S
1
Supporting Information
for
DPAGT1 Inhibitors of Capuramycin Analogues
and Their
Antimi
gratory Activities of
Solid Tumors
Katsuhiko Mitachi,
a
Rita G. Kansal
,
b
Kirk E. Hevener
,
a
Cody D. Gillman,
c
Syed
M
.
Hus
s
ain
,
b
Hyun Gi Yun
,
c
Gustav
o
A.
Miranda
-
Carboni,
d
Evan S. Glazer,
b
William M.
Clemons Jr
.,
c
and Michio Kurosu*
a
a
Department
of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health
Science Center, 881 Madison
Av
, Memphis, TN 38163, U
SA
b
Department
of
Surgery and Center for Cancer Research, College of Pharmacy
, University of
Tennessee Health Science Cent
er,
9
10
Madison S
t
,
Suite 300,
Memphis, TN 38163, U
SA
c
Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 E.
California Bld. Pasadena, CA 91125, USA
d
Department
of
Medicine, Division of Hematology
-
Oncology,
Universit
y of Tennessee Health
Science Center,
19 S.
Manassas Av
, Memphis, TN 38163, U
SA
mkurosu@uthsc.edu
Table of Contents
General
-------------------------------------------------------------------------------------
S
2
Experimental
---------------------
----------------------------------------------------------
S
3
-
S
41
References
---------------------
----------------------------------------------------------
----
S
4
2
Copies of 1H and 13C
NMR
s
pectra
--
-----------
---------------------------------------
S
4
3
-
S
14
4
S
2
General
All chemicals were purchased from commercial sources and used without further
purification unless otherwise noted. THF, CH
2
Cl
2
, and DMF were purified via Innovative
Technology's Pure
-
Solve System.
All reactions were performed under an Argon
atmosphere. All stirring was performed with an internal magnetic stirrer. Reactions were
monitored by TLC using 0.25 mm coated commercial silica gel plates (EMD, Silica Gel
60F
254
). TLC spots were visualized by U
V light at 254 nm, or developed with ceric
ammonium molybdate or anisaldehyde or copper sulfate or ninhydrin solutions by heating
on a hot plate. Reactions were also monitored by using SHIMADZU LCMS
-
2020 with
solvents: A: 0.1% formic acid in water, B: acet
onitrile. Flash chromatography was
performed with SiliCycle silica gel (Purasil 60 Å, 230
-
400 Mesh). Proton magnetic
resonance (
1
H
-
NMR) spectral data were recorded on 400, and 500 MHz instruments.
Carbon magnetic resonance (
13
C
-
NMR) spectral data were reco
rded on 100 and 125 MHz
instruments. For all NMR spectra, chemical shifts (δH, δC) were quoted in parts per million
(ppm), and
J
values were quoted in Hz.
1
H and
13
C NMR spectra were calibrated with
residual undeuterated solvent (CDCl
3
: δH = 7.26 ppm, δC =
77.16 ppm; CD
3
CN: δH = 1.94
ppm, δC = 1.32ppm; CD
3
O
D: δH =3.31 ppm, δC =49.00 ppm;
DMSO
-
d
6
: δH = 2.50 ppm,
δC = 39.52 ppm; D
2
O: δH = 4.79 ppm) as an internal reference. The following
abbreviations were used to designate the multiplicities: s = singlet, d
= doublet, dd = double
doublets, t = triplet, q = quartet, quin = quintet, hept = heptet, m = multiplet, br = broad.
Infrared (IR) spectra were recorded on a Perkin
-
Elmer FT1600 spectrometer. HPLC
analyses were performed with a Shimadzu LC
-
20AD HPLC system
.
HR
-
MS data were
obtained from a Waters Synapt G2
-
Si (ion mobility mass spectrometer with
nanoelectrospray ionization).
S
3
O
N
N
B
O
M
O
O
A
c
O
O
C
H
3
H
O
S
1
(2
R
,5
R
)
-
5
-
(3
-
((B
enzyloxy)methyl)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
2
-
(hydroxymethyl)
-
4
-
methoxytetrahydrofuran
-
3
-
yl acetate
(S1).
The title compound
was
synthesized according to the reported procedure
1
:
1
H NMR (400 MHz, Chloroform
-
d
) δ
7.71 (d,
J
= 8.2 Hz, 1H), 7.40
–
7.26 (m, 5H), 5.77 (d,
J
= 6.4 Hz, 1H), 5.75 (d,
J
= 2.6 Hz,
1H), 5.48 (d,
J
= 2.2 Hz, 2H), 5.23 (t,
J
= 5.2 Hz, 1H), 4.71 (s, 2H), 4.23
–
4.17 (m, 2H),
3.99 (dd,
J
= 12.6, 2.0 Hz, 1H), 3.78 (dd,
J
= 12.6, 2.1 Hz, 1H), 3.48 (s, 3H), 2.17 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ 170.59, 162.49, 151.01, 139.77,
137.77, 128.31 (2C),
127.70, 127.67 (2C), 102.21, 90.31, 82.65, 81.20, 72.31, 70.27, 70.01, 61.24, 58.95, 20.81
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
20
H
25
N
2
O
8
[
M
+
H
]
421.1611
, found:
421.16
41
.
(2
R
,5
R
)
-
5
-
(3
-
((
B
enzyloxy)methyl
)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
2
-
((
S
)
-
cyano(hydroxy)methyl)
-
4
-
methoxytetrahydrofuran
-
3
-
yl acetate
(13).
To a
stirred
solution of
S1
(
1.7
2
g,
4.09
mmol) and dichloroacetic acid (
0.5
1
mL,
6.1
4
mmol) in a 10:1
mixture of CH
2
Cl
2
and DMSO (
18.4
mL) was added DIC (
1.28
mL,
8.18
mmol) at 0
°
C.
After
being
stirr
ed
for
2
h
, H
2
O (
0.16
mL), TMSCN (
1.0
2
mL,
8.18
mmol) and Ti(O
i
Pr)
4
(
2.4
2
mL,
8.18
mmol)
were added to the reaction solution. After being stirred for
8
h at r.t.,
the solution was concentrated
in vacuo
. The crude mixture was suspended to a 4:1 mixture
of AcOH and H
2
O (
50
mL)
. After being stirred for
13
h at r.t., the solution was concentrated
in vacuo
.
The
residue
was
quenched with aq.
NaHCO
3
, extracted wi
th
EtOAc
. The
combined organic extracts were dried over Na
2
SO
4
and concentrated
in vacuo
.
The crude
mixture was purified by silica gel column
chromatography (hexanes/EtOAc = 2/1
-
1/2)
to
afford
0.7
0
g (
38
%) of
13
and
0.66
g (
36
%) of
epi
-
13
.
Data for
13
:
1
H NMR (400 MHz,
Chloroform
-
d
) δ 7.39
–
7.27 (m, 5H), 7.21 (d,
J
= 8.1 Hz, 1H), 5.82 (d,
J
= 8.1 Hz, 1H),
5.49 (s, 2H), 5.38 (d,
J
= 6.9 Hz, 1H), 5.35 (dd,
J
= 5.7, 2.4 Hz, 1H), 4.71 (s, 2H), 4.71
–
4.69 (m, 1H), 4.54 (dd,
J
= 6.9, 5.7 Hz, 1H), 4.36 (t,
J
=
2.2 Hz, 1H), 3.39 (s, 3H), 2.19 (s,
3H);
13
C NMR (101 MHz, CDCl
3
) δ 170.20, 161.85, 151.38, 141.95, 137.51, 128.42 (2C),
127.84, 127.66 (2C), 117.13, 103.23, 95.16, 84.08, 78.39, 72.52, 70.47, 70.31, 61.79, 59.23,
20.71
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
21
H
24
N
3
O
8
[
M
+
H
]
446.1563
, found:
446.15
68
.
Data
for
(2
R
,5
R
)
-
5
-
(3
-
((
B
enzyloxy)methyl)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
2
-
((
R
)
-
cyano(hydroxy)methyl)
-
4
-
methoxytetrahydrofuran
-
3
-
yl acetate
(
epi
-
13)
:
1
H
NMR (400 MHz, Chloroform
-
d
) δ 7.53 (d,
J
= 8.1 Hz, 1H), 7.38
–
7.27 (m, 5H), 5.80 (d,
J
= 5.5 Hz, 1H), 5.78 (d,
J
= 8.2 Hz, 1H), 5.50
–
5.43 (m, 3H), 4.82 (d,
J
= 2.6 Hz, 1H),
4.70 (s, 2H), 4.37 (dd,
J
= 4.0, 2.6 Hz, 1H), 4.19 (t,
J
= 5.4 Hz,
1H), 3.44 (s, 3H), 2.19 (s,
3H);
13
C NMR (101 MHz,
CDCl
3
) δ 170.02, 162.37, 151.07, 139.71, 137.50, 128.38 (2C),
S
4
127.85, 127.70 (2C), 116.60, 102.71, 90.45, 82.72, 80.66, 72.43, 70.43, 69.12, 60.90, 59.37,
20.73
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
21
H
24
N
3
O
8
[
M
+
H
]
446.1563
, found:
446.15
80
.
S
T
o
l
O
O
A
c
A
c
O
O
A
c
A
c
O
S
2
(2
R
,3
R
,4
S
,5
S
,6
R
)
-
2
-
(
A
cetoxymethyl)
-
6
-
(
p
-
tolylthio)tetrahydro
-
2
H
-
pyran
-
3,4,5
-
triyl
triacetate
(S2).
The title compound was synthesized according to the reported procedure
1
:
1
H NMR (400 MHz, Chloroform
-
d
) δ 7.37 (d,
J
= 8.2 Hz, 2H), 7.12 (d,
J
= 7.9 Hz, 2H),
5.50
–
5.48 (m, 1H), 5.41 (d,
J
= 1.5 Hz, 1H), 5.32 (dd,
J
= 5.6, 1.7 Hz, 2H), 4.56 (dp,
J
=
8.1, 2.5 Hz, 1H), 4.30 (dd,
J
= 12.2, 5.9 Hz, 1H), 4.10 (dd,
J
= 12.2, 2.4 Hz, 1H), 2.33 (s,
3H), 2.14 (s, 3H), 2.07 (s,
3H), 2.06 (s, 3H), 2.01 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ
170.56, 169.92, 169.81, 169.74, 138.44, 132.58 (2C), 129.93 (2C), 128.72, 85.99, 70.82,
69.34, 69.32, 66.34, 62.46, 21.13, 20.89, 20.72, 20.70, 20.65
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
21
H
27
O
9
S
[
M
+
H
]
455.1376
, found:
455.1395.
S
T
o
l
O
O
A
c
A
c
O
O
H
A
c
O
S
T
o
l
O
O
A
c
A
c
O
O
A
c
A
c
O
[
t
B
u
2
S
n
(
O
H
)
C
l
]
2
T
H
F
-
M
e
O
H
S
2
S
3
(2
R
,3
R
,4
S
,5
S
,6
R
)
-
2
-
(
H
ydroxymethyl)
-
6
-
(
p
-
tolylthio)tetrahydro
-
2
H
-
pyran
-
3,4,5
-
triyl
triacetate
(S3)
.
To a
stirred
solution of
S2
(11.1 g, 24.3 mmol) in a 1
:4 mixture of THF
and MeOH (25
mL) was added [
t
Bu
2
Sn(OH)Cl]
2
(1.39
g, 2.4
3
mmol).
After
being
stirr
ed
for
24
h
at r.t.
, the solution was concentrated
in vacuo
.
The crude mixture was purified by
silica gel column chromatography (hexanes/EtOAc = 2/1
-
1/1)
to
afford 8
.
0
1
g (80%) of
S3
:
1
H
NMR (400 MHz, Chloroform
-
d
) δ 7.36 (d,
J
= 8.1 Hz, 2H), 7.12 (d,
J
= 8.0 Hz, 2H),
5.51 (dd,
J
= 3.3, 1.6 Hz, 1H), 5.42 (d,
J
= 1.5 Hz, 1H), 5.38 (dd,
J
= 10.1, 3.3 Hz, 1H),
5.29 (t,
J
= 10.0 Hz, 1H), 4.30 (ddd,
J
= 9.7, 4.2, 2.3 Hz, 1H), 3.69 (dd,
J
= 12.
7, 2.5 Hz,
1H), 3.63 (dd,
J
= 12.8, 4.2 Hz, 1H), 2.32 (s, 3H), 2.13 (s, 3H), 2.10 (s, 3H), 2.02 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ 170.78, 169.96, 169.81, 138.43, 132.58 (2C), 129.99 (2C),
128.73, 86.04, 71.62, 70.88, 69.10, 66.51, 61.20, 21.11, 20.87,
20.75, 20.66
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
19
H
25
O
8
S
[
M
+
H
]
413.1270
, found:
413.1277.
S
T
o
l
O
O
A
c
A
c
O
O
A
c
C
l
A
c
O
S
T
o
l
O
O
A
c
A
c
O
O
H
A
c
O
C
l
C
H
2
C
O
2
H
D
I
C
,
D
M
A
P
C
H
2
C
l
2
S
3
1
4
(2
R
,3
R
,4
S
,5
S
,6
R
)
-
2
-
((2
-
C
hloroacetoxy)methyl)
-
6
-
(
p
-
tolylthio)tetrahydro
-
2
H
-
pyran
-
3,4,5
-
triyl triacetate
(14).
To a
stirred
solution of
S3
(8.0
1
g, 19
.4 mmol), chloroacetic
acid (2.75
g, 29.1 mmol) and DMAP (0.2
4
g, 1.9
4
mmol) in CH
2
Cl
2
(50
mL) was added
S
5
DIC (4.
5
6 mL, 29.1 mmol). After
being
stirr
ed
for 4
h at r.t., the reaction mixture was
concentrated
in vacuo
.
The crude
mixture was purified
by silica gel column
chromatography (hexanes/EtOAc = 3/1
-
2/1)
to
afford
8.9
2
g (94%) of
14
:
1
H NMR (400
MHz, Chloroform
-
d
) δ 7.36 (d,
J
= 8.1 Hz, 2H), 7.13 (d,
J
= 8.0 Hz, 2H), 5.49 (d,
J
= 2.2
Hz, 1H), 5.42 (d,
J
= 1.6 Hz, 1H), 5.32
(d,
J
= 6.4 Hz, 2H), 4.59
–
4.54 (m, 1H), 4.40 (dd,
J
= 12.2, 5.9 Hz, 1H), 4.21 (dd,
J
= 12.1, 2.2 Hz, 1H), 4.03 (d,
J
= 1.1 Hz, 2H), 2.33 (s,
3H), 2.14 (s, 3H), 2.08 (s, 3H), 2.02 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ 169.88, 169.81,
169.78, 166.93, 138.51
, 132.48 (2C), 129.98 (2C), 128.56, 85.84, 70.70, 69.19, 69.17,
66.19, 63.95, 40.58, 21.13, 20.87, 20.71, 20.64;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
21
H
26
ClO
9
S
[
M
+
H
]
489.0986
, found:
489.1003
.
O
N
N
B
O
M
O
O
A
c
O
O
C
H
3
O
C
N
O
O
A
c
A
c
O
O
A
c
C
l
A
c
O
1
5
H
H
g
C
l
2
a
c
e
t
a
o
d
o
x
i
m
e
E
t
O
H
-
H
2
O
O
N
N
B
O
M
O
O
A
c
O
O
C
H
3
O
O
O
A
c
A
c
O
O
A
c
C
l
A
c
O
O
N
H
2
H
S
4
(2
R
,3
S
,4
S
,5
R
,6
R
)
-
2
-
((1
R
)
-
1
-
((2
S
,5
R
)
-
3
-
A
cetoxy
-
5
-
(3
-
((benzyloxy)methyl)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
4
-
methoxytetrahydrofuran
-
2
-
yl)
-
2
-
amino
-
2
-
oxoethoxy)
-
6
-
((2
-
chloroacetoxy)methyl)tetrahydro
-
2
H
-
pyran
-
3,4,5
-
triyl
triacetate
(S4).
To a stirred solution of
15
(
0.24
g,
0.29
m
mol
) in
a 9:1 mixture of
EtOH
and
H
2
O
(
2.9
mL) were added HgCl
2
(
0.16
g,
0.59
mmol) and acetaldoxime (
0.18
m
L,
2.9
mmol)
.
After being stirred for 13
h at r
.
t
.
,
the reaction mixture was
concentrated
in vacuo
. The
residue
was
quenched with aq.
NaHCO
3
, extracted
with
CHCl
3
. The combined organic
extracts were dried over Na
2
SO
4
and concentrated
in vacuo
. The crude product was purified
by silica gel column chromatography (
hexanes/EtOAc = 1/2
-
CHCl
3
/
MeOH
= 96/4
) to
afford
S4
(
0.21
g,
87
%):
1
H NMR (400 MHz,
Chloroform
-
d
) δ 7.53 (d,
J
= 8.2 Hz, 1H),
7.39
–
7.30 (m, 5H), 6.36 (
br
s, 1H
), 6.06 (d,
J
= 8.0 Hz, 1H), 5.97 (d,
J
= 4.1 Hz, 1H), 5.76
(
br
s, 1H), 5.53
–
5.44 (m, 2H), 5.40
–
5.27 (m, 2H), 5.19 (dd,
J
= 10.0, 3.3 Hz, 1H), 5.11
(t,
J
= 5.8 Hz, 1H), 4.99 (d,
J
= 2.0 Hz, 1H), 4.70 (s, 2H), 4.68 (d,
J
= 6.5 Hz, 1H), 4.53
(dd,
J
= 6.1, 2.2 Hz, 1H), 4.42 (d,
J
= 2.2 Hz, 1H), 4.37 (dd,
J
= 12.4, 5.3 Hz, 1H), 4.21
(dd,
J
= 12.3, 2.4 Hz, 1H), 4.15 (d,
J
= 2.4 Hz, 2H), 4.01 (dd,
J
= 5.6, 4.2 H
z, 1H), 3.46 (s,
3H), 2.18 (s, 3H), 2.17 (s,
3H), 2.07 (s, 3H), 2.04 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ
170.22, 170.18, 170.04, 169.67, 169.53, 167.06, 162.40, 150.91, 137.75, 137.58, 128.34,
128.32 (2C), 127.70 (2C), 103.35, 97.10, 88.44, 80.95, 76.39, 7
2.25, 70.51, 70.33, 69.76,
68.72, 68.68, 64.98, 63.43, 59.03, 40.65, 20.76, 20.67, 20.63 (2C)
;
H
RMS (E
S
I
+
)
m
/
z
calcd
for
C
35
H
43
ClN
3
O
18
[
M
+
H
]
828.2230
, found:
828.22
46
.
S
6
O
N
N
B
O
M
O
O
A
c
O
O
C
H
3
O
O
O
A
c
A
c
O
O
A
c
C
l
A
c
O
O
N
H
2
H
S
4
O
N
N
B
O
M
O
O
A
c
O
O
C
H
3
O
O
O
A
c
A
c
O
O
H
A
c
O
O
N
H
2
T
h
i
o
u
r
e
a
T
H
F
-
M
e
O
H
H
S
5
(2
R
,3
S
,4
S
,5
R
,6
R
)
-
2
-
((1
R
)
-
1
-
((2
S
,5
R
)
-
3
-
A
cetoxy
-
5
-
(3
-
((benzyloxy)methyl)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
4
-
methoxytetrahydrofuran
-
2
-
yl)
-
2
-
amino
-
2
-
oxoethoxy)
-
6
-
(hydroxymethyl)tetrahydro
-
2
H
-
pyran
-
3,4,5
-
triyl triacetate
(S5).
To a
solution of
S4
(0.21 g, 0.26
mmol) in
a
1:1 mixture of THF and MeOH (2.6
mL)
was added
thiourea (0.059
g,
0.77
mmol). After
being stirred for
11
h at 50
°
C , the reaction mixture
was concentrated
in vacuo
. The residue was quenched with H
2
O,
extracted with
CHCl
3
.
The combined organic extracts
were
dried over Na
2
SO
4
and concentrated
in vacuo
. The
crude product was purified by silica gel column chromatography (
CHCl
3
/
MeOH
= 98/2
-
97/3
-
96/4
) to afford
S5
(
0.15
g,
75
%):
1
H
NMR (400 MHz, Chloroform
-
d
) δ 7.59 (d,
J
=
8.2 Hz, 1H), 7.40
–
7.28 (m, 5H), 6.46 (brs, 1H), 6.10 (d,
J
= 8.2 Hz, 1H), 5.93 (d,
J
= 2.7
Hz, 1H), 5.88 (brs, 1H), 5.50 (d,
J
= 9.8 Hz, 1H), 5.46 (d,
J
= 9.5 Hz, 1H), 5.41 (dd,
J
=
3.3, 1.8 Hz, 1H), 5.28
–
5.22 (m, 1H), 5.17 (dd,
J
= 10.1, 3.3 Hz, 1H
), 5.01 (dd,
J
= 7.8,
5.5 Hz, 1H), 4.99 (d,
J
= 1.9 Hz, 1H), 4.70 (s, 2H), 4.58 (dd,
J
= 7.9, 2.2 Hz, 1H), 4.43 (d,
J
= 2.2 Hz, 1H), 4.03 (dd,
J
= 5.3, 2.6 Hz, 1H), 3.70 (ddd,
J
= 9.5, 4.7, 2.9 Hz, 1H), 3.60
(dd,
J
= 5.8, 3.9 Hz, 1H), 3.49 (s, 3H), 3.38 (d,
J
= 14.3 Hz, 1H), 2.16 (s, 3H), 2.15 (s, 3H),
2.09 (s, 3H), 2.04 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ 170.42, 170.38, 170.18, 170.04,
162.54, 150.77, 137.75, 137.36, 128.32 (2C), 127.73 (2C), 127.70, 103.06, 97.25
, 88.88,
81.14, 80.39, 75.76, 72.92, 72.26, 70.25, 69.52, 68.83, 68.75, 65.42, 61.25, 58.98, 20.78,
20.74, 20.66, 20.59
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
33
H
42
N
3
O
17
[
M
+
H
]
752.2514
, found:
752.25
22
.
O
N
N
H
O
O
H
O
O
H
H
O
O
N
N
B
O
M
O
O
H
O
O
O
C
H
3
H
3
C
2
)
2
,
2
-
d
i
m
e
t
h
o
x
y
p
r
o
p
a
n
e
T
s
O
H
.
H
2
O
,
a
c
e
t
o
n
e
1
)
B
O
M
C
l
,
D
M
F
S
6
3
-
((
B
enzyloxy)methyl)
-
1
-
((3a
R
,4
R
,6
R
,6a
R
)
-
6
-
(hydroxymethyl)
-
2,2
-
dimethyltetrahydrofuro[3,4
-
d
][1,3]dioxol
-
4
-
yl)pyrimidine
-
2,4(1
H
,3
H
)
-
dione
(S6).
To
a stirred solution of uridine (14.7
g, 60 mmol) in DMF (50 mL) were added BOMCl (6.87
mL, 50 mmol) and DBU (11.2 mL,
75 mmol). After being stirred fo
r 6
h at 0 °C,
the reaction
was quenched with
1M
aq
. HCl
, extracted with
CHCl
3
. The combined organic extracts
were dried over Na
2
SO
4
and concentrated
in vacuo
.
To a stirred solution of the crude
mixture and 2,2
-
dimethoxypropane (18.4 mL, 150 mmol) in acetone (50 mL) was
added
TsOH·H
2
O (0.95 g, 5.0 m
mol). After being stirred for 8
h at r.t.,
the solution was
concentrated
in vacuo
.
The
residue
was
quenched with aq.
NaHCO
3
, extracted with
EtOAc
.
The combined organic
extracts were dried over Na
2
SO
4
and concentrated
in vacuo
.
The
crude mixture was purified by silica gel column chromatography (hexanes/EtOAc = 1/2)
S
7
to
afford 15.8 g (78
%
for 2 steps
) of
S6
:
1
H NMR (400 MHz, Chloroform
-
d
) δ 7.37
–
7.26
(m, 6H), 5.73 (d,
J
= 8.1 Hz, 1H), 5.56 (d,
J
= 2.6 Hz, 1H), 5.48 (d,
J
= 9.7 Hz, 1H), 5.44
(d,
J
= 9.7 Hz, 1H), 4.97 (dd,
J
= 6.4, 2.7 Hz, 1H), 4.94 (dd,
J
= 9.5, 3.1 Hz, 1H), 4.69 (s,
2H), 4.29 (q,
J
= 3.0 Hz, 1H), 3.91 (dd,
J
= 12.0, 2.5 Hz, 1H), 3.79 (dd,
J
= 12.0, 3.5 H
z,
1H), 1.57 (s, 3H), 1.36 (s, 3H)
;
13
C NMR (101 MHz, CDCl
3
) δ 162.50, 151.00, 141.27,
137.70, 128.30 (2C), 127.70, 127.59 (2C), 114.25, 102.01, 96.49, 86.93, 83.83, 80.23,
72.36, 70.32, 62.63, 27.21, 25.22
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
20
H
25
N
2
O
7
[
M
+
H
]
405.1662
, found:
405.1681
.
(
S
)
-
2
-
((3a
R
,4
R
,6
R
,6a
R
)
-
6
-
(3
-
((
B
enzyloxy)methyl)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
2,2
-
dimethyltetrahydrofuro[3,4
-
d
][1,3]dioxol
-
4
-
yl)
-
2
-
hydroxyacetonitrile
(20).
To a
stirred
solution of
S
6
(
2.89
g,
7.15
mmol) and dichloroacetic acid (
0.88
mL,
10.7
mmol) in a 10:1 mixture of CH
2
Cl
2
and DMSO (
31.5
mL) was added DIC (
2.24
mL,
14.3
mmol) at 0
°
C.
After
being
stirr
ed
for
2
h
, H
2
O (0.29
mL), TMSCN (
1.35
mL,
14.3
mmol)
and Ti(O
i
Pr)
4
(
4.23
mL,
14.3
mmol) were added to the reaction solution. After being stirred
for
6
h at r.t., the solution was concentrated
in vacuo
. The crude mixture was suspended to
a 4:1 mixture of AcOH and H
2
O (
50
mL). After being stirred for
9
h at r.t.,
the solution
was
concentrated
in vacuo
.
The
residue
was
quenched with aq.
NaHCO
3
, extracted with
EtOAc
.
The combined organic extracts were dried over Na
2
SO
4
and concentrated
in vacuo
.
The
crude mixture was purified by silica gel column chromatography (hexanes/EtOAc = 2
/1
-
1/
1
)
to
afford
1.18
g (
38
%) of
20
and
1.03
g (
34
%) of
epi
-
20
.
Data for
20
:
1
H NMR (400
MHz, Chloroform
-
d
) δ 7.36
–
7.30 (m, 5H), 7.15 (d,
J
= 8.0 Hz, 1H), 5.80 (d,
J
= 8.1 Hz,
1H), 5.49 (d,
J
= 9.8 Hz, 1H), 5.44 (d,
J
= 9.8 Hz, 1H), 5.35 (d,
J
= 2.8 Hz, 1H), 5.14 (dd,
J
= 6.6, 2.8 Hz, 1H), 5.07 (dd,
J
= 6.6, 3.5 Hz, 1H), 4.72
–
4.68 (m, 3H), 4.40 (dd,
J
= 3.5,
2.3 Hz, 1H), 1.57 (s, 3H), 1.36 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ 161.91, 151.68,
142.29, 137.52, 128.41 (2C), 127.87, 127.58 (2C),
117.55, 115.20, 102.97, 99.49, 86.78,
82.89, 79.50, 72.60, 70.48, 62.12, 27.09, 25.10
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
21
H
24
N
3
O
7
[
M
+
H
]
430.1614
, found:
430.1630
. Data for
(
R
)
-
2
-
((3a
R
,4
R
,6
R
,6a
R
)
-
6
-
(3
-
((
B
enzyloxy)methyl)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
2,2
-
dimethyltetrahydrofuro[3,4
-
d
][1,3]dioxol
-
4
-
yl)
-
2
-
hydroxyacetonitrile
(
epi
-
20)
:
1
H
NMR (400 MHz, Chloroform
-
d
) δ 7.36
–
7.27 (m, 5H), 7.18 (d,
J
= 8.1 Hz, 1H), 5.77 (d,
J
= 8.0 Hz, 1H), 5.49
–
5.41 (m, 3H), 5.10 (dd,
J
= 6.6, 2.9 Hz, 1H), 5.07 (dd
,
J
= 6.5, 2.3
Hz, 1H), 4.76 (d,
J
= 5.3 Hz, 1H), 4.68 (s, 2H), 4.36 (dd,
J
= 5.3, 2.9 Hz, 1H), 1.58 (s, 3H),
1.38 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ 162.20, 151.16, 141.86, 137.43, 128.40 (2C),
127.89, 127.67 (2C), 117.41, 114.95, 102.67, 98.31, 87.29,
83.40, 80.68, 72.55, 70.42,
61.78, 27.02, 25.06
;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
21
H
24
N
3
O
7
[
M
+
H
]
430.1614
, found:
430.1633
.
S
8
O
N
N
B
O
M
O
O
O
C
N
O
O
A
c
A
c
O
O
A
c
C
l
A
c
O
H
H
g
C
l
2
a
c
e
t
a
o
d
o
x
i
m
e
E
t
O
H
-
H
2
O
O
N
N
B
O
M
O
O
O
O
O
A
c
A
c
O
O
A
c
C
l
A
c
O
O
N
H
2
H
O
O
O
O
C
H
3
C
H
3
H
3
C
H
3
C
2
1
S
7
(2
R
,3
S
,4
S
,5
R
,6
R
)
-
2
-
((
R
)
-
2
-
A
mino
-
1
-
((3a
R
,4
S
,6
R
,6a
R
)
-
6
-
(3
-
((benzyloxy)methyl)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
2,2
-
dimethyltetrahydrofuro[3,4
-
d
][1,3]dioxol
-
4
-
yl)
-
2
-
oxoethoxy)
-
6
-
((2
-
chloroacetoxy)methyl)tetrahydro
-
2
H
-
pyran
-
3,4,5
-
triyl
triacetate
(S7).
To a stirred solution
of
21
(
0.3
8
g,
0.49
m
mol)
in
a 9:1 mixture of
EtOH
and
H
2
O (
4.9
mL) were added HgCl
2
(
0.27
g,
0.98
mmol) and acetaldoxime (
0.30
m
L,
4.9
mmol)
.
After being stirred for
12
h at r
.
t
.
,
the reaction mixture was
concentrated
in vacuo
.
The
residue
was
quenched with aq.
NaHCO
3
, extracted with
CHCl
3
. The combined organic
extracts were dried over Na
2
SO
4
and concentrated
in vacuo
. The crude product was purified
by silica gel column chromatography (
hexanes/EtOAc = 1/2
-
CHCl
3
/
MeOH
= 9
7
/
3
) to
afford
S7
(
0.36
g,
91
%):
1
H NMR (400
MHz, Chloroform
-
d
) δ 7.38
–
7.27 (m, 6H), 6.41
(brs, 1H), 5.86 (d,
J
= 8.1 Hz, 1H), 5.81 (brs, 1H), 5.70 (d,
J
= 2.1 Hz, 1H), 5.49 (d,
J
= 9.9
Hz, 1H), 5.46 (d,
J
= 9.8 Hz, 1H), 5.33
–
5.26 (m, 2H), 4.98 (dd,
J
= 6.3, 4.9 Hz, 1H), 4.93
–
4.91 (m, 1H), 4.89
(dd,
J
= 6.3, 2.0 Hz, 1H), 4.69 (s, 2H), 4.41 (d,
J
= 4.2 Hz, 1H), 4.33
(t,
J
= 4.6 Hz, 1H), 4.28 (dd,
J
= 12.1, 5.3 Hz, 1H), 4.18 (d,
J
= 2.5 Hz, 1H), 4.16
–
4.11
(m, 2H), 4.10 (s, 2H), 2.14 (s, 3H), 2.05 (s, 3H), 2.00 (s, 3H), 1.57 (s, 3H), 1.37 (s, 3H)
;
13
C NMR (101 MHz, CDCl
3
) δ 170.19, 170.09, 170.01, 169.63, 166.94, 162.40, 150.62,
140.32, 137.80, 128.29 (2C), 127.69, 127.65 (2C), 115.00, 102.60, 97.38, 93.65, 85.88,
84.08, 79.80, 77.63, 72.31, 70.33, 69.80, 69.05, 68.60, 65.36, 63.41, 40.56, 27.32,
25.55,
20.77, 20.67, 20.63;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
35
H
43
ClN
3
O
17
[
M
+
H
]
812.2281
, found:
812.2314
.
O
N
N
B
O
M
O
O
O
O
O
A
c
A
c
O
O
A
c
C
l
A
c
O
O
N
H
2
H
O
N
N
B
O
M
O
O
O
O
O
A
c
A
c
O
O
H
A
c
O
O
N
H
2
T
h
i
o
u
r
e
a
T
H
F
-
M
e
O
H
H
O
O
O
O
C
H
3
C
H
3
H
3
C
H
3
C
S
7
S
8
(2
R
,3
S
,4
S
,5
R
,6
R
)
-
2
-
((
R
)
-
2
-
A
mino
-
1
-
((3a
R
,4
S
,6
R
,6a
R
)
-
6
-
(3
-
((benzyloxy)methyl)
-
2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
2,2
-
dimethyltetrahydrofuro[3,4
-
d
][1,3]dioxol
-
4
-
yl)
-
2
-
oxoethoxy)
-
6
-
(hydroxymethyl)tetrahydro
-
2
H
-
pyran
-
3,4,5
-
triyl triacetate
(S8).
To a solution of
S
7
(
0.
36
g,
0.
45
mmol) in
a 1:1 mixture of THF and MeOH (
4.5
mL)
was
added thiourea (
0.10
g,
1.34
mmol
). After
being stirred for
11
h at 50
°
C
, the reaction
mixture was concentrated
in vacuo
. The residue was quenched with H
2
O,
extracted with
CHCl
3
. The combined
organic extracts were dried over Na
2
SO
4
and concentrated
in vacuo
.
The crude product was purified
by silica gel column chromatography (
CHCl
3
/
MeOH
=
98/2
-
97/3
-
96/4
) to afford
S8
(
0.25
g,
76
%):
1
H NMR (400 MHz, Chloroform
-
d
) δ 7.39
S
9
–
7.27 (m, 6H), 6.43 (brs, 1H), 5.83 (d,
J
= 8.1 Hz, 1H), 5.74 (brs, 1H), 5.68 (d,
J
=
2.1 Hz,
1H), 5.46 (s, 2H), 5.33
–
5.27 (m, 2H),
5.18 (t,
J
= 9.9 Hz, 1H), 5.08 (dd,
J
= 6.5, 4.5 Hz,
1H), 4.92
–
4.88 (m, 2H), 4.70 (s, 2H), 4.43 (d,
J
= 5.7 Hz, 1H), 4.33 (dd,
J
= 5.7, 4.5 Hz,
1H), 3.89 (dt,
J
= 10.0, 3.8 Hz, 1H), 3.50 (d,
J
= 3.9 Hz, 2H), 2.14 (s, 3H), 2.05 (s, 3H),
2.01 (s, 3H), 1.56 (s, 3H), 1.35 (s, 3H);
13
C NMR (101 MHz, CDCl
3
) δ 170.35, 170.16,
170.10, 170.09, 162.46, 150.72, 140.56, 137.69, 128.33 (2C), 127.74, 127.69 (2C), 114.84,
102.50, 97.03, 94.40, 86.72, 84.25, 8
0.13, 77.92, 72.35, 72.04, 70.23, 69.20, 68.59, 65.87,
61.25, 27.23, 25.41, 20.81, 20.73, 20.69;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
33
H
42
N
3
O
16
[
M
+
H
]
736.2565
, found:
736.2586.
O
N
N
H
O
O
O
O
O
A
c
A
c
O
O
H
A
c
O
O
N
H
2
H
i
)
S
O
3
.
p
y
,
E
t
3
N
D
M
S
O
,
C
H
2
C
l
2
i
i
)
N
a
C
l
O
2
N
a
H
2
P
O
4
.
2
H
2
O
2
-
m
e
t
h
y
l
-
2
-
b
u
t
e
n
e
t
B
u
O
H
,
H
2
O
O
N
N
H
O
O
O
N
H
2
O
H
O
O
A
c
A
c
O
O
H
O
O
O
O
O
C
H
3
C
H
3
H
3
C
H
3
C
2
2
3
1
(2
S
,3
S
,4
S
)
-
3,4
-
D
iacetoxy
-
2
-
((
R
)
-
2
-
amino
-
1
-
((3a
R
,4
S
,6
R
,6a
R
)
-
6
-
(2,4
-
dioxo
-
3,4
-
dihydropyrimidin
-
1(2
H
)
-
yl)
-
2,2
-
dimethyltetrahydrofuro[3,4
-
d
][1,3]dioxol
-
4
-
yl)
-
2
-
oxoethoxy)
-
3,4
-
dihydro
-
2
H
-
pyran
-
6
-
carboxylic acid
(31).
To a stirred solution
of
22
(
0.17
g,
0.27
mmol) and DMSO (
0.19
mL,
2.72
mmol) in a 5:1 mixt
ure of CH
2
Cl
2
and
Et
3
N (1.4 mL)
was added SO
3
·pyridine (
0.43
g,
2.72
mmol). After being stirred for
3
h at
r.t.,
the reaction mixture was added
H
2
O
(
0.27
mL) and passed through a silica gel pad
(
CHCl
3
/
MeOH =
92
/
8
).
To a stirred solution of the crude mixture in
t
BuOH (
1.0
mL) and
2
-
methyl
-
2
-
butene (
0.5
mL) was added a solution of NaClO
2
(
0.12
g,
1.36
mmol) and
NaH
2
PO
4
·2H
2
O (
0.21
g,
1.36
mmol) in H
2
O (
1.0
mL). After being stirred for
5
h at r.t., the
reaction extracted
with
CHCl
3
/MeOH (9/1).
The combined organic extracts were dried
over Na
2
SO
4
and concentrated
in vacuo
. The crude
product was purified by silica gel
column chromatography (
CHCl
3
/
MeOH
= 9/1
) to afford
31
(
0.13
g,
81
%):
1
H NMR (400
MHz, Methanol
-
d
4
) δ 7.88 (d,
J
= 8.1 Hz, 1H), 5.88 (d,
J
= 3.6 Hz, 1H), 5.84 (d,
J
= 8.1
Hz, 1H), 5.80 (dd,
J
= 2.7, 1.5 Hz, 1H), 5.53 (dd,
J
= 4.5, 2.5 Hz, 1H),
5.41 (ddd,
J
= 4.7,
3.4, 1.6 Hz, 1H), 5.28 (d,
J
= 3.3 Hz, 1H), 4.84 (d,
J
= 1.9 Hz, 1H), 4.77 (dd,
J
= 6.2, 2
.1
Hz, 1H), 4.73 (d,
J
= 2.1 Hz, 1H), 4.62 (dd,
J
= 6.1, 3.7 Hz, 1H), 2.07 (s, 3H), 2.03 (s, 3H),
1.53 (s, 3H), 1.32 (s, 3H);
13
C NMR (101 MHz, MeOD) δ 172.67, 171.86, 171.50, 168.09,
166.27, 152.17, 148.19, 142.21, 115.19, 104.36, 103.01, 98.08, 93.58, 87
.18, 86.07, 82.82,
78.22, 65.27, 65.01, 27.45, 25.47, 20.69, 20.57;
H
RMS (E
S
I
+
)
m
/
z
calcd for
C
23
H
28
N
3
O
14
[
M
+
H
]
570.1571
, found:
570.1585.
O
N
N
B
O
M
O
O
H
3
C
O
O
A
c
H
O
S
9