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Fusidienol: A novel inhibitor of Ras farnesyl-protein transferase from Fusidium griseum
TerrohedmnLencrs, Vol. 35. No. 27, pp. 46934696,
1994 Elsevier Scieuce Ltd Printed in Great Britain cKMo-4039/94$7.00+0.00
oo40-4039(94)00910-4
Fusidienol:
A Novel Inhibitor of Ras Farnesyl-Protein Transferase from Fusidium griseum
Sheo B Singh*, E. Tracy Jones, Michael A. Goetz, Gerald F. Bills, Mary Nallin-Omstead, Rosalind G. Jenkins, Russell B. Lingham, Keith C. Silverman and Jackson B. Gibbs Merck Research Laboratories, P. 0. Box 2000 Rahway, NJ 07065 and Weat Point PA 19486
Aktractt Ras (~21) protein is frequently found mutated in human crmcers and must be famesylated by famesyl protein-traosferase (FPTase) to achieve cell-transforming activity. Our continued search for inhibitors of FPTase as potential cancer chemotheqeutics led to the isolation of fusidienol from extracts of the fungus Fusidkn grisevm. Fusidienol is a novel and potent oxygen-containing [7/6/6] tricyclic hetemcycle.
Farnesyl-protein
transferase
(FTTase) catalyzes
cysteine residue near the C-terminus. activity’.
the famesylation
For Ras, post-translational
of proteins such as Ras (~21) on the
modification
is required for cell-transforming
Selective inhibitors of FFTase have the potential to be used as anticancer agents, particularly in colon
and pancreatic cancers* where the ras oncogene is mutated and believed to play a major role in tumor formation. Recently, mase
inhibitors have shown biological efficacy against ras dependent cell-transformation.3
We have recently nanomolar
potency.
pepticinnamins.5 Continued
reported the isolation
Other microbial
products
(a class of peptides), gliotoxin,6
of chaetomellic
acids4 as novel inhibitors
which have been reported lo’-desmethoxystreptonigrin,7
as inhibitors
of FPTase with of FPTase
and manumycin
are
analogs.8
screening for potent inhibitors of FPTase resulted in the isolation of fusidienol (la) from Fusidium
griseum (Deuteromycotina,
Hyphomycetes).o
Fusidienol is a novel tricyclic oxygen-containing
heterocycle with
a 7/6/6 ring system that inhibited bovine brain FPTasel” with an ICso of 300 nM while being inactive against rat liver squalene synthase and bovine brain geranyl-geranyl is non-competitive
protein transferase. Inhibition of FPTase by fusidienol
with respect to both substrates i.e. acceptor Ras-CVLS peptide and PPP. Ki values were 1.4
and 0.5 @l with respect to Ras and FPP, respectively. recombinant
Fusidienol
was less active when tested against
human PPTase enzyme to exhibiting an ICm of 2.7 pM. Fusidienol diacetate (lb) inhibited bovine
FPTase with an ICso of 1 pM.
la: R= H, Fusidienol lb: R= COCH3 4693
Fusidium griseum, grown for 11 days on medium consisting
alfalfa and corn oil, was extracted chromatography by partitioning
with methyl ethyl ketone.
of millet, yeast, sodium tartrate. sucrose, Fusidienol
(la) was initially
isolated by
OIISephadex LH-20 followed by reversed phase HPLC. Subsequent isolations were achieved
the dried methyl ethyl ketone extract with methylene
followed by silica gel chromatography
using hexane-acetone.
chloride and 50% aqueous methanol
Crystallixation
from methanol afforded yellow
granules (50 mg/L) of fusidienol (la), mp. 168-70 Oc. STRUCTURE
ELUCIDATION
Electron impact (EI) mass spectral analysis of fusidienol gave a molecular ion at m/z 3 16 for which the empirical
formula Ct&ll207
was determined
ftisidienol has 11 degree of unsaturations. reacted with BSTFA-pyridine.
by high resolution measurement.
Fusidienol
This formula indicated that
(la)” formed a his-trimethylsilyl
ether (m/z 460) when
The UV spectrum of la gave absorption bands at 230 and 327 nrn, an indication
of a highly conjugated system which was also apparent from the slightly yellow color of fusidienol. The infra red spectrum showed absorption carbonyl
bands for hydroxy (3500 cm-l), ester (1724 cm-t) and highly conjugated
(165 1 cm-l) groups. Formation
of the bis-TMS derivative
indicated
the presence
of two active
hydrogens which wes supported by acetylation with acetic anhydride and pyridine to give diacetate lb. 12C- 13 NMR analysis (Table 1) of la in C@OD displayed 16 carbons and suppotted the molecular formula derived from mass spectral analysis. The APT spectrum of fusidienol revealed the following types of carbons: a methoxy carbon (6 52.90), an oxymethylene (8 60.72), five olefinic&omatic methines and the remaining nine carbons were quatemaries
which presented a challenge in resolving the structure. Two of the carbons were
highly downiield and occupied the region of the spectrum which is normally occupied by catbonyl carbons.
Figure 1: FIMBC (“Jca=7 Hz) C.orreMhm~ of la Examination trisubstitnted olefic
of the
rH NMR
spectrum (Table 1) indicated the presence of a methoxy group, a 1,2,3-
aromatic ring, two oleftic
protons. The sub-structures
protons, and an oxymethylene
The correlation of methoxy protons with the carbonyl carbon confii bond HMBC correlations
which showed small couplings to both
were put together by HMBC correlations (Figure 1) using a n&
= 7Hx.
it to be. a methyl ester. Two and three
of H-11 to C-P, 10, 12, 13, 16, 17 and H-13 to C-11, 12, 15, 16 helped in
assembling the left hand side of the molecule which was further corroborated by respective correlations from H16. The right hand side of the molecule was similarly essembled with the help of HMBC correlations of H-3,4 and5. Unambiguous due to equivocal
assignment of chemical shifts of C-2 vs C-6 and C-3 vs C-5 was not possible in QOD
correlations.
This distinction
cormlations involving the unexchanged
became possible only when we made use of the HMBC
chelated 6-OH group. Therefore, the NMR studies were repeated in a
4695
2: 1 mixture of CD$N-CD3COCD3
(Table 1) . The HMW
experiment in this solvent gave all of the HMBC
correlationswhich were observed in CD30D with additionalcorrelationsfrom the 6-OH. The protonat 6 12.13 gave correlations to C-5 (6113.26), 6 (6161.75). 7 (6110.19) and 8 (6183.92) thereby confirming all of the carbon chemical shift assignments. Based 0~1 thesedata structurela was proposed for fusidknol. The mass spectralfragmentationsof diacetatelb (Figure 2) were in completeagreement withtheassignedsm.
m/z228 Figure 2: Mass Spectral Fragmentation
Table
1: NMR Assignment
of Fusidienol diacetate (lb)
of Fusidienol
Position
60
X!2
Mult
6H1
8H2
2
155.16
154.90
co
---
-__
3
108.11
108.23
CH
6.95, dd, 9.0, 0.5
6.95, dd, 8.4, 0.8
4
136.94
137.04
CH
7.60, t, 8.5
7.60, t, 8.4
5 6
113.37 161.94
113.26 161.75
CH C”
6.82, dd, 9.0, 0.5 ---
6.81, dd, 8.4, 1.2 -__
7
110.28
110.19
co
---
-__
8
183.95
183.92
Co
---
___
9
105.80
105.72
C“
---
-_-
10
131.34
131.24
Co
---
-_-
11
134.64
134.50
CH
7.06, s
7.00, dd, 0.8, 0.4
12
131.31
131.10
co
---
--
13
145.75
145.13
CH
6.57, t, 1.5
6.52, td, 1.2, 0.4
15
164.42
163.98
Co
---
-__
16
60.72
60.63
CH2
4.14. d, 1.5
4.13, ddd, 6.0. 1.6, 0.4
17
168.5 1
167.57
Co
---
___
19
52.90
52.93
CH3
3.79, s
3.72, s
6_OH
___
__
___
___
12.13, s
16_OH
___
___
__
___
3.61, t, 6.0
lCD3OD
at 30Oc (500MHz); kD3CN-CD3COCD3 (2:1)at 25 OC(400 MHz).
REFERENCES I.
AND NOTES
(a) Gibbs. J. B. Semin. CCM Bid. 1992, 3, 383. (b) Gibbs, J. B. Cells
1991.65.1 and refenmces cited therein. (c) Der, C.
J. and COX, A. D. Colncer Cells 1991,3.331.(d) Reiss. Y.: Goldstein,
J. L.; Se&m,
Cells 1990, 62. 81. (e) Schabcr, M. D.; O’Ham, M. B.; Garsky, V. M.; Mosser. Marshall.
M. S.: Friedman,
P. A.; Dixon,
M. C.; Casey. P. J.; and Brown, M. S. S. D., Bergstrom,
J. D.; Moores,
S. L.;
R. A. F.; and Gibbs,J. B. J. Biof. Chem. 1990,265, 14701. (f) Gibbs, J. B.;
4696
Pompliano,
D. L.; Mosser, S. D.; Rands, E.; Lingham,
R. B.; Singh, S. B.; Scolnick,
E. M.; Kohl, N. B.; and Oliff, A. J.
Biol. Chem. 1993, 268, 7617. 2. 3.
(a) Barbacid,
Rev. B&hem. 1987. 56, ‘779.(b) Rodenhuis,
M. Ann
(a) Kohl, N. E.: MOW,
S. D.; debolms. J. S.; Giuliani.
S. Semin. Gun. Biol. 1992, 3, 241.
E. A.: Pompliano,
D. L.; Graham,
E. M.; Cliff, A.; and Gibbs, J. B. Science 1993, 260. 1934. (b) James, G. J.; Goldstein, E.; Somers.
T. C.; McDowell,
R. S.; Crowley,
C. W.; Lucas, B. K.; Levinson,
S. L.; Smith, R. L.; S&rick,
J. L.; Bmwn, M. S.; Rawson,
T.
A. D.; and Marsters. J. C. Science 1993.
260, 1937. 4.
(a) Singh, S. B.; Zink, D. L.; Liesch, J. M.; Goetz, M. A.; Jenkins, R. G.; Nallin-Gmstead, F.: MO&y, R. T.: Gibbs, J. B.: Albers-Schonberg, Singb, S. B. Terrhednrn M.; Jenkins,
Microbial. 5.
I&t. 1993, 34, 6521. (d) Linghsm.
M.; Mosser, S. D.; Schaber,
Biotech
Inokoshi. J.; Van Der Pyl, D.; Nakagawa, Van Dcr Pyl, D.; Inokoshi,
7.
Liu, W. C.; Barbacid,
8.
(a) Ham M.; Akasaka,
Y.; and Takesbima, A.; Takeshima,
K.; Akinaga,
S.; Okabe, M.; Nakano,
2281. (b) Tamanoi,
(a) Fuskiium
= ATCC 74265)
griseum (MF5695
by Dr. Bary Katz of MYCCkearch, and Wilkins
Co., Baltimore, Publishing
mp. 168-70%
Cl6Hl2O7:
North Carolina.
leaf litter, near Lansing,
(b) Barron, G. L. “The Genera
F. Proc.
Michigan,
USA)
of Hyphomycetesfrom
Soil”
on land Plants, an Identification
cited theirin. (b) Chuer, C. A.; Kral, A. M.; Diel, R. E.; Prendergast,
G. C.; Powers, S.;
1993, 32, 5167.
230 (&=19815),
327
(7360) nm; IR (ZnSe): 3500, 2953,
816, 766 cm-‘:
1724. 1651, 1610,
HREIMS (m/z): 316.0600
(M+, calcd. for
6): 2.04 (3H, s), 2.34 (3H, s), 3.79 (3H, s). 4.67 (ZH, s), 6.63 (HI, s), 7.05 (IH, d, J =
s). 7.30 (lH, d. J = 8.0 Hz), 7.64 (lH, t, J = 8.5 Hz). HREIMS
400.0794),
(calcd. for Cl7Hl007:
228.0423).
1992, 45.454.
316.0583).
8.0 Hz), 7.22 (lH,
284.0313
1802. P. B.; Huang,
H.; Gomez, R.; Wood, D.; Uh. M.; and Tamanoi,
1968. (c) Ellis, M. B.; Ellis, J. P. Microfungi
W: hmax (CH30H):
lb: ‘H NMR (CD30D+CDC13,
C2OHl609:
1992.45,
A. R.; Dean, L.; Doyle, T. W.; Fernandes,
F. Trend. Biochem. Sci. (TlBS) 1993, 18, 349.
1468, 1421, 1361. 1274. 1231, 1176, 1140. 1063, 1020.896,
12.
1993, 46, 229.
Co. New York 1985.
(a) See ref. If. and 4d and references
la:
1993, 46. 222. (b) Shiomi, K.; Yang, H.;
H.; and Chnura, S. J. Antibiotics
was isolated from hardwood
Allen, C. M.; Gibbs, J. B.; and Kohl, N. E. Biochemistry 11.
L.;
D. L.; Gibbs, J. B.; and Singh, S. B. Appl.
H.; and Gmura, S. J. Antibiotics
M.; Bulgar, M.; Clsrck, J. M.; Crosswell,
Hundbook ” Macmillan 10.
C.; Sanchez,
S.; Kong, L.; Burg, R. W.; Meinz, M. S.; Huang,
H. .I. Antibiotics
I.; Sbiomi, K.; Yang, H.; Takeshima,
Narf. Acad. Sci. USA 1993.90.
Williams
K. C.; Bills, G. F.; &scales,
M. D.; Omer, C. A.; Pompliano,
S.: Manne, V.: Pimik, D. M.; Wells, J. S.; and Meyers, E. J. Antibiotics
provided
K. C.; Bills, G.
1993, 40, 370.
(a) Omura, S.; Van Der Pyl, D.; Takahashi,
6.
9.
M.; Silverman,
R. B. Tetrahedron 1993, 49, 5917. (b) Ref. If. (c)
R. B.; Silverman,
S.; Martin, I.; Pelaaz, F.; Mochales,
R. G.; Gartner,
Nallin-Gmstead,
G.: and Lingham.
326.0427).
(calcd. for Cl5Hg07:
310.0485 (calcd. for Cl7HlOG7: 284.0321).
163.0403 (calcd. for CgH703:
(Received in USA 18 April
(m/z): 400.0799
358.0693 (calcd. for C1gH1408: 358.0689), 342.0743 (cakd. for ClgH1407:
1994,
266.0198
310.0477)
(calcd. for Cl5H605:
163.0395), 137.0241 (c&d.
accepted 6 May 1994)
(M+, calcd.
298.0484 (calcd. for Clr5HlOO6: 266.0215).
for C7H503:
228.0433
137.0239).
for
342.0740), 326.0430
(calcd.
298.0477).
for Cl3Hg04:
1994 Elsevier Scieuce Ltd Printed in Great Britain cKMo-4039/94$7.00+0.00
oo40-4039(94)00910-4
Fusidienol:
A Novel Inhibitor of Ras Farnesyl-Protein Transferase from Fusidium griseum
Sheo B Singh*, E. Tracy Jones, Michael A. Goetz, Gerald F. Bills, Mary Nallin-Omstead, Rosalind G. Jenkins, Russell B. Lingham, Keith C. Silverman and Jackson B. Gibbs Merck Research Laboratories, P. 0. Box 2000 Rahway, NJ 07065 and Weat Point PA 19486
Aktractt Ras (~21) protein is frequently found mutated in human crmcers and must be famesylated by famesyl protein-traosferase (FPTase) to achieve cell-transforming activity. Our continued search for inhibitors of FPTase as potential cancer chemotheqeutics led to the isolation of fusidienol from extracts of the fungus Fusidkn grisevm. Fusidienol is a novel and potent oxygen-containing [7/6/6] tricyclic hetemcycle.
Farnesyl-protein
transferase
(FTTase) catalyzes
cysteine residue near the C-terminus. activity’.
the famesylation
For Ras, post-translational
of proteins such as Ras (~21) on the
modification
is required for cell-transforming
Selective inhibitors of FFTase have the potential to be used as anticancer agents, particularly in colon
and pancreatic cancers* where the ras oncogene is mutated and believed to play a major role in tumor formation. Recently, mase
inhibitors have shown biological efficacy against ras dependent cell-transformation.3
We have recently nanomolar
potency.
pepticinnamins.5 Continued
reported the isolation
Other microbial
products
(a class of peptides), gliotoxin,6
of chaetomellic
acids4 as novel inhibitors
which have been reported lo’-desmethoxystreptonigrin,7
as inhibitors
of FPTase with of FPTase
and manumycin
are
analogs.8
screening for potent inhibitors of FPTase resulted in the isolation of fusidienol (la) from Fusidium
griseum (Deuteromycotina,
Hyphomycetes).o
Fusidienol is a novel tricyclic oxygen-containing
heterocycle with
a 7/6/6 ring system that inhibited bovine brain FPTasel” with an ICso of 300 nM while being inactive against rat liver squalene synthase and bovine brain geranyl-geranyl is non-competitive
protein transferase. Inhibition of FPTase by fusidienol
with respect to both substrates i.e. acceptor Ras-CVLS peptide and PPP. Ki values were 1.4
and 0.5 @l with respect to Ras and FPP, respectively. recombinant
Fusidienol
was less active when tested against
human PPTase enzyme to exhibiting an ICm of 2.7 pM. Fusidienol diacetate (lb) inhibited bovine
FPTase with an ICso of 1 pM.
la: R= H, Fusidienol lb: R= COCH3 4693
Fusidium griseum, grown for 11 days on medium consisting
alfalfa and corn oil, was extracted chromatography by partitioning
with methyl ethyl ketone.
of millet, yeast, sodium tartrate. sucrose, Fusidienol
(la) was initially
isolated by
OIISephadex LH-20 followed by reversed phase HPLC. Subsequent isolations were achieved
the dried methyl ethyl ketone extract with methylene
followed by silica gel chromatography
using hexane-acetone.
chloride and 50% aqueous methanol
Crystallixation
from methanol afforded yellow
granules (50 mg/L) of fusidienol (la), mp. 168-70 Oc. STRUCTURE
ELUCIDATION
Electron impact (EI) mass spectral analysis of fusidienol gave a molecular ion at m/z 3 16 for which the empirical
formula Ct&ll207
was determined
ftisidienol has 11 degree of unsaturations. reacted with BSTFA-pyridine.
by high resolution measurement.
Fusidienol
This formula indicated that
(la)” formed a his-trimethylsilyl
ether (m/z 460) when
The UV spectrum of la gave absorption bands at 230 and 327 nrn, an indication
of a highly conjugated system which was also apparent from the slightly yellow color of fusidienol. The infra red spectrum showed absorption carbonyl
bands for hydroxy (3500 cm-l), ester (1724 cm-t) and highly conjugated
(165 1 cm-l) groups. Formation
of the bis-TMS derivative
indicated
the presence
of two active
hydrogens which wes supported by acetylation with acetic anhydride and pyridine to give diacetate lb. 12C- 13 NMR analysis (Table 1) of la in C@OD displayed 16 carbons and suppotted the molecular formula derived from mass spectral analysis. The APT spectrum of fusidienol revealed the following types of carbons: a methoxy carbon (6 52.90), an oxymethylene (8 60.72), five olefinic&omatic methines and the remaining nine carbons were quatemaries
which presented a challenge in resolving the structure. Two of the carbons were
highly downiield and occupied the region of the spectrum which is normally occupied by catbonyl carbons.
Figure 1: FIMBC (“Jca=7 Hz) C.orreMhm~ of la Examination trisubstitnted olefic
of the
rH NMR
spectrum (Table 1) indicated the presence of a methoxy group, a 1,2,3-
aromatic ring, two oleftic
protons. The sub-structures
protons, and an oxymethylene
The correlation of methoxy protons with the carbonyl carbon confii bond HMBC correlations
which showed small couplings to both
were put together by HMBC correlations (Figure 1) using a n&
= 7Hx.
it to be. a methyl ester. Two and three
of H-11 to C-P, 10, 12, 13, 16, 17 and H-13 to C-11, 12, 15, 16 helped in
assembling the left hand side of the molecule which was further corroborated by respective correlations from H16. The right hand side of the molecule was similarly essembled with the help of HMBC correlations of H-3,4 and5. Unambiguous due to equivocal
assignment of chemical shifts of C-2 vs C-6 and C-3 vs C-5 was not possible in QOD
correlations.
This distinction
cormlations involving the unexchanged
became possible only when we made use of the HMBC
chelated 6-OH group. Therefore, the NMR studies were repeated in a
4695
2: 1 mixture of CD$N-CD3COCD3
(Table 1) . The HMW
experiment in this solvent gave all of the HMBC
correlationswhich were observed in CD30D with additionalcorrelationsfrom the 6-OH. The protonat 6 12.13 gave correlations to C-5 (6113.26), 6 (6161.75). 7 (6110.19) and 8 (6183.92) thereby confirming all of the carbon chemical shift assignments. Based 0~1 thesedata structurela was proposed for fusidknol. The mass spectralfragmentationsof diacetatelb (Figure 2) were in completeagreement withtheassignedsm.
m/z228 Figure 2: Mass Spectral Fragmentation
Table
1: NMR Assignment
of Fusidienol diacetate (lb)
of Fusidienol
Position
60
X!2
Mult
6H1
8H2
2
155.16
154.90
co
---
-__
3
108.11
108.23
CH
6.95, dd, 9.0, 0.5
6.95, dd, 8.4, 0.8
4
136.94
137.04
CH
7.60, t, 8.5
7.60, t, 8.4
5 6
113.37 161.94
113.26 161.75
CH C”
6.82, dd, 9.0, 0.5 ---
6.81, dd, 8.4, 1.2 -__
7
110.28
110.19
co
---
-__
8
183.95
183.92
Co
---
___
9
105.80
105.72
C“
---
-_-
10
131.34
131.24
Co
---
-_-
11
134.64
134.50
CH
7.06, s
7.00, dd, 0.8, 0.4
12
131.31
131.10
co
---
--
13
145.75
145.13
CH
6.57, t, 1.5
6.52, td, 1.2, 0.4
15
164.42
163.98
Co
---
-__
16
60.72
60.63
CH2
4.14. d, 1.5
4.13, ddd, 6.0. 1.6, 0.4
17
168.5 1
167.57
Co
---
___
19
52.90
52.93
CH3
3.79, s
3.72, s
6_OH
___
__
___
___
12.13, s
16_OH
___
___
__
___
3.61, t, 6.0
lCD3OD
at 30Oc (500MHz); kD3CN-CD3COCD3 (2:1)at 25 OC(400 MHz).
REFERENCES I.
AND NOTES
(a) Gibbs. J. B. Semin. CCM Bid. 1992, 3, 383. (b) Gibbs, J. B. Cells
1991.65.1 and refenmces cited therein. (c) Der, C.
J. and COX, A. D. Colncer Cells 1991,3.331.(d) Reiss. Y.: Goldstein,
J. L.; Se&m,
Cells 1990, 62. 81. (e) Schabcr, M. D.; O’Ham, M. B.; Garsky, V. M.; Mosser. Marshall.
M. S.: Friedman,
P. A.; Dixon,
M. C.; Casey. P. J.; and Brown, M. S. S. D., Bergstrom,
J. D.; Moores,
S. L.;
R. A. F.; and Gibbs,J. B. J. Biof. Chem. 1990,265, 14701. (f) Gibbs, J. B.;
4696
Pompliano,
D. L.; Mosser, S. D.; Rands, E.; Lingham,
R. B.; Singh, S. B.; Scolnick,
E. M.; Kohl, N. B.; and Oliff, A. J.
Biol. Chem. 1993, 268, 7617. 2. 3.
(a) Barbacid,
Rev. B&hem. 1987. 56, ‘779.(b) Rodenhuis,
M. Ann
(a) Kohl, N. E.: MOW,
S. D.; debolms. J. S.; Giuliani.
S. Semin. Gun. Biol. 1992, 3, 241.
E. A.: Pompliano,
D. L.; Graham,
E. M.; Cliff, A.; and Gibbs, J. B. Science 1993, 260. 1934. (b) James, G. J.; Goldstein, E.; Somers.
T. C.; McDowell,
R. S.; Crowley,
C. W.; Lucas, B. K.; Levinson,
S. L.; Smith, R. L.; S&rick,
J. L.; Bmwn, M. S.; Rawson,
T.
A. D.; and Marsters. J. C. Science 1993.
260, 1937. 4.
(a) Singh, S. B.; Zink, D. L.; Liesch, J. M.; Goetz, M. A.; Jenkins, R. G.; Nallin-Gmstead, F.: MO&y, R. T.: Gibbs, J. B.: Albers-Schonberg, Singb, S. B. Terrhednrn M.; Jenkins,
Microbial. 5.
I&t. 1993, 34, 6521. (d) Linghsm.
M.; Mosser, S. D.; Schaber,
Biotech
Inokoshi. J.; Van Der Pyl, D.; Nakagawa, Van Dcr Pyl, D.; Inokoshi,
7.
Liu, W. C.; Barbacid,
8.
(a) Ham M.; Akasaka,
Y.; and Takesbima, A.; Takeshima,
K.; Akinaga,
S.; Okabe, M.; Nakano,
2281. (b) Tamanoi,
(a) Fuskiium
= ATCC 74265)
griseum (MF5695
by Dr. Bary Katz of MYCCkearch, and Wilkins
Co., Baltimore, Publishing
mp. 168-70%
Cl6Hl2O7:
North Carolina.
leaf litter, near Lansing,
(b) Barron, G. L. “The Genera
F. Proc.
Michigan,
USA)
of Hyphomycetesfrom
Soil”
on land Plants, an Identification
cited theirin. (b) Chuer, C. A.; Kral, A. M.; Diel, R. E.; Prendergast,
G. C.; Powers, S.;
1993, 32, 5167.
230 (&=19815),
327
(7360) nm; IR (ZnSe): 3500, 2953,
816, 766 cm-‘:
1724. 1651, 1610,
HREIMS (m/z): 316.0600
(M+, calcd. for
6): 2.04 (3H, s), 2.34 (3H, s), 3.79 (3H, s). 4.67 (ZH, s), 6.63 (HI, s), 7.05 (IH, d, J =
s). 7.30 (lH, d. J = 8.0 Hz), 7.64 (lH, t, J = 8.5 Hz). HREIMS
400.0794),
(calcd. for Cl7Hl007:
228.0423).
1992, 45.454.
316.0583).
8.0 Hz), 7.22 (lH,
284.0313
1802. P. B.; Huang,
H.; Gomez, R.; Wood, D.; Uh. M.; and Tamanoi,
1968. (c) Ellis, M. B.; Ellis, J. P. Microfungi
W: hmax (CH30H):
lb: ‘H NMR (CD30D+CDC13,
C2OHl609:
1992.45,
A. R.; Dean, L.; Doyle, T. W.; Fernandes,
F. Trend. Biochem. Sci. (TlBS) 1993, 18, 349.
1468, 1421, 1361. 1274. 1231, 1176, 1140. 1063, 1020.896,
12.
1993, 46, 229.
Co. New York 1985.
(a) See ref. If. and 4d and references
la:
1993, 46. 222. (b) Shiomi, K.; Yang, H.;
H.; and Chnura, S. J. Antibiotics
was isolated from hardwood
Allen, C. M.; Gibbs, J. B.; and Kohl, N. E. Biochemistry 11.
L.;
D. L.; Gibbs, J. B.; and Singh, S. B. Appl.
H.; and Gmura, S. J. Antibiotics
M.; Bulgar, M.; Clsrck, J. M.; Crosswell,
Hundbook ” Macmillan 10.
C.; Sanchez,
S.; Kong, L.; Burg, R. W.; Meinz, M. S.; Huang,
H. .I. Antibiotics
I.; Sbiomi, K.; Yang, H.; Takeshima,
Narf. Acad. Sci. USA 1993.90.
Williams
K. C.; Bills, G. F.; &scales,
M. D.; Omer, C. A.; Pompliano,
S.: Manne, V.: Pimik, D. M.; Wells, J. S.; and Meyers, E. J. Antibiotics
provided
K. C.; Bills, G.
1993, 40, 370.
(a) Omura, S.; Van Der Pyl, D.; Takahashi,
6.
9.
M.; Silverman,
R. B. Tetrahedron 1993, 49, 5917. (b) Ref. If. (c)
R. B.; Silverman,
S.; Martin, I.; Pelaaz, F.; Mochales,
R. G.; Gartner,
Nallin-Gmstead,
G.: and Lingham.
326.0427).
(calcd. for Cl5Hg07:
310.0485 (calcd. for Cl7HlOG7: 284.0321).
163.0403 (calcd. for CgH703:
(Received in USA 18 April
(m/z): 400.0799
358.0693 (calcd. for C1gH1408: 358.0689), 342.0743 (cakd. for ClgH1407:
1994,
266.0198
310.0477)
(calcd. for Cl5H605:
163.0395), 137.0241 (c&d.
accepted 6 May 1994)
(M+, calcd.
298.0484 (calcd. for Clr5HlOO6: 266.0215).
for C7H503:
228.0433
137.0239).
for
342.0740), 326.0430
(calcd.
298.0477).
for Cl3Hg04: