Diterpenes and norditerpenes from the Aristeguetia group

Phyrochemstry. Vol. 30, No. 9, pp. 2991 3MlO. 1991 Pnntcd in Great Britam.

DITERPENES

0031-9422/91 $3.00+0.00 @4 1991Pcrgamon Press plc

AND NORDITERPENES GROUP C. ZDERO,

F. BOHLMANN

FROM THE ARISTEGUETZA

and R. M.

KING*

Institute for Organic Chemistry. Technical University of Berlin, D-1000 Berlin 12, Germany; *Smithsonian Institution, Department of Botany, Washington D.C. 20560. U.S.A. (Received 17 December

1990)

Key Word Index -Aristeguetia buddleaefolia,A. glutinosa; Bodillao salicina, Grosoenoria diterpcnes; labdanes; nor-labdanes; clerodanes; tremetone derivatives.

rimbochii;

Compositae;

Abstract-A reinvestigation of Aristeguetia buddleaefoh gave 16 new labdanes and two nor-labdanes, while A. ylutinosa afforded, in addition to large amounts of 8,15-dihydroxylabdane, three new nor-labdanes. Badillao salicina gave six new cis-clerodanes and Grosvenoria rimbachii, guaianolides and tremetone derivatives, four of which were new. The structures were elucidated by high field NMR techniques and a few chemical transformations. The chemotaxonomy is discussed briefly.

INTRODUCTION

The genus Badilloa with 10 species, Aristeguetia with 21 species and Grosvenoriu with four species are placed together in one group notable for broadened strapshaped style branches in the subtribe Critoniinae [ 11. So far the chemistry does not show close relationships between the genus Grosvenoria, where one species gave gauianolides [2], and the genus Aristeguetia, where three species gave labdanes [3-77. We have, therefore, studied four further species: Badilloa salicina (Lam.) K. et R., Aristeguetia glutinosa (HBK) K. et R., again A. buddleaefolia (Benth.). K. et R. and Grosuenoria rimbachii (B. L. Robins.) K. et R.

REXJI.TS AND DSICUSSION

The extract of the aerial parts of A. buddleaefolia gave, in addition to germacrene D, spathulenol and abienol (26), the ent-labdanes 1-19, the labdanes 2&22 and 27, the nor-labdanes 2>25 and the flavono139. All acids were isolated as their methyl esters. The ent-labdanes 1, 3, 9 and 11 have been isolated previously from a Trachylobium species [8], while the enantiomer of 8 was reported from an Araucoria species [9] and that of 14 from an Agafhis species [IO]. The labdane 20 also has been reported [ 1 I]. The latter has been transformed to 24 thus establishing its absolute configuration [12]. As the lactone 24 isolated in the present study shows the same sign of optical rotation as the obtained abienol (26) compounds 2&27 are probably derived from ‘normal’labdanes. Compounds l-19, however, should be entlabdanes as in the case of 1, 3, 9 and 11 the absolute configuration was established [S]. The structures of 1-13 followed from their ‘H NMR spectra and those of the corresponding methyl esters (Tables 1 and 2). As no high field ‘H NMR data were available we have also tabled those of the known com-

pounds. Spin decoupling allowed the assignment of all significant signals. The relative position and the nature of the oxygen functions were deduced from the typical signals and the observed couplings. For some of the entlabdanes the 13C NMR data are presented (Table 3). The structures of 14 19 followed from their ‘H NMR data or from those of the corresponding methyl ester (Table 4). The configuration of the 13,14-double bond caused the typical shift differences of H- 16. Furthermore, the shifts of H- 15 differed slightly in the spectra of the E/Z isomers of the aldehydes 16a, 17a, 18 and 19. As mentioned before, the manoyloxide derivative 20 has been isolated previously [l 11. We have again established its structure. The ‘H NMR data (Table 5) indicated the presence of a manoyl oxide derivative. The stereochemistry and the position of the hydroxy group were determined by NOED. Clear effects were observed between H-20 and H-17 (So/,), between H-17, H-12 (3%) and H-14 (6%) as well as between H-16 and H-14 (10%). Accordingly, the carbinol 20 is 12z-hydroxy- 13-epimanoyl oxide. The molecular formulae of 21 and 22 and also the ‘H NMR spectra (Table 5) indicated that we were dealing with isomers of 20. The couplings of the low field signals at 63.78 and 3.92, respectively, showed that the configuration at C-l 2 must be different. However, the magnitude of the trctns-diaxial coupling was only 9.5 Hz in both cases which may be due to an oxygen function in a livemembered ring. A clear decision was possible by reaction with acetanhydride. While 20 was transformed to the acetate 2OAc both 21 and 22 were isolated unchanged under the same reaction conditions. NOED allowed the determination of the configurations of 21 and 22. In the case of 21 clear effects were observed between H-17, H-20 (5%) and H-14 (2%) as well as between H-16, H-12 (7%), H-14, (3%) and H-151 (3%). In the case of 22 effects between H-17, H-20 (5%) and H-12 (6%) as well as between H-16, H-12 (6%) and H-14 (4%) were observed.

2991

2992

K’

123 co,ti

co,11

(‘OJI

4 (‘(&II

R’

CO,tt

CtIO

(‘H,OH

5 (‘OJII

0 (‘O,II

Cll,Obl~~‘l/~0\l~l~~1

K’

I-1 c‘0:ll.l.V

(‘o~II.l.:%

RZ

(O:H

( 0:lI

c‘IIIOAi

I5

7 (‘II0

(O:lI

1. ‘H

NMR

spectral

11 CH,OH

(‘O>II

(‘H:0hlcCt120tl

IX

17

CttO.l’z

(0:tI

(‘tl:Otl

(‘ti,OH

13

Ct120H

CO,H

I ? cpr

la-7a and 8 (CDCI,.

of compounds

12

CtlIOMeCtl~OMeBu

IY

(‘tlo.l.:k’

21 22 data

IO CH,OH

C’llO.IiL

20

Table

Y CIi,OH

C’O~11c‘11>011

I6 (‘llo.I.:/

8 (‘110

400 MHz_

b-values)

H

lo

Za

311

4a

sa*

6a

7nt

8:

Multlplets

62

1.77 1.97

1.76 I .97

1.74 I.95

1.77 I.97

1.77

1.77 1.9x

1.70 1.81

1.69

1.9x

1.83

dq hr d

w

hr dt ddd

72

I .X6

I.87

I X6

I x9

1.x7

I .8X

1.92

1.92

711 13

2.30

2.3x

2.37

2.30

2.39

2.40

2.44

2.45

1.51

I.51

I.51

1.51

1.51

1.53

1.52

I.50

In

14

2.22 dd

2.40 ddd

I .oo

I.64

I .65

I .67

2.30

1.60

m

14’

2.08 dd

2 IY ddd Y 74 I

1.33

I.35

I .42

1.41

2.08

1.35

m

3.63

3.3x

4.0X

4.07

IS

3.65

16

0.93

NY6

0.87

0.8’)

0 94

0.90

0.92

0.99

7

17

4.83 4.47

4.83 4 46

4.x0 4.46

4.x3 4.49

4.83 4.47

4.83 4.48

4.86 4.50

4.86 4.52

hr 9 hr s

17’ IX

1 IX

1.17

I I5

I.IX

I.18

I.18

1.00

1.01

s

20

0.4Y

0.4Y

0.47

04’)

0.4’)

0.4Y

0.54

0.55

s

3.61

3.61

3.60

3.6 1

3.61

3.61

3.32

_

_

OMe

3.66 *OMeBu:

2.34 fq, 1.65 m. I 43 WI. I .I3

tH-I9

9.72

d.

:H-I9

9.73

d.

[I 13H~. 0.W

J[Hz].5.61=6r.68-62.7~;78-7~,7/1IJ.lJ’:-

I4 (compound

2a: 14.1%

_

S

.._

5

r (3H).

13.7x17=7/~.17=9.17IJ’,IS=2.5):

3.65

compounds

I; 13.16=6.5:compoundslnand2a: 7~ and 8: 3/1.19=

1.3.

13,14=5.5;

13,14’=8;

Diterpenes Table

2. ‘H NMR spectral

data of compounds 10

6a

I .32 dq 1.81 br d 2.35 ddd 1.90 dt

1.31 1.81 2.38 1.93

1.50 m

l

1.44 m

2.31 dd 2.08 dd

l

ddt ddt m d

br d

1.63 1.35 3.66 0.90 4.80 4.48 0.97 3.75 3.38

s

0.64

s

la

7P 13 14 14 15 16 17 17’ 18 19 19 20 OMe

0.92 4.78 4.45 0.95 3.73 3.34 0.62 3.64

br d

ddd dt

l

br d

3.39 0.88 4.79 4.48 0.97 3.75 3.39

s s

0.64 -

d

q br s

s d

m d q br s

s d

‘Overlapped multiplets. J [Hz]: 5,6a=6z,6/?=6a,7/?-13; =ll; compounds 9a and lk =14,15=14’,15-7; 14,14’-14.

C 1 2 3 4 5 6 7 8 9 10 11 I2 13 14 15 16 17 18 19 20 OMe

3.

1.35 1.81 2.38 1.94

dq

1.35 1.80 2.37 1.92 1.45 1.60 1.35 3.66 0.90 4.80 4.48 0.95 3.43 3.09 0.66 -

br d dd dt

q br s

s d br d

-

3.32 s

MHz, CDCI,,

12

11

9ll

dq

2993

group

9a, 10-12 and lJp(400

H

68

Table

from the Aristegueriu

and norditerpenes

13s m

1.36 1.82 2.39 1.93 1.45 2.32

br d

ddd d! m m m

13,16=6.5; 14,14’=14;

dq br d

ddd dl m dd

m

2.09 dd -

d

0.93

d

4

4.81

q

br s

4.48

br s

s

0.96

s

d

4.24

d

br d

3.82 br d

s

0.68

s

3.66 s -

3.28 s

6a,7a=6@,7a-3; 13,14=6; 13,14’=8;

&values)

7a,17=7/?,17=9,17-1; 19.19 compound 11: 13,14=13,14

13CNMR spectral data of compounds 2a, 3a, 7a, 8,9a, 11 and 10 (100.6 MHz, CDCI,, 2a 38.1 19.9 39.1 44.2 56.3 26.1 36.0 148.1 56.2 40.3 21.1 38.7 28.8 50.1 203.0 20.1 106.2 28.7 177.7 12.5 51.1

3s

38.2 19.9 39.5 44.2 56.4 26.2 36.3 148.2 56.3 40.3 21.0 39.1 30.2 38.7 61.0 19.8 106.2 28.7 177.7 12.5 51.0

7a 39.3 19.2 41.3 48.6 55.7 23.9 35.7 147.4 56.0 40.0 21.1 34.3 31.0 39.3 173.6 19.9 107.2 24.3 205.6 13.4 51.3

Accordingly, the isomers only differed in the configuration at C-12. Most likely hydrogen bonds between the ether oxygen and the hydroxy group were present as no NOE between H-17 and H-16 was observed. Compounds 20-22 are probably formed via the 12,13-epoxide of 26 by attack of the 8-hydroxy group. The molecular formula of 23 (C, ,H,,O,) clearly indicated that a nor-diterpene was present. Inspection of the ‘H NMR spectrum (Table 5) further showed that it is a nor-labdane with an unsaturated y-lactone ring. This was supported by the IR band at 1760 cm- I.

8

90 38.9 18.2 41.3 38.5 57.1 24.3 35.1 148.1 56.2 39.5 21.0 35.3 31.0 38.9 173.7 19.9 106.4 27.0 64.8 15.2 51.3

38.4 19.2 39.5 48.5 56.0 23.9 36.2 147.6 55.9 40.0 21.1 38.4 30.1 38.4 61.1 19.7 107.1 24.3 205.7 13.4

&values)

11

20

38.9 18.9 39.6 38.8 57.3 24.3 36.2 148.3 56.2 39.5 20.9 35.3 30.2 38.6 61.2 19.8 106.4 27.0 65.0 15.2 _

38.9 18.4 42.0 33.2 56.5 19.9 42.6 75.9 49.7 36.3 23.5 69.0 76.2 147.1 110.5 26.9 24.2 33.2 21.1 15.7

The ‘H NMR spectrum of 25 (Table 5) showed that a nor-labdane with an unsaturated y-lactone ring must be present. A broadened triplet at 62.57 was due to H-9. Irradiation of the latter collapsed the double doublets at 66.81 and 6.06 to doublets. Most likely 23-25 are formed by degradation of abienol (26). Accordingly, they should belong to the normal-labdane series as the absolute configuration of abienol seems to be secure [ 131, though theoretical calculations disagree with this result [ 143. The observed CD-curves of 23 and 25

gave

no

clear

cut

answer.

In

both

cases

negative

Diterpenes and norditerpenes from the Aristeguelia group

23 24 9.11 H

25

27

26

28

29

31

tH NMR spectrum of 301 was assigned in deuteriobenzene (Experimental), because in chloroform H-7, H-9, H11 and H-l 1’ appeared as an overlapped multiplet. The clear differences in the couplings of H-l 1 and H-11’ indicated a fixed conformation of the side chain. The presence of the exomethylene group followed from the typical signals at 64.90 and 4.77. The structure of 31~ was deduced from its molecular formula (C,,H,,O,) and its ‘H NMR spectrum in deuteriobenzene (Experimental) where all signals could be assigned by spin decoupling. A W-coupling of H-20 with H-l axial allowed the assignment of the H-l signals and consequently those of H-2 and H-3. The double doublet at 60.67 could only be due to H-5. Starting with the latter the signals of H-6 and H-7 could be assigned. As H-9 only showed a coupling with one of the protons at C-l 1 the stereochemistry of the lactone moiety must be different from that of 24. Clear NOR’s established the configurations at all chiral centres [H-17 with H-9 (6%), H-l 1(5%) and H-7e (3%), H-20 with H-l 1’(8%), OMe (3%). H-2ax (6%) and H-6ax (SO/,), H-18 with H-5 (8%), H-6e (6%) and H-3e(3%), H-9 with H-5 (4%), H-17(3%) and H-lax (6%)]. These results also established the proposed as-

2995

30

32

signment of the methyl singlets and of H-l and H-3. The absolute configuration could not be determined. The extract of the aerial parts of Badilloa salicina gave in addition to germacrene D, a-humulene and its l/?,lOaepoxide, the cis-clerodanes 3g38. The molecular formula of main constituent 33 was C,,H,,O, and addition of diazomethane afforded the methyl ester 33a the ‘H NMR spectrum (Table 6) of which required a furoclerodane with a 3,4-double bond and a carbomethoxy group at C4. These data agree with the presence of methyl hardwickioate. However, the ‘H NMR data were different. The 13C NMR spectrum shows that most likely a cisclerodane was present. This was indicated by the chemical shift of C-19 which shows a 10 ppm downfield shift compared with the shift in trans-clerodanes [20]. In the ‘H NMR spectra, the H-17 signal is slightly shifted downfield if the n-arts-isomers are compared with the cisisomers. This was supported by the observed NOES after assignment of some signals by 2D-hetero COSY and COLOC which connects C-19 with H-5, H-6 and H-10 and C-20 with H-11 and H-10. Clear NOES were observed between H-20, H-22 (5%), H-7a (4%) and H-3 (3%) as well as between H-19 and H-1/3 (6%).

C. ZDERO et al.

2996

33 34

R

=

C02H

R

=

CHO

Ohle OH

OH

0

40

41

R

aOA”g

,VOAq

R

aOAng(5OH)

46

42 POAnp

43 (SOAc)

uOAng(SOAc)

47 POAng(SOH)

The ‘H NMR spectrum of 34 (Table 6) showed that the corresponding aldehyde was present. All data were close to those of 33a, but, as in similar cases, the chemical shifts of the protons in the vicinity of the aldehyde group were slightly different. The *H NMR spectra of 3Sa and 36a (Table 6) indicated that we were dealing with the isomeric lactones derived from 33a by oxidation of the furan moiety at C-l 5 or C-16. The chemical shift of H-14 clearly allowed the assignment of the structures. The ‘H NMR spectrum of 37a (Table 6) indicated the presence of a pair of isomers which could not be separated. Only a few signals were doubled. Spin decoupling allowed the assignment of nearly all signals though some were overlapped multiplets. The resulting sequences required the presence of a cis-clerodane derived from 33a where the furan moiety was transformed to a 16.15elide. Pairs ofdoublets at 65.62 and 5.61 and of triplets at 63.84 and 3.82 agree with the presence of epimeric 14,15epoxides. Accordingly, the ‘H NMR data were similar to those of a corresponding epoxide isolated from Con.~u hypoleuca [Zl]. The ‘H NMR spectrum of 38a (Table 1) differed again from that of 33a by the absence of the furan proton signals. The signals of H-14-H-16 show that the precursor of 33a, the 15-hydroxy clerodane with a l3E

44

45

BOH

PO>lc

48 nOAy

(4OAc)

double bond was present. The configuration of the latter follows from the chemical shifts of H-14 and H-16 compared with the shifts of authentic similar compounds. The similarity of the ‘H NMR spectra of all compounds indicated that the configuration of the decalin part was the same in all cases. The aerial parts of Grosvenoria rimbachii gave germacrene D, euparin, coniferyl ferulate [22], 4-methoxy-3isovaleryl acetophenone [23], the guainaolides dehydroleucodin [24], 8-desoxycumambrin B [25], Za-acetoxy3x,4%-epoxykauniolide (49) [26,27], the germacranolide novanin [ZS] and the tremetone derivatives 4&43 [29], 44 [30] and 4548 Lactone 49 has been isolated together with an isomeric /?-epoxide from a Kaunia species 1271. Comparison of the ‘H NMR data and NOES led to a correction of the configuration of 49 at C-2. This was established by transformation to 51 by reaction with hydrochloric acid. The observed coupling of H-2 and the NOES required the presence of a 2-acetoxy derivative [H-2 with H-14 (5%). H-3 (8%) and H-6 (1%); H-3 with H-15 (3%), H-2 (9%) and OH (3%); H-5 with H-15 (4%) and H-7 (6%); H-14 with H-5 (7%) and H-3 (7%); H-15 with H-2 (8%)]. The configuration of the carbinol 50 isolated from another Kuunia species [31] also has to be corrected to 2ahydroxy. In the P-epoxide group from the Kaunia epoxide

Diterpenes

Table

6. ‘H NMR spectral

H

33a

12

2.04 1.87 2.33 2.21 6.52 2.56 1.13 1.28 1.13 1.55 1.49 1.72 1.62

lS 201 28 3 6a 6S 7 7 8 10 11 11’ 12 12’ 14

data of compounds

m hr dd m dddd I m* m m m m m m m

2.03 1.92 2.56 2.41 6.72 2.63 1.11 1.32 1.11 1.54 1.48 1.71 1.54

m hr dd dddd dddd

t m m m m m m m m

2.30 m

t

from the Arisreguetia

7.36 I

2.05 1.87 2.33 2.23 6.54 2.58 1.10 + + 1.44 1.38 + +

group

36a m m dddd dddd

2.04 1.85 2.33 2.21 6.52 2.55 1.12 1.26 1.12

t m m

m m

2.18 m

2997

33a. 34 and 3511-3&k (400 MHz, CDCI,,

35n

34:

2.30 m 7.35

and norditerpenes

37a+ m hr dd dddd dddd t m m m m

2.05 + 2.33 2.21 6.52 2.54 1.10 + + 1.55 1.42 + + + +

1.47 m

I

1.53 m 2.18 m

5.86 rr

7.10 If

15

7.21 hr s

7.22 hr s

_

4.78 dr

16 17

6.26 hr s 0.79 d

6.27 hr s 0.79 d

4.76 d 0.77 d

0.78 d

19

1.24 s

1.16 s

1.25 s

1.24 s

20

0.80 s

0.74 s

0.84 s

0.82 s

OMe

3.71 s

3.72 s

3.71 5

38a m dddd dddd

t m m

m m

i 5.62 5.61 d 0.7X d 1.23 1.22 0.82 0.81 3.71

2.02 1.84 2.39 2.20 6.51 2.54 1.10 1.24 1.10 1.47 1.42 1.55 1.36

m br dd dddd dddd t m m m m m m m m

1.87 m

3.x4 1 3.82 d

i

b-values)

s s s s

5.42 hr t 4.15 hr d 1.69 br s 0.77 d 1.24 s 0.78 s 3.71 s

*C6Ds: 2.52 df. + Overlapped multiplets. tH-13 2.74 (2.72) ddd. fH-18 9.41 s. J[Hz]: la,l/I=l5; la,2z=9; 12,2/?=4; l/j’,2a=9; 1~~.2~=1~,10~1;22,2~=21.22,3=2~,3=3.5;6cr,6~=13;6a,7a =6rx,7~=6~,7/?=3; 8,17=7; 14,15= 14,16= 1.5; compound 35x 12,14= 14,16= 1.5; compound 36~: 12,14= 14.15 =1.5; compound 37~: 12,13=4; 12’,13=8; 13,14,15=2.5; compound 3811: 14,15=7.

[27] a Za-acetoxy group may be more likely, as the observed coupling .I,,, should be the same as in 49 if a 2/jacetoxy derivative was present. The ‘H NMR spectrum of 45 shows that a 3j-methoxy derivative of 6-hydroxytremetone was present (Experimental). The chemical shifts of H-4 and H-7 differed from those reported for this ketone [32], its structure had to be

49 so

corrected to the isomer with the acetyl group at C-6 and the hydroxy group at C-S. The ‘H NMR spectra of 46 and 47 (Experimental) showed that we were dealing with 3a- and 3/Gacyloxy isomers. The nature of the ester group clearly followed from the typical ‘H NMR signals. The ‘H NMR signals of ester group of the ketone 48 (Experimental) required

R = j,c Ii = H

la-7a,9a.13a-l?‘a.29a-31a meth) lcwzr~

the presence

51

33a.35a-38a

arethec(~rrespondine

ot a 4-acetoxyangelicate.

2998

C. ZDERO et at.

The results on the chemistry of the two Aristeguetiu species again indicated that labdanes are characteristic for this genus. The chemistry of Badillou is somewhat different as here only clerodanes were isolated. However, for both species no relationships to Gror~~~oriu is indicated where again guaianolides were isolated. From the genera placed in the subtribe Critoniinae labdanes are also reported from ~p~~r~~~s~~rus [32 and refs cited therein], Bishocia [33] and Kounophyllum species [34,353 while furoclerodanes are present in Critonia [36] and co-occur with labdanes in a ~~~~~~p~~~~urn species [353. ~s4uiterpene lactones are reported from Eupntttriastrum [37], Critonia [36], Ophryosporus [2l] and Cron~u~sfj~~f~us species [38]. Of course, sesquiterpene iactones and labdanes have been isolated from genera of other subtribes of the Eupatorieae.

The plant material was collected m Sprmg 1990 in Ecuador, vouchers are deposited in the US National Herbarium. Washington. The air-dried plant material was extracted with MeOH-Et,O-petrol, (1 : I : 1: II and the extracts sepal as rcported previously [4OJ. The extract of A. ~~d~~eu~f~~~iu (350 2. collected in the Chimborazo Province, 2870 m, voucher RMK 10146) gave by CC 4 frs (I: petrol: 2: Et@-petrol, I :3; 3: Et,O-petrol, I : 1 and Et,O; 4: Et,O-MeOH, 9: I). Fr. I gaveby TLC 5 mg germacrcne D and ii. 2 10 mg abienol(26) and 4 mg spathuienol. Fr. 3 was sepd. after adding CH,N,. by medium pressure CC using Et,@petrol mixts and finally EI,O-McOH (9: I) mto IO frs (3jl .3;10). HPLC of 3/l (MeOH--H,O, 9: I, always RF X, Ilow rate, 3 mlmin- *) gave 2 mg 1Sa (K, 5.3 min), 12mg 14a (R, 5.7 mm), 10mg Ia (R, ?.Omini, 6mg 4a (R, 9.4 min), 15 mg 13a (K, 14.3 min) and a crude fr. (K, 15.3 min) which gave by TLC (I&O-petrol. I : 3) 2 mg Sa (R, 0.69). HPLC of 3:.? (MeOH-H,O, 9: I) gave 3 mg 7a (R, 4.7 min), 1mg 2a (R, 5.0 min) and 3 mg 6a (R, 7.2 min). Fr. 3.;3gave by TLC 50 mg 26. HPLC of 3/4 (MeOH-H,O, 9: I) gave 6 mg 24 (R, 2.9 min) and two mixts (3:44:1R, 3.8 min and 3;4’2 R, 6.1 min). TLC of 3/4:t gave 2 mg 1% (R, 0.51) and 2 mg 16a (R/ 0.45). TLC of 3/4:2 afforded 3 mg 21 (RI 0.62) and 3 mg 22 (R, 0.55). HPLC of 3/5 (McOH-H,O, 9:l) gave 3mg 23 (R, 2.2min), 3mg 25 (R, 2.9 mm), 60 mg 9a (R, 3.9 min). 5 mg 20 (R, 4.6 min), 5 mg tO(R, 5.7 min), 3 mg 12 (R, 7. I min) and 3 mg 27 (R, 9.8 mm). HPLC of 3/6 (MeOH-H,O, 9: 1) gave 200 mg 9a (R, 6.3 min) and of 3:7 (MeOH-H&X 9: 1) 20 mg 8a (R, 5.5 min) and 300 mg 3a (R, 7.1 min). HPLC of 3/X (MeOH- H,O, 9: 1) afforded IO0 mg 11 (R, 3.2 min) and a mrxt. (R, 7.5 min) which gave by TLC (Et&--petrol, I : I) 4 mg 19 (K, 0.65) and 4 mg 18 (R, 0.55). Fr. 3;9 gave 900 mg I1 and fr. 3?IO 100 mg 39. The extract of the aerial parts of A. ylufinosu (356 g, voucher RMK 10157) gave by CC 4 frs. TLC of fr. 1 gave 50mg germacrenc D and the second by TLC (t&O-petrol, I :9) 80 mg 32, 50 mg dammadienylaceiate, 100 mg 26 and IO mg 24. From the third fr. the acidic part was sepd by shaking with NaHCO,sol. The neutral part contained 4.5 g 28. The acids were transformed to their methyl esters by addition of CH,N, in Et,O. TLC (Et,O-petrol. 1: I) gave two hands (3:l and 3:2). HPLC (MeOH- X,0,9: I) offr. 311gave 2 mg 29a fR, 5.9 min) and 3 mg 300 (R, 9.7 min). HPLC of fr. 3&Z(McOH-.H?O. 9.1) gave 2 mg 31n (R, 1.7 min) Fr. 4 gave 2 5 p 2% The air-dried aerial parts (235 g) of Bodtllocr salicina (voucher RMK 10079) were extracted with MeOH -Et,0 petrol (1 : 1: 1). Separation of the defatted extract gave 4 CC frs. The first one afforded by TLC 2 mg germacrene D and IO mg z-humulcne.

The second fr. after addition of CH2N, and TLC gave (Et,O-petrol, I : 19) 100 mg 33a, 5 mg l&l&+epoxy-r-humulene and a crude fr. which gave by HPLC (McOH-H,O, 17.3) 5 mg 34 (R, 6.5 min). Fr. 3 contained 1.4 g 33 and fr. 4 acids. After addition of CH,N,, HPLC (MeOH--H,O, 4: I) gave 2 mg 35a (R, 5.3 min), 3 mg 27a (R, 6.3 min). 2 mg 36n (R, 6.9 min) and 5 mg 3% (R, 12.6 min). The aerial parts (250 g) of ~rosl~~~[~riuri~~u~~i~ (voucher RMK 101It) gave by CC, TLC and HPLC (s. [40]) 5 my germacrene D. 30 mg euparm, 5 mg conifcryl ferulate, 150 mg dchydroleucodm, 30 mg 49.20 mg novanin. 2 mg &desoxycumambrm 6, 2 mg 4-mcthoxy-3-isovaleryl acetophcnone, IO my 40,20mg41,5mg42,20mg43(mp75~).10mg44,10mg45 (TLC: Et,O-petrol, i :3, R,0.52), 5 mg46(HPLC: MeQH-H@. 4: 1, R, 3.0 min), 5 mg 47 (HPLC: MeOH-H,O. 4: 1. X, 3.3 min) and 2 mg48(HPLC: MeOH -H,0,4.1. R,h.7 min). To IO mg49 3 ml CHCI, saturated with HCI was added. After IO min. cvapn and TLC (Et,O-petrol. 3’ 1) gave 6 mg 51: ‘H NMR (C’DCI,): d6.01 (hr dd. H-2). 4.26 (hr dd, H-3), 3.21 (dq. H-S), 4.10 (I, H-6). 2.76 (ry, H-7), 2.13 and 0.95 (m, H-X), 2.34 (m, H-9), 6.23 and 5.50 (d, H-13), 1.77(r, H-14). 1.93(s.H-15). 2.22(d.OH), 2.lS(.s.OAc); J [Hz]: 2.3-5: 2.14=5,14=5,9-1.5: 5,6=6,7:7,8-IO; 7.8’ =7.13-3. ‘%:NMR (CDCI,, C-l C-15): 6139.8, 78.3, 83.3. 83.2. 55.2, 75.5, 49.1. 25.5, 36.0, 138.7. 129.9, 169.5, 119.4. 25.2, 24.8; OAc: 169.9, 20.6. Known compounds were identified by comparing the 400 MHz ‘H NMR spectra with those of authentic material and lit. data. I S-Oxo-cnt-lubd-8( I?)-en-19-ttic acrd (2). Isolated as irs methyl ester k IRv~~~cm~“: 3090. 1650, 900 (C=CH,). 2730. 1740 (CHO), 1730 (CO,R); MS mji (rcl. mt.): 334.251 [M]’ (6) (talc. forC,,H,,O,:334.25I).316[M-H,OJ’(8).305[M-OMeJ’ (3). 275 [3OS -CH,OJ+ (10). 27-l L305-CH%OH]’

(t?). 121 (100),81(23); [%I;;” -39(CHCI,;c1.96). 20 mg3a m 5 mlCHC1, were stirred for 2 hr with 20 mg pyridine chlorochromatc. Usual work-up gave 10 mg 2a. identical with the methyl ester of the natural product. 15-,Methoxy-ent-lahd-X( 17~en-19-oic octd (4). Isolated as its methyl ester 4p; IR v”,“;:cm‘ *: 3095, 1650, 910 (C=CH,), I740 (CO,R): MS m/z (rel. int.): 350.182 [M J ’ (6)(calc. for C,,H,,O,: 350.282). 321 [M -CHO]+ (Z), 318 [M -MeOHJ’ (7). 290[318 -CO]+ (20). 121 (100). 81 (54). 15-[2-methythutyrg/ox~J-ent-f*rbd-X( 17)~en-19-oic acid (5). Isolated as its methyl ester 5a; IR t*“,T:cm- I: 3090. 1645, 900 (C&H,) 1735 (CO,R); MS m/z (rcl. int.): 420.324 [M] ’ (22) (talc. for Cz6H1&00: 420.324), 388 [M - MeOH] ’ (S),360 [M -HCO,Me]” (52). 318 [M-RCO,HJ’ (47). 259 [318 -CO,MeJ’ (12). 180 (42). 121 (ItlO), 85 [RCO]’ (45). 57 [X5 -CO] * (84). l5-Acrrclx~-ent-krhd-8( I?)-en-19-or< ucid (6). Isolated as its methyl ester 6a; IR ~2: cm- ‘. _3090. 1650.9fxf (C=CH,). 1740 (OAc, CO,R); MS ml,- (rel. mt.): 378.277 LM]’ (23) (talc. for C,,H,,O,: 378.277). 319 [M-CO,MeJ’ (44). 318 [M-HOAcJ’ (7% 181 (80). 121 (100). 81 (60). 19-Oxo-ent-fuhd-8(17)-en-l5-oic acid (7). Isolated as irs methyl ester 7a: IRvz$cm-I: 3090, 1650. 910 (C=CHL), 2750. 1750 (CHO). 1730 (CO,R); MS m;t (rel. mt.): 334.25 t [M] * (22) (talc. for C,,H,,O,: 334.251), 316 [M -H,Of’ (8X). 305 [M -CHO]+ (86), 303 [M -0MeJ’ (24). I21 (100); [a];; -21 (CHCI,; ~4.44). I 5-fi~drouy-ent-latg~ I 7)-en-19~1 (8). I R v’,‘;:‘cm _ I: 3640 (OH), 3090, 1645,910 (GCH,), 2740. 1725 (CHO): MS m/: (rel. int.): 288.245 [M - H,O] . (20) (calf. for C,,,H3,0: 288.245). 277 [M-CHO]’ (30), 121 (100),81 (97); [z];’ -25 (CHCI,; ~1.93). 15-Mrrhoxy- 19-~ydr~~x~ent-~abd-8(I?)-ene (10). IR ~2:;:‘:’ cm“: 3640 (OH): MS m,:z (rel. int.): 322.287 [M] * (6) (talc. for

3ooo

C, ZDERO ef al.

cts-ent-Cferodo-3,13-dien-16,18-dioic arid- 16,1S-o&de (36). Isolated as its methyl ester 368; IR vz: cm-‘: 1775 (r-lactone). 1725 (C=CCO,R); MS m/z (ref. int.): 346.214 [M] + (&S)(calc. for C~LIHsoOa:346.214).314[M - MeOH]+ (fOO),29Y(h3).271(88j, 235 (94). 203 (80). 139 (84). 93 (54),91 (58). cis-ent- 14,I S-Epoxyclerod-3-en16,l&dioic acid- f6,l S-elide (37). isolated as methyl ester37a; IR 1*;5;1;: cm-l: f830~~-factone~, 1730 (C=CCO,R); MS m/z @I. int.): 362.214 [M]’ (f0) (dcalc. for C,,HJOOs: 362.214). 347 [M-MeJ’ (6), 331 [M-OMe]’ (111,33O[M -MeOH]” (1 I), 315 [347-MeOH]+ (40), 235(46). 203 (40). I39 (53). 119 (100). 93 (41), 69 (52). cis-ent-f5-H~d~oxycferoda-3,f 3-dien-f8-arc acid (38). fsofated as methyl ester %a; IR v$‘cm-I: 3620 (OH). 1720 (C=CCO,R); MS m,% (ref. int.): 334.251 [M] * (3) fcafc. for CzlH,,O,: 334.2513,319 [M-Me]+ (14). 302 [M-MeOH]” (12). 233 (SO).203 (561, 139 (83),93 (573.81 (56), 69 (661,55 (100). 6-H~dr~~xy-3~-mec~ox~cre~neco~e IR vz:-;:’cm,“: (4% 3500-2500. 1650 {PhCO, hydrogen bonded); MS m/z (ref. int.): 248.105 [MJ’ (100) (calf. for C,,H,,O,: 248.105). 233 [M -Me]+ (381, 217 [M-OMe]’ (78). 203 [233-CH,O]+ (36). 201 [233 - MeOH] ’ (27); ‘H NMR (CDCI,): 65.06 (hr d. H-ZJ, 4.69(hrd, H-3),7.74&, H-4),6.41 (s, H-7),2.56(.s, H-9), 1.70(hrs, H-l f). 5.03 and 4.92 (hr s, H-12), 3.39 (s, OMe); J [Hz]: 2,3= 2. 6-Hydroxy-3~-[S-~ydroxyan~eloyloxyJ-cremecone (46). IR v$$ cm’ I: 3600 (OH), 1725 (C=CCO,R), 3500-2700, 1640 (C=O, hydrogen bonded); MS mjz (ref. int.): 332.126 [MI’ (42) (cafe. for C,sH,OO,,: 332.126). 217 [M-OCOR]+ (82), 216 [M -RCO,H]’ (88). 201 [2f6-Me]’ (70). Y9 [RCO]” (100); ‘H NMR (CDCf, f: 65.15 (hr d, H-2), 6.35 fd. H-3). 7.90 (s, H-4). 6.48(s, H-7), 2.57(s, H-9). 1.84(hrs, H-f I), 5.34and 5,17(hrs. Hf2); OCOR; 6.41 (rq, H-3’). 2.06 (dt, H-4’), 4.19 and 4.08 (br d. H5’); J [Hz]: 2.3 =6; 3’.4’=7; 3’,5’=4’,5’= 1; 5; ,S; = 13: [z];~ - 120 (CHCI 3; c 1.06). 6-Hydroxy-3/I-[S-hydroxyunye/oyloxy)-cremecone (47). fR vgt cm-‘: 3620 (OH). 3500-2600.1650 (PhCO, hydrogen bonded). 1730 (C=CCO,R): MS m,‘; (rel. int.): 332.126 CM]‘ (11) (talc. for C,,H,,O,: 332.126). 217 [M-OCORJ’ (72). 216 [M -RCO,H]+ (fOO), 201 [216-Me]’ (87). 99 [RCO]+ (70); ‘H NMR~CDCI,):65.171d,H-2),6.!3(d,H-3),7.87(s,H4),6.47 (s, H-7). 2.56(s, H-9). 1.76 (hr s, H-f 1). 5.08 and 4.99 (br s, H-12); OCOR: 6.44 (tq. H-3’), 2.06 (dc. H-4’). 4.25 (br s. H-S’); J [Hz]. 2.3 = 2; 3’,4’= 7; 3’*5’= 4’,5’= 1, 6-Wydro.~y~3a-[4-ueefc~.~ya~geloyloxy~-tremrcone (48). IR $i!‘ cm ‘: 3620 (OH), 3SOO-2500, 1650 (PhCO, hydrogen bonded), 1730 (C=CCO,R); MS rn!z (ret. int.): 374.137 [M] + (42) (talc. for CZOH220,: 374.137), 217 [M -OCOR]” (92). 216 [M -RCO,H] A(100). 201 (216-Mel’ (84), 141 [RCO]’ (.54),99 [I41 - keteneJ + (98): ‘H NMR (CDCI,): 65.16 (hr d, H-2), 6.30 (d, H-3),7.88 (s, H-4). 6.47 Is, H-7). 2.58 fs. H-9). 1.82 (hr s, H-l 1). 5.25 and 5.13 (brs. H-12): OCOR: 6.02 (cq. H-3’). S.~(dd4, H-4;), 4.94 (ddq, H-4;), I .X6(dq. H-S’), 2.07 (s, OAc); J [Hz]: 2.3 = 6: 3’,4 =5: 3’,5’==4’,5’- 1.2 4;,4;= 16. ~~~~ff~~ed~erne~~s.-We thank the Dents&e Forschungsgemcinschaft and the Fends der Chcmischen Industrie for financial support. REFERENCES

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34. Bohfmann, F., Zdero, C.. King, R. M. and Robinson, H. f 1979) Ph.vrochemistry IS, 1234. 35. Bohlmann. F., Scheidges, C., King, R. M. and Robinson. H. (1984) Phycochemiscry 23. I 190. 36. Bohlmann, F., Abraham, W. R., King, R. M. and Robinson, H. ( 1981f P~ycochem~scry 20, 1903. 37. Jakupovic, J., Castro, V. and Bohlmann, F. (1987) Phyctr chemistry 26, 45 I. 38. Bohfmann, F., Zdero, C. and Turner, B. L. (1985) Phycarhemtscrp 24, 1263. 39. Zdero, C., Bohfmann, F. and King, R. M. (199f) Phycochemistry 30, 909. 40. Bohfmann. F., Zdero, C.. King, R. M. and Robinson, H. f 1984) Phycochemiscry 23, 1979.