Three lasiodiplodins from lasiodiplodia theobromae ifo 31059 in honour of professor G. H. Neil towers 75th birthday

Article abstract of DOI:10.1016/S0031-9422(98)00267-2

Three new fungal metabolites were isolated from the culture filtrate of the fungus, Lasiodiplodia theobromae IFO 31059 and identified as (5R) and (55) 5-hydroxylasiodiplodins and 5-oxolasiodiplodin. The first two showed weak potato micro-tuber inducing activities, although only at very high dosage levels.

Full text of DOI:10.1016/S0031-9422(98)00267-2

Phytochemistry\ Vol[ 38\ No[ 1\ pp[ 468Ð473\ 0887
Þ 0887 Elsevier Science Ltd[ All rights reserved
Printed in Great Britain
Pergamon
\
9920Ð8311:87:,Ðsee front matter
PII] S9920Ð8311"87#99156Ð1
THREE LASIODIPLODINS FROM LASIODIPLODIA THEOBROMAE
IFO 20948
IN HONOUR OF PROFESSOR G[ H[ NEIL TOWERS 64TH BIRTHDAY
HIDEYUKI MATSUURA\ KAORU NAKAMORI\ E[ A[ OMER\ CHIAKI HATAKEYAMA\ TERUHIKO YOSHIHARAꢀ
and AKITAMI ICHIHARA
Department of Bioscience and Chemistry\ Faculty of Agriculture\ Hokkaido University\ Sapporo 959\ Japan
"Received 10 October 0886^ received in revised form 14 February 0887#
Key Word Index*Lasiodiplodia theobromae^ potato micro!tuber inducing substances^ lasiodi!
plodin[
Abstract*Three new fungal metabolites were isolated from the culture _ltrate of the fungus\ Lasiodiplodia
theobromae IFO 20948 and identi_ed as "4R# and "4S# 4!hydroxylasiodiplodins and 4!oxolasiodiplodin[ The
_rst two showed weak potato micro!tuber inducing activities\ although only at very high dosage levels[ Þ 0887
Elsevier Science Ltd[ All rights reserved
INTRODUCTION
"0604 cm−0#\ aromatic ring "0596 cm−0# and ester
"0159 cm−0# groups[ Its 0H!NMR and 02C!NMR spec!
tra bore good resemblance to that of lasiodiplodin "3#\
which was isolated from Lasiodiplodia theobromae ð2Ł\
except for the 0H signals of H!3 "d 1[77 and 1[53#\ H!
5 "d 1[36# and the 02C resonance of C!4 "d 109[1#[ Both
the 02C!NMR spectral and DEPT analyses revealed
that 0 contained a ketone moiety "C!4\ d 109[1#[ Cross
peaks between H!06:H!2:H!3 were also observed in
the 0H!0H COSY spectrum[ Irradiation of H!3 "d 1[77
and 1[53# caused signal enhancements at C!2 "d 58[0#
and C!4 "d 109[1# in an INAPT experiment\ and
irradiation of H!2 "d 4[54# gave signal enhancements
at C!0 "d 057[9#\ C!3 "d 38[0# and C!06 "d 19[1#[
Accordingly\ the structure of 0 is proposed as 4!oxol!
asiodiplodin "Fig[ 0#^ Tables 0 and 1 give the complete
0H!02C NMR spectral analysis[ Its structure was also
con_rmed by HMBC analysis\ NOE experiments "Fig[
0# and by chemical conversion "Schemes 0 and 1#[
The absolute con_guration at C!2 was determined as
shown in Schemes 0 and 1[ Lasiodiplodin "3#\ whose
absolute con_guration at C!2 was previously deter!
mined as "2S# ð3Ł\ was converted to "1S# 8!"2?\4?!dime!
In a previous paper ð0Ł\ potato micro!tuber inducing
substances\ theobroxide\ jasmonic acid and mellein
from the culture _ltrates of the fungus\ Lasiodiplodia
theobromae IFO 20948 were reported[ Additional
studies led us to isolate three hitherto unreported 01
member lactones from this fungus\ and their struc!
tures were determined by means of H!NMR\ 02C!
0
NMR\ DEPT\ 1D!NMR and NOE experiments tog!
ether with the advanced Mosher method and chemical
conversions[ Potato micro!tuber inducing activities
were tested according to the method which utilises
cultures of single!node segments of potato stems in
vitro ð1Ł[
RESULTS AND DISCUSSION
The EtOAc extracts of the culture _ltrates of Lasi!
odiplodia theobromae IFO 20948 were separated by
CC on silica gel to a}ord 4!oxolasiodiplodin "0#[ Ano!
ther culture _ltrate of the same fungus was subjected
to CC on charcoal\ which following elution with
EtOH gave "4S# 4!hydroxylasiodiplodin "1# and "4R#
4!hydroxylasiodiplodin "2#[
Compound "0# was obtained as a white powder and
gave a molecular formula of C06H11O4ðMŁ¦ by high
resolution EI!mass spectrometry[ Its IR spectrum
showed bands due to hydroxyl "2289 cm−0#\ carbonyl
thoxy!1?!hydroxymethyl#
phenyl!1!nonanol
"4#
"Scheme 0#\ whose ðaŁ1D2 is −4[5> "CHCl2^ c 1[74#[
Reduction of 0 gave the 4!hydroxylasiodiplodin inter!
mediates which were converted into a compound
apparently identical to 4[ Comparison of its 0H!NMR
and IR spectrum\ and the EI!mass spectrometric frag!
mentation pattern of one substance gave good accord!
ance to 4\ although the measured ðaŁ1D4 was −7[5>
"CHCl2^ c 0[24 rather than −4[5>#[ Its absolute con!
ꢀ Author to whom correspondence should be addressed[
468

479
HIDEYUKI MATSUURA et al[
Fig[ 0[ The structures of 0 and 3 and INAPT and NOE experiments[
Table 0[ The 0H!NMR assignments for 0\ 1 and 2 "499 MHz\ CDCl2#
0H "ppm#
0
1
2
H!2
4[54 "ddq\ Jꢁ09[2\ 5[3\ 1[7 Hz#
1[77 "dd\ Jꢁ03[6\ 09[3 Hz#
1[53"dd\ Jꢁ03[6\ 1[7 Hz#
4[23 "m#
4[31 "m#
H!3a
H!3b
H!4
0[74 "ddd\ Jꢁ04[9\ 5[0\ 3[7 Hz#
1[04 "ddd\ Jꢁ04[9\ 5[2\ 2[4 Hz#
3[96 "m#
0[70 "ddd\ Jꢁ03[7\ 4[1\ 0[2 Hz#
1[04 "ddd\ Jꢁ03[7\ 09[0\ 5[4 Hz#
2[75 "m#
H!5a
H!5b
H!6a
H!6b
H!7a
H!7b
H!8a
H!8b
H!09a
H!09b
H!01
H!03
H!06
PhOCH2
1[36 "m#
1[36 "m#
0[89 "m#
0[52 "m#
0[32 "m#
0[25 "m#
0[40 "m#
0[40 "m#
1[40 "m#
1[29 "m#
5[08 "d\ Jꢁ1[0 Hz#
5[13 "d\ Jꢁ1[0 Hz#
0[32 "d\ Jꢁ5[3 Hz#
2[64 "s#
0[40 "m#
0[44 "m#
0[54 "m#
0[31 "m#
0[31 "m#
0[31 "m#
0[31 "m#
0[44 "m#
0[54 "m#
1[33 "ddd\ Jꢁ01[9\ 09[6\ 4[4 Hz#
1[54 "ddd\ Jꢁ01[9\ 09[6\ 4[4 Hz#
5[11 "d\ Jꢁ1[0 Hz#
5[14 "d\ Jꢁ1[0 Hz#
0[39 "d\ Jꢁ5[3 Hz#
2[65 "s#
0[56 "m#
0[39 "m#a
0[39 "m#a
0[18 "m#a
0[18 "m#a
0[59 "m#
0[59 "m#
1[43 "ddd\ Jꢁ02[5\ 5[9\ 5[9 Hz#
1[54 "ddd\ Jꢁ02[5\ 5[9\ 5[9 Hz#
5[11 "d\ Jꢁ1[9 Hz#
5[14 "d\ Jꢁ1[9 Hz#
0[39 "d\ Jꢁ5[4 Hz#
2[65 "s#
a exchangeable[
Scheme 0[
_guration at C!2 is proposed to be "2S# as for lasi! "0489 cm−0 for 1 and 0469 cm−0 for 2# and ester group
odiplodin 3[
"0149 cm−0# absorptions[ Additionally the 0H! and
Compounds 1 and 2 were obtained as colorless 02C!NMR spectra of 1 and 2 closely resembled that of
powders and both gave molecular formulae of lasiodiplodin "3#\ except for the signals of H!4 "d 3[96
C06H13O4ðMŁ¦ by high resolution FD! and EI!mass for 1 and d 2[75 for 2# and C!4 "d 55[6 for 1 and d 69[5
spectrometry[ The IR spectra of 1 and 2 showed for 2#[
hydroxyl
"2189 cm−0#\
conjugated
carbonyl
Therefore\ the structures of 1 and 2 were considered
"0579 cm−0 for 1 and 0569 cm−0 for 2#\ aromatic ring to be 4!hydroxylasiodiplodins[ The positions of

Three lasiodiplodins from Lasiodiplodia theobromae IFO 20948
470
Table 1[ The 02C!NMR assignments for 0\
1 and 2
"014[7 MHz\ CDCl2#
02C "ppm#
0
1
2
C!0
C!2
057[9
58[0
057[1
58[4
057[6
69[4
C!3
38[0
39[3
32[4
C!4
C!5
C!6
C!7
109[1
31[9
10\5
16[1
18[9
55[6
69[5
25[6
25[1
11[9a
13[8a
29[9
11[9a
15[8a
18[6
C!8
C!09
C!00
C!01
C!02
C!03
C!04
C!05
C!06
PhOCH2
21[5
29[9
20[0
031[9
097[6
046[8
86[9
047[1
004[8
19[1
031[8
097[2
047[0
85[8
046[3
006[9
08[8
032[9
097[2
047[2
85[8
046[6
005[2
10[3
Fig[ 2[ The structures of curvularin "6#\ dehydrocurvularin
"7# and 7!methoxycurvularin "8#[
also con_rmed by HMBC spectral analysis\ as well
as by NOE experiments "Fig[ 1#\ and chemical con!
versions[ The 0H!NMR spectra\ EI!mass and IR spec!
tra of the oxidized product 5 derived from 1 and 2
"Scheme 2# had good accordance with those of 0\ and
the ðaŁ1D4 showed ¦4[3> "CHCl2^ c 9[34#[ Therefore the
absolute con_gurations of C!2 of 1 and 2 are proposed
44[6
44[7
44[7
aExchangeable[
hydroxyl groups in 1 and 2 were determined to be at to be "2S#[ In order to determine the absolute con!
C!4 from the cross peaks "H!06:H!2:H!3:H!4\ Fig[ _gurations of the hydroxyl moieties\ 1 and 2 were
0
1# in the H!0H COSY spectra[ The complete NMR converted to MTPA esters\ 1a and 2a ""¦#!MTPA
assignments are given in Tables 0Ð1 according to the ester#\ 1b and 2b ""!#!MTPA ester#[ The advanced
results of 0H!02C COSY spectra[ These structures were Mosher method of determination of con_guration ð4Ł
Scheme 1[
Fig[ 1[ The structures of "4S#\ "4R# 4!hydrolasiodiplodins "1#\ "2# and 0H!0H NMR and NOE experiments[

471
HIDEYUKI MATSUURA et al[
Scheme 2[
was applied to these esters[ The assignments of 1a\ dehydrocurvularin "7# and 7!methoxycurvularin "8#
0
0
1b\ 2a and 2b were achieved by H!NMR and H! from Penicillium A!4!00 were reported to have unique
0H COSY spectral analyses[ The di}erences between activities ð8Ł[ That is\ they have e}ects on the spindle
chemical shifts "0H# 1a and 1b were analyzed "Table formation of the embryos of urchin cells to give barrel!
2#\ and the absolute con_guration of the hydroxyl like spindles and terminate the _rst step of the cell
group of 1 was determined to be "4S# based on the division[ Since "4R# and "4S# 4!hydroxylasiodiplodins
tendency of positive and negative values observed for have some structural similarities with curvularins\
their esters in Table 2[ According to the same method\ they might also have e}ects against the microtubules
that of 2 is proposed as "4R#[
of the cells of potato stems to induce potato micro!
Potato micro!tuber inducing activities of 0\ 1 and 2 tubers[ The fact that jasmonic acid was reported to
were also evaluated according to the method using disrupt cortical microtubules in suspension cultures
cultures of single!node segments of potato stem in of tobacco BY!1 ð09Ł and potato "cv May Queen# ð00Ł
vitro ð1Ł[ Under these conditions the activities of theo! cells is interesting^ however\ 1 and 2 only display very
broxide and jasmonic acid are observed at the concen! weak:moderate tuber!inducing properties[
trations of 09−4 M and 09−5 M\ respectively[ The
activities of 1 and 2\ on the other hand\ were only
noted at much higher concentrations "09−2 M#\
EXPERIMENTAL
although 2 had a little higher activity[ Compound 0
had no activity at the same concentrations[
General
Spectra were obtained with the following instru!
ments] IR\ Hitachi 174 spectrometer^ optical
Lasiodiplodin "3# was _rst isolated from Lasiodi!
plodia theobromae and synthesized by several groups
ð5Ð7Ł\ and lasiodiplodin analogs were isolated from
other microorganisms[ Among them\ curvularin "6#
rotations\ JASCO DIP!3 polarimeter^ NMR\ Bruker
Ã
AM!499 FT!NMR spectrometer and JEOL JNM!EX
169 FT!NMR system^ FD! and EI!MS\ JEOL JMS!
O0SG!1 and JMS!DX!299 mass spectrometers\
respectively[
Table 2[ The Dd values "d"−#!d"¦## for the esters of 1 and 2
in ppm "499 MHz#
Dd values "d"−#!d"¦##
Bioassay
0H "ppm#
1a
2b
The potato micro!tuber forming activity was exam!
ined according to the method using cultures of single!
node segments of potato stem in vitro as previously
described ð1Ł[ Brie~y\ single node segments of stem\
prepared from etiolated potato shoots "Solanum tub!
erosum L[ cv Irish Cobbler#\ were sterilized with 0)
NaClO for 0 hr[ Then three segments were planted
horizontally in a 099 ml ~ask that contained 09 ml of
basal medium "usually White|s medium# sup!
plemented with the compounds to be tested[ Five rep!
licates were prepared[ The concn of sucrose in the
medium was 1) by wt[ The medium was adjusted
to pH 4[5 and solidi_ed with 9[5) Bactoagar before
being autoclaved[ The cultures were maintained at
14>C in the dark for 2 weeks\ and the rate of tuberi!
zation was calculated as the number of tuberized lat!
erals divided by the total number of laterals that had
emerged[
H!2
9[92
9[97
9[94
−9[97
−9[94
−9[96
9[90
9[96
9[98
9[98
−9[97
9[01
H!3a
H!3b
H!4
H!5
H!6
9[92
−9[32
−9[13
−9[94
−9[31
−9[31
−9[92
9[92
−9[90
9[97
9[99
H!7
H!8a
H!8b
H!09a
H!09b
H!01
H!03
H!06
Ph!OCH2
9[95
−9[92
−9[92
−9[96
9[97
a the Dd values "d"−#!d"¦## for 1[ b the Dd values "d"−#!
d"¦## for 2[

Three lasiodiplodins from Lasiodiplodia theobromae IFO 20948
472
Cultures and isolation of 4!oxolasiodiplodin "0#
The fungus was grown in 499 ml ~asks in the
Preparation of 1a\ 1b\ 2a and 2b
Coupling reactions of 1 and 2 with "¦#! and "!#!
stationary phase at 12>C in the dark for 24 days\ with MTPA were achieved in toluene solution containing
each culture containing 049 ml of 1) potato!sucrose DMAP and DCC[ The usual work!up followed by
medium[ The culture _ltrates "about 01 L\ 79 ~asks# puri_cation employing prep[ TLC "Merck\ Me1CO!n!
were concentrated to nearly 0 L in vacuo and extracted hexane\ 3]5\ v:v# gave 1a\ 1b\ 2a and 2b[ "¦#!MTPA
with EtOAc[ Evaporation of the EtOAc extracts in ester of 1 "1a#[ EI!MS m:z "rel[ int[#] 639 ðMŁ¦ "9[0#\
0
vacuo gave a dark brown oil "5 g#[ The oil was chro! 496 "0#\ 378 "9[2#\ 078 "099#\ 094 "21#\ 66 "06#^ H
matographed on silica gel CC "C!199\ Wako!gel\ NMR "499 MHz\ CDCl2#] d 6[53 "1H\ m#\ 6[42 "1H\
MeOH!CHCl2\ 0]099\ 2]86\ 19]79\ v:v#\ and the eluents m#\ 6[37 "2H\ m#\ 6[39 "2H\ m#\ 5[51 "0H\ d\ Jꢁ0[8 Hz#\
"MeOH!CHCl2\ 2]86\ v:v# were further puri_ed by 5[43 "0H\ d\ Jꢁ0[8 Hz#\ 4[28 "0H\ m#\ 4[17 "0H\ m#\
HPLC "Inertsill PREP!ODS\ 19 mm×149 mm\ GL 2[79 "2H\ s#\ 2[57 "2H\ s#\ 2[45 "2H\ s#\ 1[69 "0H\
Sciences Inc[\ MeOH!H1O\ 39]59\ v:v\ 5[9 ml min−0
\
ddd\ Jꢁ03[9\ 5[4\ 5[4 Hz#\ 1[50 "0H\ ddd\ Jꢁ03[9\ 5[4\
UV detector 159 nm# to a}ord 0 "34 mg#[ ðaŁ1D4 ¦6> 5[4 Hz#\ 1[16 "0H\ ddd\ Jꢁ04[9\ 5[2\ 2[3 Hz#\ 0[86 "0H\
"CHCl2^ c 9[7#^ EI!MS m:z "rel[ int[#] 295 ðMŁ¦ "099#\ ddd\ Jꢁ04[9\ 4[4\ 4[4 Hz#\ 0[70Ð0[62 "1H\ m#\ 0[69Ð
180 "6#\ 154 "7#\ 135 "05#\ 119 "13#\ 194 "04#\ 066 "80#\ 0[53 "1H\ m#\ 0[42Ð0[36 "1H\ m#\ 0[39Ð0[21 "1H\ m#\
053 "33#\ 027 "20#\ 010 "3#\ 72 "02#\ 58 "00#\ 44 "7#\ 30 0[14 "2H\ d\ Jꢁ5[4 Hz#[ "−#!MTPA ester of 1 "1b#[
"6#^ EI!HR!MS m:z] 295[0358 ðMŁ¦ "calcd for EI!MS m:z "rel[ int[#] 639 ðMŁ¦ "9[0#\ 496 "0#\ 378
C06H11O4] 295[0357#^ IR nfmilamxcm−0] 2289\ 1825\ 0604\ "9[3#\ 078 "099#\ 094 "22#\ 66 "08#^ 0H!NMR "499 MHz\
0596\ 0159\ 0052^ 0H NMR] Table 0^ 02C NMR] Table CDCl2#] d 6[51 "1H\ m#\ 6[40 "1H\ m#\ 6[36 "2H\ m#\
1[
6[39 "2H\ m#\ 5[50 "0H\ d\ Jꢁ0[6 Hz#\ 5[42 "0H\ d\
Jꢁ0[7 Hz#\ 4[31 "0H\ m#\ 4[20 "0H\ m#\ 2[79 "2H\ s#\
2[57 "2H\ s#\ 2[42 "2H\ s#\ 1[62 "0H\ m#\ 1[47 "0H\ m#\
Cultures and isolation of "4S# 4!hydroxylasiodiplodin 1[21 "0H\ m#\ 1[94 "0H\ m#\ 0[23 "1H\ m#\ 0[22 "1H\ d\
"1# and "4R# 4!hydroxylasiodiplodin "2#
Jꢁ5[3 Hz#\ 0[20 "1H\ m#\ 0[14 "3H\ m#[ "¦#!MTPA
ester of 2 "2a#[ EI!MS m:z "rel[ int[#] 639 ðMŁ¦ "1#\ 496
Growth conditions were as above except that the
fungus was grown for 29 days[ The culture _ltrates
were subjected to CC on charcoal and eluted with
H1O\ EtOH and Me1CO[ The potato micro!tuber
inducing substances were present in EtOH and
Me1CO eluents[ The volatile components of the EtOH
eluents were removed in vacuo to give a dark brown
oil "01[8 g#[ A portion of the oil "4[7 g# was chro!
matographed twice on silica gel CC "C!199\ Wako!
gel\ MeOH!CHCl2!AcOH\ 1]87]0\ 4]84]0\ 09]89]0\
v:v:v# and "C!199\ Wako!gel\ Me1CO!CHCl2!AcOH\
4]84]0\ v:v:v#[ The active fractions "16 mg# were fur!
ther puri_ed by prep[ TLC "Merck\ Me1CO!CHCl2\
2]6\ Rf[ value ꢁ9[5# and HPLC "Novapak C07\
7 mm×064 mm\ Waters\ MeOH!H1O\ 39]59\ v:v\
9[4 ml min−0\ UV detector 109 nm# to a}ord "4S# 4!
hydroxylasiodiplodin "1\ 1[3 mg# and "4R# 4!hydroxyl!
asiodiplodin "2\ 1[1 mg#[ "4S# 4!hydroxylasiodiplodin
"1#[ ðaŁ1D2 ¦4[1> "CHCl2^ c 9[51#^ FD!MS m:z "rel[ int[#]
297 ðMŁ¦ "099#^ EI!MS m:z "rel[ int[#] 297 ðMŁ¦ "09#\
0
"6#\ 078 "099#\ 094 "8#^ H!NMR "499 MHz\ CDCl2#^
d 6[52 "1H\ m#\ 6[43 "1H\ m#\ 6[36 "2H\ m#\ 6[30 "2H\
m#\ 5[47 "0H\ d\ Jꢁ1[9 Hz#\ 5[42 "0H\ d\ Jꢁ1[9 Hz#\
4[31 "0H\ m#\ 4[00 "0H\ m#\ 2[79 "2H\ s#\ 2[69 "2H\ s#\
2[47 "2H\ s#\ 1[61 "0H\ m#\ 1[47 "0H\ m#\ 1[02 "0H\ m#\
1[91 "0H\ m#\ 0[57 "1H\ m#\ 0[27 "2H\ d\ Jꢁ5[4 Hz#\
0[23 "3H\ m#[ "−#!MTPA ester of 2 "2b#[ EI!MS m:z
"rel[ int[#] 639 ðMŁ¦ "0#\ 496 "4#\ 078 "099#\ 094 "06#^
0H NMR "499 MHz\ CDCl2#] d 6[52 "1H\ m#\ 6[44
"1H\ m#\ 6[36 "2H\ m#\ 6[30 "2H\ m#\ 5[48 "0H\ br s#\
5[49 "0H\ br s#\ 4[25 "0H\ m#\ 4[01 "0H\ m#\ 2[68 "2H\
s#\ 2[57 "2H\ s#\ 2[46 "2H\ s#\ 1[58 "0H\ m#\ 1[53 "0H\
m#\ 1[93 "0H\ m#\ 0[86 "0H\ m#\ 0[70Ð0[58 "2H\ m#\
0[44 "0H\ m#\ 0[32 "3H\ m#\ 0[20 "1H\ d\ Jꢁ5[3 Hz#[
Preparation of "1S# 8!"2?\ 4?!dimethoxy!1?!hydroxy!
methyl# phenyl!1!nonanol "4#
To a stirred solution of lasiodiplodin "3\ 09 mg\
189 ðM!H1OŁ¦ "7#\ 164 "5#\ 136 "03#\ 119 "19#\ 080 "08#\ 9[923 mmol# in MeOH "2 ml# was added a solution of
066 "099#\ 027 "79#\ 010 "07#\ 096 "04#\ 80 "00#\ 66 CH1N1!Et1O "2 ml#\ and the mixture was stirred at 9>C
"15#\ 44 "11#\ 30 "09#^ FD!HR!MS m:z] 297[0506 ðMŁ¦ for 1 hour[ The volatile components of the reaction
"calcd for C06H13O4] 297[0513#^ IR nfmilamxcm−0] 2189\ mixture were removed under reduced pressure to give
0
1899\ 0579\ 0489\ 0149\ 0979^ H NMR] Table 0^ 02C an oil[ To a stirred suspension of LAH "58 mg# in Et1O
NMR] Table 1[ "4R# 4!hydroxylasiodiplodin "2#[ at 9>C was added a solution of the oil in Et1O "4 ml#\
ðaŁ1D2 ¦7[7> "CHCl2^ c 9[69#^ FD!MS m:z "rel[ int[#] and the reaction was accomplished according to the
297 ðMŁ¦ "099#^ EI!MS m:z "rel[ int[#] 297 ðMŁ¦ "2#\ usual manner to give an oil\ which was puri_ed by
189 ðM!H1OŁ¦ "02#\ 164 "8#\ 136 "02#\ 119 "12#\ 080 prep[ TLC "Merck\ MeOH!CHCl2\ 01]77\ v:v# to
"08#\ 066 "099#\ 053 "17#\ 027 "44#\ 010 "03#\ 095 "01#\ a}ord "1S# 8!"2?\4?!dimethoxy!1?!hydroxymethyl#
80 "03#\ 66 "19#\ 44 "07#\ 30 "37#^ EI!HR!MS m:z] phenyl!1!nonanol "4\ 4[6 mg\ 9[907 mmol\ 42)#[ ðaŁD12
297[0506 ðMŁ¦ "calcd for C06H13O4] 297[0513#^ IR −4[5> "CHCl2^ c 1[74#^ EI!MS m:z "rel[ int[#] 209 ðMŁ¦
nmfilamxcm−0] 2189\ 1899\ 0569\ 0469\ 0149\ 0979^ 0H "43#\ 181 "05#\ 166 "03#\ 071 "099#\ 056 "51#\ 041 "61#^
NMR] Table 0^ 02C NMR] Table 1[
FD!HR!MS m:z] 209[1037 ðMŁ¦ "calcd for C07H29O3]

473
HIDEYUKI MATSUURA et al[
0
209[1034#^ IR nfmilamxcm−0] 2264\ 1818\ 0595^ H NMR at 9>C[ The reaction mixture was quenched with 1!
"499 MHz\ CDCl2#] d 5[22 "1H\ m#\ 3[55 "1H\ s#\ 2[73 propanol followed by H1O and extracted with EtOAc
"2H\ s#\ 2[79 "2H\ s#\ 1[54 "1H\ t\ Jꢁ7[9\ 6[8 Hz#\ 0[45Ð "2×09 ml#[ The combined organic layers were dried
0[29 "07H\ m#\ 0[07 "2H\ d\ Jꢁ5[1 Hz#[
over Na1SO3 and concentrated in vacuo[ The residue
was puri_ed by prep[ TLC "Merck\ MeOH!CHCl2\
4]84\ v:v# to a}ord 5 "3[4 mg\ 9[903 mmol\ 69)#[
ðaŁ1D4 ¦4[3> "CHCl2^ c 9[34#^ FD!MS m:z "rel[ int[#]
295 ðMŁ¦ "099#[
Preparation of 4 from 0
To a stirred solution of 4!oxolasiodiplodin "0\
01 mg\ 9[93 mmol# in MeOH "2 ml# was added excess
NaBH3 at 9>C\ and the mixture was stirred for 29 min[
This reaction mixture was quenched with H1O and
extracted with EtOAc[ The combined organic layers
were washed with satd aq[ NaHCO2 and dried over
Na1SO3[ The volatile components of the organic layers
were removed in vacuo to a}ord an oil\ which was
puri_ed by HPLC "Inertsill PREP!ODS\ 19 mm×
149 mm\ GL Sciences Inc[\ MeOH!H1O\ 39]59\ v:v\
5[9 ml min−0\ UV detector 109 nm# to give 4!hydroxyl!
asiodiplodins[ To a stirred solution of the 4!hydroxyl!
asiodiplodins in MeOH "4 ml# at 9>C was added a
solution of CH1N1!Et1O "2 ml#[ The volatile com!
ponents of the reaction mixture were removed in vacuo
to give an oil[ To a stirred solution of the oil in pyridine
"1 ml# was added TsCl "77[0 mg#\ and the mixture was
stirred for 03 hour at room temp[ The reaction mixture
was poured into Et1O "14 ml# and washed with 9[0 N
HCl "2×49 ml# and satd aq[ NaCl "1×49 ml#[ The
Et1O layer was dried over Na1SO3 and concentrated
up to give an oil[ To a stirred suspension of excess
LAH in Et1O at 9>C was added a solution of the oil
in Et1O\ and an usual work!up for this reaction was
employed to a}ord an oil\ which was puri_ed as before
to a}ord 4 "1[6 mg\ 9[998 mmol\ 22)#[ ðaŁ1D4 −7[5>
"CHCl2^ c 0[24#^ FD!MS m:z] 209 ðMŁ¦ "099#[
Acknowled`ements*The authors thank Mr[ K[
Watanabe and Dr[ E[ Fukushi for FD!\ EI!MS\
HMBC and INAPT measurements[
REFERENCES
0[ Nakamori\ K[\ Matsuura\ H[\ Yoshihara\ T[\ Ichi!
hara\ A[ and Koda\ Y[\ Phytochemistry\ 0883\ 24\
724[
1[ Koda\ Y[ and Okazawa\ Y[\ Plant Cell Physiol\
0877\ 18\ 858[
2[ Aldridge\ D[ C[\ Galt\ S[ and Turner\ W[B[\ J[
Chem[ Soc[ "C#\ 0860\ 0512[
3[ Lee\ K[!H[\ Hayashi\ N[\ Okano\ M[\ Hall\ I[ H[\
Wu\ R[!Y[ and McPhail\ A[ T[\ Phytochemistry\
0871\ 10\ 0008[
4[ Ohtani\ I[\ Kusumi\ T[\ Kashman\ Y[ I[ and Kak!
isawa\ H[\ J[ Am[ Chem[ Soc\ 0880\ 002\ 3981[
5[ Gerlach von\ H[ and Thalmann\ A[\ Helv[ Chim[
Acta\ 0866\ 59\ 1755[
6[ Takahashi\ T[\ Kasuga\ K[ and Tsuji\ J[\ Tetra!
hedron Lett\ 0867\ 38\ 3806[
7[ Danishefsky\ S[ and Etheredge\ S[ J[\ J[ Or`[
Chem\ 0868\ 33\ 3605[
8[ Kobayashi\ A[\ Hino\ T[\ Uneyama\ K[ and
Kawazu\ Y[\ Symposium Papers\ The Chemistry
of Natural Products\ Hiroshima\ 0874\ p[ 232[
09[ Abe\ M[\ Shibaoka\ H[\ Yamane\ H[ and Taka!
hashi\ N[\ Protoplasma\ 0889\ 045\ 0[
Preparation of 5
To a stirred solution of 1 and 2 "5[3 mg\ 9[91 mmol# 00[ Matsuki\ T[\ Tazaki\ H[\ Fujimori\ T[ and
in acetone at 9>C was added a solution of Jones
reagent "4 ml#\ and the mixture was stirred for 29 min
Hogetsu\ T[\ Biosci[ Biotech[ Biochem\ 0881\ 45\
0218[