Methylenebis(phosphate) Analogues of Pyrophosphates
J. Am. Chem. Soc., Vol. 119, No. 16, 1997 3695
8.10, 8.41 (two 1H singlets, H2, H8). 31P NMR (pyridine): δ 7.31 (br
s), 15.43 (br s).
R-anomer of Ado J1′,2′ ) 4.1 Hz), 6.05 (0.66, d, H1′ of â-anomer of
Ado, J1′,2′ ) 5.1 Hz), 8.16, 8.50 (two 1H doublets, H2, H8, Ado). 31P
NMR (D2O): δ 18.0 (brs).
Synthesis of P1,P4-Disubstituted Bicyclic Trisanhydrides (BTAs)
7. Nucleoside 5′-methylenebis(phosphonate) (4; 0.1 mmol, prepared
by reaction of 2′,3′-O-isopropylidene-5′-O-mesyladenosine, -uridine,
and -N4-acetylcytidine with the tris(tetrabutylammonium) salt of
methylenebis(phosphonic acid) in DMSO10) was dissolved or suspended
in dry pyridine (1 mL) containing DCC (103 mg, 0.33 mmol) and stirred
(2-5 h) until 31P NMR of the reaction mixture showed multisignal
resonances characteristic for BTAs 7. Treatment of 6 with DCC
afforded an identical pattern of phosphorus resonances. See Figure 3
(adenosine-BTA obtained from 4) , Figure 2 (adenosine-BTA obtained
from 6), Figure 4 (N4-acetylcytidine-BTA), Figure 5 (uridine-BTA),
and Figure 6 (adenosine-BTA, 31P NMR (P,P)-COSY spectrum).
Treatment of 4 or 6 with DIC afforded a mixture showing the same
pattern of 31P resonances.
(3) P1-(Riboflavin-5′-yl)-P2-(2′,3′-O-isopropylideneadenosin-5′-yl)-
methylenebis(phosphonate). Yield: 12.5%. 1H NMR (D2O): δ 1.40
(3H, s, Me-isopropylidene), 1.63 (3H, s, Me-isopropylidene), 2.21 (2H,
t, P-CH2-P, JP-H ) 19.8 Hz), 2.32 (3H, s, Me-flavin), 2.39 (3H, s,
Me-flavin), 3.89-4.06 (3H, m, H4′, H5′, H5′′). Deisopropylidenation
on Dowex 50/H+ gave FAD analogue 10c. 1H NMR (D2O): δ 2.30
(2H, t, P-CH2-P, JP-H ) 19.9 Hz), 2.35 (3H, s, Me-flavin), 2.41 (3H,
s, Me-flavin), 3.80-5.00 (12H, m, ribitol and ribose protons), 5.81
(1H, d, H1′, Ado, J1′,2′ ) 5.1 Hz), 7.56 and 7.67 (two 1H singlets,
flavin), 7.82 and 8.39 (two 1H singlets, H2, H8, Ado). 31P NMR
(D2O): δ 18.17 and 18.77 (AB system, JA,B ) 12.3 Hz).
(4) P1-[(N-Acetylamino)ethyl]-P2-(2′,3′-O-isopropylidene-N4-ace-
tylcytidin-5′-yl)methylenebis(phosphonate). Yield: 83%. 1H NMR
(D2O): δ 1.23 and 3.15 (Et3NH+), 1.37 and 1.57 (3H each, isopropy-
lidene), 1.89 (3H, s, N-Ac), 2.05 (2H, t , P-CH2-P, JP-H ) 19.8 Hz),
2.19 (3H, s, N4-Ac), 3.31 (2H, m, CH2-N, ethanolamine), 3.89 (3H, m,
CH2O-P, ethanolamine), 4.03-4.19 (2H, m , H5′, H5), 4.69 (1H, m,
H4′), 4.95 (1H, dd, H2′, J2′,3′ ) 6.1 Hz), 5.01 (1H, dd, H3′, J3′,4′ ) 1.4
Hz), 5.85 (1H, d, H1′, J1′,2′ ) 2.2 Hz), 7.31 and 8.29 (1H each, d H5,
H6, J5,6 ) 7.6 Hz). 31P NMR (D2O): δ 17.73 (br s).
Synthesis of P1,P2-Disubstituted Methylenebis(phosphonate)s. At
the time the formation of bicyclic anhydride 7 was completed,
compound 8, 11, 12, 13, or 14 (0.15 mmol) was added and the mixture
was heated at 65 °C for from 2 h to 2 days until the 31P NMR spectrum
of the reaction showed the formation of intermediate 9 (two multiplets
at δ 8-9 and δ 18-21). Then reaction was quenched by addition of
water, and the mixture was stirred at room temperature for 1-3 h,
filtered, and concentrated in Vacuo. The residue was chromatographed
on an HPLC column to give the following compounds.
Treatment with Dowex 50/H+ gave 10d. 1H NMR (D2O): δ 1.97
(3H, s, N-Ac), 2.18 (2H, t, P-CH2-P, JP-H ) 19.8 Hz), 3.38 (2H, t,
CH2-N, J ) 5.4 Hz), 3.95 (3H, m, CH2-OP), 4.10-4.36 (5H, m,
H2′,3′,4′,5′,5′′), 5.96 (1H, d, H1′, J1′,2′ ) 3.6 Hz), 6.10 (1H, d, H6, J5,6
) 7.6 Hz), 8.00 (1H, d, H5, J5,6 ) 7.6 Hz). 31P NMR (D2O): δ 18.01,
18.05 (AB system, JA,B ) 10.8 Hz).
(1) P1-(2′,3′-O-Isopropylidenetiazofurin-5′yl)-P2-(2′,3′-O-isopro-
pylideneadenosin-5′-yl)methylenebis(phosphonate) (10). Yield: 95%.
1H NMR (D2O): δ 1.26 (18H, t, Et3NH+), 1.38, 1.45, 1.60, and 1.67
(3H each, Me-isopropylidene of tiazofurin and Ado), 2.07 (2H, t, P-CH2-
P, JP-H ) 20.1 Hz), 3.19 (12H, q, Et3NH+), 3.88-3.98 (2H, m, H5′,5′′
of tiazofurin), 4.07-4.12 (2H, H5′,5′′ of Ado), 4.35-4.38 (1H, m, H4′
of tiazofurin), 4.59-4.62 (1H, m, H4′ of Ado), 4.89-4.95 (2H, m,
H2′, 3′ of tiazofurin), 5.09 (1H, d, H1′ of tiazofurin, J1′2′ ) 3.4 Hz),
5.22 (1H, dd, H3′, J2′,3′ ) 6.2 Hz, J3′,4′ ) 2.2 Hz), 5.36 (1H, dd, H2′,
J1′,2′ ) 3.5 Hz), 6.18 (1H, d, H1′), 8.06, 8.14, and 8.42 (three 1H singlets,
H2, H8 of Ado and H5 of tiazofurin). 31P NMR (D2O): δ 17.80 (AB
system, JP,P ) 10.0 Hz). This compound was deprotected on Dowex
(5) P1-(1,2-Dipalmitoyl-sn-glycer-3-yl)-P2-(2′,3′-O-isopropylidene-
N4-acetylcytidin-5′-yl)methylenebis(phosphonate) 10e. Yield 19%.
1H NMR (CD3OD): δ 0.89 (6H, t, CH3-palmitoyl, J ) 6.6 Hz), 1.28
(76H, m, CH2-palmitoyl), 1.34 and 1. 55 (3H each, s, CH3-isopropy-
lidene), 1.5 (4H, m, CH2-CH2COOH), 2.15 (2H, t, P-CH2-P, JP-H
)
19.8 Hz), 2.17 (3H, s, acetyl), 2.28 and 2.30 (2H each, t, CH2COOH,
J ) 7.3 and J ) 7.6 Hz), 3.18 (12H, q, CH2-N, J ) 7.3 Hz), 4.07 (2H,
m, CH2-3-glycerol), 4.19 (3H, m, H5′, H5′′, CH2-1-glycerol), 4.48 (2H,
m, H4′, CH2-1-glycerol), 4.89 (1H, dd, H3′, J2′,3′ ) 6.1 Hz, J3′,4′ ) 2.5
Hz), 5.01 (1H, dd, H2′, J1′,2′ ) 2.7 Hz), 5.24 (1H, m, CH-2-glycerol),
6.01 (1H, d, H1′, J1′,2′ ) 2.7 Hz), 7.45 (1H, d, H5, J5,6 ) 7.5 Hz), 8.41
(1H, d, H6). 31P NMR (CD3OD): δ 16.56 and 16.58 (AB system, JP,P
) 3.5 Hz).
1
50/H+ to give 10a (97%). The H NMR spectrum was identical with
that of an authentic sample.
(2) P1-(Benzyl-2,3-O-isopropylidene-â-D-ribos-5-yl)-P2-(2′,3′-O-
isopropylideneadenosin-5′-yl)methylenebis(phosphonate). Yield:
78%. 1H NMR (D2O) δ: 1.21 and 3.12 (Et3NH+), 1.23, 1.37, 1.40,
and 1.62 (3H each, Me-isopropylidene) 2.09 (2H, t, P-CH2-P, JP-H
)
Adenosine Phosphonoacetate Anhydride. Adenosine phospho-
noacetate12 (16 mg, 0.03 mmol) was dissolved in pyridine-d5 (0.5 mL)
containing DCC (6.7 mg, 0.032 mmol) and kept at room temperature
for 2 h until disappearance of the resonance signal of the starting
material at δ 11.08 and formation of new peak of the product at δ 5.38
in 31P NMR. The reaction mixture was concentrated in Vacuo, and
the residue was purified on a HPLC column to give the adenosine
phosphonoacetate anhydride (26 mg, 78%) as the bis(triethylammo-
nium) salt. 1H NMR (D2O): δ 1.19 (t, 18H, Et3N), 2.96 (q, 12H, Et3N),
3.08-3.18 (t, 2H, P-CH2-CO), 4.41-4.43 (m, 3H, H4′, 5′, 5′′), 4.52-
20.0 Hz), 3.8 (2H, m, H5,5′, ribose), 4.05 (2H, t, H5′, H5′′, Ado, J4′,5′
) J4′,5′′ ) 4.6 Hz,), 4.27 (1H, t, H4, ribose, J4,5 ) J4,5′ ) 7.7 Hz), 4.30
and 4.50 (2H, two d, PhCH2, J ) 11.4 Hz), 4.55 (1H, m, H4′, Ado),
4.56 (1H, d, H3, ribose, J2,3 ) 6.0 Hz), 4.78 (1H, H2, ribose), 5.06
(1H, s, H1, ribose), 5.12 (1H, dd, H3′, Ado, J3′,4′ ) 2.2 Hz), 5.25 (1H,
dd, H2′′, Ado, J2′,3′ ) 6.2 Hz), 6.09 (1H, d, H1′, Ado J1′,2′ ) 3.5 Hz),
7.22 (5H, m, phenyl), 8.06, 8.37 (two 1H singlets, H2, H8, Ado). 31P
NMR (D2O): δ 17.47 (d, JP-C-P ) 10.8 Hz), 17.73 (d, JP-C-P ) 10.8
Hz). Deprotection on Dowex 50/H+ afforded 10b as a 1:2 mixture of
R,â-anomers. 1H NMR (D2O): δ 2.18 (0.7 H, t, P-CH2-P, JP-H
)
4.55 (m, 1H, H3′), 4.90 (pseudo-t, 1H, H2′), 6.03 (d, 1H, H1′, J1′,2′
)
19.9 Hz, R-anomer), 2.20 (1.3H, t, P-CH2-P, JP-H ) 19.9 Hz,
â-anomer), 3.8-4.8 (10H, all H atoms of sugars except H1 of ribose
and H1′ of Ado), 5.18 (0.66H, d H1 of â-ribose, J1,2 ) 1.6 Hz), 5.30
(0.33H, d, H1 of R-ribose, J1,2 ) 4.0 Hz), 6.00 (0.33H, d, H1′ of
5.9 Hz), 8.09, 8.35 (two 1H singlets, H2, H8). 31P NMR (D2O): δ
4.91 (s). MS (ES): m/z 759 (M - H)-.
JA964058I