T. Ai et al. / Bioorg. Med. Chem. 24 (2016) 686–692
691
(d, J = 8.4 Hz, 2H), 7.60 (d, J = 7.8 Hz, 2H), 6.90 (d, J = 9.0 Hz, 2H),
1.00 (s, 9H), 0.25 (s, 6H). HRMS could not be obtained under varied
conditions.
(m, 1H), 7.05–6.97 (m, 2H), 6.46 (d, J = 15.6 Hz, 1H), 4.26–4.20
(m, 2H), 3.91–3.86 (m, 2H), 3.80–3.65 (m, 6H), 3.20–3.15 (m,
2H), 1.55 (s, 9H). HRMS (ESI+) calcd for C26H34NO6 (M+H)+
456.2381, found 456.2388.
4.2.8. (E)-tert-Butyl 3-(4-(4-hydroxybenzoyl)phenyl)acrylate
(13)
4.2.13. (E)-tert-Butyl 3-(4-(4-(2-(2-(2-(Hex-5-ynamido)ethoxy)
In a manner similar to that described for the preparation of
compound 6, bromide 12 (1.0 g, 2.47 mmol) underwent a Heck
reaction to give t-butyl ester 13 as a yellow oil (700 mg, 87%). 1H
NMR (CDCl3, 600 MHz) d 7.79–7.72 (m, 4H), 7.62 (d, J = 16.2 Hz,
1H), 7.59 (d, J = 8.4 Hz, 2H), 6.94 (d, J = 8.4 Hz, 2H), 6.46 (d,
J = 16.2 Hz, 1H), 1.55 (s, 9H). HRMS (ESIꢀ) calcd for C20H19O4
(MꢀH)ꢀ 323.1289, found 323.1295.
ethoxy)ethoxy)benzoyl)phenyl)acrylate (19)
A
solution of 18 (370 mg, 0.8 mmol), 5-hexynoic acid
(1.3 mmol) and EDCꢁHCl (1.3 mmol) in DCM (15 mL) was allowed
to stir at rt overnight and the organic phase was then washed with
water and brine. After removal of the solvent, the residue was puri-
fied by flash column chromatography (0–20% MeOH/CH2Cl2) to
afford 19 as a colorless oil (330 mg, 75%). 1H NMR (CDCl3,
600 MHz) d 7.84–7.79 (m, 2H), 7.78–7.72 (m, 2H), 7.64–7.58 (m,
2H), 7.40–7.36 (m, 1H), 7.02–6.97 (m, 2H), 6.47 (d, J = 15.6 Hz,
1H), 6.04 (br s, 1H), 4.26–4.21 (m, 2H), 3.91 (t, J = 4.8 Hz, 2H),
3.74 (t, J = 4.2 Hz, 2H), 3.67 (t, J = 4.2 Hz, 2H), 3.58 (t, J = 4.8 Hz,
2H), 3.50–3.45 (m, 2H), 3.35 (s, 1H), 2.30 (t, J = 7.2 Hz, 2H), 2.24
(dt, J = 6.6, 2.4 Hz, 2H), 1.88–1.82 (m, 2H), 1.59 (s, 9H). HRMS
(ESI+) calcd for C32H40NO7 (M+H)+ 550.2799, found 550.2796.
4.2.9. (Ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(4-
methylbenzenesulfonate) (15)28–30
To a solution of 14 (4.5 g, 30.0 mmol) and TsCl (12.59 g,
66.0 mmol) in DCM (100 mL) was added NEt3 (6.68 g, 66.0 mmol).
The reaction mixture was stirred at rt for 12 h and washed with
water. The organic phase was dried over anhydrous Na2SO4. After
removal of the solvent, the residue was purified by flash column
chromatography (0–40% EtOAc/hexanes) to afford 15 as a white
solid (12.0 g, 87%). 1H NMR (CDCl3, 600 MHz) d 7.78 (d, J = 7.8 Hz,
4H), 7.34 (d, J = 7.8 Hz, 4H), 4.19–4.10 (m, 4H), 3.70–3.62 (m,
4H), 3.57–3.48 (m, 4H), 2.44 (s, 6H). HRMS (ESI+) calcd for
4.2.14. (E)-3-(4-(4-(2-(2-(2-(Hex-5-ynamido)ethoxy)ethoxy)
ethoxy)benzoyl)phenyl)acrylic acid (20)
A solution of t-butyl ester 19 (330 mg, 0.55 mmol) in TFA/DCM
(2:1, 10 mL) was allowed to stir at rt for 1 h. After removal of the
organic solvent, the residue was purified by flash column chro-
matography (0–12% MeOH/CH2Cl2) to afford trans-cinnamic acid
20 as a white solid (30 mg, 11%). 1H NMR (DMSO-d6, 600 MHz) d
12.55 (s, 1H), 7.88–7.81 (m, 3H), 7.73 (d, J = 8.4 Hz, 2H), 7.69 (d,
J = 8.4 Hz, 2H), 7.65 (d, J = 16.2 Hz, 1H), 7.09 (d, J = 9.0 Hz, 2H),
6.65 (d, J = 16.8 Hz, 1H), 4.19 (t, J = 4.5 Hz, 2H), 3.76 (t, J = 4.5 Hz,
2H), 3.61–3.56 (m, 2H), 3.54–3.50 (m, 2H), 3.39 (t, J = 6.0 Hz, 2H),
3.17 (q, J = 6.0 Hz, 2H), 2.74 (t, J = 2.7 Hz, 1H), 2.15–2.09 (m, 4H),
1.65–1.59 (m, 2H). 13C NMR (DMSO-d6, 150 MHz) d 177.8, 163.0,
159.6, 143.3, 139.3, 137.7, 137.4, 132.2, 129.8, 129.7, 127.6,
120.5, 114.0, 82.1, 70.4, 69.9, 69.2, 69.2, 68.8, 67.5, 38.9, 34.4,
24.5, 17.2. HRMS (ESI+) calcd for C28H30NO7 (MꢀH)ꢀ 492.2022,
found 492.2027.
C
20H26NaO8S2 (M+Na)+ 481.0961, found 481.0963.
4.2.10. 2-(2-(2-Azidoethoxy)ethoxy)ethyl 4-
methylbenzenesulfonate (16)28
A
mixture of 15 (10.0 g, 21.8 mmol) and NaN3 (609 mg,
9.37 mmol) in DMF (50 mL) was allowed to stir at rt for 24 h. The
reaction mixture was washed with water and the organic phase
was dried over anhydrous Na2SO4. After removal of the solvent,
the residue was purified by flash column chromatography (0–50%
EtOAc/hexanes) to afford 16 as a light yellow oil (3.0 g, 39%). 1H
NMR (CDCl3, 600 MHz)
d 7.78 (d, J = 7.8 Hz, 4H), 7.34 (d,
J = 7.8 Hz, 4H), 4.19–4.10 (m, 4H), 3.70–3.62 (m, 4H), 3.57–3.48
(m, 4H), 2.44 (s, 6H). 13C NMR (DMSO-d6, 150 MHz) d 144.8,
132.9, 129.8, 127.9, 70.7, 70.5, 70.0, 69.3, 68.7, 50.6, 21.6. HRMS
(ESI+) calcd for C13H19NaO5S (M+Na)+ 352.0938, found 352.0937.
4.2.15. (E)-N-(2-(2-(2-(4-(4-(3-(Hydroxyamino)-3-oxoprop-1-
en-1-yl)benzoyl)phenoxy)ethoxy)ethoxy)ethyl)hex-5-ynamide
(3)
In a manner similar to that described for the preparation of
compound 4, carboxylic acid 20 (30 mg, 0.06 mmol) was converted
into THP-protected hydroxylamine 21, which was then depro-
tected with 2 M HCl in Et2O (2 mL) to give hydroxamic acid 3 as
a gray solid (14 mg, 46% for 2 steps). 1H NMR (CD3OD, 600 MHz)
4.2.11. (E)-tert-Butyl 3-(4-(4-(2-(2-(2-azidoethoxy)ethoxy)
ethoxy)benzoyl)phenyl)acrylate (17)
To a solution of 13 (324 mg, 1.0 mmol) and 16 (362 mg,
1.0 mmol) in DMF (20 mL) was added CsCO3 (390 mg, 1.2 mmol)
and the resulting mixture was stirred at 50 °C for 12 h. After
diluted with EtOAc (100 mL), the organic phase was washed with
water and dried over anhydrous Na2SO4. After removal of the sol-
vents, the residue was purified by flash column chromatography
(0–60% EtOAc/hexanes) to afford 17 as a light yellow oil (530 mg,
90%). 1H NMR (CDCl3, 600 MHz) d 7.82–7.72 (m, 4H), 7.65–7.59
(m, 2H), 7.36–7.33 (m, 1H), 7.02–6.98 (m, 2H), 6.48 (d,
J = 14.4 Hz, 1H), 4.24–4.20 (m, 2H), 3.93–3.88 (m, 2H), 3.80–3.65
(m, 6H), 3.41–3.34 (m, 2H), 1.55 (s, 9H). HRMS (ESI+) calcd for
d
7.80 (d, J = 9.0 Hz, 2H), 7.75 (d, J = 8.4 Hz, 2H), 7.71 (d,
J = 8.4 Hz, 2H), 7.64 (d, J = 16.2 Hz, 1H), 7.08 (d, J = 8.4 Hz, 2H),
6.61 (d, J = 15.0 Hz, 1H), 4.27–4.22 (m, 2H), 3.91–3.87 (m, 2H),
3.74–3.70 (m, 2H), 3.66–3.62 (m, 2H), 3.55 (t, J = 5.4 Hz, 2H), 3.36
(t, J = 5.4 Hz, 2H), 2.29 (t, J = 7.5 Hz, 2H), 2.24–2.21 (m, 1H), 2.19
(dt, J = 7.2, 2.4 Hz, 2H), 1.81–1.75 (m, 2H). 13C NMR (DMSO-d6,
150 MHz) d 171.9, 163.0, 162.7, 139.3, 137.7, 137.5, 137.4, 132.6,
130.4, 129.8, 127.9, 121.9, 114.8, 80.5, 71.9, 70.3, 70.0, 69.6, 69.2,
68.0, 38.9, 34.5, 24.7, 17.8. HRMS (ESIꢀ) calcd for C28H31N2O7
(MꢀH)ꢀ 507.2131, found 507.2132.
C
26H32N3O6 (M+H)+ 482.2286, found 482.2282.
4.2.12. (E)-tert-Butyl 3-(4-(4-(2-(2-(2-aminoethoxy)ethoxy)
ethoxy)benzoyl)phenyl)acrylate (18)
A mixture of 17 (530 mg, 1.0 mmol), PPh3 (866 mg, 3.3 mmol),
4.3. Biological assays
and H2O (200 lL, 11.0 mmol) in THF (15 mL) was stirred at 50 °C
for 4 h. The reaction mixture was diluted with EtOAc (100 mL),
and the organic phase was washed with water and dried over
anhydrous Na2SO4. After removal of the solvents, the residue was
purified by flash column chromatography (0–60% EtOAc/hexanes)
to afford 18 as a light yellow oil (370 mg, 80%). 1H NMR (CDCl3,
600 MHz) d 7.84–7.75 (m, 4H), 7.59 (d, J = 8.4 Hz, 2H), 7.40–7.36
4.3.1. RT-qPCR assay and biochemical assays
RT-qPCR experiments were performed as described previ-
ously.18 The biochemical assays against human HDAC1–11,
MMP1–3, MMP7–10, MMP12–14, and TACE were performed by
the Reaction Biology Corp. (RBC) (Malvern, PA, USA, http://www.