2742
M. Suzuki et al. / Bioorg. Med. Chem. 9 (2001) 2727–2743
Table 6 (continued)
No.
1H NMR (d; ppm)
300 MHz in DMSO-d6
13C NMR (d; ppm)
75 MHz in DMSO-d6
17gg
17hh
17ii
0.9–1.5 (m, 2H), 0.96 (d, 6H, J=7 Hz), 1.7–2.1 (m, 2H), 2.3 (m,
1H), 2.89 (d, 2H, J=7 Hz), 3.5 (m, 1H), 4.1 (m, 1H), 5.39 (dd,
1H, J=6, 16 Hz), 6.42 (d, 1H, J=16 Hz), 7.2–7.5 (m, 6H),
7.7 (m, 1H), 7.99 (d, 1H, J=8 Hz)
176.4, 161.5 (d, 1JC-F=244.0 Hz), 160.1, 145.8, 144.1,
142.3, 133.3 (d, 4JC-F=2.9 Hz), 132.2 (d, 3JC-F=8.1 Hz),
132.0 (d, 3JC-F=8.7 Hz), 130.1, 128.7, 128.6, 126.0,
125.9, 125.7, 122.9, 115.2 (2C, d, 2JC-F=24.2 Hz), 68.6,
65.6, 45.0, 43.4, 27.6, 22.6 (2C)
0.85 (t, 3H, J=7 Hz), 0.9–1.4 (m, 2H), 1.28 (d, 3H, J=6 Hz),
1.5–2.1 (m, 4H), 3.4 (m, 1H), 4.1 (m, 1H), 5.38 (dd, 1H, J=6,
16 Hz), 6.46 (d, 1H, J=16 Hz), 7.2–7.5 (m, 6H), 7.7 (m,
1H), 7.98 (d, 1H, J=8 Hz)
176.5, 164.5, 161.5 (d, 1JC-F=244.0 Hz), 146.1, 144.3,
142.3, 133.4 (d, 4JC-F=3.5 Hz), 132.2 (d, 3JC-F=7.5 Hz),
132.0 (d, 3JC-F=9.2 Hz), 130.1, 128.7, 128.6, 125.9,
125.8, 125.7, 122.8, 115.1 (2C, d, 2JC-F=21.3 Hz), 68.6,
65.7, 44.8, 43.4, 29.0, 19.9, 12.1
0.8–0.9 (m, 1H), 1.50 (s, 9H), 1.2–1.3 (m, 1H), 1.7–2.1 (m,
2H), 3.5 (m, 1H), 4.1 (m, 1H), 5.23 (dd, 1H, J=5, 16 Hz),
6.67 (d, 1H, J=16 Hz), 7.2–7.3 (m, 5H), 7.4–7.5 (m, 1H), 7.7
(m, 1H), 7.97 (d, 1H, J=8 Hz)
176.4, 165.6, 161.4 (d, 1JC-F=244.0 Hz), 146.1, 144.7,
141.2, 134.1 (d, 4JC-F=3.5 Hz), 132.2 (d, 3JC-F=8.7 Hz),
132.0 (d, 3JC-F=8.1 Hz), 130.6, 129.1, 128.7, 126.2,
125.9, 125.6, 125.0, 115.0 (2C, d, 2JC-F=20.8 Hz), 68.5,
66.0, 44.5, 43.3, 30.1 (3C)
17jj
1.0–1.5 (m, 6H), 1.7–2.1 (m, 2H), 2.5 (m, 1H), 3.5 (m, 1H),
4.1 (m, 1H), 5.61 (dd, 1H, J=6, 16 Hz), 6.49 (d, 1H,
J=16 Hz), 7.2–7.4 (m, 6H), 7.6 (m, 1H), 7.86 (d, 1H,
J=9 Hz)
176.4, 161.6 (d, 1JC-F=244.6 Hz), 160.6, 145.9, 143.6, 142.4,
3
133.1 (d, 4JC-F=2.9 Hz), 132.1 (d, JC-F=8.1 Hz), 131.8 (d,
3JC-F=8.1 Hz), 129.7, 128.8, 128.4, 125.7, 125.6 (2C), 122.9,
2
115.2 (2C, d, JC-F=21.3 Hz), 68.8, 65.7, 44.8, 43.5, 15.3,
10.7 (2C)
17kk
17ll
0.9–2.1 (m, 14H), 3.2 (m, 1H), 3.5 (m, 1H), 4.1 (m, 1H),
5.41 (dd, 1H, J=6, 16 Hz), 6.44 (d, 1H, J=16 Hz),
7.2–7.5 (m, 6H), 7.7 (m, 1H), 7.97 (d, 1H, J=8 Hz)
176.4, 164.3, 161.5 (d, 1JC-F=244.0 Hz), 146.0, 144.3, 142.2,
3
133.4 (d, 4JC-F=2.9 Hz), 132.2 (d, JC-F=8.7 Hz), 132.0 (d,
3JC-F=8.1 Hz), 129.5, 128.7, 128.6, 125.9, 125.8, 125.7, 122.6,
2
115.1 (2C, d, JC-F=21.9 Hz), 68.6, 65.7, 45.0, 43.4, 42.5, 31.9
(2C), 26.1 (2C), 25.8
0.7–0.9 (m, 1H), 1.1–1.3 (m, 1H), 1.6–2.0 (m, 2H), 3.5 (m,
1H), 3.9 (m, 1H), 5.19 (dd, 1H, J=6, 16 Hz), 6.24 (d,
1H, J=16 Hz), 7.3–7.8 (m, 12H), 8.06 (d, 1H, J=8 Hz)
176.4, 161.7 (d, 1JC-F=244.6 Hz), 158.5, 145.9, 145.1, 142.3,
140.9, 133.0 (d, 4JC-F=2.9 Hz), 132.1 (d, 3JC-F=7.5 Hz), 131.9
(d, 3JC-F=8.1 Hz), 129.8 (2C), 129.3, 129.1, 128.5, 128.0, 127.8
2
(2C), 126.9, 126.4, 125.8, 123.4, 115.5 (d, JC-F=20.8 Hz),
115.4 (d, 2JC-F=21.3 Hz), 68.7, 65.5, 44.6, 43.4
17mm
17nn
17oo
17pp
17qq
0.9–1.0 (m, 1H), 1.2–1.4 (m, 1H), 1.7–2.1 (m, 2H), 3.5 (m,
1H), 4.1 (m, 1H), 5.50 (dd, 1H, J=6, 16 Hz), 6.51 (d,
1H, J=16 Hz), 7.3–7.5 (m, 5H), 7.7 (m, 1H), 7.9 (m, 1H),
8.21 (d, 1H, J=8 Hz)
176.4, 161.9 (d, 1JC-F=244.6 Hz), 148.3, 144.8, 144.4, 144.1,
132.2 (2C, d, JC-F=9.8 Hz), 132.0 (d, JC-F=3.5 Hz), 130.5,
129.7, 129.4, 128.2 (2C), 126.1, 121.9 (q, JC-F=276.9 Hz),
119.4, 115.4 (2C, d, 2JC-F=21.4 Hz), 68.3, 65.7, 44.7, 43.4
3
4
1.1–1.2 (m, 1H), 1.3–1.6 (m, 1H), 1.8–2.1 (m, 2H), 3.6 (m,
1H), 4.08 (s, 3H), 4.1 (m, 1H), 6.08 (dd, 1H, J=6,
16 Hz), 6.30 (d, 1H, J=16 Hz), 7.18 (d, 1H, J=7 Hz), 7.2–7.5
(m, 5H), 7.81 (d, 1H, J=8 Hz)
176.6, 161.8 (d, 1JC-F=244.6 Hz), 159.6, 146.4, 144.1, 141.5,
3
132.7 (d, 4JC-F=3.5 Hz), 132.0 (d, JC-F=8.1 Hz), 131.6 (d,
3JC-F=7.5 Hz), 129.2, 126.7, 125.9, 124.9, 124.4, 120.4, 120.0,
2
115.6 (2C, d, JC-F=21.3 Hz), 69.3, 65.8, 53.5, 44.9, 43.6
1.0–1.1 (m, 1H), 1.3–1.4 (m, 1H), 1.7–2.1 (m, 2H), 2.63 (s,
3H),3.6 (m, 1H), 4.1 (m, 1H), 5.62 (dd, 1H, J=6,
16 Hz), 6.29 (d, 1H, J=16 Hz), 7.2–7.5 (m, 6H), 7.7 (m,
1H), 7.93 (d, 1H, J=8 Hz)
176.5, 161.7 (d, 1JC-F=244.6 Hz), 159.2, 146.0, 143.5, 143.2,
3
132.5 (d, 4JC-F=3.5 Hz), 132.1 (d, JC-F=8.1 Hz), 131.9 (d,
3JC-F=8.1 Hz), 129.2, 128.8, 127.6, 126.0, 125.5, 125.1, 120.6,
2
115.3 (2C, d, JC-F=21.3 Hz), 68.5, 65.7, 44.8, 43.4, 13.2
1.0–1.1 (m, 1H), 1.3–1.4 (m, 1H), 1.7–2.1 (m, 2H), 2.96
(s, 6H), 3.5–3.6 (m, 1H), 4.1 (m, 1H), 5.58 (dd, 1H, J=6,
16 Hz), 6.28 (d, 1H, J=16 Hz), 7.06 (d, 1H, J=8 Hz), 7.1–7.4
(m, 5H), 7.5 (m, 1H), 7.72 (d, 1H, J=8 Hz)
176.5, 161.5 (d, 1JC-F=244.0 Hz), 159.7, 145.7, 145.1, 139.2,
3
133.4 (d, 4JC-F=3.5 Hz), 132.0 (d, JC-F=8.1 Hz), 131.5 (d,
3JC-F=8.1 Hz), 128.8, 126.8, 125.7, 124.3, 123.5, 123.4, 122.3,
2
115.3 (2C, d, JC-F=20.2 Hz), 69.1, 65.8, 44.8, 43.5, 41.8 (2C)
1.0–1.1 (m, 1H), 1.15 (t, 3H, J=7 Hz), 1.3–1.4 (m, 1H),
1.7–2.1 (m, 2H), 2.94 (s, 3H), 3.3 (m, 2H), 3.5 (m, 1H), 4.1
(m, 1H), 5.49 (dd, 1H, J=6, 16 Hz), 6.28 (d, 1H,
J=16 Hz), 7.06 (d, 1H, J=9 Hz), 7.1–7.4 (m, 5H), 7.5 (m,
1H), 7.72 (d, 1H, J=8 Hz)
176.4, 161.5 (d, 1JC-F=244.0 Hz), 159.5, 145.6, 145.1, 139.3,
3
133.6 (d, 4JC-F=3.5 Hz), 132.0 (d, JC-F=7.5 Hz), 131.5 (d,
3JC-F=8.1 Hz), 128.7, 126.8, 125.7, 124.3, 123.4 (2C), 122.8,
2
115.2 (2C, d, JC-F=20.8 Hz), 69.1, 65.8, 47.7, 44.8, 43.5,
38.1, 12.5
rat billialy cannulated and discharged bile for over 24 h.
Liver was cut at mid-dark, then the microsomal and
supernatant fractions which could be precipitated with
40–80% saturated ammonium sulfate (sup fraction),
were prepared from liver homogenate according to the
modified method of Knauss et al.24 For the cholesterol
biosynthesis assay, microsome (1.0 mg protein) and sup
fractions (1.0 mg protein) were incubated for 2 h at
37 ꢁC in 200 mL of a reaction mixture containing ATP;
1 mM, glutathione; 6 mM, glucose-1-phosphate; 10 mM,
and; 0.25 mM, NADP; 0.25 mM, CoA; 0.04 mM and
0.2 mM sodium [2-14C] acetate (56 mCi/mmol, 0.2 mCi)
with 4 mL of test compound solution dissolved in water
or dimethyl sulfoxide. To stop the reaction and saponify
the product, 1 mL of 15% EtOH–KOH was added to
the mixture and heated at 75 ꢁC for 1 h. Lipids that were
unable to be saponified were extracted with petroleum
ether and incorporated [14C] radioactivity was counted.
Inhibitory activity of compounds was followed with
IC50 to show the rate of decreased incorporation of
[14C] acetate into lipids that cannot be saponified. Rela-
tive potencies were obtained by comparison of IC50 values