Synthesis of cyclic ADP-carbocylcic-xylose and its 3″-O-methyl analogue as stable and potent Ca2+-mobilizing agents

Article abstract of DOI:10.1080/15257770600685867

We previously showed that 3″-deoxy-cyclic ADP-carbocyclic-ribose (3″-deoxy-cADPcR, 3) is a stable and highly potent analogue of cyclic ADP-ribose (cADPR, 1), a Ca²⁺-mobilizing second messenger. From these results, we newly designed another 3″-m

Full text of DOI:10.1080/15257770600685867

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Synthesis of Cyclic ADP-Carbocylcic-
Xylose and its 3′′- O O -Methyl Analogue
as Stable and Potent Ca2+-Mobilizing
Agents
Takashi Kudoh a , Akira Matsuda a , Satoshi Shuto a , Takashi
Murayama b & Yasuo Ogawa b
a Graduate School of Pharmaceutical Sciences, Hokkaido University ,
Kita-ku, Sapporo, Japan
b Department of Pharmacology , Juntendo University School of
Medicine , Bunkyo-ku, Tokyo, Japan
Published online: 07 Feb 2007.
To cite this article: Takashi Kudoh , Akira Matsuda , Satoshi Shuto , Takashi Murayama & Yasuo Ogawa
(2006) Synthesis of Cyclic ADP-Carbocylcic-Xylose and its 3′′- O O -Methyl Analogue as Stable and
Potent Ca2+-Mobilizing Agents, Nucleosides, Nucleotides and Nucleic Acids, 25:4-6, 583-599, DOI:
10.1080/15257770600685867
To link to this article: http://dx.doi.org/10.1080/15257770600685867
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Nucleosides, Nucleotides, and Nucleic Acids, 25:583–599, 2006
Copyright ꢀ Taylor & Francis Group, LLC
C
ISSN: 1525-7770 print / 1532-2335 online
DOI: 10.1080/15257770600685867
SYNTHESIS OF CYCLIC ADP-CARBOCYLCIC-XYLOSE AND ITS
3ꢀꢀ-OO -METHYL ANALOGUE AS STABLE AND POTENT
Ca2+-MOBILIZING AGENTS
Takashi Kudoh, Akira Matsuda, and Satoshi Shuto
Graduate School of
2
Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo, Japan
Takashi Murayama and Yasuo Ogawa Department of Pharmacology, Juntendo
2
University School of Medicine, Bunkyo-ku, Tokyo, Japan
We previously showed that 3ꢁꢁ-deoxy-cyclic ADP-carbocyclic-ribose (3ꢁꢁ-deoxy-cADPcR, 3) is a stable
2
and highly potent analogue of cyclic ADP-ribose (cADPR, 1), a Ca2+-mobilizing second messenger.
From these results, we newly designed another 3ꢁꢁ-modified analogues of cADPcR and identified
the N1-“xylo”-type carbocyclic analogue, i.e., cADPcX (4), as one of the most potent cADPR-related
compounds reported so far.
Keywords cADPR; Calcium; Second messenger
INTRODUCTION
Much attention has been focused on cyclic ADP-ribose (cADPR, 1,
Figure 1), a naturally occurring metabolite of NAD+,[1] due to the bio-
logical interest.[2] cADPR has been shown to mobilize intracellular Ca2+
in various cells, and is now recognized as a general mediator involved in
Ca2+ signaling.[2] Under neutral conditions, cADPR is in a zwitterionic form
with a positive charge around the N(1)-C(6)-N6 moiety (pKa = 8.3), mak-
ing the molecule unstable. The charged adenine moiety attached to the
anomeric carbon of the N1-ribose can be an efficient leaving group. Accord-
ingly, cADPR is readily hydrolyzed at the unstable N-1-ribosyl linkage of its
adenine moiety to produce ADP-ribose (ADPR), even in neutral aqueous
Received 5 January 2006; accepted 18 January 2006.
This article is dedicated to Professor Eiko Ohtsuka on the occasion of her 70th birthday.
This report constitutes Part 239 of Nucleosides and Nucleotides. Part 238: Ichikawa, S.; Minakawa,
N.; Shuto, S.; Tanaka, M.; Sasaki, T.; Matsuda, A. Synthesis of 3ꢁ-β-carbamoylmethylcytidine (CAMC) and
its derivatives as potential Antitumor agents. Organic and Biomolecular Chemistry 2006, 4, 1284–1296.
Address correspondence to Satoshi Shuto, Graduate School of Pharmaceutical Sciences, Hokkaido
University, Kita-12, Nishi-6, Kita-ku, Sapporo, 060-0812, Japan. E-mail: shu@pharm.hokudai.ac.jp
583

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T. Kudoh et al.
FIGURE 1 cADPR (1), cADPcR (2), and the 3ꢁꢁ-modified cADPcR analogues 3–5.
solution.[3] Under physiological conditions, cADPR is also hydrolyzed at the
N-1-ribosyl linkage by cADPR hydrolase to give the inactive ADPR.[3]
cADPR analogues can be used in proving the mechanism of cADPR-
mediated Ca2+ signaling pathways and are also expected to be lead structures
for the development of drugs, since cADPR has been shown to play important
physiological roles.[2] Therefore, the synthesis of cADPR analogues has been
extensively investigated by enzymatic and chemo-enzymatic methods using
ADP-ribosyl cyclase-catalyzed cyclization. However, the analogues obtained
by these methods are limited due to the substrate-specificity of the ADP-
ribosyl cyclase.[2]
On the other hand, in the chemical synthesis of cADPR and its analogues,
construction of the large 18-membered ring structure is the key step, and
we recently developed an efficient method for forming the 18-membered
ring employing phenylthiophosphate-type substrates.[4] When these sub-
strates were activated by AgNO3 or I2 in the presence of molecular sieves
in pyridine, the corresponding 18-membered ring products were obtained
in high yields.[4b,c] Using this method, we successfully synthesized cyclic ADP-
carbocyclic-ribose (cADPcR, 2),[4c] designed as a stable mimic of cADPR, in
which the oxygen atom in the N-1-ribose ring of cADPR is replaced by a
methylene group. Biological evaluation of cADPcR showed that it actually
act as biologicically and chemically stable equivalent of cADPR.[4c]
Based on these results, we have investigated further synthetic and bi-
ological studies on N1-carbocyclic derivatives of cADPR.[5] In the course
of these studies, we describe here the synthesis and biological evaluation
of newly designed analogues of cADPcR, which are cyclic ADP-carbocyclic-
xylose (cADPcX, 4) and the corresponding 3ꢁꢁ-O-methyl derivative (3ꢁꢁ-OMe-
cADPcX, 5).
RESULTS AND DISCUSSION
Design and Synthetic Plan
We previously showed (1) that cADPcR (2) is actually resistant to both
enzymatic and chemical hydrolysis, since it has a chemically and biologically

Synthesis of Cyclic ADP-Carbocylcic-Xylose
585
SCHEME 1
stable N-alkyl linkage instead of the unstable N1-glycosidic linkage of
cADPR,[4b] (2) that cADPcR has a conformation similar to that of cADPR,[5c]
and (3) that cADPcR, like cADPR, effectively mobilizes intracellular Ca2+ in
sea urchin eggs and neuronal cells.[4c,5b,c] Furthermore, we have investigated
SAR of the N1-ribose moiety of cADPcR and clarified that modification at
the N1-ribose moiety changes the biological potency.[5b,c] Throughout these
studies, we also found that although deletion of the 2ꢁꢁ-hydroxy group re-
sulted in a marked reduction of potency, deletion of the adjacent 3ꢁꢁ-hydroxy
group (3ꢁꢁ-deoxy-cADPcR, 3) greatly potentiated the Ca2+-mobilizing abil-
ity in sea urchin eggs.[5c] These results suggest that modification at the 3ꢁꢁ-
position may improve the biological potency of cADPR and its analogues.
Thus, we newly designed another 3ꢁꢁ-modified analogues of cADPcR, which
were the N1-“xylo”-type carbocyclic analogue, i.e., cADPcX (4) and the cor-
responding O-methyl analogue 5.
As described above, we have developed an efficient total synthetic method
for cADPR analogues,[4] which we[5] and other groups[6] have effectively used
in the synthesis of a variety of cADPR analogues. Thus, we planned to syn-
thesize the target compounds based on the previous total synthetic method.
The synthetic plan is shown in Scheme 1 as a retrosynthetic analysis.
The chiral carbocyclic–xylosyl amines III, composing the N1-substituted
moiety in the targets 4 and 5, could be prepared from commercially
available (1R)-(–)-2-azabicyclo[2.2.1]hept-5-en-3-one (8). From these carbo-
cyclic amines III and the known imidazole nucleoside derivative 6,[4b] the
5ꢁ-phenylthiophosphate-type substrates II for the key intramolecular con-
densation could be prepared. Treatment of II with AgNO3/MS 3A as a
promoter[4a,b] was expected to form the cyclized products I, and subsequent
acidic treatment for deprotection would furnish the desired cADPcR ana-
logues 4 and 5.
Synthesis
The carbocyclic-xylosyl amine units 14a and 14b were synthesized from
the optically active bicyclic lactam 8, as summarized in Scheme 2. After

586
T. Kudoh et al.
SCHEME 2 Reagents and conditions: a) TIPSCl, imidazole, DMAP, DMF, rt, 91%; b) ClCH2CO2H, DIAD,
Ph3P, toluene, 0◦C, 74%; c) NaOMe, MeOH, rt, 88%; d) 1) MOMCl, i-Pr2NEt, CH2Cl2, rt, 2) TBAF, THF,
rt, 97% (13a); e) 1) MeOTf, DTBMP, CH2Cl2, rt, 2) TBAF, THF, rt, 45% (13b); f) H2O, reflux, quant.
(14a, 14b).
protection of the primary hydroxyl with a triisopropysilyl (TIPS) group of
cabocylicilc-ribose derivative 9, prepared from 8 according to the previously
reported method,[5c] treatment of the resulting 10 under Mitsunobu reac-
tion conditions with ClCH2CO2H/DIAD/Ph3P gave the corresponding 3-
position-inverted product 11, and subsequent removal the 3-O-chloroacetly
group with NaOMe/MeOH to afford the carbocyclic-xylose derivative 12.
The 3-hydroxy of 12 was protected with a MOM group under usual conditions
or methylated with MeOTf and 2,6-di-tert-butyl-4-methylpyridine (DTBMP),
followed by removal of the TIPS and BOC groups, to give the desired 3-O-
MOM-ptotected unit 14a or the 3-O-methyl unit 14b, respectively.
The target 3ꢁꢁ-modified cADPcR analogues 4 and 5 were successfully syn-
thesized from the carbocyclic amines 14a or 14b and the imidazole nucleo-
side 6, as shown in Scheme 3.
The N-1-substituted adenosine derivatives 15a and 15b were obtained
in high yield by the treatment of a mixture of 6 and either amine 14a or
amine 14b with K2CO3 in MeOH at room temperature. The 5ꢁꢁ-hydroxy
group of 15a or 15b was protected with a dimethoxytrityl (DMTr) group, and
the 5ꢁ-O-TBS group of the product was subsequently removed with TBAF to
give 16a or 16b. Treatment of 16a or 16b under the conditions reported by
Hata and co workers with an S,S ꢁ-diphenylphosphorodithioate (PSS)/2,4,6-
triisopropylbenzenesulfonyl chloride (TPSCl)/pyridine system[7] gave the
5ꢁ-bis(phenylthio)phosphate 17a or 17b, respectively. The 5ꢁꢁ-O-DMTr group
of 17a or 17b was removed to give 18a or 18b, respectively. A phosphoryl
group was introduced at the resulting 5ꢁꢁ-primary hydroxyl of 18a or 18b
by Yoshikawa’s method with POCl3/(EtO)3PO,[8] followed by treatment of
the product with H3PO2 and Et3N[9] in the presence of N-methylmaleimide
(NMM)[4c] in pyridine, to afford the corresponding 5ꢁ-phenylthiophosphate
19a or 19b, respectively, which was the substrate of the key intramolecular

Synthesis of Cyclic ADP-Carbocylcic-Xylose
587
SCHEME 3 Reagents and conditions: a) K2CO3, MeOH, rt, 83% (15a), 84% (15b); :b) 1) DMTrCl,
pyridine, rt, 2) TBAF, AcOH, THF, rt, quant. (16a), quant. (16b); c) PSS, TPSCl, py, rt, 65% (17a), 62%
(17b); d) aq. 60% AcOH, rt, 83% (18a), 88% (18b); e) 1) POCl3, (EtO)3PO, 0◦C, 2) H3PO2, Et3N, NMM,
pyridine, 0◦C, 46% (19a), 58% (19b; f) AgNO3, MS 3A, Et3N, py, rt, 46% (20a), 47% (20b); h) aq. 80%
HCO2H, then aq. NH3, rt, 90% (4), quant. (5).
condensation reaction. When a solution of 19a in pyridine was added slowly
to a mixture of a large excess of AgNO3 and Et3N in the presence of MS 3A
in pyridine at room temperature,[4b,c] the intramolecular pyrophosphate
linkage was successfully formed as the previously reported cases to give
the desired cyclization product 20a in 46% yield. The other substrate 19b
was similarly condensed and the cyclization product 20b was obtained. Fi-
nally, the protecting groups of 20a and 20b were simultaneously removed by
acidic treatment with aqueous HCO2H furnished the target cADPcX (4) and
3ꢁꢁ-OMe-cADPcX (5).

588
T. Kudoh et al.
FIGURE 2 Dose-dependent Ca2+-mobilizing activity of compounds in sea urchin egg homogenate. The
Ca2+-mobilizing activity of each compound was expressed as a percent change in ratio of fura-2 fluores-
cence (F340/F380) relative to that of 10 µM cADPR. The compounds examined are cADPR (1, filled
circles), cADPcR (2, open circles), 3ꢁꢁ-deoxy-cADPcR (3, open triangles), cADPcX (4, open squares), and
3ꢁꢁ-O-methyl-cADPcX (5, open diamonds). Data are mean SEM of 3–6 experiments.
Ca2+-Mobilizing Activity in Sea Urchin Egg Homogenate
The Ca2+-mobilizing ability of the newly synthesized compounds 4 and 5
was evaluated by the fluorometrically Ca2+-monitoring method with H. pul-
cherrimus sea urchin egg homogenate,[5b,10] and the results were compared
with those of the natural second messenger cADPR (1) and the related car-
bocyclic analogues cADPcR (2) and 3ꢁꢁ-deoxy-cADPcR (3). Both of the two
newly synthesized compounds 4 and 5 released Ca2+ from the homogenate
in a dose-dependent manner, as shown in Figure 2, where the maximal Ca2+-
mobilizing activity was almost equal to that of cADPR. Thus, 4 and 5 were
shown to be full agonists as cADPcR (2) and 3ꢁꢁ-deoxy-cADPcR (3). cADPcX
(4), the 3ꢁꢁ-epimer of cADPcR, showed marked Ca2+-mobilizing activity with
an EC50 value of 69 nM, which was similar to that of cADPcR with an EC50
value of 79 nM and was 3 times more potent than the natural ligand cADPR
(EC50 = 220 nM). The 3ꢁꢁ-O-methyl cADPcX (5) showed an EC50 value of
740 nM, which was about 10 times weaker than cADPcX. Similar to the case
of cADPcR, 3ꢁꢁ-methylation of the 3ꢁꢁ-hydroxy group of cADPcX markedly
reduced the activity.
CONCLUSION
We successfully synthesized cADPcX (4), the 3ꢁꢁ-epimer of cADPcR (2),
and the corresponding 3ꢁꢁ-O-methyl analogue (3ꢁꢁ-OMe-cADPcX, 5) and iden-
tified cADPcX as one of the most potent cADPR-related compounds reported

Synthesis of Cyclic ADP-Carbocylcic-Xylose
589
so far. Therefore, irrespective of the 3ꢁꢁ-configuration, both cADPcX and
cADPcR have strong Ca2+-mobilizing activity to suggest that the 3ꢁꢁ-hydroxy
group seems to be unnecessary in their binding with the target biomolecule,
which is in accord with the previous results on the 3ꢁꢁ-deoxy-cADPcR with
remarkable activity.[5c]
EXPERIMENTAL
General Methods
Chemical shifts are reported in ppm downfield from Me4Si (1H), MeCN
1
(13C) or H3PO4 (31P). All of the H NMR assignments described were in
agreement with COSY spectra. Thin-layer chromatography was done on
Merck coated plate 60F254. Silica gel chromatography was done on Merck
silica gel 5715. Reactions were carried out under an argon atmosphere.
(1R ,2S ,3R ,4R -1-tert -Butoxycarbonylamino-2-(methoxymethyloxy)-3-hydroxy-4
-(triisopropylsilyloxymethyl)cyclopentane ( 10). A mixture of 9 (1.20 g, 4.12
mmol), TIPSCl (2.12 ml, 9.88 mmol), imidazole (1.01 g, 14.8 mmol),
and DMAP (0.604 g, 4.94 mmol) in DMF (40 ml) was stirred at room
temperature for 4 h. After addition of MeOH (10 ml), the resulting mixture
was evaporated. The residue was partitioned between EtOAc and H2O, and
the organic layer was washed with brine, dried (Na2SO4), and evaporated.
The residue was purified by column chromatography (SiO2, 25% EtOAc
1
in hexane) to give 10 (1.68 g, 91%) as a colorless oil: H-NMR (CDCl3,
500 MHz) δ 4.98 (brs, 1H, NH), 4.78 (d, 1H, MOM-CH2, J = 6.4 Hz), 4.72
(d, 1H, MOM-CH2, J = 6.4 Hz), 4.08 (m, 1H, H-3), 3.99 (m, 1H, H-1),
3.88 (dd, 1H, H-6a, J 6a,6b = 9.6 Hz, J 6a,4 = 2.8 Hz), 3.80 (m, 1H, H-2), 3.69
(dd, 1H, H-6b, J 6b,6a = 9.6 Hz, J 6b,4 = 3.2 Hz), 3.41 (s, 3H, MOM-CH3),
2.66 (brs, 1H, OH), 2.37 (m, 1H, H-5a), 2.04 (m, 1H, H-4), 1.42 (s, 9H,
tert-Bu), 1.33 (m, 1H, H-5b), 1.07–1.18 (m, 21H, TIPS); 13C-NMR (CDCl3,
125 MHz) δ 155.10, 96.53, 84.21, 79.13, 72.64, 63.66, 55.73, 53.62, 45.40,
30.33, 28.36, 18.00, 12.05; HRMS (FAB, positive) calcd for C22H46NO6Si
448.3094 (MH+), found 448.3109.
(1R ,2S ,3S ,4R )-1-tert -Butoxycarbonylamino-2-(methoxymethyloxy)-3-(chloroa-
cethoxy)-4-(triisopropylsilyloxymethyl)cyclopentane ( 11). To a mixture of 10
(0.160 g, 0.357 mmol), chloroacetic acid (0.135 g, 1.43 mmol), and Ph3P
(0.374 g, 1.43 mmol) in toluene (2 ml), a solution of DIAD (281 µl, 1.43
mmol) in toluene (1.5 ml) was added slowly at 0◦C, and the mixture was
stirred at room temperature for 6 h and then evaporated. The residue was
purified by column chromatography (SiO2, 10% EtOAc in hexane) to give
1
11 (0.139 g, 74%) as a colorless oil: H-NMR (CDCl3, 500 MHz) δ 5.21
(dd, 1H, H-3, J 3,2 = 3.9 Hz, J 3,4 = 6.0 Hz), 4.86 (m, 1H, NH), 4.72 (d, 1H,
MOM-CH2, J = 6.6 Hz), 4.69 (d, 1H, MOM-CH2, J = 6.6 Hz), 4.05 (s, 2H,
Cl-Ac), 3.97 (m, 1H, H-1), 3.92 (m, 1H, H-2), 3.70 (m, 2H, H-6 × 2), 3.36

590
T. Kudoh et al.
(s, 3H, MOM-CH3), 2.48 (m, 1H, H-4), 2.38 (m, 1H, H-5a), 1.43 (m, 10H,
H-5b, tert-Bu), 1.09 (m, 21H, TIPS); 13C-NMR (CDCl3, 125 MHz) δ 166.33,
155.16, 95.70, 85.43, 79.78, 73.06, 70.82, 61.80, 55.48, 54.77, 41.55, 40.78,
32.43, 28.35, 17.95, 11.89; HRMS (FAB, positive) calcd for C24H47ClNO7Si
524.2810 (MH+), found 524.2816.
(1R ,2S ,3S ,4R )-1-tert-Butoxycarbonylamino-2-(methoxymethyloxy)-3-hydroxy-4
-(triisopropylsilyloxymethyl)cyclopentane ( 12). A mixture of 11 (0.173 g, 0.330
mmol) and NaOMe (1 M in MeOH, 50 µl, 50 µmol) in MeOH (3 ml) was
stirred at room temperature for 30 min. After addition of aqueous saturated
NH4Cl (1 ml) at 0◦C, the resulting mixture was extracted with EtOAc, and
the organic layer was washed with H2O and brine, dried (Na2SO4), and
evaporated. The residue was purified by column chromatography (SiO2,
25% EtOAc in hexane) to give 12 (0.130 g, 88%) as a colorless oil: 1H-NMR
(DMSO-d6, 500 MHz) δ 6.69 (d, 1H, NH, J = 7.3 Hz), 4.70 (d, 1H, OH,
J = 4.6 Hz), 4.63 (d, 1H, MOM-CH2, J = 6.4 Hz), 4.60 (d, 1H, MOM-CH2,
J = 6.4 Hz), 3.84 (m, 2H, H-6a, H-3), 3.62 (m, 1H, H-1), 3.58 (m, 2H, H-2,
H-6b), 3.22 (s, 3H, MOM-CH3), 2.02 (m, 1H, H-4), 1.96 (m, 1H, H-5a),
1.36 (s, 10H, H-5b, tert-Bu), 1.03 (m, 21H, TIPS); 13C-NMR (DMSO-d6, 125
MHz) δ 156.04, 95.36, 88.30, 78.17, 74.92, 63.31, 55.46, 55.04, 43.62, 32.28,
28.56, 18.16, 11.67; HRMS (FAB, positive) calcd for C22H46NO6Si 448.3094
(MH+), found 448.3092.
(1R ,2S ,3S ,4R -1-tert -Butoxycarbonylamino-2,3-bis (methoxymethyloxy)-4-hyd-
roxymethylcyclopentane ( 13a). A mixture of 12 (0.871 g, 1.95 mmol), MOMCl
(739 µl, 9.73 mmol) and i-Pr2NEt (3.39 ml, 19.5 mmol) in CH2Cl2 (20 ml)
was stirred at room temperature for 75 h. After addition of MeOH (10 ml),
the resulting mixture was evaporated. The residue was partitioned between
EtOAc and 0.1 M HCl, and the organic layer was washed in H2O and
brine, dried (Na2SO4), and evaporated. A mixture of the residue and TBAF
(1.0 M in THF, 3.0 ml, 3.0 mmol) in THF (20 ml) was stirred at room
temperature for 1 h and then evaporated. The residue was purified by
column chromatography (SiO2, EtOAc) to give 13a (0.634 g, 97%) as a
1
yellow oil: H-NMR (DMSO-d6, 500 MHz) δ 6.85 (d, 1H, NH, J = 7.9 Hz),
4.64 (d, 1H, MOM-CH2, J = 6.5 Hz), 4.59 (m, 3H, MOM-CH2 × 3), 4.34 (m,
1H, OH), 3.84 (m, 1H, H-3), 3.80 (m, 1H, H-2), 3.66 (m, 1H, H-1), 3.50
(m, 1H, H-6a), 3.38 (m, 1H, H-6b), 3.26 (s, 3H, MOM-CH3), 3.23 (s, 3H,
MOM-CH3), 2.06 (m, 1H, H-4), 1.93 (m, 1H, H-5a), 1.33 (s, 9H, tert-Bu),
1.32 (m, 1H, H-5b); 13C-NMR (DMSO-d6, 125 MHz) δ 155.15, 95.14, 94.64,
85.42, 80.11, 77.69, 60.05, 55.65, 54.98, 54.72, 42.69, 32.25, 28.39; FAB-MS
m/z 336 (MH+); Anal. Calcd for C15H29NO7: C, 53.72; H, 8.72; N, 4.18.
Found; C, 53.49; H, 8.57; N, 4.00.
(1R ,2S ,3S ,4R )-1-tert -Butoxycarbonylamino-2-(methoxymethyloxy)-3-methoxy-
4-hydroxymethylcyclopentane ( 13b). A mixture of 12 (0.513 g, 1.15 mmol),
MeOTf (648 µl, 5.73 mmol) and DTBMP (1.29 g, 6.30 mmol) in CH2Cl2
(11 ml) was stirred at room temperature for 12 h. After addition of aqueous

Synthesis of Cyclic ADP-Carbocylcic-Xylose
591
saturated NaHCO3 (10 ml) at 0◦C, the mixture was extracted with EtOAc,
and the organic layer was washed with H2O and brine, dried (Na2SO4), and
evaporated. The residue was purified by column chromatography (SiO2,
10% EtOAc in hexane) to give the 3-O-methylated product (0.259 g, 49%)
as a colorless oil. A mixture of the oil and TBAF (1.0 M in THF, 0.84 ml,
0.84 mmol) in THF (5 ml) was stirred at room temperature for 2 h and
then evaporated. The residue was purified by column chromatography
(SiO2, 75% EtOAc in hexane) to give 13b (0.154 g, 90%) as a yellow oil:
1H-NMR (DMSO-d6, 500 MHz) δ 6.84 (d, 1H, NH, J = 8.0 Hz), 4.65 (d, 1H,
MOM-CH2, J = 6.5 Hz), 4.58 (d, 1H, MOM-CH2, J = 6.5 Hz), 4.29 (m, 1H,
OH), 3.77 (dd, 1H, H-2, J 2,1 = 5.1 Hz, J 2,3 = 3.0 Hz), 3.65 (m, 1H, H-1),
3.55 (m, 1H, H-6a), 3.47 (dd, 1H, H-3, J 3,2 = 3.0 Hz, J 3,4 = 5.8 Hz), 3.32
(m, 1H, H-6b), 3.24 (s, 6H, OMe, MOM-CH3), 2.04 (m, 1H, H-4), 1.91 (m,
1H, H-5a), 1.36 (s, 9H, tert-Bu), 1.30 (m, 1H, H-5b); 13C-NMR (DMSO-d6,
125 MHz) δ 155.16, 94.70, 84.84, 84.45, 77.68, 59.93, 56.82, 55.69, 54.74,
42.65, 32.30, 28.39; FAB-MS m/z 306 (MH+); Anal. Calcd for C14H27NO6: C,
55.07; H, 8.91; N, 4.59. Found; C, 55.09; H, 8.76; N, 4.51.
(1R ,2S ,3S ,4R )-1-Amino-2,3-bis (methoxymethyloxy)-4-hydroxymethylcyclopen-
tane ( 14a). A solution of 13a (0.614 g, 1.83 mmol) in H2O (18 ml) was
stirred at 100◦C for 12 h and then evaporated. The residue was azeoptroped
with toluene (10 ml × 3) to give 14a (0.430 g, quant.) as a brown oil: 1H-NMR
(DMSO-d6, 500 MHz) δ 4.67 (d, 1H, MOM-CH2, J = 6.5 Hz), 4.59 (m, 3H,
MOM-CH2 × 3), 3.85 (dd, 1H, H-3, J 3,2 = 3.5 Hz, J 3,4 = 6.0 Hz), 3.57 (m,
1H, H-2), 3.50 (dd, 1H, H-6a, J 6a,6b = 10.3 Hz, J 6a,4 = 7.0 Hz), 3.37 (dd, 1H,
H-6b, J 6b,6a = 10.3 Hz, J 6b,4 = 6.5 Hz), 3.26 (s, 6H, MOM-CH3 × 2), 2.96
(m, 1H, H-1), 2.06 (m, 1H, H-4), 1.92 (m, 1H, H-5a), 1.15 (s, 1H, H-5b);
13C-NMR (DMSO-d6, 125 MHz) δ 95.24, 95.18, 89.75, 80.81, 60.41, 56.84,
54.98, 54.86, 42.57, 35.50; HRMS (FAB, positive) calcd for C10H22NO5
236.1498 (MH+), found 236.1508.
(1R ,2S,3S,4R )-1-Amino-2-(methoxymethyloxy)-3-methoxy-4-hydroxymethylcyclo-
pentane ( 14b). Compound 14b (0.239 g, quant.) was obtained from
1
13b (0.351 g, 1.15 mmol) as described for the synthesis of 14a: H-NMR
(DMSO-d6, 500 MHz) δ 4.68 (d, 1H, MOM-CH2, J = 6.6 Hz), 4.62 (d, 1H,
MOM-CH2, J = 6.6 Hz), 3.53 (m, 1H, H-2), 3.49 (m, 2H, H-3, H-6a), 3.33
(m, 1H, H-6b), 3.27 (s, 3H, MOM-CH3 or OMe), 3.25 (s, 3H, MOM-CH3
or OMe), 2.95 (m, 1H, H-1), 2.06 (m, 1H, H-4), 1.91 (m, 1H, H-5a), 1.14
(s, 1H, H-5b); 13C-NMR (DMSO-d6, 125 MHz) δ 95.30, 89.03, 85.62, 60.28,
56.96, 56.91, 54.87, 42.50, 35.44; HRMS (FAB, positive) calcd for C9H20NO4
206.1392 (MH+), found 206.1390.
N -1- {(1R ,2S ,3S ,4R )-2,3-Bis(methoxymethyloxy)-4-(hydroxymethyl)cyclopentyl }
-5ꢁ-O -(tert -butyldimethylsilyl)-2ꢁ,3ꢁ-O -isopropylideneadenosine ( 15a). A mixture
of 6 (0.765 g, 1.75 mmol), 14a (0.417 g, 1.77 mmol), and K2CO3 (12 mg,
88 µmol) in MeOH (18 ml) was stirred at room temperature for 12 h and
then evaporated. The residue was partitioned between EtOAc and H2O, and

592
T. Kudoh et al.
the organic layer was washed with brine, dried (Na2SO4), and evaporated.
The residue was purified by column chromatography (SiO2, 25% MeOH
1
in EtOAc) to give 15a (0.930 g, 83%) as a white foam: H-NMR (CDCl3,
500 MHz) δ 7.91 (s, 1H, H-2 or H-8), 7.82 (s, 1H, H-2 or H-8), ꢁ6.02 (d, 1H,
ꢁꢁ
H-1ꢁ, J 1 ,2 = 2.7 Hz), 5.37 (m, 1H, H-1 ), 5.09 (dd, 1H, H-2 , J 2 ,1 = 2.7
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Hz, J 2 ,3 = 6.1 Hz), 4.89 (dd, 1H, H-3 , J 3 ,2 = 6.1 Hz, J 3 ,4 = 2.4 Hz), 4.81
(d, 1H, MOM-CH2, J = 6.6 Hz), 4.76 (d, 1H, MOM-CH2, J = 6.6 Hz), 4.69 (d,
1H, MOM-CH2, J = 6.9 Hz), 4.65 (d, 1H, MOM-CH2, J = 6.9 Hz), 4.48 (m,
1H, H-2ꢁꢁ), 4.39 (ddd, 1H, H-4ꢁ, J 4 ,3 = 2.4 Hz, J 4 ,5 a = 6.4 Hz, J 4 ,5 b = 3.7
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Hz), 4.19 (dd, 1H, H-3ꢁꢁ, J 3 ,2 = 3.5 Hz, J 3,4 = 5.8 Hz), 3.83 (m, 2H, H-5
a, H-5ꢁꢁa), 3.77 (m, 2H, H-5ꢁ b, H-5ꢁꢁb), 3.45 (s, 3H, MOM-CH3), 3.21 (s, 3H,
MOM-CH3), 2.47 (m, 1H, H-4ꢁꢁ), 2.39 (m, 1H, H-6ꢁꢁa), 2.02 (m, 1H, H-6ꢁꢁb),
1.61, 1.39 (each s, each 3H, isopropylidene), 0.86 (s, 9H, tert -Bu), 0.046,
0.035 (each s, each 3H, TBS-Me × 2); 13C-NMR (CDCl3, 125 MHz) δ 154.45,
146.66, 140.62, 136.78, 123.40, 114.16, 96.39, 95.63, 91.26, 87.03, 85.32,
84.38, 81.76, 81.33, 63.46, 61.56, 60.08, 56.02, 55.49, 42.10, 31.22, 27.23,
25.86, 25.38, 18.32, −5.43, −5.54; FAB-MS m/z 640 (MH+); UV (MeOH)
λmax = 261, 295 (sh) nm; Anal. Calcd for C29H49N5O9Si: C, 54.44; H, 7.72;
N, 10.95. Found; C, 54.24; H, 7.66; N, 10.84.
ꢁ
ꢁꢁ ꢁꢁ
ꢁꢁ
N -1-[(1R ,2S ,3S ,4R )-2,3-bis (methoxymethyloxy)-4-(dimethoxytrityloxy methyl)
cyclopentyl]-2 ꢁ,3 ꢁ-O -isopropylideneadenosine ( 16a). A mixture of 15a (0.884 g,
1.38 mmol) and DMTrCl (1.40 g, 4.14 mmol) in pyridine (15 ml) was stirred
at room temperature for 10 min. After addition of MeOH (10 ml), the re-
sulting mixture was evaporated. The residue was partitioned between EtOAc
and H2O, and the organic layer was washed with brine, dried (Na2SO4),
and evaporated. A mixture of the residue, TBAF (1.0 M in THF, 2.76 ml,
2.76 mmol) and AcOH (79 µl, 1.38 mmol) in THF (10 ml) was stirred at
room temperature for 3 h and then evaporated. The residue was purified
by column chromatography (SiO2, 25% MeOH in EtOAc) to give 16a (1.19
1
g, quant.) as a white foam: H-NMR (CDCl3, 500 MHz) δ 8.00 (s, 1H, H-2
or H-8), 7.62 (s, 1H, H-2 or H-8), 6.82–7.43 (m, 13H, DMTr), 5.79 (m, 2H,
H-1ꢁ, H-1ꢁꢁ), 5.02 (m, 2H, H-2ꢁ, H-3ꢁ), 4.81 (d, 1H, MOM-CH2, J = 6.9 Hz),
4.67 (d, 1H, MOM-CH2, J = 6.9 Hz), 4.61 (d, 1H, MOM-CH2, J = 6.7 Hz),
4.54 (d, 1H, MOM-CH2, J = 6.7 Hz), 4.47 (m, 1H, H-4ꢁ), 4.20 (m, 1H, H-
3ꢁꢁ), 4.12 (m, 1H, H-2ꢁꢁ), 3.91 (m, 1H, H-5ꢁa), 3.80 (s, 6H, DMTr-OMe ×
2), 3.72 (m, 1H, H-5ꢁb), 3.33 (s, 3H, MOM-CH3), 3.29 (m, 1H, H-5ꢁꢁa), 3.25
(s, 3H, MOM-CH3), 3.13 (m, 1H, H-5ꢁꢁb), 2.68 (m, 1H, H-4ꢁꢁ), 2.56 (m, 1H,
H-6ꢁꢁa), 1.62, 1.35 (each s, each 3H, isopropylidene), 1.49 (m, 1H, H-6ꢁꢁ b);
13C-NMR (CDCl3, 125 MHz) δ 158.45, 146.70, 144.96, 137.95, 136.31, 136.11,
129.97, 129.93, 128.12, 127.79, 126.76, 114.20, 113.09, 113.06, 96.41, 94.93,
93.90, 86.00, 85.82, 83.61, 81.37, 63.19, 61.56, 60.37, 58.81, 55.98, 55.60,
55.20, 42.94, 35.23, 27.55, 25.18, 21.02, 14.19; HRMS (FAB, positive) calcd
for C44H54N5O11 828.3820 (MH+), found 828.3818; UV (MeOH) λmax = 260,
295 (sh) nm.

Synthesis of Cyclic ADP-Carbocylcic-Xylose
593
N -1-[(1R ,2S ,3S ,4R )-2,3-Bis (methoxymethyloxy)-4-(dimethoxytrityloxymethyl)
cyclopentyl]-5 ꢁ-O-{bis(phenylthio)phospholyl}-2 ꢁ,3 ꢁ-O-isopropylide-neadenosine
( 17a). After stirring a mixture of PSS (1.56 g, 4.09 mmol) and TPSCl
(1.12 g, 3.69 mmol) in pyridine (14 ml) at room temperature for 1 h, 16a
(1.13 g, 1.36 mmol) was added, and the resulting mixture was stirred at
room temperature for further 2 h and then evaporated. The residue was
partitioned between EtOAc and H2O, and the organic layer was washed
with brine, dried (Na2SO4), and evaporated. The residue was purified by
column chromatography (SiO2, 2% MeOH in CHCl3) to give 17a (0.890
1
g, 65%) as a white foam: H-NMR (CDCl3, 500 MHz) δ 7.95 (s, 1H, H-2 or
H-8), 7.64 (s, 1H, H-2 or H-8), 6.81-7.51 (m, 23H, DMTr, SPh × 2), 5.96 (d,
ꢁꢁ
ꢁ
1H, H-1ꢁ, J 1 2 = 2.6 Hz), 5.76 (m, 1H, H-1 ), 5.07 (dd, 1H, H-2 , J 2 ,1 = 2.6
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Hz, J 2 ,3 = 6.3 Hz), 4.85 (dd, 1H, H-3 , J 3 ,2 = 6.3 Hz, J 3 ,4 = 2.7 Hz), 4.76
(d, 1H, MOM-CH2, J = 6.8 Hz), 4.64 (d, 1H, MOM-CH2, J = 6.8 Hz), 4.57
(d, 1H, MOM-CH2, J = 6.7 Hz), 4.53 (d, 1H, MOM-CH2, J = 6.7 Hz), 4.41
(m, 1H, H-4ꢁ), 4.38 (m, 2H, H-5ꢁ × 2), 4.20 (d, 1H, H-3ꢁꢁ, J 3 ,4 = 4.0 Hz),
ꢁꢁ ꢁꢁ
4.14 (d, 1H, H-2ꢁꢁ, J 2 ,1 = 2.6 Hz), 3.79 (s, 6H, DMTr-OMe × 2), 3.30 (m,
ꢁꢁ ꢁꢁ
1H, H-5ꢁꢁa), 3.27 (s, 3H, MOM-CH3), 3.19 (s, 3H, MOM-CH3), 3.16 (m, 1H,
H-5ꢁꢁb), 2.61 (m, 1H, H-4ꢁꢁ), 2.54 (m, 1H, H-6ꢁꢁa), 1.59, 1.34 (each s, each
3H, isopropylidene), 1.54 (m, 1H, H-6ꢁꢁb); 13C-NMR (CDCl3, 125 MHz)
δ 158.43, 154.52, 146.61, 144.99, 140.49, 136.94, 136.35, 136.17, 135.34,
135.29, 135.21, 135.17, 129.98, 129.96, 129.66, 129.63, 129.46, 129.44,
128.13, 127.76, 126.72, 125.93, 125.87, 125.79, 125.74, 123.52, 114.69,
113.05, 113.02, 95.88, 94.99, 90.61, 86.00, 85.87, 84.67, 84.61, 84.45, 81.06,
80.78, 66.42, 66.36, 61.80, 58.76, 55.86, 55.51, 55.19, 42.79, 35.17, 27.13,
25.30, 21.02, 14.19; 31P-NMR (CDCl3, 202 MHz) δ 50.74 (s); HRMS (FAB,
positive) calcd for C56H63N5O12PS2 1092.3652 (MH+), found 1092.3660;
UV (MeOH) λmax = 295 (sh) nm.
N-1-[(1R,2S,3S,4R)-2,3-Bis(methoxymethyloxy)-4-(hydroxymethyl)cyclopentyl]-5 ꢁ
-O -[bis (phenylthio)phospholyl]-2 ꢁ,3 ꢁ-O -isopropylideneadeno-sine ( 18a). A so-
lution of 17a (0.916 g, 0.839 mmol) in aqueous 60% AcOH (8 ml) was
stirred at room temperature for 4 h. After addition of aqueous saturated
NaHCO3 (60 ml) at 0◦C, the mixture was extracted with EtOAc, and the
organic layer was washed with aqueous saturated NaHCO3, H2O and brine,
dried (Na2SO4), and evaporated. The residue was purified by column
chromatography (SiO2, 10% MeOH in CHCl3) to give 18a (0.551 g, 83%) as
a white foam: 1H-NMR (CDCl3, 500 MHz) δ 7.93 (s, 1H, H-2 or H-8), 7.69 (s,
1H, H-2 or H-8), 7.30–7.52 (m, 10H, SPh × 2), 5.99 (m, 1H, H-1ꢁ), 5.32 (m,
1H, H-1ꢁꢁ), 5.11 (m, 1H, H-2ꢁ), 4.90 (dd, 1H, H-3ꢁ, J 3 ,2 = 6.2 Hz, J 3 ,4 = 2.5
Hz), 4.76 (d, 1H, MOM-CH2, J = 6.5 Hz), 4.72 (d, 1H, MOM-CH2, J = 6.5
Hz), 4.66 (d, 1H, MOM-CH2, J = 6.7 Hz), 4.62 (d, 1H, MOM-CH2, J = 6.7
Hz), 4.45 (m, 1H, H-2ꢁꢁ), 4.43 (m, 2H, H-4ꢁ, H-5ꢁa), 4.38 (m, 1H, H-5ꢁb), 4.18
ꢁ
ꢁ
ꢁ
ꢁ
(m, 1H, H-3ꢁꢁ), 3.84 (dd, 1H, H-5ꢁꢁa, J 5 a,5 b = 11.2 Hz, J 5 a,4 = 3.8 Hz), 3.76
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
(dd, 1H, H-5ꢁꢁb, J 5 b,5 a = 11.2 Hz, J 5 b,4 = 5.9 Hz), 3.41 (s, 3H, MOM-CH3),
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ

594
T. Kudoh et al.
3.19 (s, 3H, MOM-CH3), 2.47 (m, 1H, H-4ꢁꢁ), 2.40 (m, 1H, H-6ꢁꢁa), 2.04 (m,
1H, H-6ꢁꢁb), 1.60, 1.36 (each s, each 3H, isopropylidene); 13C-NMR (CDCl3,
125 MHz) δ 154.27, 146.89, 137.38, 135.31, 135.27, 135.17, 135.13, 129.69,
129.66, 129.64, 129.62, 129.44, 129.42, 125.87, 125.82, 125.77, 125.72,
114.66, 96.35, 95.77, 90.74, 84.74, 84.68, 84.43, 81.64, 81.12, 66.39, 66.32,
61.49, 55.95, 55.50, 42.02, 31.10, 27.11, 25.29; 31P-NMR (CDCl3, 202 MHz)
δ 51.10 (s); FAB-MS m/z 790 (MH+); UV (MeOH) λmax = 259, 295 (sh) nm;
Anal. Calcd for C35H44N5O10PS2: C, 53.22; H, 5.61; N, 8.87. Found; C, 53.20;
H, 5.66; N, 8.68.
N-1-[(1R2S,3S,4R)-2,3-Bis(methoxymethyloxy)-4-(phosphonoxy-methyl)cyclop-
entyl]-5ꢁ-O -[(phenylthio)phospholyl]-2ꢁ,3ꢁ-O -isopropyli-deneadenosine
( 19a).
A
mixture of POCl3 (93 µl, 1.0 mmol) and 18a (79 mg, 0.10 mmol) in
PO(OEt)3 (1.0 ml) was stirred at 0◦C for 1 h. After addition of aqueous
saturated NaHCO3 (3.0 ml), the resulting mixture was stirred at 0◦C for 10
min. To the mixture was added triethylammonium acetate (TEAA, 2.0 M,
pH 7.0, 1.0 ml) buffer and H2O (5.0 ml), and the resulting solution was
applied to a C18 reversed phase column (1.1 × 14 cm). The column was
developed using a linear gradient of 0–65% MeCN in TEAA buffer (0.1 M,
pH 7.0, 400 ml). Appropriate fractions were evaporated, and excess TEAA
was removed by C18 reversed phase column chromatography (1.1 × 17 cm,
eluted with 70% aqueous MeCN). Appropriate fractions were evaporated,
and the residue was co-evaporated with pyridine (2.0 ml × 3). A mixture of
the residue, NMM (77 mg, 0.69 mmol), H3PO2 (70 µl, 1.39 mmol), and
Et3N (97 µl, 0.69 mmol) was stirred at 0◦C for 4 h under shading. After
addition of TEAA buffer (1.0 M, pH 7.0, 2.0 ml), the resulting mixture
was evaporated. The residue was partitioned between EtOAc and H2O,
and the aqueous layer was evaporated. A solution of the residue in H2O
(5.0 ml) was applied to a C18 reverse-phase column (1.1 × 16 cm), and the
column was developed using a linear gradient of 0–40% MeCN in TEAA
buffer (0.1 M, pH 7.0, 400 ml). Appropriate fractions were evaporated, and
excess TEAA was removed by C18 reversed phase column chromatography
(1.1 × 17 cm, eluted with 50% aqueous MeCN). Appropriate fractions were
evaporated, and the residue was lyophilized to give 19a (41 mg, 46%) as
1
a triethylammonium salt: H-NMR (D2O, 500 MHz) δ 8.71 (s, 1H, H-2 or
H-8), 8.40 (s, 1H, H-2 or H-8), 7.01–7.30 (m, 5H, SPh), 6.31 (d, 1H, H-1ꢁ,
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
J 1 ,2 = 2.1 Hz), 5.33 (dd, 1H, H-2 , J 2 ,1 = 2.1 Hz, J 2 ,3 = 6.0 Hz), 5.00 (m,
1H, H-1ꢁꢁ), 4.94 (dd, 1H, H-3ꢁ, J 3 ,2 = 6.0 Hz, J 3 ,4 = 1.2 Hz), 4.86 (s, 2H,
MOM-CH2), 4.72 (d, 1H, MOM-CH2, J = 7.3 Hz), 4.70 (m, 1H, H-4ꢁ), 4.68
(d, 1H, MOM-CH2, J = 7.3 Hz), 4.51 (m, 1H, H-2ꢁꢁ), 4.33 (m, 1H, H-3ꢁꢁ), 4.15
(m, 2H, H-5ꢁ × 2), 4.07 (m, 2H, H-5ꢁꢁ × 2), 3.48 (s, 3H, MOM-CH3), 3.27
(s, 3H, MOM-CH3), 3.18 (q, 6H, Et3NH-CH2 × 3, J = 7.3 Hz), 2.74 (m, 1H,
H-4ꢁꢁ), 2.68 (m, 1H, H-6ꢁꢁa), 2.08 (m, 1H, H-6ꢁꢁb), 1.62, 1.39 (each s, each 3H,
isopropylidene), 1.26 (t, 9H, Et3NH-CH3 × 3, J = 7.3 Hz); 13C-NMR (D2O,
125 MHz) δ 151.02, 146.31, 146.10, 143.91, 133.21, 133.17, 129.90, 129.85,
ꢁ
ꢁ
ꢁ
ꢁ

Synthesis of Cyclic ADP-Carbocylcic-Xylose
595
129.44, 128.28, 119.61, 115.17, 97.10, 96.75, 91.87, 87.05, 86.67, 86.59,
84.64, 81.91, 80.44, 66.27, 63.81, 56.40, 56.14, 47.12, 40.75, 40.68, 26.38,
24.73, 8.69; 31P-NMR (D2O, 202 MHz) δ 17.10 (s), 0.84 (s); HRMS (FAB,
positive) calcd for C29H42N5O14P2S 778.1919 (MH+), found 778.1934; UV
(H2O) λmax 260 nm.
2 ꢁꢁ,3 ꢁꢁ-Bis-O-methoxymethyl-cyclic ADP-carbocyclic-xylose 2 ꢁ,3 ꢁ-Acetonide ( 20a).
To a mixture of AgNO3 (310 mg, 1.83 mmol), Et3N (255 µl, 1.83 mmol), and
MS 3A (400 mg) in pyridine (70 ml), a solution of 19a (76 mg, 0.087 mmol)
in pyridine (70 ml) was added slowly over 15 h, using a syringe-pump, at
room temperature under shading. The MS 3A was filtered off with Celite and
washed with H2O. To the combined filtrate and washings was added TEAA
buffer (2.0 M, pH 7.0, 2 ml), and the resulting solution was evaporated. The
residue was partitioned between EtOAc and H2O, and the aqueous layer was
evaporated. A solution of the residue in H2O (5.0 ml) was applied to a C18
reverse-phase column (1.1 × 17 cm), and the column was developed using
a linear gradient of 0–25% MeCN in TEAA buffer (0.1 M, pH 7.0, 400 ml).
Appropriate fractions were evaporated, and excess TEAA was removed by
C18 reverse-phase column chromatography (1.1 × 17 cm, eluted with 40%
aqueous MeCN). Appropriate fractions were evaporated, and the residue was
1
lyophilized to give 20a (40 mg, 46%) as a triethylammonium salt: H-NMR
(D2O, 500 MHz) δ 9.20 (s, 1H, H-2 or H-8), 8.44 (s, 1H, H-2 or H-8), 6.41 (d,
ꢁ
1H, H-1ꢁ, J 1 ,2 = 1.6 Hz), 5.53 (dd, 1H, H-2 , J 2 ,1 = 1.6 Hz, J 2 ,3 = 6.1 Hz),
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
5.46 (dd, 1H, H-3ꢁ, J 3 ,2 = 6.1 Hz, J 3 ,4 = 2.5 Hz), 5.04 (m, 1H, H-1 ), 4.80
(s, 2H, MOM-CH2), 4.76 (d, 1H, MOM-CH2, J = 6.9 Hz), 4.67 (d, 1H, MOM-
CH2, J = 6.9 Hz), 4.63 (m, 1H, H-4ꢁ), 4.62 (m, 1H, H-2ꢁꢁ), 4.37 (m, 1H, H-3ꢁꢁ),
4.23 (m, 1H, H-5ꢁꢁa), 4.16 (m, 1H, H-5ꢁa), 4.09 (m, 2H, H-5ꢁb, H-5ꢁꢁb), 3.42
(s, 3H, MOM-CH3), 3.11 (s, 3H, MOM-CH3), 3.19 (q, 6H, Et3NH-CH2 × 3,
J = 7.3 Hz), 2.99 (m, 1H, H-6ꢁꢁa), 2.76 (m, 1H, H-4ꢁꢁ), 2.36 (m, 1H, H-6ꢁꢁb),
1.64, 1.44 (each s, each 3H, isopropylidene), 1.27 (t, 9H, Et3NH-CH3 × 3,
J = 7.3 Hz); 13C-NMR (D2O, 125 MHz) δ 151.93, 146.22, 146.28, 144.80,
119.64, 115.13, 97.69, 96.82, 92.47, 88.26, 86.92, 86.82, 85.13, 81.87, 81.29,
64.82, 64.61, 60.93, 56.25, 47.10, 37.29, 37.21, 28.70, 26.43, 24.74, 8.67; 31P-
NMR (D2O, 202 MHz) δ −9.58 (d, J = 11.2 Hz), −10.56 (d, J = 11.2 Hz);
HRMS (FAB, positive) calcd for C23H36N5O14P2 668.1729 (MH+), found
668.1730; UV (H2O) λmax 259 nm.
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Cyclic ADP-carbocyclic-xylose ( 4). A solution of 20a (31 mg, 0.040 mmol)
in aqueous 80% HCO2H (1 ml) was stirred at room temperature for 48 h
and then evaporated. After evaporation of the residue in H2O and i-PrOH,
aqueous 28% NH3 (1 ml) was added to the residue, and the mixture was
stirred at room temperature for 90 min. After evaporation of the residue in
H2O and i-PrOH, the resulting residue was dissolved in TEAB buffer (0.1 M,
pH 7.0, 600 µl), and the solution was lyophilized to give 4 (21 mg, 90%) as a
triethylammonium salt: 1H-NMR (D2O, 500 MHz, K+ salt) δ 9.11 (s, 1H, H-2
or H-8), 8.42 (s, 1H, H-2 or H-8), 6.11 (d, 1H, H-1ꢁ, J 1 ,2 = 6.0 Hz), 5.20 (dd,
ꢁ
ꢁ

596
T. Kudoh et al.
ꢁꢁ
1H, H-2ꢁ, J 2 ,1 = 6.0 Hz, J 2 ,3 = 4.8 Hz), 4.95 (m, 1H, H-1 ), 4.64 (dd, 1H,
ꢁ
ꢁ
ꢁ
ꢁ
H-3ꢁ, J 3 ,2 = 4.8 Hz, J 3 ,4 = 2.6 Hz), 4.62 (m, 1H, H-5 a), 4.51 (m, 1H, H-2 ),
4.45 (m, 1H, H-4ꢁ), 4.32 (m, 1H, H-3ꢁꢁ), 4.23 (m, 1H, H-5ꢁꢁa), 4.14 (m, 2H,
H-5ꢁb, H-5ꢁꢁb), 2.95 (m, 1H, H-6ꢁꢁa), 2.65 (m, 1H, H-4ꢁꢁ), 2.46 (m 1H, H-6ꢁꢁb);
13C-NMR (D2O, 125 MHz) δ 152.20, 146.70, 145.43 144.83, 120.40, 91.07,
85.29, 82.52, 76.53, 73.89, 71.16, 65.13, 64.20, 63.12, 39.52, 28.02; 31P-NMR
(D2O, 202 MHz) δ –9.41 (d, J = 11.4 Hz), –10.35 (d, J = 11.4 Hz); HRMS
(FAB, positive) calcd for C16H24N5O12P2 540.0891 (MH+), found 540.0875;
UV (H2O) λmax 259 nm.
ꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
ꢁ
N -1-[(1R ,2S ,3S ,4R )-2-(Methoxymethyloxy)-3-methoxy-4-(hydroxymethyl)cyclo-
pentyl]-5ꢁ-O-(tert-butyldimethylsilyl)-2ꢁ,3ꢁ-O-isopropylideneadenosine ( 15b). Com-
pound 15b (0.565 g, 84%) was obtained from 6 (0.480 g, 1.10 mmol) and
1
14b (0.227 g, 1.11 mmol) as described for the synthesis of 15a: H-NMR
(CDCl3, 500 MHz) δ 7.91 (s, 1H, H-2 or H-8), 7.83 (s, 1H, H-2 or H-8),
ꢁꢁ
ꢁ
6.02 (d, 1H, H-1ꢁ, J 1 ,2 = 2.6 Hz), 5.54 (m, 1H, H-1 ), 5.11 (dd, 1H, H-2 ,
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
J 2 ,1 = 2.6 Hz, J 2 ,3 = 6.1 Hz), 4.89 (dd, 1H, H-3 , J 3 ,2 = 6.1 Hz, J 3 ,4 = 2.5
Hz), 4.74 (d, 1H, MOM-CH2, J = 6.8 Hz), 4.65 (d, 1H, MOM-CH2, J = 6.8
Hz), 4.39 (ddd, 1H, H-4ꢁ, J 4 ,3 = 2.5 Hz, J 4 ,5 a = 3.7 Hz, J 4 ,5 b = 6.3 Hz),
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
4.35 (m, 1H, H-2ꢁꢁ), 3.90 (dd, 1H, H-5ꢁꢁa, J 5 a,4 = 3.0 Hz, J 5 a,5 b = 11.3
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
Hz), 3.84 (dd, 1H, H-5ꢁa, J 5 a,4 = 3.8 Hz, J 5 a,5 b = 11.2 Hz), 3.79 (m, 2H,
H-3ꢁꢁ, H-5ꢁb), 3.76 (m, 1H, H-5ꢁꢁb), 3.50 (s, 3H, MOM-CH3 or OMe), 3.25
(s, 3H, MOM-CH3 or OMe), 2.42 (m, 2H, H-4ꢁꢁ, H-6ꢁꢁa), 2.08 (m, 1H,
H-6ꢁꢁb), 1.61, 1.39 (each s, each 3H, isopropylidene), 0.86 (s, 9H, tert-Bu),
0.048, 0.037 (each s, each 3H, TBS-Me × 2); 13C-NMR (CDCl3, 125 MHz)
δ 154.48, 146.35, 140.70, 136.89, 123.26, 114.17, 95.33, 91.27, 87.04, 86.99,
85.28, 83.40, 81.34, 63.45, 61.54, 59.37, 57.70, 55.51, 42.02, 31.65, 27.22,
25.86, 25.37, 18.32, −5.43, −5.54; FAB-MS m/z 610 (MH+); Anal. Calcd
for C28H47N5O8Si: C, 55.15; H, 7.77; N, 11.48. Found; C, 54.95; H, 7.59; N,
11.26.
ꢁ
ꢁ
ꢁ
ꢁ
N -1-[(1R ,2S ,3S,4R )-2-(Methoxymethyloxy)-3-methoxy-4-(dimethoxytrityloxym-
ethyl) cyclopentyl]-2 ꢁ,3 ꢁ-O-isopropylideneadenosine ( 16b). Compound 16b
(0.697 g, quant.) was obtained from 15b (0.527 g, 0.864 mmol) as described
for the synthesis of 16a: 1H-NMR (CDCl3, 500 MHz) δ 7.92 (brs, 1H, H-2 or
H-8), 7.62 (brs, 1H, H-2 or H-8), 6.82-7.45 (m, 13H, DMTr), 5.78 (m, 2H,
H-1ꢁ, H-1ꢁꢁ), 5.02 (m, 2H, H-2ꢁ, H-3ꢁ), 4.82 (d, 1H, MOM-CH2, J = 6.7 Hz),
4.68 (d, 1H, MOM-CH2, J = 6.7 Hz), 4.49 (m, 1H, H-4ꢁ), 4.11 (m, 1H, H-2ꢁꢁ),
3.91 (m, 1H, H-3ꢁꢁ), 3.80 (s, 7H, H-5ꢁa, DMTr-OMe × 2), 3.75 (m, 1H, H-5ꢁb),
3.37 (s, 3H, MOM-CH3 or OMe), 3.35 (s, 3H, MOM-CH3 or OMe), 3.25 (m,
1H, H-5ꢁꢁa), 3.19 (m, 1H, H-5ꢁꢁb), 2.56 (m, 1H, H-4ꢁꢁ), 2.48 (m, 1H, H-6ꢁꢁa),
1.62, 1.36 (each s, each 3H, isopropylidene), 1.46 (m, 1H, H-6ꢁꢁb); 13C-NMR
(CDCl3, 125 MHz) δ 180.59, 165.74, 163.74, 152.56, 152.49, 144.47, 142.10,
141.56, 140.21, 136.11, 124.30, 122.87, 122.84, 99.99, 98.40, 88.24, 87.29,
86.38, 85.52, 82.67, 59.34, 56.69, 54.38, 51.96, 49.88, 49.24, 33.87, 23.65,

Synthesis of Cyclic ADP-Carbocylcic-Xylose
597
18.31, 14.03, 11.04, 9.88, 4.19, −3.60, −21.04; HRMS (FAB, positive) calcd
for C43H52N5O10 798.3714 (MH+), found 798.3721.
N -1-[(1R ,2S ,3S ,4R )-2-(Methoxymethyloxy)-3-methoxy-4-(dimethoxytrityloxym-
ethyl) cyclopentyl]-5 ꢁ-O-[bis(phenylthio)phospholyl]-2 ꢁ,3 ꢁ- O -isopropylideneadenosine
( 17b). Compound 17b (0.539 g, 62%) was obtained from 16b (0.659
1
g, 0.826 mmol) as described for the synthesis of 17a: H-NMR (CDCl3,
500 MHz) δ 7.91 (s, 1H, H-2 or H-8), 7.64 (s, 1H, H-2 or H-8), 6.81–7.51
(m, 23H, DMTr, SPh x 2), 5.96 (d, 1H, H-1ꢁ, J 1 ,2 = 2.6 Hz), 5.76 (m, 1H,
ꢁ
ꢁ
H-1ꢁꢁ), 5.08 (dd, 1H, H-2ꢁ, J 2 ,1 = 2.6 Hz, J 2 ,3 = 6.3 Hz), 4.86 (dd, 1H,
ꢁ
ꢁ
ꢁ
ꢁ
H-3ꢁ, J 3 ,2 = 6.3 Hz, J 3 ,4 = 2.7 Hz), 4.79 (d, 1H, MOM-CH2, J = 6.8 Hz),
4.63 (d, 1H, MOM-CH2, J = 6.8 Hz), 4.41 (m, 3H, H-4ꢁ, H-5ꢁ × 2), 4.11 (m,
1H, H-2ꢁꢁ), 3.79 (s, 6H, DMTr-OMe × 2), 3.78 (m, 1H, H-3ꢁꢁ), 3.33 (s, 3H,
MOM-CH3 or OMe), 3.30 (s, 3H, MOM-CH3 or OMe), 3.25 (m, 1H, H-5ꢁꢁa),
3.18 (m, 1H, H-5ꢁꢁb), 2.54 (m, 1H, H-4ꢁꢁ), 2.47 (m, 1H, H-6ꢁꢁa), 1.59, 1.35
(each s, each 3H, isopropylidene), 1.48 (m, 1H, H-6ꢁꢁb); 13C-NMR (CDCl3,
125 MHz) δ 158.38, 154.52, 146.79, 145.20, 140.52, 136.97, 136.42, 136.38,
135.34, 135.29, 135.22, 135.18, 130.04, 130.02, 129.65, 129.63, 129.43,
128.15, 127.69, 126.63, 125.94, 125.89, 125.82, 125.77, 123.51, 114.69,
113.00, 112.98, 95.09, 90.65, 85.79, 85.23, 84.71, 84.64, 84.45, 81.04, 66.45,
66.38, 61.23, 58.69, 57.39, 55.56, 55.19, 43.12, 35.07, 27.13, 25.30, 11.44;
31P-NMR (CDCl3, 202 MHz) δ 48.14 (s); FAB-MS m/z 1062 (MH+); UV
(MeOH) λmax = 295 (sh) nm; Anal. Calcd for C55H60N5O11PS2: C, 62.19; H,
5.69; N, 6.59. Found; C, 62.34; H, 5.83; N, 6.34.
ꢁ
ꢁ
ꢁ
ꢁ
N -1-[(1R ,2S ,3S ,4R )-2-(Methoxymethyloxy)-3-methoxy-4-(hydroxymethyl)cyclop-
entyl]-5 ꢁ-O -{bis (phenylthio)phospholyl}-2 ꢁ,3 ꢁ-O -isopropylideneadenosine ( 18b).
Compound 18b (0.312 g, 88%) was obtained from 17b (0.496 g, 0.467
1
mmol) as described for the synthesis of 18a: H-NMR (CDCl3, 500 MHz)
δ 7.92 (s, 1H, H-2 or H-8), 7.69 (s, 1H, H-2 or H-8), 7.31–7.5ꢁꢁ 2 (m, 10H,
SPh × 2), 5.99 (d, 1H, H-1ꢁ, J 1 ,2 = 2.3 Hz), 5.48 (m, 1H, H-1 ), 5.12 (dd,
ꢁ
ꢁ
1H, H-2ꢁ, J 2 ,1 = 2.3 Hz, J 2 ,3 = 6.2 Hz), 4.91 (dd, 1H, H-3 , J 3 ,2 = 6.2 Hz,
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
J 3 ,4 = 2.4 Hz), 4.71 (d, 1H, MOM-CH2, J = 6.7 Hz), 4.62 (d, 1H, MOM-CH2,
J = 6.5 Hz), 4.43 (m, 2H, H-4ꢁ, H-5ꢁa), 4.39 (m, 2H, H-2ꢁꢁ, H-5ꢁb), 3.90 (dd,
ꢁꢁ
1H, H-5ꢁꢁa, J 5 a,5 b = 11.4 Hz, J 5 a,4 = 2.8 Hz), 3.79 (m, 1H, H-3 ), 3.76 (dd,
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ
1H, H-5ꢁꢁb, J 5 b,5 a = 11.4 Hz, J 5 b,4 = 4.7 Hz), 3.46 (s, 3H, MOM-CH3 or
OMe), 3.21 (s, 3H, MOM-CH3 or OMe), 2.41 (m, 2H, H-4ꢁꢁ, H-6ꢁꢁa), 2.06 (m,
1H, H-6ꢁꢁb), 1.60, 1.36 (each s, each 3H, isopropylidene); 13C-NMR (CDCl3,
125 MHz) δ 154.37, 146.65, 140.61, 137.38, 135.34, 135.30, 135.21, 135.17,
129.70, 129.68, 129.66, 129.63, 129.44, 125.92, 125.86, 125.81, 125.76,
123.57, 114.70, 95.54, 90.76, 86.98, 84.79, 84.73, 84.47, 83.46, 81.14, 66.44,
66.38, 61.55, 60.36, 59.69, 57.74, 55.55, 41.90, 31.44, 27.13, 25.30, 21.01,
14.18; 31P-NMR (CDCl3, 202 MHz) δ 51.05 (s); FAB-MS m/z 760 (MH+);
UV (MeOH) λmax = 259, 295 (sh) nm; Anal. Calcd for C34H42N5O9PS2: C,
53.75; H, 5.57; N, 9.22. Found; C, 53.59; H, 5.60; N, 8.97.
ꢁꢁ
ꢁꢁ
ꢁꢁ
ꢁꢁ

598
T. Kudoh et al.
N -1-[(1R ,2S ,3S ,4R )-2-(Methoxymethyloxy)-3-methoxy-4-(phosphonoxymethyl)
cyclopentyl]-5 ꢁ-O -[(phenylthio)phospholyl]-2 ꢁ, 3 ꢁ-O -isopropylideneadenosine ( 19b).
Compound 19b (90 mg, 58%) was obtained from 18b (140 mg, 0.184
mmol) as described for the synthesis of 19a: 1H-NMR (D2O, 500 MHz) δ 8.65
(s, 1H, H-2 or H-8), 8.39 (s, 1H, H-2 or H-8), 7.12–7.30 (m, 5H, SPh), 6.30
ꢁ
(d, 1H, H-1ꢁ, J 1 ,2 = 2.2 Hz), 5.32 (dd, 1H, H-2 , J 2 ,1 = 2.2 Hz, J 2 ,3 = 5.9
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Hz), 5.01 (m, 1H, H-1ꢁꢁ), 4.94 (dd, 1H, H-3ꢁ, J 3 ,2 = 5.9 Hz, J 3 ,4 = 1.6 Hz),
4.74 (d, 1H, MOM-CH2, J = 7.1 Hz), 4.69 (m, 2H, H-4ꢁ, MOM-CH2), 4.50
(m, 1H, H-2ꢁꢁ), 4.15 (m, 1H, H-5ꢁa), 4.11 (m, 1H, H-5ꢁb), 4.03 (m, 2H,
H-5ꢁꢁ × 2), 3.99 (m, 1H, H-3ꢁꢁ), 3.54 (s, 3H, MOM-CH3 or OMe), 3.26 (s, 3H,
MOM-CH3 or OMe), 3.18 (q, 6H, Et3NH-CH2 × 3, J = 7.3 Hz), 2.71 (m, 1H,
H-4ꢁꢁ), 2.60 (m, 1H, H-6ꢁꢁa), 2.06 (m, 1H, H-6ꢁꢁb), 1.62, 1.38 (each s, each
3H, isopropylidene), 1.26 (t, 9H, Et3NH-CH3 × 3, J = 7.3 Hz); 13C-NMR
(D2O, 125 MHz) δ 150.82, 146.42, 146.30, 143.88, 133.18, 133.14, 129.92,
129.87, 129.42, 128.25, 119.82, 119.61, 115.15, 97.02, 91.86, 86.61, 86.53,
84.62, 84.33, 81.89, 66.24, 63.67, 58.06, 56.12, 47.10, 40.69, 40.63, 26.39,
24.73, 8.68; 31P-NMR (D2O, 202 MHz) δ 16.70 (s), 0.55 (s); HRMS (FAB,
positive) calcd for C28H40N5O13P2S 748.1813 (MH+), found 748.1804; UV
(H2O) λmax 260 nm.
ꢁ
ꢁ
ꢁ
ꢁ
2 ꢁꢁ-O -Methoxymethyl-3 ꢁꢁ-O-methyl-cyclic ADP-carbocyclic-xylose 2 ꢁ,3 ꢁ-Acetonide
( 20b). Compound 20b (37 mg, 47%) was obtained from 19b (90 mg, 0.11
mmol) as described for the synthesis of 20a: 1H-NMR (D2O, 500 MHz) δ 9.10
(s, 1H, H-2 or H-8), 8.45 (s, 1H, H-2 orꢁH-8), 6.41 (s, 1H, H-1ꢁ), 5.55 (d, 1H,
H-2ꢁ, J 2 ,3 = 6.1 Hz), 5.44 (dd, 1H, H-3 , J 3 ,2 = 6.1 Hz, J 3 ,4 = 2.4 Hz), 5.02
(m, 1H, H-1ꢁꢁ), 4.75 (d, 1H, MOM-CH2, J =ꢁ6.8 Hz), 4.67 (d, 1H, MOM-CH2,
J = 6.8 Hz), 4.64 (m, 1H, H-4ꢁ), 4.57 (m, 1H, H-2ꢁꢁ), 4.15 (m, 2H, H-5ꢁa, H-
5ꢁꢁa), 4.09 (m, 3H, H-5ꢁb, H-5ꢁꢁb, H-3ꢁꢁ), 3.48 (s, 3H, MOM-CH3 or OMe), 3.16
(s, 3H, MOM-CH3 or OMe), 3.19 (q, 6H, Et3NH-CH2 × 3, J = 7.3 Hz), 2.94
(m, 1H, H-6ꢁꢁa), 2.88 (m, 1H, H-4ꢁꢁ), 2.37 (m, 1H, H-6ꢁꢁb), 1.64, 1.44 (each
s, each 3H, isopropylidene), 1.27 (t, 9H, Et3NH-CH3 × 3, J = 7.3 Hz); 13C-
NMR (D2O, 125 MHz) δ 152.18, 145.29 144.63, 119.65, 115.10, 97.24, 92.46,
87.79, 86.93, 86.83, 85.04, 84.82, 81.87, 64.60, 64.25, 58.47, 56.24, 47.10,
36.34, 36.26, 28.41, 26.42, 24.73, 8.67; 31P-NMR (D2O, 202 MHz) δ −9.56
(d, J = 11.4 Hz), −10.46 (d, J = 11.4 Hz); HRMS (FAB, positive) calcd for
C22H34N5O13P2 638.1623 (MH+), found 638.1627; UV (H2O) λmax 259 nm.
3 ꢁꢁ-O -Methyl-cyclic ADP-carbocyclic-xylose ( 5). Compound 5 (30 mg,
quant.) was obtained from 20b (37 mg, 0.050 mmol) as described for the
synthesis of 4: 1H-NMR (D2O, 500 MHz, K+ salt) δ 9.04 (s, 1H, H-2 or H-8),
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
8.41 (s, 1H, H-2 or H-8), 6.10 (d, 1H, H-1ꢁ, J 1 ,2 = 6.1 Hz), 5.21 (m, 1H,
H-2ꢁ), 4.95 (m, 1H, H-1ꢁꢁ), 4.62 (m, 1H, H-5ꢁa), 4.60 (m, 1H, H-3ꢁ), 4.56 (m,
1H, H-2ꢁꢁ), 4.44 (m, 1H, H-4ꢁ), 4.11 (m, 3H, H-5ꢁb, H-5ꢁꢁ × 2), 4.03 (m, 1H,
H-3ꢁꢁ), 3.52 (s, 3H, OMe), 2.94 (m, 1H, H-6ꢁꢁa), 2.90 (m, 1H, H-4ꢁꢁ), 2.52
(m 1H, H-6ꢁꢁ b); 13C-NMR (D2O, 125 MHz) δ 152.19, 146.69, 145.43, 144.52,
120.42, 91.00, 85.77, 85.34, 81.35, 73.82, 71.18, 65.14, 63.94, 63.46, 58.59,
ꢁ
ꢁ

Synthesis of Cyclic ADP-Carbocylcic-Xylose
599
36.81, 27.72; 31P-NMR (D2O, 202 MHz) δ −9.70 (d, J = 11.4 Hz), −10.32
(d, J = 11.4 Hz); HRMS (FAB, positive) calcd for C17H26N5O12P2 554.1048
(MH+), found 554.1035; UV (H2O) λmax 259 nm.
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