Full Papers
doi.org/10.1002/cmdc.202000801
ChemMedChem
Procedure for Grubbs metathesis and hydrogenolysis/hydro-
genation reactions to generate library compounds 1a–1ab.
Toluene (1.00 mL, purged with N2 for one h) was added to Grubbs
second-generation catalyst (1.00 mg, 1.25 μmol). The purple
solution was transferred to a flask containing a mixture of 103
(2.50 mg, 4.01 μmol) and olefin (80.0 μmol). The resulting mixture
was stirred for 3 h before being concentrated to a crude oil. The oil
was purified using pipette column chromatography (25–50% EtOAc
in hexanes) to separate the Grubbs catalyst and the olefin dimer
from the cross metathesis product. The resulting purified oil was
dissolved in 1 mL of CH2Cl2 and 5% Pd/C (2.00 mg) was added to
the solution. The reaction mixture was vigorously stirred under
1 atm of H2 gas for 15 min before being evacuated and purged
with nitrogen gas. The Pd/C was removed by vacuum filtration
through a pad of celite using CH2Cl2 as an eluent. The filtrate was
concentrated to give the anhydride analogue. The library analogues
2.52 (dd, J=17.0, 3.2, 1H), 2.43 (d, J=1.1, 1H), 2.28 (q, J=7.0, 2H),
2.07 (s, 3H), 1.94 (quint, J=7.1, 2H), 1.70 (td, J=8.0, 2.3, 1H), 1.64 (s,
3H), 1.31 (d, J=6.3, 3H), 1.05 (d, J=7.1, 3H), 0.97 (d, J=6.0, 3H); 13C
NMR (CDCl3, 125 MHz): δ=214.8, 170.7, 167.3, 167.1, 166.6, 145.8,
140.0, 135.2, 133.2, 131.9, 131.2, 130.1, 129.1, 129.0, 128.6, 128.4,
128.2, 127.3, 127.1, 75.4, 73.2, 67.3, 66.5, 66.2, 64.2, 52.2, 47.3, 42.8,
40.8, 28.7, 28.2, 18.3, 15.1, 13.8, 9.9; IR (thin film): 3479 (br), 2933,
2860, 1767, 1713, 1383, 1279, 1115 cmÀ 1; HRMS (ESI): m/z calcd for
C50H56O12 [M+Na]+ 871.3669; found: 871.3655.
1
2
3
4
5
6
7
8
9
(6S,7S,10R,11S,12R)-6,10-Dihydroxy-12-(((R)-3-hydroxy-3-(4-meth-
yl-2,5-dioxo-2,5-dihydrofuran-3-yl)propanoyl)oxy)-7,11-dimethyl-
8-oxotridecyl benzoate (1d). To a solution of 1-((2R,3S,4R,7S,8S,E)-
13-(benzoyloxy)-4,8-dihydroxy-3,7-dimethyl-6-oxotridec-9-en-2-yl)
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
3,4-dibenzyl
(R,Z)-2-hydroxypent-3-ene-1,3,4-tricarboxylate
(3.55 mg, 4.60 μmol) in 5 mL of CH2Cl2 was added 5% Pd/C
(10.0 mg). The reaction mixture was vigorously stirred under 1 atm
of H2 gas for 15 min before being evacuated and purged with
nitrogen gas. The Pd/C was removed by vacuum filtration through
a pad of celite using CH2Cl2 as an eluent. The filtrate was
concentrated to yield 2.31 mg of the anhydride analogue 1d (88%),
which was sufficiently pure for the biological assays: 1H NMR (CDCl3,
500 MHz): δ=8.09 (d, J=8.0, 2H), 7.61 (t, J=8.0, 1H), 7.49 (t, J=8.0,
2H), 5.08 (quint, J=6.2, 1H), 4.37–4.41 (m, 4H), 3.80 (t, J=7.9, 1H),
3.03–3.20 (br s, 1H), 2.93 (dd, J=16.3, 3.5, 1H), 2.80–2.88 (m, 2H),
2.69 (quint, J=7.2, 1H), 2.50 (dd, J=13.2, 2.5, 2H), 3.32 (s, 3H), 1.84
(quint, J=5.6, 2H), 1.74 (dt, J=7.3, 2.0, 1H), 1.41–1.70 (m, 9H), 1.36
(d, J=6.3, 3H), 1.14 (d, J=7.1, 3H), 0.98 (d, J=7.1, 3H); 13C NMR
(CDCl3, 125 MHz): δ=215.9, 170.1, 166.8, 165.7, 164.9, 143.1, 142.1,
133.0, 130.4, 126.6, 128.4, 73.6, 66.7, 64.9, 63.9, 52.7, 46.7, 42.7, 40.7,
34.4, 29.8, 28.8, 26.0, 25.0, 18.5, 13.7, 10.4, 10.2; IR (thin film): 3462
(br), 2976, 2937, 1767, 1714, 1277 cmÀ 1; HRMS (ESI): m/z calcd for
C30H40O11 [M+Na]+ 599.2468; found: 599.2485.
1
were characterized and tested for purity by LC–MS, as H NMR and
13C NMR data were difficult to extrapolate due to complex mixtures
of the anhydride and diacid forms that are formed with exposure to
moisture.
1-((2R,3S,4R,7S,8S,E)-13-(Benzoyloxy)-4,8-dihydroxy-3,7-dimethyl-
6-oxotridec-9-en-2-yl) 3,4-dibenzyl (R,Z)-2-hydroxypent-3-ene-
1,3,4-tricarboxylate. Toluene (5.00 mL, purged with N2 for one h)
was added to Grubbs second-generation catalyst (5.00 mg, 6.25
μmol). The purple solution was transferred to a flask containing a
mixture of 15 (12.5 mg, 20.0 μmol) and 3d (76.1 mg, 400 μmol). The
resulting mixture was stirred for 3 h before being concentrated to a
crude oil. The oil was purified using column chromatography (25–
50% EtOAc in hexanes) to furnish 14.2 mg of 1-((2R,3S,4R,7S,8S,E)-
13-(benzoyloxy)-4,8-dihydroxy-3,7-dimethyl-6-oxotridec-9-en-2-yl)
3,4-dibenzyl (R,Z)-2-hydroxypent-3-ene-1,3,4-tricarboxylate (92%):
1H NMR (CDCl3, 500 MHz): δ=8.08 (d, J=7.0, 2H), 7.49 (t, J=7.0,
2H), 7.30–7.40 (m, 10H), 5.79 (dt, J=15.2, 6.6, 1H), 5.51 (dd, J=15.2,
7.6, 1H), 5.51 (dd, J=15.2, 7.6, 1H), 5.09–5.20 (m, 2H), 5.05–5.09 (m,
2H), 5.03 (dd, J=7.8, 6.4, 1H), 4.37 (dt, J=6.4, 2.2, 1H), 4.21 (t, J=
8.1, 1H), 3.73 (d, J=5.1, 1H), 3.05 (br s, 1H), 2.99 (dd, J=16.0, 10.3,
1H), 2.82 (dd, J=16.9, 9.6, 1H), 2.68 (quint, J=7.0, 1H), 2.64 (dd, J=
6.1, 3.5, 1H), 2.52 (dd, J=16.5, 2.8, 1H), 2.43 (br s, 1H), 2.27 (q, J 7.1,
1H), 2.08 (s, 3H), 1.91 (quint, 7.5, 2H), 1.71 (dt, J=7.1, 1.8, 1H), 1.31
(d, J=7.1, 3H), 0.97 (d, J=7.0, 3H); 13C NMR (CDCl3, 125 MHz): δ=
214.8, 170.7 167.3, 167.1, 166.7, 140.0, 135.2, 133.2, 133.0, 131.8,
131.2, 130.3, 129.6, 128.6, 128.5, 128.4, 75.3, 73.2, 67.3, 66.5, 66.2,
64.2, 52.2, 47.3, 42.8, 40.9, 28.7, 28.1, 22.7, 18.3, 15.1, 14.2, 13.5, 9.9;
IR (thin film): 3454 (br), 3035, 2958, 1714, 1454, 1275, 1171,
1070 cmÀ 1; HRMS (ESI): m/z calcd for C44H52O12 [M+Na]+ 795.356;
found: 795.3343.
(6S,7S,10R,11S,12R)-6,10-Dihydroxy-12-(((R)-3-hydroxy-3-(4-meth-
yl-2,5-dioxo-2,5-dihydrofuran-3-yl)propanoyl)oxy)-7,11-dimethyl-
8-oxotridecyl [1,1’-biphenyl]-4-carboxylate (1e). To a solution of
1-((2R,3S,4R,7S,8S,E)-13-(([1,1’-biphenyl]-4-carbonyl)oxy)-4,8-dihy-
droxy-3,7-dimethyl-6-oxotridec-9-en-2-yl) 3,4-dibenzyl (R,Z)-2-hy-
droxypent-3-ene-1,3,4-tricarboxylate (3.31 mg, 3.90 μmol) in 5 mL
of CH2Cl2 was added 5% Pd/C (10.0 mg). The reaction mixture was
vigorously stirred under 1 atm of H2 gas for 15 min before being
evacuated and purged with nitrogen gas. The Pd/C was removed
by vacuum filtration through a pad of celite using CH2Cl2 as an
eluent. The filtrate was concentrated to yield 2.53 mg of the
anhydride analogue 1e (99%), which was sufficiently pure for the
1
biological assays: H NMR (CDCl3, 500 MHz): δ=8.15 (d, J=8.5, 2H),
7.71 (d, J=8.5, 2H), 7.67 (d, J=7.1, 2H), 7.52 (t, J=7.4, 2H), 7.45 (t,
J=7.3, 1H), 5.23 (d, J=5.6, 1H), 5.08 (quint, J= 6.4, 1H), 4.40 (t, J=
5.9, 2H), 4.36 (d, J=6.4, 1H), 3.81 (t, J=8.2, 1H), 3.10 (br s, 1H), 2.93
(dd, J=16.9, 3.6, 1H), 2.84 (quint, J=9.3, 2H), 2.32 (s, 3H), 1.86 (
quint, J=6.9, 2H), 1.73 (quint, J=7.9, 1H), 1.59–1.69 (m, 6H), 1.42–
1.58 (m, 5H), 1.34 (d, J=6.3, 3H), 1.15 (d, J=7.1, 3H), 0.99 (d, J=7.1,
3H); 13C NMR (CDCl3, 125 MHz): δ=215.9, 170.2, 166.7, 165.7, 164.9,
130.1, 129.2, 129.0, 128.2, 127.3, 127.1, 73.6, 66.7, 64.9, 63.9. 52.7,
46.5, 42.7, 40.7, 34.4, 29.8, 28.8, 26.0, 25.0, 18.5, 13.7, 10.5, 10.2; IR
(thin film): 3480 (br), 3020, 2978, 1769, 1714, 1280 cmÀ 1; HRMS (ESI):
m/z calcd for C36H44O11 [M+Na]+ 675.2781; found: 675.2780.
1-((2R,3S,4R,7S,8S,E)-13-(([1,1’-Biphenyl]-4-carbonyl)oxy)-4,8-dihy-
droxy-3,7-dimethyl-6-oxotridec-9-en-2-yl) 3,4-dibenzyl (R,Z)-2-hy-
droxypent-3-ene-1,3,4-tricarboxylate. Toluene (5.00 mL, purged
with N2 for one h) was added to Grubbs second-generation catalyst
(5.00 mg, 6.25 μmol). The purple solution was transferred to a flask
containing a mixture of 15 (12.5 mg, 20.0 μmol) and 3e (106 mg,
400 μmol). The resulting mixture was stirred for 3 h before being
concentrated to a crude oil. The oil was purified using column
chromatography (25–50% EtOAc in hexanes) to furnish 10.5 mg of
pure 1-((2R,3S,4R,7S,8S,E)-13-(([1,1’-biphenyl]-4-carbonyl)oxy)-4,8-di-
hydroxy-3,7-dimethyl-6-oxotridec-9-en-2-yl) 3,4-dibenzyl (R,Z)-2-hy-
droxypent-3-ene-1,3,4-tricarboxylate (62%): 1H NMR (CDCl3,
500 MHz): δ=8.15 (d, J=8.2, 2H), 7.71 (d, J=8.2, 2H), 7.67 (d, J=
8.3, 2H), 7.46 (t, J=7.7, 1H), 7.30–7.42 (m, 10H), 5.80 (dt, J=15.4,
6.7, 1H), 5.52 (dd, J=15.3, 7.7, 1H), 5.17 (quint, J=5.1, 1H), 5.11–
5.16 (m, 1H), 5.04–5.09 (m, 1H), 5.03 (quint, J=7.5, 1H), 4.41 (td, J=
11.0, 2.7, 1H), 4.32–4.44 (m, 2H), 4.22 (t, J=7.3, 1H), 3.72 (d, J=5.4,
1H), 3.05 (d, J=2.3, 1H), 2.98 (dd, J=16.0, 10.0, 1H), 2.82 (dd, J=
16.9, 9.5, 1H), 2.69 (quint, J=7.2, 1H), 2.64 (dd, J=16.9, 4.4, 1H),
General methods for the malachite green assay. All of the
reagents to perform the assays were acquired from Upstate
Biotechnology with the exception of PP1 and PP2A, which were
purchased from New England Biolabs. Tautomycetin was obtained
from Tocris Bioscience. The enzyme dilution buffer was composed
of the following: 50 mM Tris·HCl (pH 7.0), 0.1 mM Egtazic acid
(EDGT), 0.1% β-mercaptoethanol, and 1 mg/mL bovine serum
albumin. The assay buffer contained the 50 nM Tris·HCl and
100 μM CaCl2. The malachite green solution A was composed of
ChemMedChem 2020, 15, 1–13
10
© 2020 Wiley-VCH GmbH
��
These are not the final page numbers!