1462
A. Hirose et al. / Tetrahedron 70 (2014) 1458e1463
4.3. Calculations
2a4H-3, H-34H3-13, H-34H3-15, H-54H3-134H3-14, H-
6a
4H3-13, H-6b4H3-14, H-6b4H3-15, H-7a4H9, H-7b4H12
Calculations were performed with Spartan 10 (Wavefunction,
Irvine, CA) using PC (operating system: Windows7 Professional,
CPU: AMD Phenom(tm)ꢂ4 970 processor 3.50 GHz, RAM 32 GB).
Model X was subjected to conformational analyses with AM1 by
rotating C3eO, C9eC11, and C11eC60 linkages to provide 18 ten-
tative stable conformers. These were further optimized with den-
sity functional EDF2 employing 6-31G* basis set. Chemical shift
calculations were performed also with EDF2/6-31G* based on the
optimized conformations. Theoretical chemical shifts were ob-
tained by considering Boltzmann distribution. Similar calculations
were also performed for its C50,C60-diastereomer.
(4.81 ppm), H-94H-50, H-114H-12 (4.48 ppm), H-12 (4.48 ppm)4
H50, H-204H-70 (5.07 ppm), H-404Hꢁ50, H-2004H-400, H-3004H-400.
4.4.3. Neomacrophorin III (3). The 1H and 13C NMR data are shown
25
in Fig. 1. [
a
]
ꢁ150 (c 0.5.8, CHCl3), IR (film) 3340, 2965, 1727,
D
1677 cmꢁ1. UV (8.0ꢂ10ꢁ5 mol/L, CH3CN,
3
) l 239 (sh, 6000), ECD
(8.0ꢂ10ꢁ5 mol/L, CH3CN)
D
3
ꢁ3.2 (339 nm), ꢁ0.19 (286 nm), ꢁ6.1
(243 nm), þ8.23 (215 nm), þ1.54 (200 nm), ESIMS m/z 467.2723
(calcd for C26H39O7, MþH: 463.2696), HMBC: H-1
H-1 /C-2, C-10, H-2 /C-1, C-3, H-2 /C-1, H-3/C-2, C-4, C-14,
H-5/C-10, C-15, H-6 /C-5, C-7, H-7 /C-8, H-7 /C-5, C-6, C-8,
a/C-2 and C-10,
b
a
b
b
a
b
C-9, C-12, H-9/C-8, C-10, C-11, C-60, H-11/C-10, C-10, C-50, C-60,
each H-12 (4.43, 4.83 ppm)/C-7, C-8, C-9, C-10, H3-13/C-3, C-4, C-
5, C-14, H3-14/C-4, C-5, C-13, H3-15/C-1, C-5, C-9, C-10, H-20/C-
60, C-30, C-70, H-40/C-30, C-50, C-60, H-50/C-11, C-30, C-40, C-60, H-70
(4.80 ppm)/C-20, C-30, C-40 C-100, H-70 (5.07 ppm)/C-20,,C-30, C-40,
C-100, H-200(2.50 ppm)/C-100, C-300, C-400, H-200(2.57 ppm)/C-100, C-
300, C-400, H-300/C-100, C-200, C-400, H3-400/C-200, C-300.
4.4. Isolation
Trichoderma sp. 1212-03 was cultured in potatoedextrose me-
dium (200 mL in 500 mL baffled Erlenmeyer flaskꢂ5) on a rotary
shaker (110 rpm) at 26 ꢀC for 14 days. After filtration in suction, the
filtrate was extracted with EtOAc (300 mLꢂ4) and the organic layer
was dried over MgSO4 and concentrated under reduced pressure to
give crude extract (235 mg). That was purified with silica gel col-
umn chromatography. The fraction eluted with acetone/CHCl3
(10:90) was recovered and concentrated to give a residue (80.3 mg).
Further silica gel chromatography [AcOEt/hexane¼30:70, then
50:50] gave neomacrophorin I (1, 14.3 mg), II (2, 39.5 mg), III (3,
4.5. Benzoylation of (1)
A solution of 1 (5.0 mg, 11
mmol) in CH2Cl2 (500
mL) and pyridine
(8.0 L) was stirred with benzoyl chloride (6.0 mg, 43
m
mmol) at
room temperature for 100 min. Et2O (2.0 mL) and saturated aque-
ous CuSO4 solution (1.0 mL) were added and the resulting mixture
was stirred vigorously for 1 min. After aqueous solution was re-
moved by pipetting, MgSO4 was added. The mixture was filtered
and concentrated in vacuo. Purification with silica gel column
chromatography (AcOEt/benzene¼20:80) gave dibenzoate 1a
(3.0 mg) and monobenzoate 1b (3.0 mg) both as oil. The amounts of
7.2 mg), and trichodermin (20.4 mg). The 1H NMR spectra and the
25
specific rotation value of trichodermin ([
a
]
D
ꢁ11 (c 0.87, CHCl3))
20
showed good accordance with the data in literature ([a]
D
ꢁ10.2).6,29
4.4.1. Neomacrophorin I (1). The 1H and 13C NMR data are shown in
Fig. 1. [
a
]
25 ꢁ53 (c 0.58, MeOH), IR (film) 3440, 1745, 1687 cmꢁ1. UV
1a and 1b were estimated based on UV absorption ( 15,000,
3
(8.0ꢂ10ꢁD5 mol/L, CH3CN,
)
l
218 (8800), 260 (sh, 2800), ECD
230 nm) for a benzoyl group.
(8.0ꢂ10ꢁ5 mol/L, CH3CN)
D
3
3 ꢁ0.64 (386 nm), þ2.9 (261 nm), ꢁ2.2
(238 nm), ꢁ0.16 (228 nm), ꢁ5.04 (214 nm), ꢁ2.8 (204 nm), ESIMS m/
4.5.1. Physical data of 1a. 1H NMR (CDCl3)
d 0.74, 0.84, 0.97 (each
z 477.2493 (calcd for C26H37O8, MþH: 477.2488), HMBC: H-1
C-10, H-1 /C-2, C-10, H-2 /C-1, C-3, H-2 /C-1, H-3/C-2, C-4,
C-14, H-5/C-10, C-15, H-6 /C-5, C-7, H-7 /C-8, H-7 /C-5, C-6,
a/C-2,
3H, s), 1.55 (3H, d, J¼6.5 Hz), 1.96 (2H, m), 2.24 (1H, dd, J¼2.4,
16.2 Hz), 2.30 (1H, dd, J¼10.6, 16.2 Hz), 2.39 (1H, ddd, J¼2.4, 4.2,
13.0 Hz), 3.43 (1H, s), 3.44 (1H, t, J¼2.8 Hz), 4.32 (1H, br s), 4.79 (1H,
br s), 4.88 (1H, dd, J¼1.9, 16.5 Hz), 4.98 (1H, dd, J¼1.9, 16.5 Hz), 5.49
(1H, d, J¼3.4 Hz), 5.77 (1H, dq, J¼3.4, 6.5 Hz), 6.54 (1H, t, J¼1.9 Hz),
7.45, 7.50 (each 2H, t, J¼7.8 Hz), 7.58, 7.63 (each 1H, t, J¼7.8 Hz),
8.02, 8.14 (each 2H, dd, J¼1.0, 7.8 Hz). ESIMS (rel int. %) m/
z¼707.2814 (10, C40H44NaO10 [MþNa]þ: 707.2832), 702.3248 (100,
calcd for C40H48NO10 [MþNH4]þ: 702.3278), 667.2872 (10,
b
a
b
b
a
b
C-8, C-9, C-12, H-9/C-8, C-10, C-11, C-60, H-11/C-10, C-10, C-60,
each H-12 (4.37, 5.81 ppm)/C-7, C-8, C-9, C-10, H3-13/C-3, C-4, C-
5, C-14, H3-14/C-4, C-5, C-13, H3-15/C-1, C-5, C-9, C-10, H-20/C-
11, C-60, C-30, C-70, H-50/C-11, C-30, C-40, C-70, H-70 (4.89 ppm)/C-20,
C-30, C-40 C-100, H-70 (5.11 ppm)/C-20, C-40, C-100, H-200/C-100, C-300,
C-400, H-300/CeC100, C200, C400, H3eC400/C-200, C-300, NOE: H-
1
b
4H3ꢁ15, H-2
b
4Hꢁ3, H-2
b
4H3ꢁ14, H-2
a
4Hꢁ3, H-34H3-13,
C
40H43O9 [MꢁOH]þ: 667.2907).
H-34H3-15, H-54H3-134H3-14, H-6
a
4H3-13, H-6
b
4H3-14, H-
6b
4H3-15, H-7
a
4H9, H-7
b
4H12 (4.81 ppm), H-94H-50, H-
4.5.2. Physical data of 1b. 1H NMR (CDCl3)
d 0.74, 0.84, 0.97 (each
114H-12 (4.37 ppm), H-12 (4.37 ppm)4H50, H-204H-70
(5.11 ppm), H-2004H-400, H-3004H-400.
3H, s), 1.40 (3H, d, J¼6.6 Hz), 1.92 (1H, ddd, J¼2.1, 4.2, 14.1 Hz), 2.00
(2H, m), 2.28 (1H, dd, J¼2.6, 16.1 Hz), 2.33 (1H, dd, J¼10.4, 16.1 Hz),
2.40 (1H, ddd, J¼2.0, 3.7, 13.0 Hz), 3.44 (1H, t, J¼2.7 Hz), 3.50 (1H, s),
4.35 (1H, br s), 4.46 (1H, br dq, J¼3.2, 6.6 Hz), 4.80 (1H, br s), 4.90
(1H, dd, J¼1.9, 16.6 Hz), 5.14 (1H, dd, J¼1.9, 16.6 Hz), 5.28 (1H, d,
J¼3.2 Hz), 6.25 (1H, t, J¼1.9 Hz), 7.49 (2H, t, J¼7.8 Hz), 7.63 (1H, t,
J¼7.8 Hz), 8.11 (2H, dd, J¼1.0, 7.8 Hz). ESIMS (rel int. %) m/
z¼563.2624 (30, C33H39O8 [MꢁOHꢁ]þ: 563.2645), 598.2991 (100,
4.4.2. Neomacrophorin II (2). The 1H and 13C NMR data are shown in
Fig. 1. [
a
]
25 ꢁ33 (c 0.24, MeOH), IR (film) 3440, 1733, 1683 cmꢁ1. UV
(8.0ꢂ10ꢁD5 mol/L, CH3CN,
3 )
l
284 (sh, 1000), ECD (8.3ꢂ10ꢁ5 mol/L,
CH3CN)
D
3
ꢁ3.2 (338 nm), ꢁ0.36 (282 nm), ꢁ5.4 (243 nm), þ4.3
(215 nm), þ2.0 (200 nm), ESIMS m/z 479.2645 (calcd for C26H39O8,
/C-2, C-10, H-1 /C-2, C-10, H-
/C-1, H-3/C-2, C-4, C-14, H-5/C-10, C-15, H-
/C-8, H-7 /C-5, C-6, C-8, C-9, C-12, H-9/C-8,
MþH: 479.2645), HMBC: H-1
a
b
C
33H44NO9 [MþNH4]þ: 598.3016), 603.2562 (10, C33H40NaO9
2a/C-1, C-3, H-2
b
[MþNa]þ: 603.2570).
6b/C-5, C-7, H-7
a
b
C-10, C-11, C-60, H-11/C-10, C-10, C-60, each H-12 (4.48,
4.81 ppm)/C-7, C-8, C-9, C-10, H3-13/C-3, C-4, C-5, C-14, H3-
14/C-4, C-5, C-13, H3-15/C-1, C-5, C-9, C-10, H-20/C-60, C-30, C-
40, H-40/C-10, C-20, C-30, C-50, C-60, C-70, H-50/C-11, C-30, C-400, C-60,
H-70 (4.76 ppm)/C-20, C-30, C-40 C-100, H-70 (5.07 ppm)/C-2 ,,C-30,
C-40, C-100, H-200/C-100, C-300, C-400, H-300/C-100, C-200, C-400, H3-
4.6. Chiral investigation of the C100eC400 moiety of 3
A solution of 3 (0.5 mg) in MeOH was stirred with NaOMe
(0.2 mg) at room temperature. After consumption of the starting 3
based on TLC analysis, acetic acid (1.0 mL) was added to neutralize
the solution. The mixture was sampled (0.1 mL) and diluted with
MeOH, which was subjected to gas chromatography equipped with
400/C-200, C-300, NOE: H-1
b4H3-15, H-2b4H-3, H-2b4H3-14, H-