. T w o other diterpene D and F were proved also to have antiinflam m atory activity.

In contrast w ith these results, diterpenes w ith pimarane skeleton ( A and C ) were proved not to have such activities as antiinflam m ation and antihypertension.

These diterpenes w ill be described here.

Compound A걸

황

* Presented on Oct. 5, 1974 at the symposium on **Triterpenoids** organized by Natural Products Research Institute, Seoul National University.

129

130 H an • K im • O h ： Diterpenoids of Siegesbeckia Vol. 19, No. 3

Table I

Com pound F o rm ula M p W d

H O H ^ C

C20H34O4

C O O H

t2bH3E04

192^3 -22

260^^2

- 8 9

'• C O O H

C 2 6 H 4 4 O 0 - H 2 O

225

6 - 4 8

C 2oH 3 2 D 3

2 0 7 .5 •105.8

''C O O H Ca)H3o04

2 5 8

6 0 >133,25

separated butyl alcohol fraction and on standing over night the butanol solution deposited a crystalline solid, w hich was filtered off. Recrystallization of the crude crystals from boiling water gave needles of compound A ( I ) , C20H34O4, mp 192시 193° (ethyl alcohol), M d =

—22° (c, 1.0 in dioxane), IR 도; 3325 (O H ), 1654 (double bond), N M R (C₅D₅N ) d\ 1. 26 (3H, s, CHs), 1.14 (3H, s, C H3), 0.80 (3H, s, C H3), 5.15 ( I H , bs, - C - C H = C

- C H2- ) ; M S (m /e) 338 (M+), 320 (M+ - H2O ), 289, 277, 259, 241.

Acethylation w ith acetic anhydride in pyridine at room temperature afforded a tetraacetate ( I I ) , C₂₈H₄₂O₈, IR 호; 1725, 1660, 1250, while treatment w ith acetone and anhydrous C11S O4 at room temperature yielded a monoacetonide ( I I I ) , C23H38O4, m p 164-^165°,

—44° (c, 1. 0 in C H C I3)*.

* O ptical rotations were measured in chloroform solution unless otherwise stated.

September, 1975 J. Pharm . Soc. Korea 131

O x idation of I w ith H IO₄ in methanol gave formaldehyde and the aldehyde ( I V ) , C₁₉H₃₀O₃, m p 127^-129°, I R 3400 (O H ), 2700 (C H O ), 1718 (C H O ), 166D (double bond), N M R (C D C I3) d; 1,04 s, 0.70 s, (C H3), 3.69 d, 3.34 d ( J = l l c/s, Cis—

H2), 5.22 bs ( C u - H ) , 9.0 (C H O ),

Spectral features of compound A and its derivatives I I , I I I and IV , together w ith isolation of pimanthrene (V ) by selenium dehydrogenation, suggest for it partial structure.

Partial structure of compound A.

In accord w ith this, oxidation of monoacetonide ( I I I ) w ith Sarett reagent gave a ketoalde- hyde acetonide ( V I), C23H34O4, m p 116시 117% C«Od=—46°, IR iVaxCm■고; 1720 ( C = 0 ) , 165S (double bond). Its N M R spectrum (Table I I ) shows, among other, two lines at 3.06 d a n d at 2. 83 d ( / = 1 3 . 7 c/s, IH , part A of an A B system) w hich can be attributed to one proton of a methylenic group both allylic and adjacent to a carbonyl function. T h is fact locates the secondary hydroxyl function at Ce-

By treatment w ith ^-toluenesulfonyl chloride in pyridine, the monoacetonide ( I I I ) is con­

verted into a mixture of a monotoluene-/>-sulfonate ( V I I ) , C30H44O6S, a ditoluene-/>-sulfonate ( V I I I ) , C37H50OSS2, and a cyclic ether ( I X ) , C23H36O3, w hich was resolved by chromato­

graphy on silica gel. In the N M R spectrum of I X the tertiary oxymethylene signals appear as an A B system centered at 3.90 d and 3.23 8 C J = 8 c/s), this coupling constant idicates the presence of a five membered heterocyclic ring®\ thus suggesting the tertiary hydroxym ethyl group in compound A is at C₄ or at Ciq.

Since the chemical shifts of the methyl groups of compound A and its derivatives (Table I I ) are in accordance w ith those of diterpenoids w ith a pimarane skeleton, assuming a chair conformation for ring B in order to explain the shielding effect®그 on C20, the tertiary hydro-

Table II—Chemical shifts of the derivatives of compound A (5 values in CDCI3}.

C om pound Ci3 Oio C4 C(18)-H2 C(14)-H

I I 0.95 s 0.91s 1.04 s 4.10 d 3 .9 4 d ( 7 = 1 0 .8 ：> 5.15 bs

I I I 0 ,92 s 0 .78 s 1.04 s 3.73 d 3 ,3 4 d C /= 1 2 ) 5. 08 bs

I V 1.04 s 0.70 s 1.07 s 3.69 d 3.34 (!(]/= 11) 5. 22 bs

V I 0.93 s 0 . 68 s 1.24 s -CHO 9 .7 s 5.22 bs

I X 0 .95 s 0 .8 8 s 1 .0 0 s 3 .9 0 d 3 .^3 dCJ=S') ^{5. 04 bs}

X I 1.16 s 0 .51s 1 .3 1 s — — 5.45 bs

X I I I 0 .93 s 1.0 1s 1.04 s ? 3 . 3 4 d C / = l l ) 5.12 bs

132 H an • K im • O h : Diterpenoids of Siegesheckia Vol. 19, No. 3 xymethyl group should be at C4.

The assignment of configuration at' C4 rests upon the chemical shift and m ultiplicity of the aldehydic proton of V I (singlet at 9. 7 d X

In a study*^^ of the N M R spectra of diterpenoids containing an aldehyde at C4 and an axial proton C3 it has been shown that equatorial aldehydic protons appear at 9.12-^9. 3 5 as singlets w hile axial aldehydic protons at 9 .7 ^ 9 . 9 5 as doublets (/<C3 c/s) (Scheme 1). The value of the chemical shift of the aldehydic proton in V I therefore can be explained through a down- field shift due to the carbonyl ic function at Ce- Since it resonates at 10. 1^ 10. 4 J 유^® 그 in d i­

terpenoids containing an axial aldehydic group at C4 and a keto group at Cs, the aldehydic function in V I can be assumed, equatorial.

E Q u o t o r lo l P P m ( C C | j )

O H C

9.13

O H C

9.19..

O H C

9.12 s.

O H C

9 2 3 s.

O H C

9 .2 9

ppm (CCjj!

#

CHO

O H C 10.4s.

" C H O

9.93 c.

vl=LO

Scheme 1— D ifferent between chemical shifts of axial aldehyde and that of equatorial aldehyde.

#

# September, 1975 J. Pharm. Soc. Korea 133 Furthermore, aldehyde (IV ) w ith Jones reagent in acetone gave a ketodiacid ( X ) , C19H26 O₅, which yields the corresponding dimethylester ( X I ) , C₂₁H₃₀O₅, by treatment w ith diazo­

methane. Saponification of X I w ith aqueous 2. 5N N a O H in ethylene glycolmonomethylether in ₃ hr at 150"\ standard conditions for C₄ equatorial 'carbomethoxyl groups, afforded an isomeric ketodiacid ( X I I ) which exhibits an U V absorption m axim um at 240nm (Scheme2).

H0H2C^ '

C H O

J O N E S R E A G E N T

OH'

f ^f-COOMe 사

MeOOC

Scheme 2—Standard condition for C4—COOMe(eq/

The configuraion at Ce is provided by obtaining a Ce epimeric monoacetonide ( X I I I ) ,

^ 23H3s0 4, on NaBHU reduction of V I in aqueous tetrahydrofuran. T h is monoacetonide ( X I I I ) is reconverted in V I by Sarett oxidation. The conclusion is, therefore, that the Ce hydroxyl group in X I I I must have the axial configuration피그 since the complex hydride reduction of V I is sterically controlled by the 1, 3-axial substituents in C4 and in Cio.

The stereoconfiguration at C13 was established to be ^-orientation (same as in darutigenol 호1그) by comparing the difference ( ^ R M —286, Table I I I ) of molecular rotation MCD] between the hydroxy aldehyde (IV ) and the nor-alcohol ( X I V ) both derived from compound A w ith

Table I I I —Molar rotatory power of compound A and pimaric acid.

Compound Derivatives MCD]

'tm ' ■ ■

Compd. A Hydroxy-aldehyde (IV ) -361 -286

Nor-alcohol (X IV ) -75 —

Pimaric acid Aldehyde acid (X V I) +359 +307

Nor-acid (X V II) +52 —

-- ---

134 H an • K im • O h : Diterpenoids of Siegesheckia Vol. 19, No. 3；

# that ( J R M +307, Table I I I ) between the aldehyde acid (V I) and the nor-acid ( X V I I ) both, derived from pimaric acid of which the configuration at C13 was already know n as the a- orientation (Scheme 3).

'COOH M (D J » +3 5 9*

X V I

H O H ^ C

X I V

Scheme 3—Syntheses of XVI, XV II, IV and XIV.

#

The shielding effect on the tertiary methyl group at Cio in the derivative of compound A having a carbonyl function at C13 suggests a cis relation between these two centers.

Therefore it is suggested that the compound A has its methyl group at Cio in the a - orientation.

Derivatives of compound A are listed in Scheme 4.

Com pound C고M3그一Dried plant was extracte4 w ith cold methanol. The extract was repeat­

edly extracted w ith E t2 0 and then butanol. The butanol extract was dissolved in water and the solution saturated w ith N a O H was added to it. O n standing over night, a considerable quantity of crystalline solid was precipitated.

Crystallization of the crude crystals from hot water and alumina column chromatography of the crystals gave needles of the compound C ( I ) , C26H440s*H2 0, m p 22 5^6°, C«Od=*—48° ( c = l . 0, M e O H ), I R 3330 (O H ), 1650 (double bond), N M R (C D3O D ) 8; 5.2 ( I H , bs, - C - C H = C - C H2- ) , 0.83 (3H, s, — C - C H3), 1.05 (3H , s ,- C H3).

Acethylation of I w ith acetic anhydride and pyridine at room temperature afforded a hexa- acetate ( I I ) CasHseO^, m p 128°, M d = - 4 3 . 2° (c, 1.0, M e O H ), I R iWxCirTi; 1735, 1240 (ester), N M R (C D3O D ) 8; 5. 3 ( I H , bs. — C - C U = C - C U 2- ) y 2. 03시2. 07 ( - 0 - C 0 - CH s), 0.83 (3H, s, - C H s ), 1.03 (3H, s, - C H3).

H ydrolysis of I w ith beta-glucosidase in aqueous solution at 36° for 60 hrs gave 1 mole of glucose (glucoosazone, m p 208°) and an aglucone ( I I I ) , C₂₀H₃₄O₃, m p 163°, C^^Dd— ~10**

September, 1975 J. Pharm. Soc. Korea 135

M eOOC

#

^cOH^

Scheme

4

책Cc, 1.0, [M eO H ), I R Vmaxcm■효; 3450 (O H ), 1650 (double bond), N M R (C D3O D ) 3; 5.18 ( I H , i s , C - C H：그C - C H2- ) , 0.83 (3H, s, C - C H g ).

Oxidation of I I I w ith sodium melaperiodate in ethylalcohol solution gave formaldehyde together w ith nor-aldehyde (IV ), C19H30O2, m p 114"니5°, M d = —77.1® (c, 1.0, E tO H ), I R v„axcm-i； 3360, 2720 (C H O ), 1730 (C H O ), N M R (C D₃O D ) d; 5.2 ( I H , bs, C - C H = C

—C H2—), 0. 88 (3H , s, — C H3), 1. 0 (3H , s, — C H3), 9.25 ( I H , s, —C H O )

Reduction of the nor-aldehyde (IV ) by Huang-M inlon method gave a mono-nor-alcohol X₆육hydroxy-nor-darutene) (V ), C19H₃₂O ; m p 137. 5 ° , I R iVaxCin■호; 3200 ( O H ) ,

H an • K im • O h ： Diterpenoids of Siegesqeckia ^{Vol. 19,}

#

Scheme

5

N M R (C D C I3) 크; 5.2 (IH , bs, C14- H ) , 0.8 (s, C H3), 0.92 (s, C H3), 1.0 (s, CHg).

O xidation of mono-nor-alcohol (V ) w ith Sarett reagent gave 6-keto-nor-darutene (V I)^

C19H30O ; m p 97—8% I R iw c m - i; 1695 ( C = 0 ) , N M R (C D C I3) 0.95 s, 0.97 s. 1.0 1.08 s, ( —C H3), 5.27 (bs, C u —H ), 2.89 d, 2.66 d ( / — 12. 75 c/s, IH , part A of an A B . system).

Reduction of the ketone (V I) by H uang-M inlon method gave a nor-darutene ( V I I) , C19H32, m p 42°, 28®, identical w ith authentic hydrocarbon (nor-darutene) ( V I I I ) , C19H32, m p 42®, C a]o=—28. 72°, derived from compound A (Scheme 5).

Spectral feature of compound ( I ) , its aglucone ( I I I ) and derivatives ( I I ) and (IV ), toge­

ther w ith isolation of pimanthrene by selenium dehydrogenation and synthesis of nor-darutenerr

C^HOH CHgOH

Partial structure of aglucone (III). ^#

September, 1975 J. Pharm . Soc. Korea 137

( V I I ) , suggest for aglucone ( I I I ) partial structure.

In the N M R spectrum two lines at 2.87 r and 2.66 z ( / = 1 2 . 75 c/s, IH , part A o f a n A B system) can be attributed to one proton of a methylenic group both allylic and adjacent to a carbonyl function, moreover the aglucone acetonide ( X ) was identical w ith authentic sample prepared from the compound A , X I in w hich the secondary hydroxyl function was established to be located at Ce (Scheme 6). These facts indicate that the secondary hydroxyl fu n c tio n is flt Cg*

T rithylation of I w ith triphenylchloromethane and pyridine at room temperature fo r 8 days affords ditrithylether ( I X ) , C64H72O8, m p M d = —91. 85°. Oxidation of I w ith so­

dium metaperiodate in ethylalcohol solution gave formaldehyde ( X I ) together w ith nor-aldehyde ( X I I I ) (Scheme 6).

These facts (form ation of formaldehyde and ditrithylether) indicate that the glucose m oiety is combined w ith the Ce hydroxyl group of the aglucone.

Com pound B효분그一Dried crushed plant collected near the Seoul, Korea, was extracted w ith cold ether. The extract was repeatedly washed w ith S % N aHCOa and extracted w ith N aO H . The solution was washed out w ith a small volume of ether and when acidified w ith 1 Q % H C l, a precipitate of a acid was formed.

-CHOH (iHgOTRY

TRtTTHYU

CHO

XIll

OGl 0-GI

#

o X

X

Scheme

6

H an • K im - O h ： Diterpenoids of Siegesbeckia Vol. 19, No. 3 Crystallization of the precipitate from methanol—— ethylacetate mixture gave a prism of comp­

ound B ( I ) C20H32O4, m p 260--2% M S (m /e ); 336 (M+), 318, 305, 287, 259, 241, M d = - 8 8°, I R 3470 (O H ), 1700 (C O O H ), N M R d; 8.7, 8.55 (C H3).

Pimanthrene ( I I ) and retene ( I I I ) was obtained by selenium dehydrogenation of the com­

p ound B methylester(IV), C21H34O4, m p 153^4°, IR iVaxCrrT호; 3620 (O H ), 3570 (O H ), 1730 (C O O C H3), N M R 8; 6 .36(s, C0 2M e), 6.3 (s, — C H2O H ), which was prepared by the m ethylation w ith diazomethane, was undepressed by admixture w ith an authentic sample (w h ic h was given by professor P .R . Jefferies),

Compound B methylester easily formed a monoacetate ( V I), C23H36O5, mp 121 시2"", N M R T； 5,27 (q, *7=12.4 c/s, —C H2—O A c) was obtained. Periodic acid oxidation of the methyl­

ester gave nor-ketoester ( V I I ) C20H30O3, m p 142사3°, IR 1노axcm-i; 17 4 0, 17 2 5 ( - C O - 0 - C H3), The I R spectral absorption of ketoester at 1740 cm~효 indicates the presence of a five membered ketone ring.

The triol ( V I I I ) , C20H34O3, m p 222시223° was obtained by reduction of methylester w ith lith iu m alum inium hydride.

Pinacolic rearrangement of the methylester gave the aldehyde which was identified as its 2, 4-dinitrophenylhydra2one ( I X ) , C27H56O6N4, mp 183사5° by comparison w ith an authentic sample (Scheme 7). Formation of the nor-ketoester and the results of oinacnlic rearrangement

Scheme 7~ D erivatives of com pound B.

September, 1975 J. Pharm. Soc. Korea 139

indicate that a glycolic hydroxy group exist in the side chain. W ith these results obtained by the above experiments and observations, compound B was identified as 16 ,17-dihyroxy-16^-

( —)-kauran-19-oic acid, already isolated by P .R . Jefferies호®그 from B e y e r i a species.

Compound D호®그—— The filtrate, obtained by filtration of precipitate of the com oound B as show n above was recovered w ith of H C l into ether. The ether extract was c h ro m ato ­ graphed on silica gel. E lution w ith C H C I3—-M eO H (9 ； 1) mixture gave needles of c o m p o u n d D (I), ^C20H32O3, m p 207.5", M d ^ - 1 0 5 . ₈°, I R ^maxcnTi; 3440 (O H ), 1700 ( C O O H ) .

#

#

N M R <5; 8.75 (s, C4- C H3), 9.05 (s, C10- C H3).

Acetate ( I I ) , 0 22^ 3404, m p 170°, I R iWxCm■호; 1745, 1695 (ester carbonyl), 1236 (acetoxy) and methylester acetate ( IV ), C23H36O4, m p 105°, I R VmaxCnT호; 1738 (ester), 1236 (acetoxy) were prepared by acethylation of compound D or methylester w ith acetic anhydride and anhydrous sodium acetate, respectively.

Methylester ( I I I ) , C21H34O3, m p 113. 5*^ was prenared by methylation of compound D w ith diazomethane. Methylester-2, 4-dinitrophenylhydrazone ( V I), m p 183사 4° was prepared by the usual way from aldehyde (V ) obtained by Sarett oxidation of the methyl ester ( I I I ) . M elting point of this phenylhydrazone was undepressed by admixture w ith an authentic sample prepared from the compound B w ith pinacolic rearrangement (Scheme 8). The chemical shifts (singlet) at ₈. 75 t and 9. 05 1: were observed, respectively.

According to N M R studies about kaurane derivatives by Tiers호구그, in —C O O H group is sub­

stituted at C4 axially, the chemical shift of C H3 proton at C4 and Cio shows 8.76 z and 9.06

^ 9 . 07 T, respectively as shown in the Table IV .

Based upon these N M R data observed and the results of the Tiers N M R studies on the k a ­ urane derivatives (Table IV ), it was established that the —C O O H groups is located at C4.

W ith these results obtained by the above experiments and observations, compound D wa&

identified as 17-hydroxy-16jS-( —) -kauran- 19-oic acid ( I ) .

Com pound F고®그一 The soluble fraction in of N a O H solution was acidified w ith 5% of H C l and resulting precipitate was filtered. C H C I3 was added into the filtrate and the resulting precipitate were chromatographed on silicagel. E lution w ith C H C I3— M eO H mixture (8 ； 1>

gave needles of compound F ( I ) , C20H30O4, m p 258시260®, M S (m /e), 334 (M ■느), M d = — 133.25°, I R V 2650, 1701, 1245 ( - C O O H ) , N M R d\ 8.77, 9.05.

Dimethylester ( I I ) , C22H34O4, m p 110°, M d = ^ —96. 8°, was obtained by usual way. 16우 ( —) -Kauran-17,19-diol ( I I I ) , C20H34O2, mp 179°, M d = —49 .1 ° , was prepared by reduction of the methylester ( I I ) w ith L iA lH4.

M elting point of the diol was undepressed by admixture w ith the authentic sample ( I I I ) prepared by reduction of the compound D methylester w ith U A IH4 (Scheme 9). A nd alsa the I R spectra of both are coincidental.

Table IV— C hem ical shifts o f m ethyl groups in kaurane derivatives ( r values in C D C I3).

A x ia l C4 g roup C4—~CH3 C io ~ C H3

140 H an • K im ■ O h : Diterpenoids of Siegesheckia Vol. 19, No. 3

CHs 9.15, 9.19 8.99— 9. 00

C H2O H 9 .0 4 ^ 9 . 05 9.01

C H2OAC 9.06 8 .9 6— 8. 98

C H O 9.01 9.14

C O O M e 8.83 9 .1 8 ^ 9 .1 9

C O O H 8 .76^8.77 ^{9. 06-신. 07}

C H2OTS 9 . 10--9.11 9 .1 4 ^ 9 .1 8

Sam ple ₈.75 9.05

# September, 1975 Pharm. Soc. Korea 141

^COOH

' C O O H

출

C H ^ O H

Ml, 111'

C H3O H

CH2N2

''C O O H

CH^Oh

S c h e m e 9—— D e r i v a t i v e s o f c o m p o u n d F ( I ) .

In the N M R spectra, the chemical shifts at 8. 77 8 and 9. 66 8 was observed, respectively. T h e chemical shifts indicate the location of —C O O H group at C4 as in the case of com pound D .

W ith these results, the compound F was identified as 16/3-(—)-kauran-17,19-diacid ( I ) . In order to examine the effect of the compound B on blood pressure of renal hypertensive rats, the follow ing method was used. In order to make artificial hypertension, we selected 12.

rats weighing 90^115g, and applied the figure of eight ligature method to the left k idney of each rat, and two weeks later, the other side kidney was ectomized. Ten weeks later ₆ ani mals were given 50mg of the compound per kilogram body weight per day by oral a d m in i stration for 10 days and 6 animals were given 50 m g of water alone as a control group Blood pressures were measured under light ether anesthesia and body weights were recorded Their results are shown in the Fig. 1. As shown in the F ig . 1, the rats received the com pound showed decreased blood pressure. The mean maximal decrease was 23 m m H g d u rin g _{1 0} days of administration.

A nd also, in order to test the effect of this compound B on antiinflam m ation, the fo llo w in g two methods were used. One is protein heat coagulation method as prelim inary test and the other is rat-paw edema test.

In the case of heat coagulation test, in a small glass tube, 2. 7 m l of 0. \ % album in so lu tio n and 0. 3 m l of sample solution were added and warmed exactly 3 minutes at 67° in w ater bath. A fter cooling, the turbidity was determined by colorimetric method. The rate of inhi-

142 H an • K im * O h ： Diterpenoids of Siegesbeckia Vol. 19, No. 3

00-

---

9 0 -^ — SOO-

§ 80

70 — doo.

350.

3 00 7S0~

200- / J O . / 0 0

A n t ih y p e r t e n s iv e A c t i v i t y

- 1 2 -!0 - B - 6 - c -2 0 I 2 3 4 S 6 7 d Q t O

( EHS) ( PAYS)

Fig. 1— Antihypertensive effects o f com pound B. T he d rug was administered on O day.

Table V —In h ib ito ry effecs o f c o m po un d B and D on heat coagulation o f bovine serum album in.

T u r b id ity on the conc. (M o le ) o f I n h ib itio n ( % ) on the conc (M o le ) of

■urug

I X10-* 3 X 1 0■결 1X10-3 I X10■설 3X 10■센 1X10-3

C o n tro l 0.52 — 0.70 ^{— —} —

C o m p o u n d B 0.27 — 0.30 48.0 — 57.1

/>-Methoxycinnamic acid 0.48 0.42 7 ,5 — 40.0

C o n tro l 0 .6 6 1.43 — — — —

C o m p o u n d D 0.45 0.77 — 31.8 46.2

Table V I— In h ib ito ry effects o f com pound B on rat-paw edema.

P hlog istic agent

M ean volum e of edema ( m l)

P value

C o n tro l Exp.

10% yeast-saline 0.972 0.806 17.1 P <0 . 001

3 5% formalin-saline 0. 603 0.372 38.3 P C O .O l

10% egg white-saline 0. 587 0. 273 53.5 P <0. 01

Bition was measured from the turbidity. The antiinflammatory activity of compound D was tested by the same method as mentioned above.

#

m

The results are shown in the Table V . As shown in the Table V , heat coagulation o f album in was remarkably inhibited by compound B compared w ith other antiphlogistic agent such as ^-methoxycinnamic acid.

The remarkable antiinflammatory activity of compound B by rat-paw edema test was showiv in the Table V I.

Rat-paw edema test was undertaken as follows. A fter 1 hour of oral administration of 300 m g of compound B per kg body weight of rats, yeast-saline, formaline-saline or egg-white-saline as a phlogistic agent was injected into the plantar surface of the paw of each rat. A n d then the mean volume of edema paw surface was measured, and antiinflam m atory activity was.

estimated from the mean value of the volum e compared w ith that of the control.

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September, 1975 J- Pharm. Soc. Korea 143