Synthesis and biological activity of cyclic ADP-carbocyclic-ribose analogs: Structure-activity relationship and conformational analysis of N-1-carbocyclic-ribose moiety

Article abstract of DOI:10.1081/NCN-200060167

Several cyclic ADP-carbocyclic-ribose analogs 3-10 modified in the N-1-carbocyclic-ribose moiety were synthesized. Their Ca²⁺-releasing activity was estimated in sea urchin eggs to show that the 3″-deoxy analog 6 shows 5 times more potent activ

Full text of DOI:10.1081/NCN-200060167

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SYNTHESIS AND BIOLOGICAL ACTIVITY OF CYCLIC ADP-
CARBOCYCLIC-RIBOSE ANALOGS: STRUCTURE-ACTIVITY
RELATIONSHIP AND CONFORMATIONAL ANALYSIS OF
N-1-CARBOCYCLIC-RIBOSE MOIETY
Takashi Kudoh a , Masayoshi Fukuoka a , Satoshi Shuto a & Akira Matsuda a
a Graduate School of Pharmaceutical Sciences, Hokkaido University , Sapporo, Japan
Published online: 15 Nov 2011.
To cite this article: Takashi Kudoh , Masayoshi Fukuoka , Satoshi Shuto & Akira Matsuda (2005) SYNTHESIS AND BIOLOGICAL
ACTIVITY OF CYCLIC ADP-CARBOCYCLIC-RIBOSE ANALOGS: STRUCTURE-ACTIVITY RELATIONSHIP AND CONFORMATIONAL
ANALYSIS OF N-1-CARBOCYCLIC-RIBOSE MOIETY, Nucleosides, Nucleotides and Nucleic Acids, 24:5-7, 655-658, DOI: 10.1081/
NCN-200060167
To link to this article: http://dx.doi.org/10.1081/NCN-200060167
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Nucleosides, Nucleotides, and Nucleic Acids, 24 (5--7):655–658, (2005)
Copyright D Taylor & Francis, Inc.
ISSN: 1525-7770 print/ 1532-2335 online
DOI: 10.1081/NCN-200060167
SYNTHESIS AND BIOLOGICAL ACTIVITY OF CYCLIC
ADP-CARBOCYCLIC-RIBOSE ANALOGS: STRUCTURE-ACTIVITY
RELATIONSHIP AND CONFORMATIONAL ANALYSIS OF
N-1-CARBOCYCLIC-RIBOSE MOIETY
Takashi Kudoh, Masayoshi Fukuoka, Satoshi Shuto, and Akira Matsuda
5
Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan
5
Several cyclic ADP-carbocyclic-ribose analogs 3--10 modified in the N-1-carbocyclic-ribose moiety
were synthesized. Their Ca2+-releasing activity was estimated in sea urchin eggs to show that the 3@-
deoxy analog 6 shows 5 times more potent activity than cADPcR, but the 2@,3@-didieoxy-2@,3@-
unsunsaturated analog 3 has very weak activity. We also calculated their stable conformation and
found that 3 and 6 were significantly different in their stable conformation.
Keywords CyclicADP-Carbocyclic-Ribose, Ca2+-ReleasingActivity, ConformationalAnalysis
INTRODUCTION
Cyclic ADP-carbocyclic-ribose (cADPcR, 1) was designed as a stable mimic of
cyclic ADP-ribose (cADPR, 2), an intracellular Ca2+-mobilizing second messen-
ger.[1] cADPcR has a carbocyclic-ribose bound to N-1 position of adenine moiety
instead of D-ribose in cADPR, the synthesis of which had previously been
completed using effective formation of the pyrophosphate linkage.[2] cADPcR is
actually very stable under chemical and biological conditions and exhibits 3 times
more potent Ca2+-releasing activity than cADPR in sea urchin egg homogenates.
In this study, we focused on SAR of the N-1-carbocyclic-ribose moiety of
cADPcR. Only a few analogs modified in this moiety have been synthesized,
because of the difficulty in their synthesis by the classical enzymatic method.[3--5]
So we designed and synthesized the eight cADPcR analogs 3--10 modified in the
N-1-carbocyclic-ribose moiety (Figure 1). We estimated their Ca2+-releasing
activity and analyzed their stable conformation.
We are grateful to Dr. T. Murayama, Faculty of Medicine, Juntendo University, for biological evaluations.
Address correspondence to Satoshi Shuto, Graduate School of Pharmaceutical Sciences, Hokkaido
University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
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T. Kudoh et al.
FIGURE 1
SYNTHESIS
Scheme 1 shows the synthetic route of the cADPcR analog 3. The optically
active cyclopentenylamine units 11, corresponding to the carbocyclic-ribose
moiety, were condensed with imidazole nucleoside 12 to construct the N-1-cyclo-
pentenylenyladenosine derivative 13. The 5@-hydroxy group of 13 was protected
by MMTr group, and 5’-O-TBS protection was removed to give 14. Bisphenylth-
iophosphate group was introduced to the 5’-position of 14 with S,S’-diphenylphos-
phorodithioate (PSS) and TPSCl, and then the 5@-O-MMTr group was removed to
give 15. The 5@-hydroxy group of 15 was phosphorylated by Yoshikawa’s method,
and one phenylthio group was removed by hypophosphorous acid to give the
cyclization substrate 16. The intramolecular condensation of two phosphates group
was successfully proceeded by a silver nitrate/MS3A/pyridine systems to give the
cyclic product 17. Finally, remaining isopropylidene protection of hydroxy groups
was removed by aqueous formic acid to complete the synthesis of 3. Similarly, the
other analogs 4--10 were synthesized using the corresponding optically active
cyclopentylamine derivatives.
BIOLOGICAL ACTIVITY
We estimated Ca2+-releasing activity of these analogs in sea urchin egg
homogenates. The EC50 values of analogs were summarized in Figure 2. In this

T. Kudoh et al.
657
SCHEME 1 (a) K2CO3, MeOH, 82%; (b) MMTrCl, pyridine, 84%; (c) TBAF, AcOH, THF, 96%; (d) PSS, TPSCl,
pyridine, 54%; (e) 80% AcOH aq., 77%; (f) POCl3, (EtO)3 PO, 0°C; (g) H3PO2, Et3N, N-methylmaleimide, pyridine,
0°C!r.t., 2 steps 64%; (h) AgNO3, Et3N, MS3A, pyridine, 84%; (i) 60% HCOOH aq., quant.
FIGURE 2 Ca2+ releasing activity in sea urchin egg homogenates. Maximum Ca2+ releasing by cADPR is 100%.

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T. Kudoh et al.
FIGURE 3 Conformational analysis by molecular dynamics: simulated annealing with NOESY constraints.
system, the EC50 value of cADPR was 0.22 mM, and cADPcR was 0.079 mM. As a
result, 3@-deoxy-cADPcR (6) showed 5 times more potent activity than cADPcR.
Xylose type analog 9 had potent activity equal as cADPcR, and 2@-deoxy (5), 2@,3@-
dideoxy (4), 3@-O-methyl (8,10) analogs showed moderate activity. However, the
unsaturated analog 3 showed very weak activity.
CONFORMATIONAL ANALYSIS
We calculated the stable conformation of cADPR, cADPcR and the cADPcR
analogs 6 and 3 by molecular dynamics with a simulated annealing method, based
on the NOE constraints of the intramolecular proton pairs measured in D2O.
Analogs with strong Ca2+-mobilzing activity, such as cADPR, cADPcR, and 6 had
similar syn conformations around N-9-glycosidic linkage, while the almost inactive
analog 3 showed highanti conformation (Figure 3).
REFERENCES
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2. Shuto, S.; Fukuoka, M.; Manikowsky, A.; Ueno, Y.; Nakano, T.; Kuroda, R.; Kuroda, H.; Matsuda, A. Total
synthesis of cyclic ADP-carbocyclic-ribose, a stable mimic of Ca2+-mobilizing second messenger cyclic ADP-
ribose. J. Am. Chem. Soc. 2001, 123, 8750–8759.
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