Journal of Medicinal Chemistry
Article
N-(4-(5-Amino-2,3,4,5-tetrahydro-1H-benzo[b]azepine-1-
carbonyl)phenyl)-[1,1′-biphenyl]-2-carboxamide (26). To a
stirred solution of compound 25 (53 mg, 0.12 mmol) in MeOH
(2.5 mL) was added NH4OAc (532 mg, 6.91 mmol) and NaBH3CN
(36 mg, 0.58 mmol). Following microwave irradiation (2 × 5 min at
100 °C, 1 bar), the mixture was washed, sequentially, with a saturated
solution of NaHCO3 and NaCl and concentrated in vacuo. The
residue was purified by RP-HPLC then freeze-dried to provide
compound 26 as a white powder (31 mg, 57%). Rf = 0.1 (EtOAc/
MeOH/Et3N, 9/1/0.01, v/v/v); mp 177−178 °C. 1H NMR (400
MHz, DMSO-d6): δ 10.30 (bs, 1H), 8.74 (bs, 2H), 7.58−7.53 (m,
2H), 7.49−7.44 (m, 2H), 7.39−7.29 (m, 9H), 7.14−7.09 (m, 3H),
6.76 (d, J = 7.1 Hz, 1H), 4.73 (bs, 1H), 4.58 (bs, 1H), 2.82 (t, J = 11.3
Hz, 1H), 2.19 (bs, 1H), 1.92 (bs, 1H), 1.65 (bs, 2H). 13C NMR (75
MHz, CD3OD): δ 171.3, 171.2, 142.7, 142.4, 141.5, 141.4, 137.8,
132.6, 131.5, 131.4, 130.1, 129.8, 129.6, 129.5, 129.1, 129.0, 128.7,
128.6, 125.7, 120.2, 53.3, 48.3, 36.7, 27.3. RP-HPLC purity: 91%; tR =
12.51. HRMS for C30H28N3O2: calcd 462.2176, found 462.2171.
5-(N-(1-(4-([1,1′-Biphenyl]-2-ylcarboxamido)benzoyl)-
2,3,4,5-tetrahydro-1H-benzo[b]azepin-5-yl)sulfamoyl)-2-(6-(di-
ethylamino)-3-(diethyliminio)-3H-xanthen-9-yl)-
benzenesulfonate (27). To a stirred solution of compound 26 (12
mg, 0.03 mmol) in a CH2Cl2/DMF (1/1, v/v) mixture was added p-
purity: 95%; tR = 15.54. HR-MS for C62H65N6O9S2: calcd 1101.4249,
found: 1101.4245.
5-((4-(3-((1-(4-([1,1′-Biphenyl]-2-ylcarboxamido)benzoyl)-
2,3,4,5-tetrahydro-1H-benzo[b]azepin-5-yl)amino)-3-
oxopropyl)piperazin-1-yl)sulfonyl)-2-(6-(diethylamino)-3-(di-
ethyliminio)-3H-xanthen-9-yl)benzenesulfonate (29). Red pow-
der (24 mg, 67%); mp 233−234 °C. 1H NMR (400 MHz, CD3OD): δ
8.61 (s, 1H), 8.02 (d, J = 8.1 Hz, 1H), 7.53−7.21 (m, 16H), 7.10−6.94
(m, 10H), 6.83−6.67 (m, 4H), 6.59 (d, J = 7.5 Hz, 1H), 5.30 (d, J =
10.0 Hz, 1H), 4.54 (sl, 1H), 3.65−3.54 (m, 8H), 3.27 (sl, 3H), 3.04
(sl, 1H), 2.87−2.28 (m, 7H), 2.05 (sl, 2H), 1.66 (sl, 2H), 1.30−1.26
(m, 12H). 13C NMR (75 MHz, CD3OD): δ 173.5, 171.2, 170.9, 159.1,
157.4, 156.9, 147.6, 141.7, 141.2, 139.9, 139.0, 137.4, 135.9, 133.6,
133.5, 132.3, 132.0, 131.2, 130.3, 129.8, 129.5, 129.3, 128.8, 128.6,
128.6, 128.4, 128.3, 125.6, 119.9, 115.1, 114.9, 96.9, 54.9, 53.0, 51.6,
47.2, 46.8, 34.0, 32.4, 26.3, 13.0. RP-HPLC purity: >95%; tR = 14.20.
HR-MS for C64H68N7O9S2: calcd: 1142.4514, found: 1142.4495.
5-(N-(2-(2-(2-(3-((1-(4-([1,1′-Biphenyl]-2-ylcarboxamido)-
benzoyl)-2,3,4,5-tetrahydro-1H-benzo[b]azepin-5-yl)amino)-3-
oxopropoxy)ethoxy)ethoxy)ethyl)sulfamoyl)-2-(6-(diethylami-
no)-3-(diethyliminio)-3H-xanthen-9-yl)benzenesulfonate (30).
1
Red powder (15 mg, 40%); mp 182−183 °C. H NMR (400 MHz,
CDCl3): δ 8.83 (s, 1 H), 8.01 (d, J = 7.9 Hz, 1H), 7.73 (d, J = 7.9 Hz,
1H), 7.46 (d, J = 7.6 Hz, 1H), 7.41−7.27 (m, 10H), 7.21−7.18 (m,
3H), 7.06 (t, J = 7.7 Hz, 2H), 6.85−6.73 (m, 4H), 6.64 (sl, 2H), 6.47
(d, J = 7.6 Hz, 1H), 5.18 (sl, 1H), 4.50 (sl, 1H), 3.85 (t, J = 6.2 Hz,
2H), 3.71−3.65 (m, 10H), 3.56−3.46 (m, 10H), 3.25 (sl, 2H), 2.92
(sl, 1H), 1.90 (sl, 2H), 1.63 (sl, 1H), 1.54 (sl, 1H), 1.27−1.24 (m,
12H). 13C NMR (75 MHz, CDCl3): δ 158.4, 158.0, 155.8, 142.7,
140.9, 139.9, 139.1, 133.8, 133.4, 130.8, 130.5, 130.2, 129.8, 129.5,
129.1, 128.8, 128.3, 128.2, 127.9, 127.9, 127.6, 127.4, 127.3, 125.1,
118.8, 114.5, 113.9, 95.9, 77.4, 70.5, 70.4, 70.4, 70.3, 69.9, 67.9, 50.9,
46.9, 46.1, 43.3, 36.8, 31.2, 25.6, 12.8. RP-HPLC purity: >95%; tR =
15.54. HR-MS for C66H73N6O12S2: calcd 1205.4722, found:
1205.4703.
LRB sulfonyl chloride (23 mg, 0.04 mmol) in the presence of Hunig’s
̈
base (23 μL, 0.13 mmol) and a catalytic amount of 4-DMAP.
Following 1 h at room temperature, the resulting mixture was
concentrated under reduced pressure. The residue was purified by
flash chromatography on a C18 column (prepacked columns, 25−40
μm, Merck) using a linear gradient (95% C to 100% D in 30 min, flow
rate of 15 mL·min−1) of solvent D (100% CH3CN) in solvent C
(100% H2O). After freeze-drying, compound 27 was obtained as a red
powder (16 mg, 62%); mp 233−235 °C. 1H NMR (400 MHz,
CDCl3): δ 8.92 (s, 1 H), 8.04 (d, J = 8.0 Hz, 1 H), 7.72 (s, 1 H), 7.55
(d, J = 7.9 Hz, 1 H), 7.38 (d, J = 7.3 Hz, 2H), 7.27−7.25 (m, 5 H),
7.19 (d, J = 8.2 Hz, 2 H), 7.12 (d, J = 8.2 Hz, 2 H), 7.02 (d, J = 8.2 Hz,
2 H), 6.96 (t, J = 10.5 Hz, 2 H), 6.81 (t, J = 7.5 Hz, 1 H), 6.64−6.57
(m, 5 H), 6.46 (d, J = 7.4 Hz, 1 H), 4.86 (d, J = 9.5 Hz, 1 H), 4.34 (t, J
= 9.4 Hz, 1 H), 3.54−3.43 (m, 8 H), 3.00 (t, J = 9.4 Hz, 1 H), 2.09 (sl,
2 H), 1.83 (sl, 1 H), 1.23−1.18 (m, 12 H). 13C NMR (75 MHz,
CDCl3): δ 167.5, 158.1, 158.0, 155.7, 140.3, 140.1, 137.6, 135.3, 133.8,
133.6, 133.5, 131.2, 130.6, 130.5, 130.2, 129.2, 129.1, 128.8, 128.8,
128.4, 127.9, 127.6, 127.5, 127.4, 126.5, 118.9, 114.5, 114.5, 113.8,
N1-(1-(4-([1,1′-Biphenyl]-2-ylcarboxamido)benzoyl)-2,3,4,5-
tetrahydro-1H-benzo[b]azepin-5-yl)-N4-(prop-2-yn-1-yl)-
succinamide (39). A solution of compound 26 (54 mg, 0.117 mmol)
in DMF (0.6 mL) was slowly added to a solution of compound 31 (15
mg, 0.10 mmol) dissolved in DMF (0.6 mL) in the presence of
PyBOP (61 mg, 0.12 mmol) and Hunig’s base (51 μL, 0.29 mmol).
̈
Following 16 h stirring at room temperature, the mixture was
concentrated in vacuo. The residue was dissolved in EtOAc (5 mL)
and washed sequentially with a saturated solution of NaHCO3 and
brine. The organic layer was concentrated in vacuo and the residue
purified by flash chromatography. Compound 39 was obtained as a
95.7, 54.6, 46.5, 46.0, 33.2, 25.1, 12.8. RP-HPLC purity: 95%; tR
=
15.09. HRMS for C57H56N5O8S2: calcd 1002.3570, found 1002.3568.
General Procedure for the Coupling of para-LRB Spacers 15,
17, and 16. To a solution of p-LRB spacer carboxylic acid (1 equiv)
in dry DMF (4 M) was added PyBOP (1 equiv) and compound 26 (1
equiv). The mixture was stirred for 5 min at room temperature before
1
clear oil (47 mg, 87%). H NMR (400 MHz, CD3OD): δ 7.50 (t, J =
7.6 Hz, 2H), 7.36−7.41 (m, 4H), 7.15−7.28 (m, 10H), 6.95 (t, J = 7.8
Hz, 1H), 5.24−5.27 (m, 1H), 3.93 (t, J = 2.2 Hz, 2H), 3.27 (m, 4H),
2.60−2.67 (m, 2H), 2.52−2.55 (m, 3H), 1.97−206 (m, 2H), 1.60−
1.67 (m, 2H), 1.25 ppm (s, 1H). 13C NMR (75 MHz, CD3OD): δ
141.9, 141.9, 141.4, 141.4, 141.4, 141.4, 140.4, 137.8, 132.5, 131.4,
131.3, 130.3, 129.7, 129.6, 129.5, 129.5, 128.9, 128.6, 128.5, 128.4,
125.6, 120.1, 80.7, 72.3, 51.8, 48.0, 32.7, 31.9, 30.8, 29.6, 26.6. RP-
HPLC purity: >95%; tR = 14.18. ES-MS for C37H35N4O4: calcd 599.26,
found 599.25.
Fluorescein Labeled Compound (40). To a solution of
compound 39 (23 mg, 0.04 mmol) and compound 37 (26 mg, 0.04
mmol) in a DMF/H2O (9/1, v/v, 0.8 mL) mixture was added 10% of
sodium ascorbate (1 mg, 0.005 mmol), dissolved in the minimum of
water, 10% of TBTA (2 mg, 0.005 mmol) dissolved in the minimum of
DMF, and finally CuSO4·5H2O (0.6 mg, 0.004 mmol). After 5 h at
room temperature, the mixture was concentrated in vacuo to a residue
that was purified by RP-HPLC on a C18 Symmetry Shield column (19
mm × 300 mm) following a linear gradient from 20% to 70% of
solvent B in solvent A in 40 min (10 mL·min−1, detection at 220 and
254 nm). Following a freeze-drying step, compound 40 was obtained
addition of Hunig’s base (5 equiv). The resulting mixture was stirred at
̈
room temperature for 2 h, reduced in vacuo, and subsequently purified
by flash chromatography on a C18 column (prepacked columns, 25−
40 μm, Merck) using a linear gradient (95% C to 100% D in 30 min,
flow rate of 15 mL·min−1) of solvent D (100% CH3CN) in solvent C
(100% H2O).
5-(N-(5-((1-(4-([1,1′-Biphenyl]-2-ylcarboxamido)benzoyl)-
2,3,4,5-tetrahydro-1H-benzo[b]azepin-5-yl)amino)-5-
oxopentyl)sulfamoyl)-2-(6-(diethylamino)-3-(diethyliminio)-
3H-xanthen-9-yl)benzenesulfonate (28). Red powder (5 mg,
50%), mp 202−203 °C. 1H NMR (400 MHz, CDCl3): δ 8.65 (s, 1 H),
7.98 (d, J = 8.0 Hz, 1 H), 7.57 (d, J = 7.9 Hz, 1 H), 7.40 (d, J = 7.7 Hz,
1 H), 7.32 (d, J = 6.8 Hz, 4 H), 7.23−7.16 (m, 7 H), 7.11−7.04 (m, 5
H), 6.84 (t, J = 7.5 Hz, 1 H), 6.76 (td, J = 9.5, 2.3 Hz, 2 H), 6.63 (sl, 2
H), 6.46 (s, J = 7.5 Hz, 1 H), 5.19 (d, J = 10.4 Hz, 1 H), 4.46 (sl, 1 H),
3.53−3.46 (m, 8 H), 3.30−3.29 (m, 1 H), 2.95 (sl, 1 H), 2.27 (t, J =
7.6 Hz, 2 H), 1.99 (sl, 1 H), 1.88 (sl, 1 H), 1.75−1.66 (m, 2 H), 1.59−
1.51 (m, 4 H), 1.35−1.30 (m, 4 H), 1.24−1.20 (m, 12 H). 13C NMR
(75 MHz, CDCl3): δ 169.9, 158.9, 157.9, 155.7, 146.8, 145.8, 142.9,
140.6, 139.9, 138.7, 135.8, 133.6, 132.9, 130.5, 130.3, 129.5, 128.7,
128.6, 128.4, 127.8, 127.6, 127. 5, 126.8, 124.3, 118.7, 114.2, 113.8,
95.8, 54.4, 46.8, 45.9, 42.6, 42.6, 35.6, 31.2, 29.1, 22.9, 12.6. RP-HPLC
as an orange-colored powder (12 mg, 26%); mp 154−155 °C; tR
=
14.48. HRMS for C66H64N9O12S: calcd 1206.4395, found 1206.4412.
General Procedure for the Preparation of Compounds 41,
42, and 43 by Solution-Phase “Click” Chemistry. To a solution of
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dx.doi.org/10.1021/jm3006146 | J. Med. Chem. 2012, 55, 8588−8602