7-halogenated 7-deazapurine 2'-c-methylribonucleosides

Article abstract of DOI:10.1135/cccc2011124

7-Halogenated 7-deazapurine 2'-C-methylribonucleosides related to adenosine, inosine and guanosine were synthesized employing a "one-pot" glycosylation protocol (Vorbrueggen glycosylation). The immunosine 2'-C-methylribonucleoside 4 was prepared by the sa

Full text of DOI:10.1135/cccc2011124

7-Deazapurine 2′-C-Methylribonucleosides
1413
7-HALOGENATED 7-DEAZAPURINE 2′-C-METHYLRIBONUCLEOSIDES
a4,##
Frank SEELAa1,b,*, Simone BUDOWa2, Kuiying XUa3,#, Xioahua PENG
and
c
Henning EICKMEIER
a
Laboratory of Bioorganic Chemistry and Chemical Biology, Center for Nanotechnology,
Heisenbergstrasse 11, 48149 Münster, Germany; e-mail: frank.seela@uni-osnabrueck.de,
1
2
3
4
sbudow@uni-osnabrueck.de, kuiyingxu@yahoo.de, pengx@uwm.edu
b
c
Laboratorium für Organische und Bioorganische Chemie, Institut für Chemie,
Universität Osnabrück, Barbarastrasse 7, 49069 Osnabrück, Germany
Anorganische Chemie II, Institut für Chemie, Universität Osnabrück,
Barbarastrasse 7, 49069 Osnabrück, Germany; e-mail: heickmei@uni-osnabrueck.de
Received June 20, 2011
Accepted July 27, 2011
Published online November 30, 2011
This article is dedicated to Professor Antonín Holý, a good friend and an excellent scientist, on the
occasion of his 75th birthday.
7-Halogenated 7-deazapurine 2′-C-methylribonucleosides related to adenosine, inosine and
guanosine were synthesized employing a “one-pot” glycosylation protocol (Vorbrüggen
glycosylation). The immunosine 2′-C-methylribonucleoside 4 was prepared by the same
glycosylation strategy. By this route, the synthesis of the anti-HCV active compound
7-fluoro-7-deaza-2′-C-methyladenosine (1b) was notably simplified, while other nucleosides
were synthesized for the first time. A single-crystal X-ray analysis of 7-fluoro-7-deaza-
2′-C-methylinosine (2b) was performed showing that the glycosylic bond adopts an anti
orientation (χ = –139.9(2)°), while the sugar moiety has an N conformation (C3′-endo; P =
4.3(1)°, τm = 42.4(1)°).
Keywords: Nucleosides; 2′-C-Methylribonucleosides; 7-Deazapurines; Pyrrolo[2,3-d]pyrimi-
dine; Glycosylation; Single-crystal X-ray analysis.
Hepatitis C virus (HCV) infections still represent an unsolved health prob-
lem inducing chronic hepatitis, liver cirrhosis, and hepatocellular carci-
noma1. Significant effort has been devoted to the development of active
compounds of therapeutic value for efficient HCV infection treatment1–3
.
Virally encoded polymerases, which are essential for HCV replication, were
established as drug targets. Among the nucleoside inhibitors, 2′-C-
#
Current affiliation: Department of Chemistry, Roy and Diana Vagelos Laboratories, Univer-
sity of Pennsylvania, PA, USA
##
Current affiliation: Department of Chemistry and Biochemistry, University of Wiscon-
sin-Milwaukee, 3210 North Cramer Street, Milwaukee, Wisconsin 53211, USA
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431
© 2011 Institute of Organic Chemistry and Biochemistry
doi:10.1135/cccc2011124

1414
Seela, Budow, Xu, Peng, Eickmeier:
methylribonucleosides showed excellent chain-terminating properties, thus
affecting RNA replication. In this context, 2′-C-methyladenosine and 2′-C-
methylguanosine were identified as potent inhibitors of HCV RNA replica-
tion in vitro4. However, both compounds also suffer from significant draw-
backs. 2′-C-Methyladenosine was found to serve as a substrate for adenosine
deaminase and purine nucleoside phosphorylase, indicating that its re-
stricted oral bioavailability results from metabolic conversion4–6. For 2′-C-
methylguanosine, cellular uptake and intracellular metabolism to the corre-
sponding triphosphate were limited5,6. Nucleosides employing modified
nucleobases were expected to circumvent these drawbacks. Recently, the
triphosphate of 7-deaza-2′-C-methyladenosine (1a) was identified as a po-
tent inhibitor of HCV replication7 and its 7-fluorinated derivative 1b was
even found to be the most potent compound6 (purine numbering is used
throughout the manuscript).
Due to our continuous efforts on the synthesis of 7-deazapurine nucleo-
sides as structural analogues of natural purine nucleosides, we became in-
terested in synthesizing novel 7-deazapurine 2′-C-methylribonucleosides
and to study their conformational properties. Moreover, we devoted con-
siderable effort towards the simplification of synthetic protocols of already
reported compounds with notable anti-HCV activity making them more
easily accessible.
In this work, we focus on the synthesis and properties of 7-halogenated
7-deazapurine 2′-C-methylribonucleosides related to adenosine, inosine
and guanosine (Fig. 1). The immunosine 2′-C-methylribonucleoside 4 was
prepared as well. A single-crystal X-ray analysis of compound 2b was per-
formed and conformational properties were studied in solution and in solid
state.
O
4
NH2
O
O
7
R
5
R
7
I
1
6
4a
5
S
6
3 HN
1
HN
HN
N
6
O
8
2
2
2
9
N 7
N
N3
N
4
5
N
7a
H2N
HO
N
H2N
HO
N
N
3
4
1
O
O
O
O
HO
HO
Me
OH
Me
OH
Me
OH
Me
OH
HO
HO
HO
HO
1a-1c
2a-2c
3
4
systematic numbering
a: R = H
b: R = F
c: R = Cl
a: R = H
b: R = F
c: R = Cl
purine numbering
systematic numbering
FIG. 1
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

7-Deazapurine 2′-C-Methylribonucleosides
1415
RESULT AND DISCUSSION
Synthesis of 7-Deazapurine 2′-C-Methylribonucleosides
Several strategies were developed for the synthesis of 7-deazapurine ribo-
nucleosides8. Among them, the Silyl–Hilbert–Johnson reaction using a Lewis
acid as catalyst (Vorbrüggen glycosylation9) performed in one step was
established as a versatile protocol for the synthesis of a broad range of
7-deazapurine ribonucleosides8. Wolfe and co-workers used the “one-pot”
protocol for the synthesis of pyrimidine and purine 2′-C-methylribo-
nucleosides10. In their work, the heterocyclic moiety was silylated with BSA
[N,O-bis(trimethylsilyl)acetamide] in MeCN and glycosylated with 2-C-
methyl-1,2,3,5-tetra-O-benzoyl-D-ribofuranose (6) in the presence of SnCl4
or trimethylsilyl trifluoromethanesulfonate (TMSOTf)10. Later, Ding et al.
used a similar procedure for the synthesis of 2′-C-methyltoyocamycin and
7-functionalized 7-deaza-2′-C-methylinosine (MeCN, TMSOTf)11. However,
all efforts to employ 6-chloro-7-deazapurine (5a) as nucleobase and the
2-C-methyl sugar donor 6 10 in the “one-pot” glycosylation reaction
failed12. A corresponding observation was made in the series of ribo-
nucleosides by Seela and co-workers as well as by others8,13. On the con-
trary, in the laboratories of Merck, the nucleobase anion glycosylation
(6-chloro-7-deazapurine (5a) as nucleobase and 3,5-bis-O-(2,4-dichloro-
phenylmethyl)-2-C-methyl-1-O-methyl-α-D-ribofuranose as sugar compo-
nent) was used to synthesize the 7-nonhalogenated 6-chloro-7-deazapurine
2′-C-methylribonucleoside as key intermediate6. Later it was found that the
above mentioned nucleobase anion glycosylation reaction proceeds in fact
via an α-epoxide intermediate as sugar donor12.
As we envisaged, the synthesis of 7-halogenated 7-deazapurine 2′-C-
methylribonucleosides, we intended to employ the 7-fluorinated nucleo-
base 5b 14 or the corresponding 7-chloro derivative 5c 15 together with the
commercially available sugar precursor 1,2,3,5-tetra-O-benzoyl-2-C-methyl-
D-ribofuranose10 (6) in the glycosylation reaction. From our experiences in
the synthesis of 7-halogenated 7-deazapurine ribonucleosides, we were con-
fident that the “one-pot” glycosylation protocol (Vorbrüggen glycosyl-
ation) would proceed smoothly with the halogenated nucleobases 5b and
5c. It should be noted that the 7-fluorinated 7-deazapurine derivative 1b
was also prepared under Mitsunobu conditions (triphenylphosphine, di-
ethyl azodicarboxylate in THF) employing 5b as nucleobase and 2,3,5-tri-
O-benzoyl-2-C-methyl-D-ribofuranose as sugar precursor6. However, under
the reported conditions, compound 1b was only obtained in low overall
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

1416
Seela, Budow, Xu, Peng, Eickmeier:
yield of 5% 6. The same authors reported the synthesis of the 7-chloro ana-
logue 1c from 1a employing N-chlorosuccinimide to introduce the chloro
substituent6.
Our synthesis route towards 7-halogenated 7-deazapurine 2′-C-methyl-
ribonucleosides (1b, 1c, 2b, 2c, 8b, 8c) using the “one-pot” glycosylation
protocol and Vorbrüggen conditions is summarized in Scheme 1. Com-
pound 5b was first silylated with BSA (1.2 equiv.) in anhydrous acetonitrile
at room temperature. Then, the sugar 6 was added in portions together
with TMSOTf as catalyst, and the reaction was allowed to proceed over-
night at elevated temperature. It was found that under the acidic condi-
tions due to TMSOTf and elevated temperature (60 °C), the sugar precursor
6 was easily decomposed when long reaction times were applied. Con-
sequently, it was necessary to use an excess of 6 (totally 2.5 equiv.)
which had to be added in portions to the reaction mixture. By this method,
the protected glycosylation product 7b was obtained in 60% yield;
the formation of other isomers was not observed. Deprotection of 7b in
0.5 M NaOMe/MeOH afforded the deblocked nucleoside 8b with simulta-
neous displacement of the 6-chloro substituent by a 6-methoxy group.
O
OCH3
R
R
HN
N
2 M NaOH,
reflux, 3 h
N
N
N
N
2b: 80%
2c: 75%
O
HO
O
HO
Me
OH
Me
OH
8b, 8c
HO
HO
2b, 2c
0.5 M
NaOMe/MeOH,
reflux, 2 h
8b: 80%
8c: 79%
Cl
N
R
NH2
R
N
Cl
N
R
N
N
N
BSA,
TMSOTf,
60°C, 24 h
H
NH3/MeOH,
120 °C, 20 h
N
N
5a-5c
N
O
HO
+
1b: 70%
1c: 72%
7b: 60%
7c: 70%
O
BzO
Me
OH
1b, 1c
OBz
Me
OBz
O
BzO
Me
HO
BzO
OBz
BzO
7b, 7c
6
a: R = H; b: R = F; c: R = Cl
SCHEME 1
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

7-Deazapurine 2′-C-Methylribonucleosides
1417
Treatment of 8b with 2 M aq. NaOH yielded 7-fluoro-7-deaza-2′-C-
methylinosine (2b) in 80% yield. Treatment of compound 7b with
methanolic ammonia (autoclave) led to the deprotection of the sugar
moiety and concomitant conversion of the chloro substituent to an amino
group yielding 7-fluoro-7-deaza-2′-C-methyladenosine (1b) in a good 42%
yield over two steps. Thus, the “one-pot” procedure reported herein repre-
sents a significant improvement over the earlier reported protocol employ-
ing Mitsunobu conditions6 and allows the convenient synthesis of the
HCV-active compound 1b as well as its analogues 2b, 7b and 8b.
By the same glycosylation strategy, the 7-chloro intermediate 7c was ob-
tained in even higher yield (70%). From the protected 7c, the 7-chloro-
6-methoxy compound 8c (79%), the inosine derivative 2c (75%) and the
adenosine derivative 1c (72%) were obtained in good yields. During all the
reactions, the 7-fluoro or 7-chloro substituent was not affected.
In the series of halogenated 7-deaza-2′-C-methylguanosines, the
7-fluorinated derivative has been reported recently16. In our work, the
“one-pot” method was applied to the synthesis of the 7-iodo-7-deaza-
2′-C-methylguanosine 3, the 6-methoxy analogue 12 and its 2,6-diamino
derivative 11 (Scheme 2). For this, the silylated 2-pivaloylamino-6-chloro-
7-iodo-7-deazapurine aglycon 9 17 was employed in the glycosylation reac-
tion together with the protected sugar 6 (TMSOTf, MeCN, 50 °C, 72 h) to
give the fully protected nucleoside 10 in 75% yield. The protected com-
pound 10 was transformed to the 2,6-diamino-7-iodo-7-deazapurine 2′-C-
methylribonucleoside18 (11) by treatment with aq. NH3/dioxane at 120 °C
NH2
Cl
I
I
BSA,
TMSOTf,
50 °C, 72 h
Cl
N
N
N
I
aq. NH3, dioxane
120 °C, 24 h
N
N
N
H2N
HO
N
O
PivHN
BzO
N
+
6
75%
85%
N
O
PivHN
H
Me
OH
Me
OBz
9
HO
BzO
10
11
OCH3
O
I
I
N
HN
0.5 M NaOMe/MeOH,
reflux, 3 h
2 M NaOH, dioxane,
reflux, 3 h
N
N
H2N
HO
N
H2N
HO
N
O
10
88%
O
85%
Me
OH
Me
OH
HO
HO
12
3
SCHEME 2
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

1418
Seela, Budow, Xu, Peng, Eickmeier:
in a sealed vessel (85% yield), while the 6-methoxy compound 12 was ob-
tained in 88% yield by refluxing 10 in 0.5 M NaOMe/MeOH. Nucleoside 12
was further converted (2 M NaOH, dioxane, reflux) to the 7-iodinated 2′-C-
methylguanosine analogue 3 (85% yield).
Among the guanosine analogues, immunosine (7-thia-8-oxoguanosine,
TOG, isatoribine) possesses in vivo activity against a variety of DNA and
RNA viruses19,20. It was found that immunosine exhibits a stimulatory ef-
fect on both cellular and humoral components of the immune response.
The observed antiviral effect has been attributed primarily to the induction
of α-interferon20,21. In this regard, we became interested in synthesizing the
2′-C-methylribonucleoside derivative of immunosine (4). A patent22 de-
scribes the synthesis of 4, using the same glycosylation conditions reported
by Wolfe et al.10 (BSA as silylating agent, MeCN, glycosylation of 13 and 6
in the presence of SnCl4). By this route, the 2′-C-methylribonucleoside 4
was obtained in 14% overall yield over two steps22.
However, we favoured the “one-pot” glycosylation protocol as an im-
proved synthesis strategy as shown in Scheme 3. The nucleobase 13
employed in the glycosylation reaction was prepared in 5 steps from
2,4-diamino-5-hydroxypyrimidine according to Baker and Chatfield23. Due
to the high reactivity of the 5-aminothiazolo[4,5-d]pyrimidine-2,7(3H,6H)-
dione (systematic numbering) moiety (13) compared to the inertness of the
7-deazapurines in Lewis acid catalyzed glycosylation reactions, the glyco-
sylation reaction was carried out at room temperature. For that, the
immunosine base 13 was first silylated with hexamethyldisilazane (HMDS)
in the presence of ammonium sulfate and pyridine under reflux. Then, the
silylated 13 was reacted with 1 equiv. of the 2-C-methyl sugar 6 in the pres-
ence of TMSOTf (1.4 equiv.) at room temperature to give 14. It was found
that excessive sugar caused multiple glycosylations confirmed by NMR
spectra which showed more than one set of sugar signals. During the
nucleobase anion glycosylation of 2′-deoxyimmunosine a similar observa-
tion was made24. An excess of more than 1.3 equivalents of the 2′-deoxy
sugar component led to the introduction of two sugar residues (one at each
O
O
OBz
Me
OBz
O
BzO
HMDS,
reflux,
(NH4)2SO4
S
N
S
N
O
HN
HN
O
O
1 M NaOMe/MeOH,
rt, 16 h
S
BzO
HN
H2N
H2N
BzO
N
H2N
HO
N
6
O
Triflate
54.5%
O
71%
N
H
O
N
Me
OBz
Me
OH
13
BzO
HO
4
14
SCHEME 3
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

7-Deazapurine 2′-C-Methylribonucleosides
1419
lactam moiety). This was attributed to the two immunosine lactam
deprotonation sites (pKa values of 7.2 and 10.0)24. After work-up, the
2′-C-methyl compound 14 was obtained in 54.5% yield. The benzoylated
14 was deprotected with 1 M NaOMe/MeOH to afford the free nucleoside 4
in 71%. By our route, the overall yield of 2′-C-methylimmunosine (4) over
two steps was improved to 37%.
Physical Properties of 7-Deazapurine 2′-C-Methylribonucleosides
All compounds were characterized by 1H and 13C NMR spectra and by
DEPT-135 (distortionless enhancement by polarization transfer) NMR as
well as by ESI-TOF mass spectrometry or elemental analysis (see Experimen-
tal). The 13C NMR chemical shifts are compiled in Table I. The sugar car-
bons were assigned according to ref.10, and the nucleobase carbons were
assigned according to refs13,17,24–26 19F NMR spectra were taken for the fluo-
.
rinated compounds 1b, 2b, 7b and 8b. The position of the fluorine
substituent at C-7 or C-8 for 2′-C-methylribonucleosides 1b, 2b, 7b and 8b
was assigned from J(F,C) coupling constants given in Table I (values in pa-
rentheses; purine numbering is used throughout discussion). J(F,C-7) has a
1
2
value of about 245–250 Hz showing J coupling, while the J coupling con-
stants of J(F,C-8) and J(F,C-5) show values of about 25 and 15 Hz, respec-
3
tively. In some case, J couplings with C4 and C6 were observed (J(F,C-4)
and J(F,C-6) around 3 Hz).
Single-Crystal X-ray Analysis of Compound 2b
Studies conducted towards the structure-activity relationship of purine
ribonucleosides as inhibitors of HCV replication showed that, the ribo
conformation of the 2′-hydroxy group is of particular importance for the
activity of nucleosides4. Moreover, effective inhibition seems to require a
sensitive balance of the bulkiness of the substituent at C2′; between too
little (2′-H) and too much (2′-C-ethyl) steric bulkiness. In this regard, the
size of the 2′-C-methyl substituent appeared to be nearly optimal4. Indeed,
2′-C-methylribonucleosides demonstrated promising activity in cell-based
HCV RNA replication assays. On the other hand, modification of the
nucleobase appeared to be tolerated to some extent by the HCV RNA poly-
merase (HCV NS5B)4,6
.
For a series of nucleosides, the preferred sugar conformation in solution
1
was calculated on the basis of H NMR coupling constants (J1′2′, J2′3′, J3′4′).
However, this calculation method cannot be applied to 2′-C-methylribo-
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

1420
Seela, Budow, Xu, Peng, Eickmeier:
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

7-Deazapurine 2′-C-Methylribonucleosides
1421
nucleosides. The 2′-C-methyl substituent excludes the calculation of the J1′2′
and J2′3′ coupling constants, leaving only J3′4′ as a clue to the sugar pucker-
ing. For compound 2b, the J3′4′ value of 7.8 Hz, obtained from 1H NMR
measurements in D2O, points to a predominant N sugar pucker27.
To obtain additional information on the conformation, we determined
the solid-state structure of compound 2b by single-crystal X-ray crystallog-
raphy. To the best of our knowledge, only three 2′-C-methylribonucleoside
crystal structures have been reported in the literature so far. These are
2′-C-methyltubercidin (1a)6, 2′-C-methylsangivamycin (15)28 and 2′-C-
methyluridine (16)29,30 (Fig. 2).
O
NH2
NH2
O
F
CONH2
HN
N
N
HN
N
N
N
N
N
O
N
O
O
N
O
HO
O
HO
HO
HO
Me
OH
Me
OH
Me
OH
Me
OH
HO
HO
1a
HO
HO
2b
15
16
FIG. 2
2′-C-methylribonucleosides analyzed by single-crystal X-ray analysis
The crystal structure of 7-fluoro-7-deazainosine 2′-C-methylribonucleo-
side (2b) is shown in Fig. 3 and characteristic crystal data are summarized
in Table II. In the crystal structure of 2b, the glycosylic bond adopts a tor-
sion angle of χ = –139.9(2)°. According to the syn/anti definition of the tor-
sion angle χ (O4′–C1′–N9–C4) referring to the orientation of the nucleobase
relative to the sugar moiety31, the glycosylic bond of 2b is in the anti
TABLE II
Selected crystal data obtained from the single-crystal X-ray analysis of compound 2b
Empirical formula
C
12H14FN3O5
Unit cell dimensions a = 4.8617(2) Å
b = 16.0982(6) Å
c = 16.8400(6) Å
Formula weight
Crystal size
299.26
Volume
V = 1317.98(9) Å3
0.20 × 0.18 × 0.10 mm Z value
Z = 4
Crystal system
Space group
orthorhombic
Calculated density
1.508 g cm–3
P212121
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

1422
Seela, Budow, Xu, Peng, Eickmeier:
FIG. 3
ORTEP presentation of the single crystal X-ray structure of 7-fluoro-7-deaza-2′-C-methylino-
sine (2b), showing the crystallographic numbering scheme. Displacement ellipsoids are drawn
at the 50% probability level and H atoms are shown as spheres of arbitrary size
FIG. 4
The crystal packing of 2b, showing the intermolecular hydrogen-bonding network (projection
parallel to the bc plane)
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

7-Deazapurine 2′-C-Methylribonucleosides
1423
conformation. Also, the 2′-C-methylribonucleosides 1a, 15 and 16 show
anti conformation6,28–30. The glycosylic bond length (N9–C1′) of 2b is
1.462(2) Å, which is slightly longer than that of 1a (1.460(2) Å)6 but shorter
than N9–C1′ of 16 (1.479(2) Å)28–30
.
The sugar moiety of compound 2b shows an N conformation (unsym-
3
metrical twist of C3′-endo–C2′-exo, T2), with a pseudorotation phase angle
of P = 4.3(1)° and a maximum puckering amplitude of τm = 42.4(1)°. This is
in line with the N sugar conformations (C3′-endo) found for the reported
2′-C-methylribonucleosides 1a, 15 and 16 6,28–30 and is rational for 2′-C-
methylribonucleosides, which have the preference to keep the bulky
methyl group in the equatorial position4. The conformation around the
C4′–C5′ bond of 2b is +sc (gauche, gauche) with the torsion angle
γ(O5′–C5′–C4′–C3′) = 65.2(2)°, which is similar to that of compound 16
(γ = 60.8°)29.
In the solid state, the 7-deazapurine moiety of 2b is nearly planar. The
deviations of the ring atoms from the least-squares plane
(N9/C8–C4/N3/C2/N1) are in the range of –0.0150 Å (atom C6) to 0.0118 Å
(atom N1), with a rms deviation of 0.0090 Å. The 7-fluoro substituent and
atom O6 lie 0.0089 and 0.0500 Å, respectively, below this plane.
The crystal structure of the 7-fluorinated 2′-C-methylribonucleoside 2b is
stabilized by three-dimensional hydrogen bond network involving the H at-
oms of the heteroatoms (N1–H1···O6i, O2′1–H2′O···O6ii, O3′–H3′O···O2′1iii,
O5′–H5′O···O5′iv; for symmetry codes see Table III, Fig. 4). The fluoro sub-
stituent (F-7) does not take part in the hydrogen bonding as observed in
other fluorinated compounds32.
TABLE III
Hydrogen bond geometry of 2b
D–H···A
D–H, Å
H···A, Å
D···A, Å
D–H···A, °
N1–H1···O6i
0.88
0.84
0.84
0.84
1.91
1.87
1.96
2.06
2.792(2)
2.702(2)
2.794(2)
2.863(1)
176.5
168.7
169.9
160.1
O2′1–H2′O···O6ii
O3′–H3′O···O2′1iii
O5′–H5′O···O5′iv
Symmetry codes: (i) x + 1/2, –y + 3/2, –z + 2; (ii) –x + 1, y – 1/2, –z + 3/2; (iii) x – 1, y, z;
(iv) x + 1/2, –y + 3/2, –z + 1.
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

1424
Seela, Budow, Xu, Peng, Eickmeier:
CONCLUSION
A series of 7-deazapurine 2′-C-methylribonucleosides carrying a 7-fluoro,
7-chloro and 7-iodo substituent were prepared employing a “one-pot”
glycosylation protocol. The immunosine 2′-C-methylribonucleoside 4 was
prepared by the same strategy. The “one-pot” procedure reported herein
represents a significant improvement over the earlier reported protocol6
for the synthesis of 7-fluoro-7-deaza-2′-C-methyladenosine (1b) and allows
the convenient synthesis of the HCV-active compound 1b as well as its
7-fluoro and 7-chloro analogues 1c, 2b, 2c, 7b, 7c, 8b and 8c. Also, the
7-iodinated guanosine derivatives 3, 11 and 12 as well as the 2′-C-methyl-
immunosine 4 were obtained in good yields. Moreover, from 7-fluoro-
7-deaza-2′-C-methylinosine (2b), a single-crystal X-ray analysis was per-
formed showing that the glycosylic bond has an anti conformation (χ =
–139.9(2)°), while the sugar moiety adopts an N conformation (C3′-endo;
P = 4.3(1)°, τm = 42.4(1)°).
EXPERIMENTAL
General
All chemicals are purchased from Acros, Fluka, or Sigma–Aldrich and were used without fur-
ther purification. Thin layer chromatography (TLC) was performed on TLC aluminium
sheets covered with silica gel 60 F254 (0.2 mm, VWR International). Flash chromatography
(FC): silica gel 60 (40–60 µM, VWR International, Darmstadt, Germany) at 0.4 bar. Melting
point (m.p.): Electrothermal 9200, uncorrected. UV-spectra were recorded on a U-3000
UV-Vis spectrophotometer (Hitachi, Japan); λmax (ε) in nm, ε in l mol–1 cm–1. NMR spectra:
Avance DPX 250 spectrometer (Bruker, Germany) measured at 250.13 MHz for 1H,
62.90 MHz for 13C and 235.33 MHz for 19F, and DPX 300 spectrometer (Bruker, Germany)
measured at 300.15 MHz for 1H, 75.48 MHz for 13C and 282.40 MHz for 19F. Chemical
shifts are given in ppm (δ-scale) and coupling constants (J) in Hz. For NMR spectra recorded
in DMSO, the chemical shift of the solvent peak was set to 2.50 ppm for 1H and 39.50 ppm
for 13C. Elemental analyses were performed by the Mikroanalytisches Laboratorium Beller,
Göttingen, Germany. ESI-TOF mass spectra were performed on a MicroTOF Bruker Daltonics
mass spectrometer in the electropositive mode.
Single-Crystal X-ray Analysis of 2b
The solid state structure of compound 2b, C12H14FN3O5, was determined by single-crystal
X-ray analysis. Crystals of 2b were grown by slow evaporation from methanol. The obtained
crystals of 2b (colorless, block) were orthorhombic with the space group P212121 and cell
dimensions of a = 4.8617(2) Å, b = 16.0982(6) Å, c = 16.8400(6) Å, V = 1317.98(9) Å3, and
Z = 4. The calculated density was 1.508 g cm–3
.
For the diffraction experiment, a single-crystal was mounted on a MiTeGen Micro-
Mountsfibre in a thin smear of oil. All diffraction measurements were made using
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

7-Deazapurine 2′-C-Methylribonucleosides
1425
monochromatized MoKα radiation (λ = 0.71073 Å) on a CCD area detector equipped
diffractometer (Bruker, Germany), at T = 100(2) K, with a θ range 1.75–28.52°. 1836 inde-
pendent reflections out of 26.587 reflections were measured and 1655 reflections observed at
the I > 2σ(I) level (Rint = 0.0397).
The known configuration of the parent precursor was used to define the enantiomer
employed in the refined model. In the absence of suitable anomalous scattering, Friedel
equivalents could not be used to determine the absolute structure. Refinement of the Flack
parameter33 led to inconclusive values for this parameter [0.0 (8)]. Therefore, Friedel equiva-
lents (1171) were merged before the final refinement. All H atoms were found in a differ-
ence Fourier synthesis. In order to maximize the data/parameter ratio, the H atoms were
placed in geometrically idealized positions C–H = 0.95–1.00 Å and N–H = 0.88 Å (AFIX 93)
and constrained to ride on their parent atoms with Uiso(H) = 1.2Ueq(C) = Ueq(N). The OH
groups were refined as rigid groups allowed to rotate but not tip (AFIX 147) with O–H =
0.84 Å and U(H) = 1.5Ueq(O). The structure was refined using full-matrix least-squares on F2
using 194 parameters and all independent reflections. The refinement converged with agree-
ment statistics of R [F2 > 2σ(F2)] = 0.031, wR = (F2) 0.077, S = 1.05.
CCDC 836029 contains the supplementary crystallographic data for this paper. These data
can be obtained free of charge via www.ccdc.cam.ac.uk/conts/retrieving.html (or from
the Cambridge Crystallographic Data Centre, 12, Union Road, Cambridge, CB2 1EZ, UK;
fax: +44 1223 336033; or deposit@ccdc.cam.ac.uk).
4-Chloro-5-fluoro-7-(2,3,5-tri-O-benzoyl-2-C-methyl-β-D-ribofuranosyl)-
7H-pyrrolo[2,3-d]pyrimidine (7b)
4-Chloro-5-fluoro-7H-pyrrolo[2,3-d]pyrimidine14 (5b; 343 mg, 2 mmol) was suspended in
dry acetonitrile (15 ml) and N,O-bis(trimethylsilyl)acetamide (BSA; 0.6 ml, 2.42 mmol) was
added to the mixture. The mixture became clear, and after 10 min 1,2,3,5-tetra-O-benzoyl-
2-C-methyl-D-ribofuranose10 (6; 1.74 g, 3 mmol) and trimethylsilyl trifluoromethane-
sulfonate (TMSOTf; 0.54 ml, 2.80 mmol) were added. The mixture was stirred at r.t. for 10
min, and then stirring was continued at 60 °C. Another portion of compound 6 (1.16 g, 2
mmol) and TMSOTf (0.34 ml, 1.76 mmol) was added after 6 h. The reaction mixture was
stirred overnight. The mixture was diluted with dichloromethane (100 ml) and washed with
5% aq. NaHCO3. The organic layer was dried (Na2SO4) and filtered. The filtrate was evapo-
rated to dryness and the residue was applied to FC (silica gel, column 5 × 15 cm, eluted with
CH2Cl2). The main zone afforded 7b as a colorless foam (0.75 g, 60%). TLC (silica gel,
CH2Cl2): RF 0.25. UV (MeOH): λmax (ε) = 227 (48 800), 274 (5 400), 282 (5 000). 1H NMR
(250 MHz, DMSO-d6): 1.59 s, 3 H (CH3); 4.73–4.86 m, 3 H (H-4′, 2 × H-5′); 5.89–5.90 m, 1 H
(H-3′); 7.04 s, 1 H (H-1′); 7.31–8.04 m, 15 H (H-arom); 8.12 d, 1 H, J(C,F) = 1.75 (H-6);
8.88 s, 1 H (H-2). 19F NMR (235 MHz, DMSO-d6): –169.24 (F-5). MS ESI-TOF calculated for
C
33H25ClFN3O7Na [M + Na]+: 652.1263; found: 652.1262.
4,5-Dichloro-7-(2,3,5-tri-O-benzoyl-2-C-methyl-β-D-ribofuranosyl)-
7H-pyrrolo[2,3-d]pyrimidine (7c)
4,5-Dichloro-7H-pyrrolo[2,3-d]pyrimidine15 (5c; 200 mg, 1.06 mmol) was suspended in dry
acetonitrile (10 ml) and N,O-bis(trimethylsilyl)acetamide (BSA; 0.42 ml, 1.70 mmol) was
added to the mixture. The mixture became clear and after 10 min 1,2,3,5-tetra-O-benzoyl-
2-C-methyl-D-ribofuranose10 (6; 1.0 g, 1.72 mmol) and trimethylsilyl trifluoromethane-
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1426
Seela, Budow, Xu, Peng, Eickmeier:
sulfonate (TMSOTf; 0.3 ml, 1.6 mmol) were added. The mixture was stirred at 60 °C. After
5 h, compound 6 (800 mg, 1.38 mmol) and TMSOTf (0.23 ml, 1.19 mmol) were added. The
reaction mixture was stirred overnight. The mixture was diluted with dichloromethane
(50 ml) and washed with 5% aq. NaHCO3. The organic layer was dried (Na2SO4) and fil-
tered. The filtrate was evaporated to dryness, and the residue was applied to FC (silica gel,
column 5 × 15 cm, eluted with CH2Cl2) affording 7c as a colorless foam (0.48 g, 70%). TLC
(silica gel, CH2Cl2): RF 0.26. UV (MeOH): λmax (ε) = 230 (26 300), 275 (2 500), 282 (2 300).
1H NMR (300 MHz, DMSO-d6): 1.58 s, 3 H (CH3); 4.77–4.87 m, 3 H (H-4′, 2 × H-5′);
5.92–5.93 m, 1 H (H-3′); 7.03 s, 1 H (H-1′); 7.37–8.04 m, 15 H (H-arom); 8.29 s, 1 H (H-6);
8.88 s, 1 H (H-2). MS ESI-TOF calculated for C33H25Cl2N3O7Na [M + Na]+: 668.0967; found:
668.0960.
5-Fluoro-7-(2-C-methyl-β-D-ribofuranosyl)-4-methoxy-
7H-pyrrolo[2,3-d]pyrimidine (8b)
Compound 7b (630 mg, 1 mmol) was suspended in 0.5 M NaOMe/MeOH (50 ml), and the
mixture was refluxed for 2 h. Then, the solvent was evaporated and the residue was ad-
sorbed on silica gel and applied to FC (silica gel, column
8 × 5 cm, eluted with
CH2Cl2/MeOH, 30:1→20:1). The main zone afforded 8b as a colourless solid. Recrystal-
lization from CH2Cl2 gave 8b as colourless crystals (250 mg, 80%). M.p. 102 °C. TLC (silica
gel, CH2Cl2/MeOH, 10:1): RF 0.34. UV (MeOH): λmax (ε) = 221 (19 000), 261 (sh, 4 700), 280
(5 700). 1H NMR (300 MHz, DMSO-d6): 0.67 s, 3 H (CH3); 3.64–3.70 2 m, 1 H (H-5′b);
3.81–3.88 m, 2 H (H-5′a, H-4′); 3.92–3.97 m, 1 H (H-3′); 4.06 s, 3 H (OMe); 5.17 d, 1 H,
J(3′,OH) = 6.30 (3′-OH); 5.20 s, 1 H (2′-OH); 5.23 t, 1 H, J(5′,OH) = 4.80 (5′-OH); 6.25 d, 1 H,
J(H,F) = 1.5 (H-1′); 7.79 d, 1 H, J(H,F) = 1.8 (H-6); 8.44 s, 1 H (H-2). 19F NMR (282 MHz,
DMSO-d6): –167.73 (F-5). MS ESI-TOF calculated for C13H16FN3O5Na [M + Na]+: 336.0966;
found: 336.0964.
5-Chloro-7-(2-C-methyl-β-D-ribofuranosyl)-4-methoxy-
7H-pyrrolo[2,3-d]pyrimidine (8c)
Compound 7c (323 mg, 0.5 mmol) was suspended in 0.5 M NaOMe/MeOH (25 ml), and the
mixture was refluxed for 2 h. Then, the mixture was evaporated, and the residue was ad-
sorbed on silica gel and applied to the top of a column (silica gel, column 8 × 5 cm, eluted
with CH2Cl2/MeOH, 30:1→20:1). The main zone afforded 8c as colourless solid. Recrystal-
lization from CH2Cl2 gave 8c as colourless crystals (130 mg, 79%). M.p. 98 °C. TLC (silica
gel, CH2Cl2/MeOH, 10:1): RF 0.36. UV (MeOH): λmax (ε) = 222 (18 300), 261 (sh, 4 200), 281
(4 900). 1H NMR (300 MHz, DMSO-d6): 0.68 s, 3 H (CH3); 3.64–3.71 2 m, 1 H (H-5′b);
3.82–3.89 m, 2 H (H-5′a, H-4′); 3.94–3.99 m, 1 H (H-3′); 4.06 s, 3 H (OMe); 5.18 d, 1 H,
J(3′,OH) = 6.30 (3′-OH); 5.22 s, 1 H (2′-OH); 5.26 t, 1 H, J(5′,OH) = 4.80 (5′-OH); 6.21 s, 1 H
(H-1′); 7.99 s, 1 H (H-6); 8.47 s, 1 H (H-2). MS ESI-TOF calculated for C13H16ClN3O5Na [M +
Na]+: 352.0676; found: 352.0682.
5-Fluoro-7-(2-C-methyl-β-D-ribofuranosyl)-3,7-dihydro-
4H-pyrrolo[2,3-d]pyrimidin-4-one (2b)
A solution of 8b (156 mg, 0.5 mmol) in 2 M NaOH (3 ml) was heated under reflux for 3 h.
The mixture was cooled, diluted with water (15 ml) and neutralized with 1 M AcOH to pH 7.
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7-Deazapurine 2′-C-Methylribonucleosides
1427
The solution was filtered, and the filtrate was applied to a column (15 × 2 cm, Serdolit AD-4,
resin 0.1–0.2 mm). The column was washed with H2O (200 ml), and the product was eluted
with H2O/i-PrOH (95:1, 300 ml). The product containing fractions were collected, and the
solvent was evaporated yielding 2b as a colorless solid. Recrystallization from methanol af-
forded 2b as colorless needles (120 mg, 80%). M.p. 269 °C. TLC (silica gel, CH2Cl2/MeOH,
10:1): RF 0.10. UV (MeOH): λmax (ε) = 216 (21 600), 264 (8 200). 1H NMR (250 MHz,
DMSO-d6): 0.71 s, 3 H (CH3); 3.63–3.67 m, 1 H (H-5′b); 3.80–3.97 m, 3 H (H-5′a, H-4′, H-3′);
5.20 br s, 3 H (2′-OH, 3′-OH, 5′-OH); 6.10 d, 1 H, J(H,F) = 1.63 (H-1′); 7.49 d, 1 H, J(H,F) =
1.63 (H-6); 7.93 s, 1 H (H-2). 19F NMR (235 MHz, DMSO-d6): –166.13 (F-5). MS ESI-TOF cal-
culated for C12H14FN3O5Na [M + Na]+: 322.0815; found: 322.0808.
5-Chloro-3,7-dihydro-7-(2-C-methyl-β-D-ribofuranosyl)-
4H-pyrrolo[2,3-d]pyrimidin-4-one (2c)
A solution of 8c (70 mg, 0.21 mmol) in 2 M NaOH (3 ml) was heated under reflux for 3 h.
The mixture was cooled, diluted with water (10 ml) and neutralized with 1 M HCl to pH 7.
The solution was evaporated to dryness, and the residue was applied to FC (silica gel, eluted
with CH2Cl2/MeOH, 10:1→5:1). Compound 2c was obtained as a colorless solid. Recrystal-
lization from methanol afforded 2c as colourless needles (50 mg, 75%). M.p. 250 °C (dec.).
TLC (silica gel, CH2Cl2/MeOH, 10:1): RF 0.11. UV (MeOH): λmax (ε) = 218 (15 600), 264
(6 700). 1H NMR (300 MHz, DMSO-d6): 0.71 s, 3 H (CH3); 3.62–3.67 2 m, 1 H (H-5′b);
3.80–3.94 m, 3 H (H-5′a, H-4′, H-3′); 5.15 d, 1 H, J(3′,OH) = 3.60 (3′-OH); 5.18 s, 1 H
(2′-OH); 5.21 t, 1 H, J(5′,OH) = 4.65 (5′-OH); 6.06 s, 1 H (H-1′); 7.68 s, 1 H (H-6); 7.94 s, 1 H
(H-2); 12.12 br s, 1 H (NH). MS ESI-TOF calculated for C12H14ClN3O5Na [M + Na]+:
338.0520; found: 338.0523.
4-Amino-5-fluoro-7-(2-C-methyl-β-D-ribofuranosyl)-
7H-pyrrolo[2,3-d]pyrimidine (1b)
Compound 7b (900 mg, 1.43 mmol) was suspended in methanolic ammonia (saturated at
0 °C, 80 ml) and stirred at 120 °C for 20 h. The mixture was evaporated to dryness and the
residue was applied to FC (silica gel, column 5 × 8 cm, eluted with CH2Cl2/MeOH,
50:1→10:1) yielding 1b as a colorless solid. Compound 1b 6 was crystallized from methanol
affording 1b as colorless crystals (300 mg, 70%). M.p. 219 °C. TLC (silica gel,
CH2Cl2/CH3OH, 10:1): RF 0.13. UV (MeOH): λmax (ε) = 211 (23 600), 281 (8 600). 1H NMR
(250 MHz, DMSO-d6): 0.68 s, 3 H (CH3); 3.62–3.68 m, 1 H (H-5′b); 3.80–3.83 m, 2 H (H-5′a,
H-4′); 3.90–3.93 m, 1 H (H-3′); 5.10–5.17 m, 3 H (2′-OH, 3′-OH, 5′-OH); 6.16 d, 1 H, J(H,F) =
1.65 (H-1′); 7.00 br s, 2 H (NH2); 7.49 d, 1 H, J(H,F) = 1.62 (H-6); 8.07 s, 1 H (H-2). 19F NMR
(235 MHz, DMSO-d6): –168.00 (F-5). MS ESI-TOF calculated for C12H15FN4O4Na [M + Na]+:
321.0970; found: 321.0971.
4-Amino-5-chloro-7-(2-C-methyl-β-D-ribofuranosyl)-
7H-pyrrolo[2,3-d]pyrimidine (1c)
Compound 1c was prepared as described for 1b with 7c (200 mg, 0.31 mmol). After FC (sil-
ica gel, eluted with CH2Cl2/MeOH, 50:1→10:1), 1c 6 was obtained as a colorless solid
(70 mg, 72%). M.p. 153 °C (dec.). TLC (silica gel, CH2Cl2/MeOH, 10:1): RF 0.15. UV (MeOH):
λmax (ε) = 210 (20 300), 281 (7 700). 1H NMR (300 MHz, DMSO-d6): 0.69 s, 3 H (CH3);
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1428
Seela, Budow, Xu, Peng, Eickmeier:
3.63–3.68 2 m, 1 H (H-5′b); 3.80–3.85 m, 2 H (H-5′a, H-4′); 3.92 d, 1 H, J(3′,OH) = 6.88
(H-3′); 5.12 d, 1 H, J(3′,OH) = 6.76 (3′-OH); 5.14 s, 1 H (2′-OH); 5.20 t, 1 H, J(5′,OH) = 4.62
(5′-OH); 6.13 s, 1 H (H-1′); 6.87 br s, 2 H (NH2); 7.74 s, 1 H (H-6); 8.11 s, 1 H (H-2). MS
ESI-TOF calculated for C12H15ClN4O4Na [M + Na]+: 337.0674; found: 337.0682.
4-Chloro-5-iodo-2-pivaloylamino-7-[(2,3,5-tri-O-benzoyl)-2-C-methyl-β-D-ribofuranosyl]-
7H-pyrrolo[2,3-d]pyrimidine (10)
Into a stirred suspension of 4-chloro-5-iodo-2-pivaloylamino-7H-pyrrolo[2,3-d]pyrimidine17
(9; 757 mg, 2.0 mmol) in anhydrous MeCN (14 ml), N,O-bis(trimethylsilyl)acetamide (BSA;
1.0 ml, 4.04 mmol) was added at room temperature. After stirring for 5 min, trimethylsilyl
trifluoromethanesulphonate (TMSOTf; 0.75 ml, 3.9 mmol) was added and 1,2,3,5-tetra-
O-benzoyl-2-C-methyl-D-ribofuranose10 (6; 3.02 g, 5.2 mmol) was introduced in three por-
tions (once per 24 h). In total, the reaction was stirred at 50 °C for 72 h, cooled to room
temperature, and diluted with CH2Cl2 (80 ml). The solution was washed with aq. saturated
NaHCO3 and brine, dried over Na2SO4, and evaporated under reduced pressure to give a
syrup, which was applied to FC (silica gel, column 4 × 12 cm, eluted with CH2Cl2/petroleum
ether, 2:1→100:0). The main zone afforded compound 10 as a yellowish foam (1.26 g, 75%).
TLC (silica gel, CH2Cl2/MeOH, 99:1): RF 0.44. 1H NMR (300 MHz, DMSO-d6): 1.27 s, 9 H (3 ×
CH3); 1.66 s, 3 H (CH3); 4.74–4.85 m, 3 H (H-4′, 2 × H-5′); 6.15 br s, 1 H (H-3′); 6.87 s, 1 H
(H-1′); 7.36–8.05 3 m, 15 H (arom-H); 8.08 s, 1 H (H-6); 10.42 s, 1 H (NH). For C38H34ClIN4O8
(837.06) calculated: 54.53% C, 4.09% H, 6.69% N; found: 54.63% C, 4.15% H, 6.54% N.
5-Iodo-7-(2-C-methyl-β-D-ribofuranosyl)-
7H-pyrrolo[2,3-d]pyrimidin-2,4-diamine (11)
A suspension of 10 (418 mg, 0.50 mmol) in dioxane (30 ml) and 25% aq. NH3 (70 ml) was
introduced into an autoclave and stirred at 120 °C for 24 h. The reaction mixture was evap-
orated under reduced pressure to give a syrup, which was adsorbed on silica gel, and applied
on the top of a column (silica gel, column 3 × 8 cm, CH2Cl2/MeOH, 95:5). Fractions con-
taining compound 11 were collected and evaporated to dryness to give 11 as a yellowish
solid (179 mg, 85%). TLC (silica gel, CH2Cl2/MeOH, 9:1): RF 0.17. UV (MeOH): λmax (ε) =
232 (31 100), 270 (8 500), 289 (7 600). 1H NMR (300 MHz, DMSO-d6): 0.73 s, 3 H (CH3);
3.57–3.64 2 m, 1 H (H-5′b); 3.76–3.81 m, 2 H (H-5′a, H-4′); 3.90 dd, 1 H, J(3′,OH) = 6.60,
J(3′,4′) = 8.70 (H-3′); 4.88 s, 1 H (2′-OH); 5.11 t, 1 H, J(5′,OH) = 4.80 (5′-OH); 5.16 d, 1 H,
J(3′,OH) = 6.60 (3′-OH); 5.85 br s, 2 H (NH2); 5.95 s, 1 H (H-1′); 6.19 s, 2 H (NH2); 7.34 s,
1 H (H-6). For C12H16IN5O4 (421.19) calculated: 34.22% C, 3.83% H, 16.63% N; found:
34.19% C, 3.90% H, 16.45% N.
2-Amino-5-iodo-4-methoxy-7-(2-C-methyl-β-D-ribofuranosyl)-
7H-pyrrolo[2,3-d]pyrimidine (12)
A solution of 10 (837 mg, 1.0 mmol) in 0.5 M NaOMe/MeOH (20 ml) was heated under re-
flux for 3 h. The reaction mixture was neutralized with glacial acetic acid, the crude product
was adsorbed on silica gel, and applied on the top of a column (silica gel, column 3 × 8 cm,
CH2Cl2/MeOH, 95:5). Fractions containing compound 12 were collected and evaporated to
dryness to give 12 as a colorless solid (384 mg, 88%). TLC (silica gel, CH2Cl2/MeOH, 9:1):
RF 0.36. UV (MeOH): λmax (ε) = 234 (29 700), 267 (6 500), 289 (6 200). 1H NMR (250 MHz,
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

7-Deazapurine 2′-C-Methylribonucleosides
1429
DMSO-d6): 0.71 s, 3 H (CH3); 3.64–3.80 2 m, 4 H (2 × H-5′, H-4′, H-3′); 3.92 s, 3 H (OMe);
4.92 s, 1 H (2′-OH); 5.13–5.19 m, 2 H (5′-OH, 3′-OH); 5.98 s, 1 H (H-1′); 6.40 s, 2 H (NH2);
7.45 s, 1 H (H-6). For C13H17IN4O5 (436.20) calculated: 35.80% C, 3.93% H, 12.84% N; found:
35.69% C, 3.83% H, 12.63% N.
2-Amino-5-iodo-7-(2-C-methyl-β-D-ribofuranosyl)-3,7-dihydro-
4H-pyrrolo[2,3-d]pyrimidin-4-one (3)
Compound 12 (174 mg, 0.40 mmol) was dissolved in 2 M NaOH (40 ml) and 1,4-dioxane
(6 ml). The mixture was stirred under reflux for 3 h. After neutralization with 2 M HCl, the
volume was reduced by 50%. The solution was applied to a Serdolit AD-4 column (3 ×
12 cm, resin 0.1–0.2 mm; Serva, Germany). Salt was removed by elution with H2O (150 ml),
and the product was eluted with H2O/i-PrOH (9:1→5:1). Fractions containing compound 3
were combined, and the solvent was evaporated to afford 3 as a colorless solid (144 mg,
85%). TLC (silica gel, CH2Cl2/MeOH, 5:1): RF 0.33. UV (MeOH): λmax (ε) = 227 (18 600), 267
(10 500), 289 (6 600). 1H NMR (300 MHz, DMSO-d6): 0.74 s, 3 H (CH3); 3.57–3.64 2 m, 1 H
(H-5′b); 3.75–3.81 m, 2 H (H-5′a, H-4′); 3.88 dd, 1 H, J(3′,OH) = 6.15, J(3′,4′) = 8.85 (H-3′);
4.87 s, 1 H (2′-OH); 5.10 t, 1 H, J(5′,OH) = 4.80 (5′-OH); 5.19 d, 1 H, J(3′,OH) = 6.30 (3′-OH);
5.86 s, 1 H (H-1′); 6.35 br s, 2 H (NH2); 7.28 s, 1 H (H-6); 10.44 s, 1 H (NH). For C12H15IN4O5
(422.18) calculated: 34.14% C, 3.58% H, 13.27% N; found: 34.26% C, 3.69% H, 13.20% N.
5-Amino-3-(2,3,5-tri-O-benzoyl-2-C-methyl-β-D-ribofuranosyl)-
thiazolo[4,5-d]pyrimidin-2,7(3H,6H)-dione (14)
A mixture of dry 5-aminothiazolo[4,5-d]pyrimidin-2,7(3H,6H)-dione23 (13; 184 mg, 1.0 mmol),
hexamethyldisilazane (HMDS; 3.3 ml), ammonium sulfate (2 mg), and pyridine (0.33 ml)
was heated under reflux overnight with the exclusion of moisture. Excess HMDS was
removed by evaporation to provide the silylated derivative as a syrup. To the silylated inter-
mediate were added dry acetonitrile (10 ml) and 1,2,3,5-tetra-O-benzoyl-2-C-methyl-
D-ribofuranose10 (6; 580 mg, 1.0 mmol). To the formed suspension, trimethylsilyl
trifluoromethanesulfonate (0.31 ml, 1.4 mmol) was added. The suspension became a clear
solution, and the reaction mixture was stirred at ambient temperature for 16 h. The solution
was evaporated to dryness, and the residual syrup was dissolved in EtOAc (30 ml). The solu-
tion was washed with 5% aq. NaHCO3 solution (2 × 10 ml), and dried (Na2SO4). The solvent
was evaporated to yield a syrup. The syrup was dissolved in dichloromethane (2 ml) and ap-
plied to the top of a column (silica gel, 12 × 2.5 cm, eluted with CH2Cl2/EtOAc, 4:1→2:1).
Compound 14 22 (350 mg, 54.5%) was obtained as a colorless foam. TLC (silica gel,
CH2Cl2/MeOH, 10:1): RF 0.45. NMR data in analogy to ref.26 1H NMR (300 MHz, DMSO-d6):
.
1.80 s, 3 H (CH3); 4.55–4.60 m, 2 H (2 × H-5′); 4.76–4.82 m, 1 H (H-4′); 6.13–6.27 m, 1 H
(H-3′); 6.68 s, 1 H (H-1′); 7.06 br s, 2 H (NH2); 7.40–8.09 m, 15 H (H-arom); 11.40 br s, 1 H
(NH). MS ESI-TOF calculated for C32H26N4O9SNa [M + Na]+: 665.1313; found: 665.1326.
5-Amino-3-(2-C-methyl-β-D-ribofuranosyl)-
thiazolo[4,5-d]pyrimidin-2,7(3H,6H)-dione (4)
To a suspension of 14 (220 mg, 0.34 mmol) in methanol (2.6 ml), 1 M NaOMe/MeOH
(1.75 ml) was added. The mixture became clear and was stirred at r.t. for 16 h. The reaction
mixture was neutralized with Dowex-50+ resin and filtered. The filtrate was evaporated to
Collect. Czech. Chem. Commun. 2011, Vol. 76, No. 12, pp. 1413–1431

1430
Seela, Budow, Xu, Peng, Eickmeier:
dryness. The residue was triturated with ether (2 × 2.5 ml) to yield 4 22 as a slight yellow
solid (80 mg, 71%). NMR data in analogy to ref.26 1H NMR (300 MHz, DMSO-d6): 1.05 s,
.
3 H (CH3); 3.60 br s, 2 H (2 × H-5′); 3.74–3.80 m, 1 H (H-4′); 3.96 br s, 1 H (H-3′); 4.60 s,
1 H (3′-OH); 4.80 s, 1 H (2′-OH); 5.25 br s, 1 H (5′-OH); 5.98 s, 1 H (H-1′); 7.08 br s, 2 H
(NH2); 11.02 br s, 1 H (NH). MS ESI-TOF calculated for C11H14N4O6SNa [M + Na]+: 353.0526;
found: 353.0541.
We would like to thank Dr. P. Leonard for helpful discussions and support. We are grateful for
critical reading of the manuscript by Mr. S. S. Pujari. Financial support from ChemBiotech, Münster,
Germany is highly appreciated.
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