Nucleosides and Nucleotides. 173. Synthesis of Cyclic IDP-carbocyclic-ribose, a Stable Mimic of Cyclic ADP-ribose. Significant Facilitation of the Intramolecular Condensation Reaction of N-1-(Carbocyclic-ribosyl)inosine 5′,6″-Diphosphate Derivatives by an 8-Bromo-Substitution at the Hypoxanthine Moiety

Article abstract of DOI:10.1021/jo9717797

Cyclic ADP-ribose (cADPR, 1) is a general mediator involved in cellular Ca²⁺ signaling. However, both the biological and chemical instability of cADPR limit studies on its physiological role. We designed cyclic ADP-carbocyclic-ribose (3) and its inosine congener 4 as stable mimics of cADPR and successfully synthesized 4. Starting with cyclopentadiene, the optically active carbocyclic unit 8 was constructed via enzymatic optical resolution. S_N2 reactions of 8 with inosine derivative 7 and the 8-bromoinosine derivative 25 gave the N-1-substituted derivatives 6 and 26, which were converted to the corresponding diphosphate derivatives 5 and 22. The intramolecular condensation reactions between the two phosphate groups of 5 and 22 were investigated. Although the reaction with inosine derivative 5 did not produce any of the cyclization product 20, treatment of the corresponding 8-bromoinosine derivative 22 with EDC gave the desired intramolecular condensation product 29 in 23% yield. Thus, the significant effect of the 8-bromo group at the hypoxanthine moiety in facilitating the key intramolecular condensation reaction between the phosphate groups of the substrate 22 was recognized. This is possibly due to conformational restriction of the molecule in a syn-form around its glycosyl linkage. The 8-bromo and isopropylidene groups were removed in succession to give the target compound 4. This is the first total synthesis of this type of cyclic nucleotide.

Full text of DOI:10.1021/jo9717797

1
986
J . Org. Chem. 1998, 63, 1986-1994
Nu cleosid es a n d Nu cleotid es. 173. Syn th esis of Cyclic
IDP -ca r bocyclic-r ibose, a Sta ble Mim ic of Cyclic ADP -r ibose.
Sign ifica n t F a cilita tion of th e In tr a m olecu la r Con d en sa tion
Rea ction of N-1-(Ca r bocyclic-r ibosyl)in osin e 5′,6′′-Dip h osp h a te
Der iva tives by a n 8-Br om o-Su bstitu tion a t th e Hyp oxa n th in e
Moiety
Satoshi Shuto,* Michiyo Shirato, Yuji Sumita, Yoshihito Ueno, and Akira Matsuda*
Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060, J apan
Received September 24, 1997
Cyclic ADP-ribose (cADPR, 1) is a general mediator involved in cellular Ca2+ signaling. However,
both the biological and chemical instability of cADPR limit studies on its physiological role. We
designed cyclic ADP-carbocyclic-ribose (3) and its inosine congener 4 as stable mimics of cADPR
and successfully synthesized 4. Starting with cyclopentadiene, the optically active carbocyclic unit
8
N
was constructed via enzymatic optical resolution. S 2 reactions of 8 with inosine derivative 7
and the 8-bromoinosine derivative 25 gave the N-1-substituted derivatives 6 and 26, which were
converted to the corresponding diphosphate derivatives 5 and 22. The intramolecular condensation
reactions between the two phosphate groups of 5 and 22 were investigated. Although the reaction
with inosine derivative 5 did not produce any of the cyclization product 20, treatment of the
corresponding 8-bromoinosine derivative 22 with EDC gave the desired intramolecular condensation
product 29 in 23% yield. Thus, the significant effect of the 8-bromo group at the hypoxanthine
moiety in facilitating the key intramolecular condensation reaction between the phosphate groups
of the substrate 22 was recognized. This is possibly due to conformational restriction of the molecule
in a syn-form around its glycosyl linkage. The 8-bromo and isopropylidene groups were removed
in succession to give the target compound 4. This is the first total synthesis of this type of cyclic
nucleotide.
In tr od u ction
Cyclic ADP-ribose (cADPR, 1), a recently discovered
2
cyclic nucleotide, has been shown to mobilize intracel-
lular Ca2 in various cells, indicating that it is a general
+
mediator involved in Ca signaling.3 It is also known
2+
that cADPR mobilizes Ca2 more actively than inositol
+
1
,4,5-triphosphates (IP
3
) by a mechanism completely
The structure of cADPR has been
investigated, and the structure shown in Figure 1 was
3
3
independent of IP .
4
4
c
recently confirmed by X-ray crystallographic analysis.
In cells, cADPR is synthesized from NAD+ by ADP-
ribosyl cyclase and acts as a transient second messenger;
it is hydrolyzed promptly by cADPR hydrolase to give
ADP-ribose and inactivated under physiological condi-
tions.3 cADPR is also known to be readily hydrolyzed
nonenzymatically at the unstable N-1-glycosidic linkage
F igu r e 1.
of its adenine moiety to give ADP-ribose, even in neutral
aqueous solution (t1/2 ) 40 h).5 Although further inten-
sive studies of cADPR is needed because of its biological
importance, this biological as well as chemical instability
of cADPR limits studies of its physiological role, at least
to some extent. Therefore, stable analogues of cADPR
that have a Ca -mobilizing activity in cells similar to
that of cADPR are urgently required.
Recently, the synthesis of cADPR analogues and their
biological effects have been studied extensively.
(
1) Part 172: Shuto, S.; Kanazaki, M.; Ichikawa, S.; Minakawa, N.;
Matsuda, A. J . Org. Chem., in press.
2) Clapper, D. L.; Walseth, T. F.; Dargie, P. J .; Lee, H. C. J . Biol.
Chem. 1987, 262, 9561-9568.
(
2+
(
3) (a) Galione, A. Trends Pharmacol. Sci. 1992, 13, 304-306. (b)
Galione, A. Science 1993, 259, 325-326. (c) Lee, H. C.; Galione A.;
Walseth, T. F. Vitamines Hormones 1994, 48, 199-257. (d) J acobson,
M. K.; Ame, J . C.; Lin, W. S.; Coyle, D. L.; J acobson, E. L. Receptor
6,7
One
1
995, 5, 43-49. (e) Lee, H. C. Cell. Signalling 1994, 6, 591-600.
analogue, cyclic aristeromycin-diphosphate-ribose (2),
reported by Potter and co-workers, may be a useful
biological tool, since it acts as a poorly hydrolyzable Ca
mobilizing mimic of cADPR. However, 2 is hydrolyzed
(
4) (a) Lee, C. H.; Walseth, T. F.; Bratt, G. T.; Hayes, R. N.; Clapper,
6i
D. L. J . Biol. Chem. 1989, 264, 1608-1615. (b) Kim, H.; J acobson, E.
L.; J acobson, M. K. Biochem. Biophys. Res. Commun. 1993, 194, 1143-
2
+
-
1
1
7
147. (c) Lee, H. C.; Aarhus, R.; Levitt, D. Nature Struct. Biol. 1994,
, 143-144. (d) Gu, Q.-M.; Sih, C. J . J . Am. Chem. Soc. 1994, 116,
481-7486. (e) Wada, T.; Inageda, K.; Aritomo, K.; Tokita, K.; Nishina,
H.; Takahashi, K.; Katada, T.; Sekine, M. Nucleosides Nucleotides,
(5) Lee, H. C.; Aarhus, R. Biochim. Biophys. Acta 1993, 1164, 68-
74.
1
995, 14, 1301-1341.
S0022-3263(97)01779-9 CCC: $15.00 © 1998 American Chemical Society
Published on Web 02/27/1998

Carbocyclic Analogue of cADP-ribose
J . Org. Chem., Vol. 63, No. 6, 1998 1987
Sch em e 1
Sch em e 2
by enzymes, although the rate is considerably slower
than that of cADPR.6i We designed cyclic ADP-carbocy-
clic-ribose (3) and its inosine congener 4 (cIDP-carbocy-
clic-ribose), as stable mimics of cADPR, in which an
oxygen in the ribose ring of cADPR is replaced by a
methylene group. The mimics 3 and 4 should be com-
pletely resistant to both enzymatic and chemical hydroly-
sis, because they lack the unstable N-1-glycosidic linkage
of cADPR. These analogues preserve all the functional
groups of cADPR, except for this ring oxygen, and the
conformation of these molecules is similar to that of
cADPR. Therefore, we expect that these analogues would
effectively mobilize intracellular Ca2 , in the same way
as cADPR, so that they could be used as pharmacological
tools for proving the mechanism of cADPR-modulated
to attempt a total synthesis of 3 and 4. In this paper,
we describe the synthesis of the inosine congener 4.
9
Resu lts a n d Discu ssion
Our synthetic plan is shown in Scheme 2. Cyclization
of the 18-membered ring is carried out by intramolecular
condensation between the two phosphate groups of
diphosphate 5, which can be prepared from the N-1-
(carbocyclic-ribosyl)inosine derivative 6. Compound 6 is
N
obtained in an S 2 reaction between carbocyclic unit 8
+
(6) (a) Walseth, T. F.; Lee, H. C. Biochim. Biophys. Acta 1993, 1178,
35-242. (b) Lee, H. C.; Aarhus, R.; Walseth, T. F. Science 1993, 261,
52-355. (c) Graeff, R. M.; Walseth, T. J .; Fryxell, K.; Branton, W. D.;
2
3
Lee, H. C. J . Biol. Chem. 1994, 269, 30260-30267. (d) Zhang, F.-J .;
Sih, C. J . Bioorg. Med. Chem. Lett. 1995, 5, 1701-1706. (e) Zhang,
F.-J .; Gu, Q.-M.; J ing, P. C.; Sih, C. J . Bioorg. Med. Chem. Lett. 1995,
2
+
Ca signaling pathways.
cADPR and its analogues, including 2, have been
synthesized by enzymatic or chemoenzymatic methods.6
As shown in Scheme 1, ADP-ribosyl cyclase from Aplysia
california mediates the intramolecular ribosylation of
5
, 2267-2272. (f) Zhang, F.-J .; Yamada, S.; Gu, Q.-M.; Sih, C. J . Bioorg.
,7
Med. Chem. Lett. 1996, 6, 1203-1208. (g) Zhang, F.-J .; Sih, C. J .
Bioorg. Med. Chem. Lett. 1996, 6, 2311-2316. (h) Ashamu, G. A.;
Galione, A.; Potter, B. V. L. J . Chem. Soc., Chem. Commun. 1995,
1
359-1356. (i) Bailey, B. C.; Fortt, S. M.; Summerhill, R. J .; Galione,
+
+
NAD and some modified NAD (prepared chemically or
A.; Potter, B. V. L. FEBS Lett. 1996, 379, 227-230. (j) Bailey, V. C.;
Sethi, J . K.; Fortt, S. M.; Galione, A.; Potter, B. V. L. Chem. Biol. 1997,
4, 51-60. (k) Bailey, V. C.; Sethi, J . K.; Galione, A.; Potter, B. V. L. J .
Chem. Soc., Chem. Commun. 1997, 695-696.
enzymatically) at the N-1-position of the purine moiety,
to yield cADPR or the corresponding analogues.6 Al-
though the specificity of ADP-ribosyl cyclase is somewhat
loose, the analogues that can be obtained by this method
are limited by the substrate specificity of the enzyme. A
(7) Yamada, S.; Gu, Q.-M.; Sih, C. J . J . Am. Chem. Soc. 1994, 116,
1
0787-10788.
(8) Synthesis of cADPR via N-1-phosphoribosyladenosine 5′-triph-
osphate with the enzymes of the histidine biosynthetic pathway has
been reported (see ref 4d). However, this method is not applicable to
synthesizing various cADPR analogues.
+
chemical intramolecular ribosylation reaction of NAD
mediated by metal halides, such as NaBr, which yields
(9) During preparation of this paper, two communications on
7
cADPR, has also been reported. This synthesis does not
approaches to carbocyclic analogues of cADP-ribose were published.
(a) Synthesis of 1-[(3-hydroxymethyl)cyclopentenyl]inosine diphosphate
offer any advantages over the enzymatic method.6
Accordingly, it is important to develop a general method
for synthesizing cADPR analogues.8 Both the enzymatic
and chemical intramolecular ribosylation reactions using
f,g
3
1 was described: Fortt, S.; Potter, B. V. L. Tetrahedron Lett. 1997,
8, 5371-5374. (b) Synthesis of N -carbocyclic-ribosyladenosine deriva-
1
3
tives, 33 and 34, was described: Hutchinson, E. J .; Taylor, B. F.;
Blackburn, G. M. J . Chem. Soc. Chem. Commun. 1997, 1859-1860.
+
NAD or its analogues as substrates proceed via nucleo-
philic substitution at the anomeric 1′′-position of a
nicotinamide riboside moiety via an oxocarbenium inter-
mediate, such as A (Scheme 1), or an enzymatically
stabilized equivalent. Our targets 3 and 4 could not be
synthesized by the previous methods using enzymatic or
chemical intramolecular ribosylation reaction, because
they lack N-1-glycosidic linkage. We therefore decided

1
988 J . Org. Chem., Vol. 63, No. 6, 1998
Shuto et al.
Sch em e 3
reduction at the 1-position occurred to give the (1S)-
alcohol 17 in 88% yield. The stereochemistries of 15 and
1
1
7 were confirmed from J values in H NMR and NOE
1
4
experiments. The (1S)-alcohol 17 was converted to the
corresponding 1-O-triflate 8. This was unstable and used
for the next reaction immediately without purification.
S
N
2 reactions between the carbocyclic unit 8 and 2′,3′-
O-isopropylidene-5′-O-TBS-inosine 7 in the presence of
1
5,16
a base were investigated under various conditions.
When the reactions were performed using organic bases,
6
such as DBU, or in polar solvents, such as DMF, the O -
substituted derivative 18 was a major product. The use
of relatively softer bases and nonpolar solvents increased
the yield of the desired product 6. The optimum result
occurred when the reaction was performed with equimo-
2 3
lar 8 and 7, in the presence of K CO in DME at 50 °C.
The product included the N-1-(carbocyclic-ribosyl) deriva-
6
tive 6 in 48% yield and the O -substituted derivative 18
in 7% yield. Adding 18-crown-6 to the reaction system
did not improve the yield of 6. The IR spectrum of 6
-
1
showed a strong νCdO absorption peak at 1703 cm
caused by the C-6 carbonyl, which is typical for the 1H-
hypoxanthine structure. Similar absorption around 1700
-
1
13
and the protected inosine derivative 7. Carbocyclic unit
cm was not observed in the spectrum of 18. The
C
1
0
8
can be prepared from an optically active cyclopentene
NMR chemical shift pattern of the base moiety of 6 was
analogous to those of inosine and N-1-ribosyl inosine
derivatives, but the shift pattern of the base moiety of
18 was significantly different. These results suggest
N-1- and O -substituted structures for 6 and 18, respec-
derivative 9, which can also be synthesized from cyclo-
pentadiene.11
1
7
The synthesis of carbocyclic unit 8 was shown in
Scheme 3. Starting with cyclopentadiene, an optically
active diol 912 was prepared via optical resolution with
Pseudomonas fluorescens lipase using the method previ-
ously reported by MacKeith and co-workers.11 The
hydroxyls of 9 were successively protected with dimeth-
ylthexylsilyl (DTS) and acetyl groups to give 11 in 69%
6
tively. The regio- and stereochemistries of 6 were further
confirmed by NOE experiments. When the H-2 of 6 was
irradiated, 4.2% NOE at the H-2′′ of the carbocyclic
moiety was observed. No NOE at the H-2′′ was produced
by irradiation at the H-2 of 18.
Simultaneous removal of the TBS and DTS groups by
treating 6 with TBAF in THF produced 19. The two
phosphate groups were introduced by Yoshikawa’s
method:18 treatment of 19 with POCl in PO(OEt) at 0
°C and subsequent C-18 column chromatography gave
the diphosphate derivative 5 as a triethyammonium salt
in 37% yield. The intramolecular condensation reaction
between the two phosphate groups of 19 was investigated
with 1-(3-(dimethylamino)propyl)-3-ethylcarbodiimide hy-
drochloride (EDC) in N-methylpyrrolidone (MPD) under
various conditions. However, the desired cyclization
product 20 was not obtained.
2
+
yield. Treatment of 11 with Pd in the presence of
1
3
p-benzoquinone in THF produced the desired allylic
rearrangement product 12 in a mixture with 11 (60%,
1
2:11 ) 10:1). When the mixture of 12 and 11 was
treated with OsO in the presence of morpholine N-oxide
NMO) in acetone and t-BuOH, oxidation proceeded in a
3
3
4
(
highly stereoselective manner. The desired cis-diol 13
was isolated in a pure form in 55% yield from 11,
following silica gel column chromatography. After pro-
tecting the cis-diol of 13 with an isopropylidene group,
the 1-O-acetyl group was removed with K
to give (1R)-alcohol 15. The 1-hydroxyl moiety of 15 was
oxidized with PDC/molecular sieves 4 A in CH Cl to
give16 in 92% yield. When the ketone 16 was treated
with NaBH in MeOH at -20 °C, highly stereoselective
2 3
CO in MeOH
2
2
Sih and co-workers attempted to synthesize cADPR by
intramolecular condensation of N-1-phosphoribosyl-AMP
21) with EDC, but it was unsuccessful (<1%), similar
to our result. It is presumed that the conformation of
4
4d
4d
(
(10) Although synthesis of an optically active 1-trifluoromethane-
sulfonyloxy cyclopentane derivative similar to 8 via resolution with
(
+)-N,S-dimethyl-S-phenylsulfoximine or electric eel acetylcholinest-
(14) 15: J 1,2 ) about 0 Hz; NOE, irradiated H-1, observed H-5
(3.2%), H-2 (2.2%). 17: J 1,2 ) 5.4 Hz; NOE, irradiated H-1, observed
H-5â (4.2%), H-2 (5.1%).
(15) Reactions of 7 with the corresponding 1-O-mesylate of 17 was
also tried. However, 6 was not obtained at all.
R
erase has been reported (Schmitt, L.; Caperelli, C. A. Nucleosides
Nucleotides 1995, 14, 1929-1945, and references therein), the synthetic
route is different from our route.
(
11) MacKeith, R. A.; McCague, R.; Olivo, H. F.; Palmer, C. F.;
Roberts, S. M. J . Chem. Soc., Perkin Trans 1 1993, 313-314.
12) Compound 34, a precursor for 9, was converted to the benzoate
5, and its optical purity was determined as 92% ee by chiral HPLC
Chiralcel-OJ , Daicel Chemical).
(16) Sekine et al. investigated N-1 selective ribosylation of sugar-
protected inosines under various conditions, but the yields were very
low: Arimoto, K.; Urashima, C.; Wada, T.; Sekine, M. Nucleosides
Nucleotides 1996, 15, 1-16.
(
3
(
1
3
(
17) C NMR data of base moieties: inosine (Kalinowski, H.-O.;
Berger, S.; Braun, S. Carbon-13 NMR spectroscopy; J ohn Miley &
Sons: 1988); δ 156.9 (C6), 148.5 (C4), 146.2 (C2), 140.2 (C8), 124.6
(
C5). 1-(5-O-Trityl-2, 3-O-isopropylidene-R-D-ribofuranosyl)-2′,3′,5′-O-
tris(tert-butyldimethylsilyl)inosine (ref 16); δ 155.2 (C6), 146.1 (C2),
1
45.1 (C4), 139.5 (C8), 123.4 (C5). 6; δ 156.5 (C6), 146.5 (C2), 146.3
C4), 138.4 (C8), 125.2 (C5). 18; δ 160.1 (C6), 152.5 (C2), 151.9 (C4),
(
(
13) (a) Oehlschlager, A. C.; Mishra, P.; Dhami, S. Can. J . Chem.
141.0 (C8), 122.5 (C5).
1
984, 62, 791-797. (b) Medich, J . R.; Kunnen, K. B.; J ohnson, C. R.
Tetrahedron Lett. 1987, 28, 4131-4134.
(18) Yoshikawa, M.; Kato, T.; Takenishi. T. Bull, Chem. Soc. J pn.
1969, 42, 3505-3508.

Carbocyclic Analogue of cADP-ribose
J . Org. Chem., Vol. 63, No. 6, 1998 1989
phoramidite method. Treatment of 28 with (2-cyanoet-
hoxy)(N,N-diisopropylamino)chlorophosphine and i-Pr -
2
NEt in MeCN afforded the corresponding 5′,6′′-bis(phos-
phoramidite) compound which was successively treated
with I
2
3
/tetrazole in aqueous pyridine-THF and NH /
MeOH to give diphosphate 22, the substrate for cycliza-
tion, in a pure form after DEAE-Sephadex column chro-
matography. The intramolecular condensation reaction
of 22 was investigated with EDC in MPD. At room
temperature, the reaction barely proceeded. However,
when 22 was treated with 2.0 equiv of EDC in MPD at
5
0 °C, the desired cyclization product 29 was obtained
F igu r e 2.
as a sodium salt in 23% yield after purification by ion-
exchange column chromatography. The cyclic structure
of 29 was confirmed by the following data: (1) The
molecular-ion peaks corresponding to 29 were observed
at m/z 697 and at m/z 699 in a FAB mass spectrum; (2)
Its 31P NMR spectrum showed two signals, observed at
the molecule determines whether the intramolecular
condensation reaction between the two phosphate groups
of 5 and 21 occurs. The syn-anti conformation around
the glycosyl linkages of the nucleosides has been recog-
nized as an important determinant of the three-dimen-
sional structure of these molecules.19 Although a syn
conformation in 5 and 21, in which the two phosphate
moieties are near each other, may be preferable for
occurrence of the desired condensation reaction, the
predominance of anti- over syn-conformers is well-known
for natural nucleosides and their analogues. This may
be why the intramolecular condensation reactions of 5
and 21 were unsuccessful. Introducing a bulky substitu-
ent into the 8-position of purine nucleosides is known to
-
10.67 and -10.95 ppm, which are typical chemical
shifts for a pyrophosphate moiety, with a coupling
constant (J ) 15.0 Hz) similar to that of cADPR (-9.92
and -10.67 ppm, J ) 14.6 Hz);4 (3) It was completely
resistant to alkaline phosphatase which hydrolyzes phos-
phomonoester linkages.22 These results suggest that, as
expected, the conformation of 22 may be restricted in a
syn-form to facilitate the intramolecular condensation
reaction between the two phosphates. Next, the 8-bromo
group of 29 was removed reductively by catalytic hydro-
genation with Pd-carbon in MeOH to give 30 in 77%
yield. Finally, 30 was treated with aqueous formic acid,
and the target cIDP-carbocyclic-ribose (4) was isolated
in 67% yield.
e
restrict the conformation in a syn-form, through the steric
repulsion for the ribose moiety.1
9,20
We therefore de-
signed an 8-bromo-substituted substrate 22 for the
intramolecular condensation reaction. As shown in
Figure 2, the 8-bromo group may restrict the conforma-
tion of 22 to the syn-form and facilitate the desired
intramolecular condensation reaction between the two
phosphate groups. After cyclization, it can be readily
removed by catalytic hydrogenation.
In conclusion, we designed carbocyclic analogues 3 and
as stable mimics of cADPR and successfully synthesized
4
the inosine congener 4. This is the first total synthesis
of a cADPR related compound and may lead to the
development of a general method for synthesizing these
compounds, hopefully including 3, our next target. In
this study, we found a significant effect of the 8-bromo
group in the hypoxanthine moiety. This is probably due
to conformational restriction of the molecule in a syn-
form around its glycosyl linkage, which facilitates the key
intramolecular condensation reaction between the phos-
phate groups of the N-1-(carbocyclic-ribosyl)inosine diphos-
phates.
The 8-bromo-substituted substrate 22 was synthesized
following the steps in Scheme 6. 8-Bromo-2′,3′-O-isopro-
pylideneadenosine (23), prepared by a previously re-
ported method,21 was treated with NaNO
in aqueous
2
AcOH to give the corresponding inosine derivative 24.
The 5′-hydroxy group of 24 was protected with a TBS
group to give the 8-bromoinosine unit 25. The S 2
N
reaction between 25 and the carbocyclic unit 8 was
performed in a manner similar to that used for preparing
Synthesis of 3 and the biological evaluation of 4 are
now under investigation.
6
to gave the desired N-1-substituted 26 in 44% yield,
6
along with the corresponding O -substituted isomer 27
9%). The structures of 26 and 27 were confirmed by
(
analyses similar to those described above, used to deter-
mine the structures of 6 and 18. The silyl protecting
groups of 26 were removed with TBAF to give 28. First,
the phosphorylation of the two hydroxy groups of 28 was
Exp er im en ta l Section
Melting points are uncorrected. 1H, 13C, and 31P NMR
1
spectra were recorded at 270, 400, and 500 MHz ( H), at 67.8
13
31
and 125 MHz ( C), and at 125 MHz ( P). Chemical shifts
performed with POCl
3
/PO(OEt)
3
. Although phosphory-
1
13
3
are reported in ppm downfield from TMS ( H and C) or H -
3
1
lation at the two hydroxyl groups proceeded under the
conditions, the reaction was somewhat tricky and not
reproducible: replacement of the 8-bromo group with a
chloro atom was sometimes observed which was recog-
nized from the mass spectrometric analysis. The desired
diphosphate 22 was successfully obtained by a phos-
4
PO ( P), and J values are given in hertz. Mass spectra were
obtained by the electron ionization (EI) or fast atom bombard-
ment (FAB) methods. Thin-layer chromatography was done
on Merck coated plate 60F254. Silica gel chromatography was
done with Merck silica gel 5715. Reactions were carried out
under an argon atmosphere.
1S,5R)-1-Hydr oxy-5-[[(dim eth ylth exylsilyl)oxy]m eth yl]-
-cyclop en ten e (10). To a stirring solution of 9 (5.00 g, 43.8
mmol), DMAP (300 mg, 2.50 mmol), and Et N (15.0 mL, 108
(
1
1
2
(
19) Saenger, W. Principals of nucleic acid structure; Springer-
Verlag: 1983.
20) (a) Travale, S. S.; Sobell, M. J . Mol. Biol. 1970, 48, 109-123.
b) Ikehara, M.; Uesugi, S.; Yoshida, K. Biochemistry 1972, 11, 830-
3
(
(
(22) Compound 29 (0.1 OD) was incubated with calf intestinal
alkaline phosphatase (6 units) in Tris-HCl buffer (500 mM, pH 8.0,
8
36.
(
21) Ikehara, M.; Uesugi, S.; Kaneko, M. Tetrahedron 1970, 26,
2
100 µL) containing 10 mM of MgCl at 37 °C for 12 h, and the reaction
mixture was analyzed by HPLC.
4
251-4259.

1
990 J . Org. Chem., Vol. 63, No. 6, 1998
Shuto et al.
Sch em e 4
Sch em e 5
(1R,2R,3R,4R)-1-Acetoxy-4-[[(dim eth ylth exylsilyl)oxym -
eth yl]cyclop en ta n e-2,3-d iol (13). A mixture of 11 (7.01 g,
3.5 mmol), p-benzoquinone (891 mg, 7.96 mmol), and PdCl
MeCN) (180 mg, 0.690 mmol) in THF (250 mL) was stirred
2
2
-
(
2
at 50 °C for 30 min. After cooling to room temperature, the
mixture was evaporated under reduced pressure, and the
residue was purified by column chromatography (SiO , 5% of
2
EtOAc in hexane) to give 12 as a mixture with 11 (4.20 g, 60%,
1
2:11 ) 10:1, from 1H NMR spectrum) as an oil, which was
1
directly used in the next reaction: H NMR (CDCl
3
, 270 MHz)
δ 6.05-6.02 (m, 1 H), 5.83 (dt, 1 H, J ) 2.1, 5.6), 5.67-5.62
m, 1 H), 3.51 (d, 2 H, J ) 6.8), 2.84-2.77 (m, 1 H), 2.42 (ddd,
H, J ) 8.0, 8.0, 14), 2.00 (s, 3 H), 1.63 (m, 1 H), 1.50 (ddd, 1
H, J ) 8.0, 8.0, 14), 0.88 (d, 6 H, J ) 6.8), 0.82 (s, 6 H), 0.09
(
1
2 2
mmol) in DMF (70 mL) and CH Cl (150 mL) was added DTSCl
(
3.8 mL × 5, at 8 h intervals, total 96.6 mmol) at room
temperature, and the mixture was stirred at room temperature
for tolal 40 h. The resulting mixture was partitioned between
+
13
(
s, 6 H); MS (FAB) m/z 331 (MH ). C NMR (CDCl
MHz) δ 170.9, 138.5, 130.3, 79.7, 66.6, 47.4, 33.1, 20.4, 20.3,
0.1, 20.0, 18.6, 18.5, 18.4, -1.5, -3.5.
A solution of OsO in t-BuOH (5 mg/mL, 3 mL) was added
3
, 67.8
CHCl
dried (Na
residue was purified by column chromatography (SiO
of EtOAc in hexane) to give 10 (8.00 g, 71%) as an oil: 1H NMR
CDCl , 270 MHz) δ 5.97-5.92 (m, 1 H), 5.86-5.80 (m, 1 H),
.90-4.83 (m, 1 H), 3.85 (dd, 1 H, J ) 4.6, 10.0), 3.76 (dd, 1
3
(500 mL) and brine (300 mL). The organic layer was
SO ) and evaporated under reduced pressure. The
, 5-50%
2
2
4
2
4
to a solution of the above oil (4.20 g) and NMO (1.98 g, 16.9
mmol) in acetone (60 mL) and t-BuOH (10 mL), and the
resulting mixture was stirred at room temperature for 8 h.
(
3
4
H, J ) 7.5, 10.1), 2.48-2.38 (m, 1 H), 2.35-2.29 (m, 1 H), 2.23-
2 2 4
After Na S O (1.0 g) and talc (2.0 g) were added, the resulting
2
0
.13 (m, 1 H), 1.62 (m, 1 H, J ) 6.8), 0.88 (d, 6 H, J ) 6.8),
mixture was stirred at room temperature for 20 min, and the
insoluble materials were filtered off. The filtrate was evapo-
rated under reduced pressure, and the residue was purified
.86 (s, 6 H), 0.11, 0.10 (each s, each 3 H); MS (EI) m/z 256
+
(M ). This oil was directly used in the next reaction.
by column chromatography (SiO
to give 13 (4.30 g, 55% from 11) as an oil; H NMR (CDCl
2
, 40% of EtOAc in hexane)
(
1S,5R)-1-Acetoxy-5-[[(dim eth ylth exylsilyl)oxy]m eth yl]-
-cyclop en ten e (11). A mixture of 10 (4.11 g, 16.0 mmol),
N (7.80 mL, 56.0 mmol), and
O (4.9 mL, 52 mmol) in MeCN (300 mL) was stirred at 0
C for 1 h. After EtOH (50 mL) was added, the resulting
1
,
2
3
5
3
00 MHz) δ 4.96-4.92 (m, 1 H), 3.96 (dd, 1 H, J ) 5.3, 9.4),
DMAP (100 mg, 0.82 mmol), Et
Ac
°
3
.95 (dd, 1 H, J ) 4.1, 9.4), 3.72 (dd, 1 H, J ) 4.7, 9.9), 3.55
2
(
(
1
dd, 1 H, J ) 6.3, 9.9), 3.27 (br s, 1 H), 2.84 (br s, 1 H), 2.31
ddd, 1 H, J ) 8.4, 8.4, 13.7), 2.20-2.12 (m, 1 H), 2.05 (s, 3 H),
solution was evaporated under reduced pressure, and the
residue was partitioned between EtOAc (300 mL) and water
.61 (m, 1 H), 1.39 (ddd, 1 H, J ) 4.6, 8.3, 13.7), 0.87 (d, 6 H,
13
J ) 6.8), 0.83 (s, 6 H), 0.09 (s, 6 H). C NMR (CDCl
3
, 125
(
150 mL). The organic layer was washed with brine (150 mL),
dried (Na SO ), and evaporated under reduced pressure. The
residue was purified by column chromatography (SiO , 5% of
EtOAc in hexane) to give 11 (4.60 g, 98%) as an oil: 1H NMR
MHz) δ 171.8, 79.9, 76.8, 74.7, 64.5, 44.3, 34.1, 29.4, 25.1, 21.0,
2
4
1
8
2
0.3, 20.1, 18.5, 18.5, -3.6, -3.7; [R]
D
-0.679 (c 0.588,
2
+
CHCl ); MS (FAB) m/z 333 (MH ). Anal. Calcd for C H O -
3
16 32
5
Si: C, 57.80; H, 9.70. Found: C, 57.70; H, 9.52.
(
(
(
3
CDCl , 500 MHz) δ 6.12-6.06 (m, 1 H), 5.85 (m, 1 H), 5.68
ddd, 1 H, J ) 6.9, 6.9, 2.1), 3.76 (dd, 1 H, J ) 7.3, 9.9), 3.57
dd, 1 H, J ) 7.2, 9.9), 2.59-2.42 (m, 1 H), 2.40-2.35 (m, 1
(1R,2R,3R,4R)-1-Acetoxy-2,3-(isop r op ylid en ed ioxy)-4-
[[(d im et h ylt h exylsilyl)oxy]m et h yl]cyclop en t a n e (14).
A mixture of 13 (5.0 g, 15.0 mmol), dimethoxypropane (18.8
H), 2.28-2.15 (m, 1 H), 2.01 (s, 3 H), 1.60 (m, 1 H), 0.86 (d, 6
H, J ) 6.8), 0.83 (s, 6 H), 0.080, 0.070 (each s, each 3 H);
NMR (CDCl
1
3
C
mL, 153 mmol), and TsOH‚H
(120 mL) was stirred at room temperature for 3 h. After being
neutralized with aqueous saturated NaHCO , the mixture was
2
O (290 mg, 1.53 mmol) in acetone
3
, 67.8 MHz) δ 170.6, 137.1, 129.6, 78.6, 61.6, 43.3,
19
3
-
4.6, 34.2, 24.9, 21.1, 20.4, 20.2, 20.2, 18.5, -3.30, -3.50; [R]
D
3
+
96.7 (c 0.390, CHCl
3
); MS (EI) m/z 213 (M - thexyl). Anal.
H O Si: C, 64.38; H, 10.13. Found: C, 64.36;
evaporated under reduced pressure, and the residue was
partitioned between EtOAc (300 mL) and water (150 mL). The
organic layer was washed with brine (150 mL), dried (Na -
2
Calcd for C16
H, 10.09.
30 3

Carbocyclic Analogue of cADP-ribose
J . Org. Chem., Vol. 63, No. 6, 1998 1991
Sch em e 6
SO
4
), and evaporated under reduced pressure. The residue
, 15% of EtOAc
added, and the resulting mixture was further stirred at room
temperature for 3 h. The reaction mixture was diluted with
was purified by column chromatography (SiO
in hexane) to give 14 (5.1 g, 91%) as an oil; H NMR (CDCl
2
1
3
,
Et
evaporated under reduced pressure, and the residue was
purified by column chromatography (SiO , 10% of EtOAc in
hexane) to give 16 (3.66 g, 92%) as an oil; H NMR (CDCl
2
O (700 mL) and filtered through Celite. The filtrate was
5
1
6
1
6
1
2
00 MHz) δ 5.05-5.00 (m, 1 H), 4.50 (d, 1 H, J ) 6.1), 4.47 (d,
H, J ) 6.1), 3.52 (dd, 1 H, J ) 7.0, 10.1), 3.52 (dd, 1 H, J )
.4, 10.1), 2.32-2.25 (m, 2 H), 2.05 (s, 3 H), 1.69 (m, 1 H),
.61 (m, 1 H), 1.46, 1.28 (each s, each 3 H), 0.88 (d, 6 H, J )
2
1
3
,
500 MHz) δ 4.61 (d, 1 H, J ) 5.4), 4.22 (d, 1 H, J ) 5.4), 3.81
(dd, 1 H, J ) 2.6, 9.8), 3.61 (dd, 1 H, J ) 2.9, 9.8), 2.71 (dd, 1
H, J ) 9.1, 18.2), 2.50 (br d, 1 H, J ) 9.1), 2.08 (br d, 1 H,
H-5b, J ) 18.2), 1.56 (m, 1 H, J ) 6.8), 1.42, 1.34 (each s, each
3 H), 0.83 (d, 6 H, J ) 6.8), 0.80, 0.79 (s, each 3 H), 0.07, 0.06
.8), 0.84 (s, 6 H), 0.09 (s, 6 H); 13C NMR (CDCl
, 125 MHz) δ
70.0, 111.1, 84.8, 81.7, 79.8, 63.4, 47.1, 34.2, 31.6, 26.8, 25.1,
3
1
8
4.3, 21.1, 20.3, 20.3, 18.4, -3.51, -3.55; [R]
D
-23.0 (c 0.482,
); MS (FAB) m/z 373 (MH ). Anal. Calcd for C19
Si: C, 61.25; H, 9.74. Found: C, 61.06; H, 9.77.
1R,2S,3R,4R)-2,3-O-Isop r op ylid en e-4-[[(d im et h ylt h -
exylsilyl)oxy]m eth yl]cyclopen tan e-1,2,3-tr iol (15). A mix-
ture of 14 (5.00 g, 13.4 mmol) and K CO (500 mg, 3.6 mmol)
+
CHCl
3
36 5
H O -
(each s, each 3 H). 13C NMR (CDCl
, 125 MHz) δ 212.9, 111.0,
81.8, 79.0, 65.1, 39.1, 37.2, 34.0, 26.8, 25.2, 24.6, 20.2, 20.0,
3
(
2
0
18.4, 18.3, -3.7, -3.8; [R]
D
-84.7 (c 0.885, CHCl
3
); MS (FAB)
+
m/z 329 (MH ). Anal. Calcd for C17H O Si: C, 62.15; H, 9.82.
32 4
2
3
in MeOH (120 mL) was stirred at room temperature for 1 h.
After being neutralized with 1 M AcOH in benzene, the
mixture was evaporated under reduced pressure. The residue
was purified by column chromatography (SiO
in hexane) to give 15 (4.00 g, 90%) as an oil; H NMR (CDCl
Found: C, 62.13; H, 9.66.
(1S,2S,3R,4R)-2,3-O-Isop r op ylid en e-4-[[(d im eth ylth ex-
ylsilyl)oxy]m eth yl]cyclop en ta n e-1,2,3-tr iol (17). A mix-
2
, 15% of EtOAc
ture of 16 (3.60 g, 11.0 mmol) and NaBH (393 mg, 10.4 mmol)
4
1
3
,
in MeOH (100 mL) was stirred at -20 °C for 30 min. After
being neutralized with 1 M AcOH in benzene, the mixture was
evaporated under reduced pressure. The residue was purified
5
00 MHz) δ 4.55 (d, 1 H, H-3, J 2,3 ) 5.7), 4.39 (d, 1 H, H-2, J 2,3
)
5.7), 4.07 (d, 1 H, H-1, J 1,5R ) 5.2), 4.07 (br s, 1 H, OH), 3.83
(
dd, 1 H, H-6a, J 4,6a ) 3.4, J 6a,6b ) 10.1), 3.62 (dd, 1 H, H-6b,
by column chromatography (SiO , 10% of EtOAc in hexane)
2
1
J
4,6b ) 2.9, J 6a,6b ) 10.1), 2.45 (ddd, 1 H, H-5R, J 1,5R ) 5.2, J 4,5R
to give 17 (3.17 g, 88%) as an oil; H NMR (CDCl +D O, 500
3
2
)
1
1
9.6, J 5R,5â ) 14.4), 2.31 (br d, 1 H, H-4, J 4,5R ) 9.6), 1.64 (m,
H, thexyl CH), 1.55 (br d, 1 H, H-5â, J 5R,5â ) 14.4), 1.42,
MHz) δ 4.49 (dd, 1 H, H-3, J 2,3) 5.9, J 3,4 ) 1.0), 4.44 (dd, 1 H,
H-2, J 1,2 ) 5.4, J 2,3 ) 5.9), 4.20 (ddd, 1 H, H-1, J 1,2 ) 5.4, J 1,5â
) 7.3, J 1,OH ) 8.3), 3.60 (dd, 1 H, H-6a, J 6a,4 ) 4.4, J 6a,b )10.3),
3.47 (dd, 1 H, H-6b J 6b,4 ) 4.9, J 6a,b )10.3), 2.20 (dddd, 1 H,
H-4, J 3,4 ) 1.0, J 4,5R ) 5.4, J 4,6a ) 4.4, J 4,6b ) 4.9,), 1.853 (d, 1
H, H-5â, J 1,5â ) 7.3), 1.852 (d, 1 H, H-5R, J 4,5R ) 5.4), 1.60 (m,
1 H, thexyl CH), 1.40, 1.35 (each s, each 3 H), 0.88 (d, 6 H,
thexyl CH , J ) 6.8), 0.87 (s, 6 H, thexyl CH ), 0.08 (s, each 6
.28 (each s, each 3 H, isopropyl CH
3
), 0.88 (d, 6 H, thexyl
), 0.16, 0.15 (each s,
CH
3
, J ) 6.8), 0.87 (s, 6 H, thexyl CH
3
3
each 3 H, SiCH ), the assignments were in agreement with
COSY spectrum; NOE (CDCl
observed H-2 (5.1%), H-5â (4.2%); C NMR (CDCl
δ 109.8, 88.3, 83.9, 76.4, 65.7, 47.6, 35.8, 33.9, 26.7, 25.4, 24.1,
3
, 500 MHz) irradiated H-1,
1
3
3
, 125 MHz)
3
3
1
8
2
(
0.2, 20.1, 18.4, -3.5, -3.56; [R]
D
+2.41 (c 0.935, CHCl
FAB) m/z 331 (MH ). Anal. Calcd for C17 Si: C, 61.77;
H, 10.37. Found: C, 61.65; H, 10.32.
2R,3R,4R)-2,3-(Isop r op ylid en ed ioxy)-4-[[(d im eth ylth -
exylsilyl)oxy]m eth yl]cyclop en ta n on e (16). A mixture of
5 (4.00 g, 12.1 mmol), PDC (9.10 g, 24.2 mmol), and molecular
sieves 4 A powder (4.0 g) in CH Cl (300 mL) was stirred at
room temperature. After 3 h, PDC (1.00 g, 2.60 mmol) was
3
); MS
H, SiCH ), the assignments were in agreement with COSY
3
+
H
34
O
4
spectrum and the J values were determined by decouplings
of H-1 and H-4; NOE (CDCl , 500 MHz) irradiated H-1,
3
(
observed H-2 (2.2%), H-5a (3.2%); 13C NMR (CDCl
3
, 125 MHz)
δ 111.3, 83.0, 80.0, 71.8, 64.4, 44.0, 35.7, 34.1, 26.2, 25.1, 24.3,
2
0
1
20.3, 20.2, 18.5, 18.4, -3.63; [R]
D
-11.1 (c 0.842, CHCl
3
). MS
+
2
2
(FAB) m/z 331 (MH ). Anal. Calcd for C17H O Si: C, 61.77;
34 4
H, 10.37. Found:C, 61.51; H, 10.30.

1
992 J . Org. Chem., Vol. 63, No. 6, 1998
-[(1R,2S,3R,4R)-2,3-(Isop r op ylid en ed ioxy)-4-[[(d im e-
Shuto et al.
1
3.93 (m, 1 H), 3.82-3.77 (m, 3 H), 2.46-2.31 (m, 3 H) 1.64,
t h ylt h exylsilyl)oxy]m et h yl]cyclop en t yl]-5′-O-(ter t-b u -
t yld im et h ylsilyl)-2′,3′-O-isop r op ylid en ein osin e (6) a n d
O -[(1R,2S,3R,4R)-2,3-(Isop r op ylid en ed ioxy)-4-[[(d im eth -
1.54, 1.38, 1.29 (each s, each 3 H); 13C NMR (CDCl
3
, 67.9 MHz)
δ 156.34, 147.0, 146.1, 139.9, 126.3, 114.3, 113.2, 93.4, 86.1,
83.8, 83.3, 82.2, 81.3, 66.0, 63.9, 63.0, 46.5, 32.7, 27.7, 27.5,
6
ylth exylsilyl)oxy]m eth yl]cyclopen tyl]-5′-O-(ter t-bu tyldim -
eth ylsilyl)-2′,3′-O-isop r op ylid en ein osin e (18). To a solu-
tion of 17 (500 mg. 1.52 mmol) and DMAP (555 mg, 4.52
25.2; HRMS (FAB, positive) calcd for C22
found 479.2168; UV (MeOH) λmax 253 nm, 247 nm, sh 265 nm.
-[(1R,2S,3R,4R)-2,3-(Isopr opyliden edioxy)-4-(ph osph o-
n ooxym et h yl)cyclop en t yl]-2′,3′-O-isop r op ylid en e-5′-O-
p h osp h on oin osin e (5). POCl (54 µL, 0.58 mmol) was added
to a solution of 19 (56 mg, 0.12 mmol) in PO(OEt) (3 mL) at
°C, and the mixture was stirred at the same temperature
for 4 h. The reaction was quenched by aqueous saturated
NaHCO (5 mL), and the resulting mixture was washed with
CHCl
column (1.8 × 14 cm). After washing with water (100 mL),
the column was developed using a linear gradient of 0.1 N
triethylammonium acetate (TEAA) buffer (pH 7.0) to 0.5 M
TEAA buffer (pH 7.5)/MeCN (1:1) (300 mL). Appropriate
fractions were evaporated under reduced pressure, and excess
TEAA was coevaporated with water. The residue was freeze-
30 4 8
H N O 479.2141,
1
mmol) in CH
at -20 °C, and the resulting mixture was stirred at room
(25
2 2
Cl (10 mL) was added TfCl (660 µL, 6.2 mmol)
3
temperature for 30 min. Ice-water (20 mL) and CHCl
3
3
mL) were added, and the mixture was partitioned. The
organic layer was washed with aqueous HCl (0.5 N, 10 mL)
0
2 4
and brine (15 mL), dried (Na SO ), and evaporated under
3
reduced pressure to give 8 as an oil. The oil was dissolved in
DME (0.5 mL), which was directly used the next reaction
without purification due to the instability of 8.
3
(20 mL × 2). The aqueous layer was applied to a C-18
A suspension of 7 (642 mg, 1.52 mmol) and K
.9 mmol) in DME (2 mL) was heated under reflux for 1 h. To
the mixture, was added the above prepared solution of 8 at
0 °C, and the resulting mixture was stirred at the same
2 3
CO (399 mg,
2
6
temperature for 13 h. EtOAc (30 mL) and water (15 mL) were
added, and the resulting mixture was partitioned. The organic
dried to give triethylammonium salt of 5 (43 mg, 37%) as a
1
solid: H NMR (D
8
1
(
(
2
O, 500 MHz) δ 8.53 (s, 1 H, H-8 or H-2),
layer was washed with brine (15 mL), dried (Na
evaporated under reduced pressure. The residue was purified
by column chromatography (SiO , 25% of EtOAc in hexane)
to give 6 (534 mg, 48%) as a solid and 18 (78 mg, 7%) as an
2 4
SO ), and
.45 (s, 1 H, H-8 or H-2), 6.30 (d, 1 H, H-1′, J ) 2.4), 5.43 (dd,
H, H-2′, J ) 2.4, 3.0), 5.21-5.12 (m, 3 H, H-1′′, 2′′, 3′), 4.84
2
dd, 1 H, H-3′′, J ) 5.9, 6.7 Hz), 4.68 (m, 1 H, H-4′), 4.07-3.98
+
m, 4 H, H-5′a, 5′b, 6′′a, 6′′b), 3.22 (q, 12 H, CH
3
CH
.7), 2.59-2.55 (m, 1 H, H-4′′), 2.50-2.26 (m, 2 H, H-5′′), 1.68,
.61, 1.46, 1.38 (each s, isopropyl CH ), 1.30 (t, 18 H, CH
N , J ) 5.7), the assignments were in agreement with
2
N , J )
1
oil. 6: H NMR (CDCl
(
3
, 500 MHz) δ 8.02 (s, 1 H, H-8), 7.99
5
1
s, 1 H, H-2), 6.12 (d, 1 H, H-1′, J ) 2.6), 5.08 (dd, 1 H, H-2′,
3
3
-
J ) 2.6, 6.1), 4.98 (dd, 1 H, H-2′′, J ) 4.9, 6.8), 4.92 (dd, 1 H,
H-3′, J ) 2.5, 6.1), 4.85-4.81 (m, 1 H, H-1′′), 4.62 (m, 1 H,
H-3′′), 4.41 (m, 1 H, H-4′), 3.86 (dd, 1 H, H-5′a, J ) 3.7, 11.2),
+
CH
COSY spectrum; C NMR (CDCl
2
13
3
, 67.9 MHz) δ 160.7, 150.3,
17.6, 117.0, 93.5, 88.2, 88.1, 87.0, 86.4, 84.2, 84.1, 68.5, 68.4,
7.6, 67.6, 65.0, 49.4, 47.0, 46.9, 35.4, 29.2, 28.8, 27.1, 10.9;
1
6
3
9
1
0
.81-3.75 (m, 2 H, H-5′b, 6′′a), 3.70 (dd, 1 H, H-6′′b, J ) 5.5,
.9), 2.37-2.24 (m, 3 H, H-4′′, 5′′a,b), 1.62 (m, 1 H, thexyl CH),
31
P NMR (125 MHz, D
negative) calcd for C22
2
O) δ 0.77 (s), 0.42 (s); HRMS (FAB,
.64, 1.55, 1.40, 1.29 (each s, each 3 H, isopropyl CH
3
), 0.89-
), 0.10-0.00 (m, 12 H, SiCH ),
H
31 4 14 2
N O P
637.1310, found 637.1314;
.85 (m, 21 H, t-Bu, thexyl CH
3
3
UV (MeOH) λmax 253 nm, sh 270 nm.
the assignments were in agreement with COSY spectrum;
NOE (CDCl , 400 MHz) irradiated H-2, observed H-1′′ (13.5%),
H-2′′ (4.2%), H-1′ (0.6%), H-2′ (0.6%); C NMR (67.8 Mz,
CDCl ) δ 156.5, 146.6, 146.3, 138.4, 125.2, 114.2, 113.1, 91.1,
7.0, 85.4, 83.4, 81.2, 81.0, 63.9, 63.4, 46.6, 34.2, 33.8, 27.7,
7.2, 25.9, 25.3, 25.1, 20.3, 18.5, 18.3, -3.6; HRMS (FAB,
Si 735.4181, found 735.4185; UV
MeOH) λmax 248, 253 nm, sh 265 nm; IR (CHCl
8
-Br om o-2′,3′-O-isop r op ylid en ein osin e (24). NaNO (18
2
3
2
1
1
3
g, 240 mmol) was added to a solution of 23 (8.00 g, 20.7 mmol)
in AcOH (200 mL) and water (30 mL), and the resulting
mixture was stirred at room temperature for 6 h. After the
mixture was evaporated under reduced pressure, the residue
was dissolved in EtOH (100 mL), and the solution was
evaporated under reduced pressure. The residue was parti-
3
8
2
positive) calcd for C36
H
63
N
4
O
8
2
-
1
(
(
8
(
(
3
) 1703 cm
2 2
H O: C, 58.11; H,
1
tioned between CHCl
organic layer was washed with aqueous saturated NaHCO
100 mL) and brine (150 mL), dried (Na SO ), and evaporated
3
(300 mL) and water (150 mL), and the
ν
CdO). Anal. Calcd for C36
.53; N, 7.53. Found: C, 58.13; H, 8.37; N, 7.50. 18: H NMR
CDCl , 500 MHz) δ 8.58 (s, 1 H, H-2), 8.16 (s, 1 H, H-8), 6.22
d, 1 H, H-1′, J ) 2.5), 5.66-5.64 (m, 1 H, H-1′′), 5.23 (dd, 1
H
62
N
4
O
8
Si
2
‚ /
1
3
(
2
4
3
under reduced pressure. The residue was treated with aque-
ous EtOH to give 24 (4.7 g, 59%) as crystals: mp 225 °C (dec);
H, H-2′, J ) 2.5, 6.1), 4.96 (dd, 1 H, H-3′, J ) 2.4, 6.1), 4.82 (d,
1
4
1
H NMR (CDCl
.11 (d, 1 H, J ) 4.8), 5.25 (dd, 1 H, J ) 4.8, 5.9), 5.07 (dd, 1
H, J ) 1.7, 5.9), 5.00 (br s, 1 H), 4.49 (dd, 1 H, J ) 1.7, 1.7),
3
, 270 MHz) δ 12.9 (br s, 1 H), 8.31 (s, 1 H),
H, H-2′′, J ) 6.0), 4.65 (dd, 1 H, H-3′′, J ) 2.1, 6.0), 4.44-
.42 (m, 1 H, H-4′), 3.88 (dd, 1 H, H-5′a, J ) 3.8, 11.2), 3.77
6
(
dd, 1 H, H-5′b, J ) 4.2, 11.2), 3.74 (dd, 1 H, H-6′′a, J ) 7.8,
3
1
1
.95 (dd, 1 H, J ) 1.7, 12.6), 3.79 (dd, 1 H, J ) 1.7, 12.6), 1.67,
.39 (each s, each 3 H); 13C NMR (DMSO-d
, 67.8 MHz) δ; 157,
49, 146, 126, 126, 114, 93.4, 85.7, 82.9, 81.3, 63.0, 27.5, 25.5;
1
0.3), 3.62 (dd, 1 H, H-6′′b, J ) 6.7, 10.3), 2.51-2.45 (m, 1 H,
H-5′′a), 2.38 (m, 1 H, H-4′′), 1.94-1.90 (m, 1 H, H-5′′b), 1.64,
.51, 1.41, 1.31 (each s, each 3 H, isopropyl CH ), 1.57 (m, 1
H, thexyl CH), 0.84 (m, 21 H, t-Bu, thexyl CH ), 0.03 (m, 12
H, SiCH ), the assignments were in agreement with COSY
spectrum; NOE (CDCl , 400 MHz) irradiated H-2, observed
H-1′′ (0.8%), H-2′′ (0.6%), H-2′ (0.3%); C NMR (67.8 Mz,
CDCl ) δ 160.1, 152.5, 151.9, 141.0, 122.5, 114.5, 111.3, 91.8,
7.5, 85.4, 85.1, 82.3, 82.10, 81.7, 63.8, 47.8, 34.5, 32.2, 27.5,
7.1, 26.2, 25.7, 25.3, 24.6, 20.6, 18.8, 18.6, -3.2, -3.4; HRMS
Si 735.4181, found
35.4169; UV (MeOH) λmax 253 nm; IR (CHCl ) νCdO (around
6
1
3
+
MS (FAB) m/z 331 (MH ); UV (MeOH) λmax 253 nm, sh 275
nm. Anal. Calcd for C13 15BrN : C, 40.33; H, 3.90; N,
4.47. Found: C, 40.23; H, 3.93; N, 14.52.
8-Br om o-5′-O-(ter t-b u t yld im et h ylsilyl)-2′,3′-O-isop r o-
3
H
4 5
O
3
1
3
1
3
p ylid en ein osin e (25). To a solution of 24 (2.50 g, 7.57 mmol)
and imidazole (1.35 g, 19.8 mmol) in DMF (50 mL) was added
TBSCl (1.49 g, 9.9 mmol) at 0 °C, and the mixture was stirred
at the same temperature for 20 min. Ice-water (50 mL) and
EtOAc (250 mL) were added, and the resulting mixture was
partitioned. The organic layer was washed with water (50 mL
3
8
2
(
7
1
FAB, positive) calcd for C36
H
63
N
4
O
8
2
3
-
1
700 cm ) was not observed.
-[(1R ,2S ,3R ,4R )-2,3-(I s o p r o p y lid e n e d io x y )-4-(h y -
1
x 5) and brine (50 mL), dried (Na
reduced pressure. The residue was purified by column chro-
matography (SiO , 50% of EtOAc in hexane) to give 25 (2.83
g, 75%) as a solid: 1H NMR (CDCl
, 270 MHz) δ 8.21 (s, 1 H),
2 4
SO ), and evaporated under
d r o x y m e t h y l)c y c lo p e n t y l]-2′,3′-O -i s o p r o p y li d e n e -
in osin e (19). A mixture of 6 (222 mg, 0.30 mmol), and TBAF
2
(1 M in THF, 640 µL, 0.64 mmol) in THF (3 mL) was stirred
3
at room temperature for 5 h. The resulting mixture was
evaporated under reduced pressure, and the residue was
6.16 (d, 1 H, J ) 2.0), 5.61 (dd, 1 H, J ) 2.0, 6.6), 5.05 (dd, 1
H, J ) 4.0, 6.6), 4.24 (ddd, 1 H, J ) 4.0, 5.9, 6.6), 3,73 (dd, 1
purified by column chromatography (SiO
to give 19 (153 mg, quant) as a white solid; H NMR (CDCl
2
, 9% EtOH in CHCl
3
)
,
H, J ) 6.6, 20.5), 3.70 (dd, 1 H, J ) 5.9, 20.5), 1.60, 1.39 (each
1
13
3
3
s, each 3 H), 0.83 (s, 9 H), -0.63 (s, 6 H); C NMR (CDCl ,
5
5
00 MHz) δ 8.07 (s, 1 H), 7.91 (s, 1 H), 5.89 (d, 1 H, J ) 4.1),
67.8 MHz) δ 157.9, 149.6, 145.7, 126.9, 125.4, 114.4, 91.5, 87.9,
.09-5.05 (m, 3 H), 5.02 (dd, 1 H J ) 1.8, 10.0), 4.74 (dd, 1 H,
83.1, 81.7, 76.5, 63.1, 27.2, 25.8, 25.5, 18.3; HRMS (FAB,
J ) 5.9, 6.0), 4.71-4.66 (m, 1 H), 4.49 (d, 1 H, J ) 1.5), 3.96-
4 5
positive) calcd for C19H30BrN O Si 501.1169, found 501.1176;

Carbocyclic Analogue of cADP-ribose
J . Org. Chem., Vol. 63, No. 6, 1998 1993
UV (MeOH) λmax 253, sh 275. Anal. Calcd for C19
H
29BrN
4
O
5
-
ing 5′,6′′-bis(phosphoramidite) of 28 (135 mg, 79%) as a foam:
3
1
Si: C, 45.51; H, 5.83; N, 11.17. Found: C, 45.50; H, 5.92; N,
P NMR (D
2
O, 125 MHz) δ 149.02 (s), 148.36 (s), 148.13 (s);
1
0.99.
-Br om o-1-[(1R,2S,3R,4R)-2,3-(isop r op ylid en ed ioxy)-4-
[(dim eth ylth exylsilyl)oxy]m eth yl]cyclopen tyl]-5′-O-(ter t-
bu tyldim eth ylsilyl)-2′,3′-O-isopr opyliden ein osin e (26) an d
HRMS (FAB, positive) calcd for C40H64BrN O P
8 10 2
957.3404,
8
found 957.3422, which was immediately used in the next
reaction.
[
To a mixture of the above form (135 mg, 0.14 mmol) and
6
8
-Br om o-O -[(1R,2S,3R,4R)-2,3-(isop r op ylid en ed ioxy)-4-
1
H-tetrazole (30 mg, 0.43 mmol) in aqueous MeCN (94%, 3.2
mL) was added a solution of I (3% in water:pyridine:THF )
:19:76, 2 mL), and the resulting mixture was stirred at room
temperature for 15 min. After the reaction was quenched by
aqueous Na (1 mL), CHCl (2 mL) was added, and the
resulting mixture was partitioned. To the aqueous layer was
added a saturated NH in MeOH (30 mL), and the mixture
[
[(dim eth ylth exylsilyl)oxy]m eth yl]cyclopen tyl]-5′-O-(ter t-
2
bu t yld im et h ylsilyl)-2′,3′-O-isop r op ylid en ein osin e (27).
Compound 26 (solid, 320 mg, 44%), along with the correspond-
5
6
ing O -regioisomer 27 (solid, 65 mg, 9%), was obtained from 8
2
S
2
O
3
3
(
412 mg, 0.90 mmol) as described for 6, with 25 (460 mg, 0.90
1
mmol) instead of 7. 26: H NMR (CDCl
1
2
3
, 500 MHz) δ 7.91 (s,
H, H-2), 6.14 (d, 1 H, H-1′, J ) 2.2), 5.56 (dd, 1 H, H-2′, J )
.2, 6.3), 5.04 (dd, 1 H, H-3′, J ) 3.9, 6.3), 4.96 (dd, 1 H, H-2′,
3
was allowed to stand at room temperature for 2 days. The
resulting solution was evaporated under reduced pressure. The
residue was dissolved in water (150 mL) and then pH of the
J ) 4.9, 6.6), 4.77-4.75 (m, 1 H, H-1′′), 4.61 (dd, 1 H, H-3′′, J
4.9, 5.3), 4.23-4.20 (m, 1 H, H-4′), 3.78 (dd, 1 H, H-5′, J )
.4, 9.8), 3.74-3.68 (m, 3 H, H-5′, 6′′a, 6′′b), 2.31-2.30 (m, 3
H, 4′′, 5′′a, 5′′b), 1.63 (m, 1 H, thexyl CH), 1.61, 1.59, 1.39,
.26 (each s, each 3 H, isopropyl CH ), 0.89-0.85 (m, 21 H,
t-Bu, thexyl CH ), 0.09-0.02 (m, 12 H, SiCH ), the assign-
ments were in agreement with COSY spectrum; NOE (CDCl
00 MHz) irradiated H-2, observed H-1′′ (14.7%), H-2′′ (6.0%),
)
3
solution was adjusted about 6 with AcOH, and the solution
-
was applied to a DEAE-Sephadex A-25 column (HCO
3
form,
3
.5 × 8 cm). The column was developed using a linear gradient
of 0 to 0.4 M triethylammonium bicarbonate (TEAB) buffer
pH 8.1, 400 mL) and was further developed with 0.4 M TEAB
1
3
3
3
(
3
,
buffer (pH 8.1, 600 mL). Appropriate fractions were evapo-
rated under reduced pressure, and excess TEAB was coevapo-
4
1
3
H-2′ (0.6%); C NMR (CDCl
3
, 67.8 MHz) δ 155.1, 147.7, 146.2,
rated with water. The residue was freeze-dried to give 22 (75
1
6
2
26.3, 125.5, 114.4, 113.1, 91.1, 87.5, 83.2, 81.6, 81.0, 64.6, 63.4,
31
mg) as triethylammonium salts: P NMR (D O, 125 MHz) δ
2
3.0, 46.5, 34.2, 33.5, 27.7, 27.2, 25.9, 25.7, 25.6, 25.4, 25.3,
0
.85 (s), 0.68 (s); HRMS (FAB, negative) calcd for C22
30
H -
+
5.1, 20.3, 18.5, 18.3, -3.6, -3.7; MS (FAB) m/z 813 (MH );
BrN 715.0417, found 715.0398. The countercations were
4
14 2
O P
UV (MeOH) λmax 253, 258 nm, sh 275 nm; IR (CHCl
3
) 1707
+
exchanged for sodium with a Diaion WK-20 resin column (Na
form, 1.2 × 5 cm, developed by water). The eluent was
evaporated under reduced pressure, and the residue was
-
1
cm (νCdO). Anal. Calcd for C36
.55; N, 6.88. Found: C, 52.91; H, 7.48; N, 6.72. 27: H NMR
CDCl , 500 MHz) δ 8.48 (s, 1 H, H-2), 6.21 (d, 1 H, H-1′, J )
H62BrN O Si : C, 53.12; H,
4 8 2
1
7
(
3
freeze-dried to give 22 (white solid, 64 mg, 34% from 28) as
2
5
4
3
5
.0), 5.75 (dd, 1 H, H-2′, J ) 2.0, 6.3), 5.61-5.60 (m, 1 H, H-1′′),
.15 (dd,1 H, H-3′, J ) 2.1, 6.2), 4.77 (d, 1 H, H-2′′, J ) 6.1),
.63 (dd, 1 H, H-3′′, J ) 2.1, 6.2), 4.27 (m, 1 H, H-4′), 3.76-
.61 (m, 4 H, H-5′a, 5′b, 6′′a, 6′′b), 2.48-2.31 (m, 3 H, H-5′′a,
′′b, 4′′), 1.62, 1.50. 1.40, 1.31 (each s, each 3 H, isopropyl
1
sodium salts: H NMR (D
2
O, 500 MHz) δ 8.46 (s, 1 H, H-2),
6
5
4
.34 (d, 1 H, H-1′, J ) 2.3), 5.69 (dd, 1 H, H-2′, J ) 2.3, 6.5),
.28 (dd, 1 H, H-3′, J ) 4.5, 6.5), 5.14-5.08 (m, 2 H, H-1′′, 2′′),
.82 (m, 1 H, H-3′′), 4.46 (m, 1 H, H-4′), 4.03-3.82 (m, 4 H,
H-5′a, 5′b, 6′′a, 6′′b), 2.50 (m, 1 H, H-4′′), 2.41 (m, 1 H, H-5′′a),
.27 (m, 1 H, 5′′b), 1.66, 1.60, 1.44, 1.37 (each s, each 3 H,
isopropyl CH ), the assignments were in agreement with COSY
spectrum; 13C NMR (D
O, 125 MHz) δ 159.6, 151.1, 150.4,
30.0, 126.9, 118.3, 116.9, 92.9, 89.0, 86.2, 85.6, 84.5, 84.0, 67.5,
6.4, 65.3, 47.2, 35.6, 29.1, 28.8, 27.1, 27.0; 31P NMR (D
O,
CH
thexyl CH
in agreement with COSY spectrum; NOE (CDCl
irradiated H-2, observed H-1′′ (0.8%), H-2′ (0.6%), H-3′ (0.4%);
3
), 1.57 (m, 1 H, thexyl CH), 0.91-0.78 (m, 21 H, t-Bu,
), 0.09-0.06 (m, 12 H, SiCH ), the assignments were
, 400 MHz)
2
3
3
3
3
2
1
6
1
λ
1
3
C NMR (CDCl , 125 MHz) δ 169.6, 158.9, 152.7, 152.2, 130.5,
3
2
1
7
2
8
22.6, 114.4, 111.5, 111.3, 91.9, 88.2, 85.0, 83.1, 82.6, 82.0, 80.4,
0.1, 63.7, 63.3, 59.6, 47.7, 47.2, 34.5, 32.1, 27.5, 27.0, 26.1,
25 MHz) δ 4.26 (s, two signals were coincident); UV (MeOH)
max 255 nm, sh 280 nm.
5.7, 25.4, 25.4, 24.6, 20.6, 18.7, -3.3, -5.2; MS (FAB) m/z
+
8-Br om o-cyclic IDP -ca r b ocyclic-r ib ose Dia cet on id e
13 (MH ); UV (MeOH) λmax 260, sh 265 nm; IR (CHCl
3
), νCdO
-
1
(29). Compound 22 (sodium salts, 56 mg, 0.074 mmol) was
dissolved in MPD (11 mL) by heating. EDC (21 mg, 0.11
mmol) was added to the solution of 22, and the mixture was
stirred at 50 °C for 60 h. After cooling the mixture with ice-
bath, the mixture was diluted with water (90 mL). The
solution was applied to a DEAE-Sephadex A-25 column
(
around 1700 cm ) was not observed. Anal. Calcd for
Si : C, 53.12; H, 7.55; N, 6.88. Found: C, 53.22;
H, 7.47; N, 6.62.
-Br om o-1-[(1R,2S,3R,4R)-2,3-(isop r op ylid en ed ioxy)-4-
h yd r oxym e t h yl)cyclop e n t yl]-2′,3′-O-isop r op ylid e n e -
C
36
H
61BrN
4
O
8
2
8
(
in osin e (28). Compound 28 (solid, 1.22 g, 89%) was obtained
from 26 (2.00 g, 2.46 mmol) as described for 19: H NMR
-
1
(HCO3 form, 1.8 × 8 cm). The column was washed with 0.1
M TEAB buffer (pH 7.5, 100 mL) and developed using a linear
gradient of 0.1 to 0.4 M TEAB buffer (pH 7.5, 300 mL).
Appropriate fractions were evaporated under reduced pres-
sure, and excess TEAB was coevaporated with water. Coun-
tercations were exchanged for sodium with a Diaion WK-20
(
CDCl
3
, 500 MHz) δ 8.09 (s, 1 H), 6.08 (d, 1 H, J ) 5.1), 5.15
(
dd, 1 H, J ) 5.1, 5.6), 5.04-4.99 (m, 2 H), 4.74-4.70 (m, 3
H), 3.93-3.75 (m, 4 H), 2.45-2.28 (m, 3 H), 2.01 (s, 2 H), 1.66,
1
3
1
.54, 1.38, 1.29 (each s, each 3 H); C NMR (CDCl
δ 155.0, 147.2, 147.0, 126.1, 126.0, 93.2, 85.5, 83.2, 82.9, 82.1,
1.1, 65.9, 63.7, 63.0, 46.3, 32.6, 27.6, 27.6, 25.4, 25.2; HRMS
FAB, positive) calcd for 557.1248, found
57.1257; UV (MeOH) λmax 253, sh 275 nm. Anal. Calcd for
29BrN : C, 47.41; H, 5.24; N, 10.05. Found: C, 47.11;
H, 5.51; N, 9.75.
-Br om o-1-[(1R,2S,3R,4R)-2,3-(isop r op ylid en ed ioxy)-4-
p h osp h on ooxym e t h yl)cyclop e n t yl]-2′,3′-O-isop r op y-
3
, 67.8 MHz)
+
8
(
5
C
resin column (Na form, 1.2 × 5 cm, developed by water). The
C
22
H
30BrO
8
N
4
eluate was evaporated under reduced pressure, and the residue
was freeze-dried to give 29 (solid, 13 mg, 23%) as sodium
salts: 1H NMR (D
O, 500 MHz) δ 8.52 (s, 1 H, H-2), 6.39 (d, 1
22
H
4
O
8
2
H, H-1′J ) 1.3), 5.83 (dd, 1 H, H-2′, J ) 1.3, 6.2), 5.53 (dd, 1
H, H-3′, J ) 2.2, 6.2), 4.88 (d, 1 H, H-2′′, J ) 9.3), 4.69 (m, 2
H, H-1′′, 3′′), 4.63-4.57 (m, 1 H, H-4′), 4.18-4.15 (m, 1 H′,
H-5′a), 4.04-3.97 (m, 2 H, H-6′′a, 6′′b) 3.81-3.76 (m, 1 H′,
H-5′b), 2.93-2.82 (m, 2 H, H-5′′a, 4′′), 2.64 (d, 1 H, H-5′′b, J )
8
(
lid en e-5′-O-p h osp h on oin osin e (22). (2-Cyanoethoxy)(N,N-
diisopropylamino)chlorophosphine (161 µL, 0.72 mmol) and
i-Pr
2
NEt (188 µL, 1.1 mmol) were added to a solution of 28
16.4), 1.66, 1.60, 1.48, 1.39 (each s, each 3 H, isopropyl CH
the assignments were in agreement with COSY spectrum;
3
3
),
C
1
(100 mg, 0.18 mmol) in CH
2
Cl (5 mL) at room temperature,
2
and the mixture was stirred at the same temperature for 15
min. After the reaction was quenched by aqueous saturated
NMR (D
117.12 113.7, 93.5, 90.5, 89.5, 86.5, 85.6, 84.8, 68.1, 66.9, 61.8,
47.4, 29.6, 28.59, 26.9, 26.3; 31P NMR (D
2
O, 125 MHz) δ 160.3, 150.4, 148.5, 141.5, 125.2,
NaHCO
mixture was partitioned. The organic layer was washed with
brine (5 mL), dried (Na SO ), and evaporated under reduced
pressure. The residue was purified by column chromatography
neutral SiO , 33% of EtOAc in hexane) to give the correspond-
3
(1 mL), CHCl
3
(30 mL) was added, and the resulting
2
O, 125 MHz) δ -10.67
(d, J ) 15.0), -10.95 (d, J ) 15.0); MS (FAB, negative) m/z
-
2
4
677, 699 (M ); UV (MeOH) λmax 256 nm, sh 280 nm. Anal.
1
Calcd for C22
H
27BrN
4
O
8
P
2
Na
2
‚ /14Et
3
N: C, 35.55; H, 3.66; N,
(
2
7.54. Found: C, 35.89; H, 3.76; N, 7.59.

1
994 J . Org. Chem., Vol. 63, No. 6, 1998
Shuto et al.
Cyclic IDP -ca r bocyclic-r ibose d ia ceton id e (30).
A
a linear gradient of 0.1 to 0.4 M TEAB buffer (pH 7.5, 200
mL). Appropriate fractions were evaporated under reduced
pressure, and excess TEAB was coevaporated with water. The
residue was freeze-dried to give 4 (triethylammonium salts,
82 OD254 units, 67%). The triethylammonium salts were
dissolved in water (1 mL) and desalted with a C-18 column
(1.8 × 16 cm, developed by water). Countercations were
mixture of 29 (162 OD254 units) and Pd-C (10%, 1.5 mg) in
EtOH/aqueous NaHCO (0.5 M) (1:2, 6 mL) was stirred under
atmospheric pressure of H at room temperature for 17 h. The
3
2
catalyst was filtered off with Celite, and the filtrate was
evaporated under reduced pressure. The residue was dissolved
in water (50 mL), and pH of the mixture was adjusted to about
+
6
with AcOH. The solution was applied to a DEAE-Sephadex
exchanged for sodium with a Diaion WK-20 resin column (Na
A-25 column (HCO3- form, 1.8 × 4 cm), and the column was
form, 1.0 × 9 cm, developed by water). The eluent was
developed using a gradient of 0.1 to 0.4 M TEAB buffer (pH
evaporated under reduced pressure, and the residue was
1
7
.5, 200 mL). Appropriate fractions were evaporated under
freeze-dried to give 4 as sodium salts: H NMR (D
2
O, 500
reduced pressure, and excess TEAB was coevaporated with
water. The residue was freeze-dried to give 30 (triethylam-
monium salt, 125 OD254 units, 77%). The countercations were
MHz) δ 9.02 (s, 1 H, H-2), 8.19 (s, 1 H, H-8), 6.02 (d, 1 H, H-1′,
J ) 6.5), 5.20-5.17 (m, 2 H, H-2′, 1′′), 4.66-4.62 (m, 2 H, H-2′′,
3′), 4.43 (m, 1 H, H-4′), 4.27-4.10 (m, 5 H, H-3′′, 5′a, 5′b, 6′′a,
6′′b), 2.95 (m, 1 H, H-5′′a), 2.48 (m, 1 H, H-4′′), 2.25 (m, 1 H,
+
exchanged for sodium with a Diaion WK-20 resin column (Na
form, 1.2 × 5 cm, developed by water). The eluent was
H-5′′b); 13C NMR (D
2
O, 125 MHz) δ 161.3, 150.1, 145.2, 126.8,
evaporated under reduced pressure, and the residue was
93.1, 87.6, 87.5, 81.2, 75.9, 75.1, 73.7, 67.8, 67.6, 61.5, 49.5,
1
31
freeze-dried to give 30 as sodium salts: H NMR (D
2
O, 500
44.9, 29.9, 11.0; P NMR (D
-10.51 (d, J ) 10.7); HRMS (FAB, negative) calcd for
Na 561.0398, found 561.0358; UV (MeOH) λmax
2
O, 125 MHz) δ -9.16 (d, J ) 10.7),
MHz) δ 8.55 (s, 1 H, H-2), 8.19 (s, 1 H, H-8), 6.36 (d, 1 H, H-1′,
J ) 1.6), 5.75 (dd, 1 H, H-2′, J ) 1.6, 6.2), 5.32 (dd, 1 H, H-3′,
J ) 2.3, 6.2), 4.91 (d, 1 H, H-2′′, J ) 9.3), 4.73-4.69 (m, 2 H,
H-1′′, 3′′), 4.59 (m, 1 H, H-4′), 4.18-4.03 (m, 3 H, H-5′a, 6′′a,
16 21 4 13 2
C H N O P
250 nm.
6
′′b), 3.80-3.75 (m, 1 H, H-5′b), 2.93-2.83(m, 2 H, H-5′′a,
Ack n ow led gm en t . This investigation was sup-
ported in part by a Grant from the Ministry of Educa-
tion, Science, Sports, and Culture of J apan. We are
grateful to Prof. Kazuyoshi Ikeda (Hokkaido University)
and Dr. Tamotsu Yamamoto (Asahi Chemical Industry)
for their helf for mass spectra measurements.
H-4′′), 2.65 (d, 1 H, H-5′′b, J ) 19.6), 1.67, 1.61, 1.48, 1.40 (each
s, each 3 H), the assignments were in agreement with COSY
1
3
spectrum; C NMR (D
2
O, 125 MHz) δ 161.3, 149.5, 148.1,
45.0, 126.8, 117.2, 113.66, 94.2, 90.5, 89.0, 86.6, 85.6, 84.7,
1.2, 68.1, 67.9, 67.1, 47.5, 29.7, 28.6, 27.0, 26.3, 22.8; 31P NMR
O, 125 MHz) δ -10.42 (d, J ) 15.3), -10.82 (d, J ) 15.3);
HRMS (FAB, positive) calcd for C22 Na 665.0999,
found 665.1209; UV (MeOH) λmax 250, 256 nm.
Cyclic IDP -ca r bocyclic-r ibose (4). A solution of 30
triethylammonium salts, 122 OD254 units) in aqueous HCO
60%, 3 mL) was stirred at room temperature for 6 h. After
1
7
(D
2
H
29
N
4
O
13
P
2
2
1
Su p p or tin g In for m a tion Ava ila ble: H NMR spectral
charts of 4, 5, 19, 22, 29, and 30 (6 pages). This material is
contained in libraries on microfiche, immediately follows this
article in the microfilm version of the journal, and can be
ordered from the ACS; see any current masthead page for
ordering information.
(
(
2
H
the solvent was evaporated under reduced pressure, the
residue was dissolved in water (100 mL), and the resulting
solution was applied to a DEAE-Sephadex A-25 column
-
(HCO
3
form, 1.8 × 4 cm). The column was developed using
J O9717797