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 ðM٦ "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 ðM٦ "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 ðM٦ "calcd for EI!MS m:z "rel[ int[#] 639 ðM٦ "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 ðM٦ "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 ðM٦ "099#^ EI!MS m:z "rel[ int[#] 297 ðM٦ "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 ðM٦ "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!H1O٦ "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 ðM٦ 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 ðM٦ "099#^ EI!MS m:z "rel[ int[#] 297 ðM٦ "2#\ usual manner to give an oil\ which was puri_ed by
189 ðM!H1O٦ "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 ðM٦ "calcd for C06H13O4] 297[0513#^ IR −4[5> "CHCl2^ c 1[74#^ EI!MS m:z "rel[ int[#] 209 ðM٦
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 ðM٦ "calcd for C07H29O3]