Recognition of Adenine Nucleotides by P2Y1-R. 1
J ournal of Medicinal Chemistry, 1999, Vol. 42, No. 26 5333
heated under reflux for 2 days. The mixture was concentrated
under reduced pressure and the residue was treated with
MeOH (5 mL). The suspension was filtered and the filtrate
was evaporated to give the product in a quantitative yield as
a beige solid: 1H NMR (DMSO-d6, 600 MHz) δ 7.91 (s, 1H,
H-8), 6.84 (br s, 2H, NH2), 6.24 (br s, 1H, NHCH2), 5.71 (d,
J ) 6 Hz, 1H, H-1′), 5.37 (br s, 1H, OH-2′), 5.13 (br d, J ) 4
Hz, 2H, OH-3′ & OH-5′), 4.60 (br s, 1H, H-2′), 4.13 (br q, J )
3 Hz, 1H, H-3′), 3.89 (q, J ) 3 Hz, 1H, H-4′), 3.63 (dd, J ) 12,
4 Hz, 1H, H-5′), 3.52 (dd, J ) 12, 4.5 Hz, 1H, H-5′), 3.23 (q,
J ) 6 Hz, 2H, NHCH2), 1.48 (quint, J ) 7 Hz, 2H, NHCH2CH2),
1.31 (sex, J ) 7 Hz, 2H, CH2CH3), 0.89 (t, J ) 7 Hz, 3H, CH3);
13C NMR (DMSO-d6, 300 MHz) δ 158.9 (s, C-2), 155.6 (s, C-6),
151.5 (s, C-4), 136.6 (d, C-8), 113.4 (s, C-5), 87.2 (d, C-1′), 85.3
(d, C-2′), 72.9 (d, C-3′), 70.7 (d, C-4′), 61.8 (t, C-5′), 40.7 (t,
NHCH2), 31.4 (d, NHCH2CH2), 19.7 (d, CH2CH3), 13.9 (q, CH3).
and filtered through a short column of silica (washed with
chloroform/methanol 4:1). The pure white solid obtained after
evaporation was dried in vacuo for 2 days to give 0.244 g (92%)
of 8: 1H NMR (DMSO-d6, 200 MHz) δ 8.22 (s, 1H, H-8), 7.35
(bs, 2H, NH2), 5.81(d, J ) 6 Hz, 1H, H-1′), 5.45 (d, J ) 6 Hz,
1H, OH-2′), 5.20 (d, J ) 5 Hz, 1H, OH-3′), 5.07 (t, J ) 5.5 Hz,
1H, OH-5′), 4.68-4.56 (m, 1H, H-2′), 4.19-4.09 (m, 1H, H-3′),
3.97-3.87 (m, 1H, H-4′), (H-5′ is hidden by the water peak),
3.15-3.03 (m, 2H, SCH2), 1.73-1.56 (m, 2H, SCH2CH2), 1.53-
1.32 (m, 2H, CH2CH3), 0.92 (t, J ) 7 Hz, 3H, CH3); 13C NMR
(DMSO-d6, 300 MHz) δ 163.7 (s, C-2), 155.4 (s, C-6), 150.0 (s,
C-4), 138.7 (d, C-8), 116.8 (s, C-5), 87.3 (d, C-1′), 85.3 (d, C-4′),
73.0 (d, C-2′), 70.4 (d, C-3′), 61.5 (t, C-5′), 31.1 (t, SCH2), 29.7
(t, SCH2CH2), 21.3 (t, CH2CH3), 13.5 (q, CH3); MS (CI/CH4)
m/z 356 MH+.
8-(n -Bu tylth ioeth er )a d en osin e (11). The compound was
prepared as described for 2-thioadenosine and obtained in 90%
yield as a white solid: mp 171.5 °C; H NMR (DMSO-d6, 200
2-Br om o-2′,3′,5′-t r i-O-(ter t-b u t yld im et h ylsilyl)a d en o-
sin e (4). 2-Bromo-adenosine (0.4 g, 1.16 mmol), tert-butyldi-
methylsilyl chloride (1.73 g, 10 equiv), and imidazole (1.6 g,
20 equiv) were dissolved in dry THF:DMF solution (5:2 mL).
The reaction mixture was stirred under nitrogen at RT
overnight and then concentrated under reduced pressure. The
residue was dissolved in CHCl3, washed twice with aqueous
NaHCO3 and once with water, and then dried over Na2SO4.
The residue obtained after evaporation was purified on a silica
gel column and eluted with CHCl3:MeOH 10:1. The product
was obtained as a white solid (0.722 g, 90% yield, mp 148-
151 °C): 1H NMR (CDCl3, 200 MHz) δ 8.50 (s, 1H, H-8), 5.96
(d, J ) 4 Hz, 1H, H-1′), 4.60-4.51 (m, 1H, H-2′), 4.27 (t, J )
4.5 Hz, 1H, H-3′), 4.21-4.11 (m, 1H, H-4′), 4.09 (dd, J ) 11, 4
Hz, 1H, H-5′), 3.80 (dd, J ) 11, 2 Hz, 1H, H-5′), 0.97 (s, 9H,
TBDMS), 0.92 (s, 9H, TBDMS), 0.86 (s, 9H, TBDMS), 0.18 (s,
3H, TBDMS), 0.17 (s, 3H, TBDMS), 0.10 (s, 3H, TBDMS), 0.09
(s, 3H, TBDMS), 0.04 (s, 3H, TBDMS), -0.05 (s, 3H, TBDMS);
13C NMR (CDCl3, 300 MHz) δ 154.1 (s, C-6), 148.9 (s, C-2),
146.3 (s, C-4), 137.4 (d, C-8), 114.4 (s, C-5), 89.9 (d, C-1′), 84.9
(d, C-4′), 76.0 (d, C-2′), 70.2 (d, C-3′), 61.5 (t, C-5′), 26.2 (q, 3C,
TBDMS), 25.8 (q, 3C, TBDMS), 25.7 (q, 3C, TBDMS), 18.6 (s,
TBDMS), 18.0 (s, TBDMS), 17.0 (s, TBDMS), -4.2 (q, TBDMS),
-4.8 (q, TBDMS), -4.8 (q, TBDMS), -4.9 (q, TBDMS), -5.1
(q, TBDMS), -5.4 (q, TBDMS).
1
MHz) δ 8.06 (s, 1H, H-2), 7.29 (br s, 2H, NH2), 5.77 (d, J ) 6
Hz, 1H, H-1′), 5.68 (dd, J ) 8.5, 3.5 Hz, 1H, OH-5′), 5.42 (d,
J ) 6.5 Hz, 1H, OH-2′), 5.21 (d, J ) 4.5 Hz, 1H, OH-3′), 4.99
(q, J ) 6 Hz, 1H, H-2′), 4.19-4.11 (m, 1H, H-3′), 4.00-3.92
(m, 1H, H-4′), 3.74-3.45 (m, 2H, H-5′), (SCH2 is hidden by
the water peak), 1.77-1.60 (m, 2H, SCH2CH2), 1.52-1.32 (m,
2H, CH2CH3), 0.90 (t, J ) 7 Hz, 3H, CH3); 13C NMR (DMSO-
d6, 300 MHz) δ 154.4 (s, C-6), 151.2 (d, C-2), 150.3 (s, C-4),
148.6 (s, C-8), 119.5 (s, C-5), 88.8 (d, C-1′), 86.5 (d, C-4′), 71.2
(d, C-2′), 70.9 (d, C-3′), 62.1 (t, C-5′), 32.0 (t, SCH2CH2), 30.8
(t, SCH2), 21.1 (t, CH2CH3), 13.3 (q, CH3); MS (CI/CH4) m/z
356 MH+.
8-(n -Bu tyla m in o)a d en osin e (13) was prepared as de-
scribed for 2. The reaction was completed after 1 day. The
product was obtained in a quantitative yield as a beige solid:
1H NMR (DMSO-d6, 200 MHz) δ 7.88 (s, 1H, H-2), 6.86 (t, J )
5 Hz, 1H, NH), 6.48 (br s, 2H, NH2), 5.90 (d, J ) 7.5 Hz, 1H,
H-1′), 5.87 (t, J ) 5 Hz, 1H, OH-5′), 5.23 (d, J ) 7 Hz, 1H,
OH-2′), 5.15 (d, J ) 4 Hz, 1H, OH-3′), 4.61 (q, J ) 6.5 Hz, 1H,
H-2′), 4.13-4.04 (m, 1H, H-3′), 3.97-3.91 (m, 1H, H-4′), 3.64-
3.56 (m, 2H, H-5′), (NHCH2 is hidden by the water peak),
2.54-1.46 (m, 2H, NHCH2CH2), 1.43-1.23 (m, 2H, CH2CH3),
0.90 (t, J ) 7 Hz, 3H, CH3); 13C NMR (DMSO-d6, 300 MHz) δ
152.4 (s, C-6), 152.0 (d, C-8), 150.2 (s, C-4), 149.3 (d, C-2), 117.0
(s, C-5), 86.8 (d, C-1′), 86.3 (d, C-4′), 71.6 (d, C-2′), 71.2 (d, C-3′),
62.1 (t, C-5′), 42.6 (t, NHCH2), 31.2 (t, NHCH2CH2), 20.2 (t,
CH2CH3), 114.3 (q, CH3); HRMS calcd for C14H23N6O4 339.1780,
found 339.1800.
8-Br om o-2′,3′,5′-t r i-O-(ter t-b u t yld im et h ylsilyl)a d en o-
sin e (15). Product 15 was obtained as described for 4 starting
from 8-bromo-adenosine (0.3 g, 0.87 mmol). TLC on a silica
gel plate (CHCl3:MeOH 6:1) indicated the formation of 15
(Rf ) 0.47) and a byproduct 16 (Rf ) 0.85). The products were
separated on a silica gel column. Product 15 was obtained in
78% yield (0.47 g): 1H NMR (CDCl3) δ 8.24 (s, 1H, H-2), 6.02
(br s, 2H, NH2), 5.94 (d, J ) 6 Hz, 1H, H-1′), 5.51 (dd, J ) 6,
4 Hz, 1H, H-2′), 4.57 (dd, J ) 4, 2 Hz, 1H, H-3′), 4.13-4.01
(m, 2H, H-4′ and H-5′), 3.77-3.65 (m, 1H, H-5′), 0.96 (s, 9H,
TBDMS), 0.84 (s, 9H, TBDMS), 0.79 (s, 9H, TBDMS), 0.15 (s,
6H, TBDMS), 0.02 (s, 3H, TBDMS), -0.02 (s, 3H, TBDMS),
-0.06 (s, 3H, TBDMS), -0.34 (s, 3H, TBDMS); 13C NMR
(CDCl3) δ 154.3 (s, C-6), 152.5 (d, C-2), 150.8 (s, C-4), 128.8 (s,
C-8), 120.5 (s, C-5), 90.6 (d, C-1′), 85.6 (d, C-4′), 72.2 (d, C-2′),
71.8 (d, C-3′), 62.3 (t, C-5′), 25.9 (q, 3C, TBDMS), 25.8 (q, 3C,
TBDMS), 25.7 (q, 3C, TBDMS), 18.3 (s, TBDMS), 18.1 (s,
TBDMS), 17.8 (s, TBDMS), -4.5 (q, TBDMS), -4.6 (q, 2C,
TBDMS), -5.2 (q, TBDMS), -5.4 (q, TBDMS), -5.6 (q,
TBDMS).
Product 16 was obtained in 17% yield (0.117 g): 1H NMR
(CDCl3) δ 8.25 (s, 1H, H-2), 5.92 (d, J ) 5.5 Hz, 1H, H-1′),
5.52 (dd, J ) 5.5, 4.5 Hz, 1H, H-2′), 5.18 (br s, 1H, NHTBDMS),
4.66 (dd, J ) 4.5, 3 Hz, 1H, H-3′), 4.15-4.02 (m, 2H, H-4′ &
H-5′), 3.76-3.62 (m, 1H, H-5′), 0.96 (s, 9H, TBDMS), 0.86 (s,
9H, TBDMS), 0.82 (s, 9H, TBDMS), 0.81 (s, 9H, TBDMS), 0.35
(s, 6H, TBDMS), 0.16 (s, 6H, TBDMS), 0.00 (s, 3H, TBDMS),
-0.04 (s, 6H, TBDMS), -0.31 (s, 3H, TBDMS); 13C NMR
2-(n -Bu t oxy)a d en osin e (6). Product 4 (0.79 mmol) was
dissolved in Et3N (4 mL). Sodium butoxide (0.48 mmol Na
dissolved in 5 mL n-butanol) was added, and the solution was
stirred under reflux for 7 days. The mixture was concentrated
under reduced pressure. CHCl3 was added to the residue, and
the suspension was filtered using a membrane filter. The
filtrate was evaporated to dryness and then dissolved in THF
(1 mL). Tetrabutylammonium fluoride (1 M in THF, 3 equiv,
2.4 mL) was added, and the solution was stirred for 1.5 h. The
residue obtained after evaporation was chromatographed on
a
silica gel column (CHCl3:MeOH 15:1). Product 6 was
obtained in 26% yield: 1H NMR (CD3OD) δ 8.12 (s, 1H, H-8),
5.91 (d, J ) 5.5 Hz, 1H, H-1′), 4.75 (t, J ) 5.5 Hz, 1H, H-2′),
4.35 (dd, J ) 5.5, 3.5 Hz, 1H, H-3′), 4.29 (t, J ) 6.5 Hz, 2H,
OCH2), 4.13 (q, J ) 3.5 Hz, 1H, H-4′), 3.88 (dd, J ) 12.5, 3
Hz, 1H, H-5′), 3.74 (dd, J ) 12.5, 3.5 Hz, 1H, H-5′), 1.82-1.30
(m, 4H, OBu), 0.97 (t, J ) 7 Hz, 3H, CH3); 13C NMR (CD3OD,
300 MHz) δ 163.3 (s, C-2), 158.2 (s, C-6), 152.2 (s, C-4), 140.4
(d, C-8), 116.8 (s, C-5), 90.6 (d, C-1′), 87.4 (d, C-4′), 75.2 (d,
C-2′), 72.3 (d, C-3′), 68.3 (t, OCH2), 63.3 (t, C-5′), 32.3 (t,
OCH2CH2), 20.3 (t, CH2CH3), 14.2 (q, CH3); MS (CI/NH3) m/z
340 MH+.
2-(n -Bu tylth ioeth er )aden osin e (8). A suspension of 2-thiol-
adenosine (0.223 g, 0.747 mmol, in 10 mL MeOH) was
dissolved in 0.25 M NaOH (3.35 mL). The clear, yellow solution
was stirred at room temperature for 1 h. After freeze-drying,
the thiolate sodium salt obtained as a yellowish solid was
dissolved in dry DMF (5 mL) and 1-butyl bromide (89 µL, 1.1
equiv) was added. The solution was stirred under nitrogen at
RT for 4 h. The solvent was evaporated in vacuo and the yellow
residue was coevaporated repeatedly with MeOH, until the
residue turned into a yellow solid. The solid was triturated
with petroleumether/ether 1:1, then dissolved in MeOH (2 mL),