Cell-type Selective cADPR Analogues
A R T I C L E S
(dd, 1H, H-2′, J2′,1′ ) 1.5, J2′,3′ ) 4.4 Hz), 4.52 (dd, 1H, H-3′, J3′,2′
4.4, J3′,4′ ) 6.3 Hz), 4.32 (m, 1H, H-4′), 4.13 (d, 1H, H-5′a, J5′a,5′b
11.2 Hz), 3.93 (m, 2H, H-5′′ × 2), 3.86 (d, 1H, H-5′b, J5′b,5′a
)
)
)
29 mg, 41 µmol) as described for the synthesis of 8, using aqueous
80% HCO2
H instead of aqueous 60% HCO H: 1H NMR (D2O,
2
400 MHz, J values were determined by spin decoupling) δ 9.05
(s, 1H, H-2), 8.39 (s, 1H, H-8), 6.08 (d, 1H, H-1′, J1′,2′ ) 6.1 Hz), 5.19
11.2 Hz), 3.25 (m, 1H, H-4′′), 3.20 (q, 6H, Et3NH-CH2 × 3, J )
7.3 Hz), 2.94 (ddd, 1H, H-6′′a, J6′′a,6′′b ) 15.2, J6′′a,1′′ ) 7.8, J6′′a,4′′
9.5 Hz), 2.07 (ddd, 1H, H-6′′b, J6′′b,6′′a ) 15.2, J6′′b,4′′ ) 2.0, J6′′b,1′′
)
)
(dd, 1H, H-2′, J2′,1′ ) 6.1, J2′,3′ ) 4.7 Hz), 4.86 (dd, 1H, H-1′′, J1′′,2′′
)
2.9, J1′′,6′′a ) 9.3 Hz), 4.64 (dd, 1H, H-3′, J3′,2′ ) 4.7, J3′,4′ ) 2.6 Hz),
4.54 (ddd, 1H, H-2′′, J2′′,1′′ ) 2.9, J2′′,3′′a ) 5.6, J2′′,3′′b ) 5.4 Hz), 4.51
1.5 Hz), 1.28 (t, 9H, Et3NH-CH3 × 3, J ) 7.3 Hz); 13C NMR (D2O,
68 MHz) δ 150.37, 147.73, 144.62, 143.54, 141.86, 126.98, 120.15,
90.80, 83.71, 83.56, 74.26, 69.85, 67.16, 67.09, 65.54, 63.85, 47.05,
46.24, 46.10, 32.08, 8.65; 31P NMR (D2O, 108 MHz) δ -8.94, -10.80,
J ) 14.3 Hz; HRMS (FAB, positive) calcd for C16H22N5O10P2 506.0842
(MH+), found 506.0803; UV (H2O) λmax ) 259 nm.
(m, 1H, H-5′a), 4.42 (m, 1H, H-4′), 4.20 (dd, 1H, H-5′′a, J5′′a,5′′b
)
10.3, J5′′a,4′′ ) 2.9 Hz), 4.13 (m, 1H, H-5′b), 4.02 (dd, 1H, H-5′′b, J5′′b,5′′a
) 10.3, J5′′b,4′′ ) 3.9 Hz), 2.93 (ddd, 1H, H-6′′a, J6′′a,6′′b ) 16.6, J6′′a,1′′
) 9.3, J6′′a,4′′ ) 10.0 Hz), 2.80 (dddddd, 1H, H-4′′, J4′′,3′′a ) 6.1, J4′′,3′′b
) 7.8, J4′′,5′′a ) 2.9, J4′′,5′′b ) 3.9, J4′′,6′′a ) 10.0, J4′′,6′′b ) 2.2 Hz), 2.50
(dd, 1H, H-6′′b, J6′′b,6′′a ) 16.6, J6′′b,4′′ ) 2.2 Hz), 2.17 (ddd, 1H, H-3′′a,
J3′′a,3′′b ) 14.2, J3′′a,2′′ ) 5.6, J3′′a,4′′ ) 6.1 Hz), 1.99 (ddd, 1H, H-3′′b,
J3′′b,3′′a ) 14.2, J3′′b,2′′ ) 5.4, J3′′b,4′′ ) 7.8 Hz); 13C NMR (D2O,
125 MHz) δ 151.86, 146.68, 145.36, 120.66, 90.88, 85.36, 85.27, 78.35,
73.68, 71.16, 67.81, 65.26, 37.53, 37.45, 35.66, 30.75; 31P NMR (D2O,
202 MHz) δ -9.44 (d, J ) 11.4 Hz), -10.42 (d, J ) 11.4 Hz); HRMS
(FAB, positive) calcd for C16H24N5O11P2 524.0948 (MH+), found
524.0977; UV (H2O) λmax 260 nm.
2′′,3′′-Dideoxy-cyclic ADP-Carbocyclic Ribose (7). Compound
7 (44 mg, quant.) was obtained from 33b (see Supporting Information,
48 mg, 73 µmol) as described for the synthesis of 8: 1H NMR (D2O,
400 MHz, J values were determined by spin decoupling) δ 9.04
(s, 1H, H-2), 8.37 (s, 1H, H-8), 6.07 (d, 1H, H-1′, J1′,2′ ) 5.4 Hz), 5.17
(dd, 1H, H-2′, J2′,1′ ) 5.4, J2′,3′ ) 4.4 Hz), 4.99 (m, 1H, H-1′′), 4.63
(dd, 1H, H-3′, J3′,2′ ) 4.4, J3′,4′ ) 2.4 Hz), 4.39 (m, 1H, H-4′), 4.36
(m, 1H, H-5′a), 4.12 (m, 2H, H-5′′ × 2), 3.97 (m, 1H, H-5′b), 3.19
(q, 6H, Et3NH-CH2 × 3, J ) 7.3 Hz), 2.66 (m, 2H, H-6′′a, H-4′′),
2.46 (m, 1H, H-6′′b), 2.30 (m, 1H, H-2′′a), 2.15 (m, 1H, H-2′′b), 1.95
(m, 1H, H-3′′a), 1.82 (m, 1H, H-3′′b), 1.27 (t, 9H, Et3NH-CH3 × 3,
J ) 7.3 Hz); 13C NMR (D2O, 68 MHz) δ 155.727, 145.987, 142.358,
140.615, 123.221, 90.535, 85.036, 84.874, 73.572, 71.254, 67.337,
65.270, 58.586, 46.403, 39.143, 38.999, 33.644, 31.830, 26.726, 9.566;
31P NMR (D2O, 108 MHz) δ -8.98 (d, J ) 13.4 Hz), -10.23 (d, J )
13.4 Hz); HRMS (FAB, positive) calcd for C16H24N5O10P2 508.0998
(MH+), found 508.1006; UV (H2O) λmax 259.
2′′-O-(Methoxymethyl)-3′′-O-methyl-cyclic ADP-Carbocyclic
Ribose (6). Compound 6 (7.6 mg, 56%) was obtained from 54 (see
Supporting Informatin, 12 mg, 16 µmol) as described for the synthesis
of 7, using aqueous 80% HCO2H instead of aqueous 60% HCO2H:
1H NMR (D2O, 400 MHz, J value was determined by spin decoupling)
δ 9.04 (s, 1H, H-2), 8.31 (s, 1H, H-8), 6.04 (d, 1H, H-1′, J1′,2′
)
6.3 Hz), 5.14 (dd, 1H, H-2′, J2′,1′ ) 6.3, J2′,3′ ) 4.9 Hz), 4.98 (ddd, 1H,
H-1′′, J1′′,2′′ ) 3.9, J1′′,6′′a ) 10.2, J1′′,6′′b ) 2.9 Hz), 4.63 (dd, 1H, H-3′,
J3′,2′ ) 4.9, J3′,4′ ) 2.4 Hz), 4.53 (m, 2H, H-2′′, H-5′a), 4.41 (m, 1H,
H-4′), 4.20 (m, 2H, H-5′′ × 2), 4.14 (m, 1H, H-5′b), 3.92 (dd, 1H,
H-3′′, J3′′,2′′ ) 4.4, J3′′,4′′ ) 5.9 Hz), 3.43 (s, 3H, Me), 2.99 (ddd, 1H,
H-6′′a, J6′′a,6′′b ) 16.6, J6′′a,1′′ ) 10.2, J6′′a,4′′ ) 11.2 Hz), 2.65 (br s, 1H,
2′′-Deoxy-Cyclic ADP-Carbocyclic Ribose (3). Compound 3
(4.1 mg, quant.) was obtained from 52 (see Supporting Information,
4.5 mg, 6.8 µmol) as described for the synthesis of 8: 1H NMR (D2O,
400 MHz, J value was determined by spin decoupling) δ 9.07 (s, 1H,
H-4′′), 2.37 (ddd, 1H, H-6′′b, J6′′b,6′′a ) 16.6, J6′′b,4′′ ) 3.9, J6′′b,1′′
)
H-2 or H-8), 8.38 (s, 1H, H-2 or H-8), 6.08 (d, 1H, H-1′, J1′,2′
)
2.9 Hz); 13C NMR (D2O, 125 MHz) δ 152.81, 145.02, 144.46, 121.18,
90.89, 85.37, 85.28, 83.19, 76.27, 73.84, 71.18, 65.62, 65.30, 63.55,
57.88, 40.97, 40.90, 28.19; 31P NMR (D2O, 202 MHz) δ -9.41 (d,
J ) 11.4 Hz), -10.53 (d, J ) 11.4 Hz); HRMS (FAB, negative) calcd
for C17H24N5O12P2 552.0897 [(M-H)-], found 552.0903; UV (H2O)
λmax 260 nm.
5.9 Hz), 5.18 (dd, 1H, H-2′, J2′,1′ ) 5.9, J2′,3′ ) 4.9 Hz), 5.08 (m, 1H,
H-1′′), 4.64 (dd, 1H, H-3′, J3′,2′ ) 4.9, J3′,4′ ) 3.0 Hz), 4.44 (m, 3H,
H-4′, H-5′a, H-5′′a), 4.17 (m, 3H, H-3′′, H-5′b, H-5′′b), 2.96 (dd, 1H,
H-6′′a, J6′′a,6′′b ) 17.2, J6′′a,4′′ ) 11.2 Hz), 2.49 (ddd, 1H, H-6′′b, J6′′b,6′′a
) 17.2, J6′′b,1′′ ) 8.4, J6′′b,4′′ ) 3.6 Hz), 2.35 (m, 3H, H-4′′, H-2′′ × 2);
13C NMR (D2O, 125 MHz) δ 144.61, 90.75, 85.19, 73.70, 72.08, 71.09,
65.23, 64.68, 59.11, 46.16, 40.31, 30.22; 31P NMR (D2O, 202 MHz)
δ -9.18, -10.39, J ) 11.4 Hz; HRMS (FAB, positive) calcd for
C16H24N5O11P2 524.0948 (MH+), found 524.0932; UV (H2O) λmax
259 nm.
2′′-O-(Methoxymethyl)-3′′-deoxy-cyclic ADP-Carbocyclic Ribose
(5). Compound 5 (13 mg, quant.) was obtained from 53 (see Supporting
Information, 13 mg, 18 µmol) as described for the synthesis of 8:
1H NMR (D2O, 500 MHz) δ 9.18 (s, 1H, H-2 or H-8), 8.41 (s, 1H,
H-2 or H-8), 6.08 (d, 1H, H-1′, J1′,2′ ) 5.3 Hz), 5.14 (dd, 1H, H-2′,
J2′,1′ ) 5.3, J2′,3′ ) 4.8 Hz), 4.98 (m, 1H, H-1′′), 4.88 (d, 1H, MOM-
CH2, J ) 7.2 Hz), 4.77 (d, 1H, MOM-CH2, J ) 7.2 Hz), 4.63 (dd, 1H,
H-3′, J3′,2′ ) 4.8, J3′,4′ ) 2.2 Hz), 4.52 (m, 1H, H-5′a), 4.43 (m, 2H,
H-4′, H-2′′), 4.22 (m, 1H, H-5′′a), 4.13 (m, 1H, H-5′b), 4.10 (m, 1H,
H-5′′b), 3.43 (s, 3H, MOM-CH3), 3.20 (q, 6H, Et3NH-CH2 × 3, J )
7.3 Hz), 2.98 (m, 1H, H-6′′a), 2.81 (m, 1H, H-4′′), 2.50 (m, 1H, H-6′′b),
2.19 (m, 1H, H-3′′a), 2.06 (m, 1H, H-3′′b), 1.28 (t, 9H, Et3NH-CH3
× 3, J ) 7.3 Hz); 13C NMR (D2O, 125 MHz) δ 151.45, 147.02, 145.60,
120.51, 97.15, 90.90, 86.02, 85.48, 85.39, 73.84, 71.19, 67.53, 66.34,
65.30, 56.19, 47.15, 37.25, 37.18, 33.72, 30.72, 8.68; 31P NMR (D2O,
202 MHz) δ -9.48 (s), -10.45 (s); HRMS (FAB, positive) calcd for
C18H28N5O12P2 568.1210 (MH+), found 568.1212; UV (H2O) λmax
260 nm.
Acknowledgment. This investigation was supported by a
Grant-in-Aid for Scientific Research on “Exploitation of Mul-
tielement Cyclic Molecules” (S.S.) from the Ministry of
Education, Culture, Sports, Science and Technology, Japan, and
a Grant-in-Aid for Creative Scientific Research (Grant 13NP0401,
A.M.) from the Japan Society for the Promotion of Science.
We thank the Wellcome Trust for a Project Grant (055709 to
B.V.L.P.) and a Biomedical Research Collaboration Grant
(068065 to B.V.L.P. and A.H.G.). This study was also partially
supported by the Deutsche Forschungsgemeinschaft (Grant GU
360/0-1 and 9-2 to A.H.G.), Werner-Otto-Stiftung (to A.H.G.),
and Gemeinnu¨tzige Hertie-Stiftung (Grant 1.01.1/04/010 to
A.H.G.). We also thank Mr. Kohei Nozawa for his kind
suggestion on the NOE-based calculations.
Supporting Information Available: Experimental procedures
for the synthesis of compounds other than the final compounds
3-8, biological evaluations, and calculations and Figure S1
(PDF). This material is available free of charge via the Internet
3′′-Deoxy-cyclic ADP-Carbocyclic Ribose (4). Compound 4
(22 mg, 87%) was obtained from 53 (see Supporting Information,
JA050732X
9
J. AM. CHEM. SOC. VOL. 127, NO. 24, 2005 8855