Synthesis and cytostatic activity of substituted 6-phenylpurine bases and nucleosides: Application of the Suzuki-Miyaura cross-coupling reactions of 6-chloropurine derivatives with phenylboronic acids

Article abstract of DOI:10.1021/jm991167+

The Suzuki-Miyaura reaction of protected 6-chloropurine and 2-amino-6- chloropurine bases and nucleosides with substituted phenylboronic acids led to the corresponding protected 6-(substituted phenyl)purine derivatives 6-9. Their deprotection yielded a se

Full text of DOI:10.1021/jm991167+

J . Med. Chem. 2000, 43, 1817-1825
1817
Syn th esis a n d Cytosta tic Activity of Su bstitu ted 6-P h en ylp u r in e Ba ses a n d
Nu cleosid es: Ap p lica tion of th e Su zu k i-Miya u r a Cr oss-Cou p lin g Rea ction s of
6-Ch lor op u r in e Der iva tives w ith P h en ylbor on ic Acid s
Michal Hocek,*,† Anton´ın Holy´,† Ivan Votruba,† and Hana Dvorˇa´kova´‡
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
CZ-16610 Prague 6, Czech Republic, and Laboratory of NMR Spectroscopy, Prague Institute of Chemical Technology,
CZ-16628 Prague 6, Czech Republic
Received September 24, 1999
The Suzuki-Miyaura reaction of protected 6-chloropurine and 2-amino-6-chloropurine bases
and nucleosides with substituted phenylboronic acids led to the corresponding protected
6-(substituted phenyl)purine derivatives 6-9. Their deprotection yielded a series of substituted
6-phenylpurine bases and nucleosides 10-13. Significant cytostatic activity (IC50 0.25-20 µmol/
L) in CCRF-CEM, HeLa, and L1210 cell lines was found for several 6-(4-X-substituted phenyl)-
purine ribonucleosides 12 (X ) H, F, Cl, and OR), while the 6-phenylpurine and 2-amino-6-
phenylpurine bases 10 and 11, as well as 2-amino-6-phenylpurine ribosides 13, were entirely
inactive against these cell lines.
In tr od u ction
its water-soluble 5′-phosphate (NSC 312887, fludarabine
phosphate).15 Also the influence of introduction of alkyl
substituents at position 2 of the purine bases has been
examined, but no cytostatic activity was noted among
these compounds; only the antihypertensive activity of
2-octynyladenosine was reported.16
Purine bases and their nucleosides constitute an
important group of antineoplastic and antileukemic
agents (for reviews, see ref 1). Thus, olomoucin and its
congeners exhibit their cytostatic activity by virtue of
inhibiting the cyclin-dependent protein kinases.2 In the
search for novel antimetabolites, purine bases as well
as the carbohydrate residue in the nucleoside molecules
were modified by structural alterations. An exhaustive
review covers numerous cytostatic aza and deaza ana-
logues of purine nucleosides.3-5 The most attractive
compound of this group is 3-deazaguanine (NSC 261726)6
and its ribonucleoside,7 but an important antineoplastic
activity was also encountered among 7-deazapurine
nucleoside antibiotics (tubercidin, toyocamycin, sangi-
vamycin) and their sugar-modified analogues.3,8
Replacement of the oxygen atom at position 6 in
guanine and hypoxanthine by sulfur gave clinically
widely used anticancer drugs thioguanine (NSC752)9
and 6-mercaptopurine (NSC755).10 Structurally related
nucleoside sulfinosine [2-amino-9-(â-D-ribofuranosyl)-
purine-6-sulfinamide] and its congeners11 also exhibit
unique antitumor properties.
Substitution at position 2 of the adenine ring by a
halogen atom gives rise to biologically active nucleo-
sides: while 2-chloroadenosine (an adenosine receptor
agonist) causes solely a selective depletion of natural
killer cells via signal transduction,12 substantial thera-
peutic success was achieved with 2-chloro-2′-deoxyade-
nosine13 (NSC 105014-F, cladribine) which is clinically
used in the treatment of hairy cell leukemia, human
primary lymphoma, chronic lymphocytic leukemia, and
myeloid leukemia.14 A closely related drug in clinical
use for treatment of lymphoproliferative diseases is 9-(â-
D-arabinofuranosyl)-2-fluoroadenine (fludarabine) and
Most of the above substitutions maintain the amino
or oxo (thioxo) group at position 6 of the purine base
which is essential for the formation of hydrogen bonds
important for the interactions with polymerases and
other key enzymes of nucleic acid metabolism. The
cytostatic activity was also observed in 2-amino-6-
methoxypurine arabinoside, which was investigated as
a possible tool for combating T-cell malignancies,17 and
the 6-alkylmercaptopurine derivatives exhibiting cer-
tain cytostatic activity;18 nonetheless, these compounds
might be eventually converted to guanine nucleosides
by the action of adenosine aminohydrolase.
Such a catabolic reaction is indeed excluded in 6-alkyl-
purine derivatives. The parent compound of this group,
6-methylpurine, is known for its cytotoxicity; its libera-
tion from the 2′-deoxyribonucleoside by purine nucleo-
side phosphorylases is used for detection of mycoplasma
in cell cultures.19 It is highly potent and toxic to
nonproliferating and proliferating tumor cells. Recently,
the use of cytotoxic bases liberated by purine nucleoside
phosphorylases such as 6-methylpurine was proposed
as a novel principle in the gene therapy of cancer.20
Peculiarly little attention was aimed at the investiga-
tion of other 6-alkylpurine derivatives. Recently, cyto-
kinin activity was reported in some 6-(arylalkynyl)-,
6-(arylalkenyl)-, and 6-(arylalkyl)purines.21 We have
recently described the cytostatic activity of 6-(trifluo-
romethyl)purine riboside.22 The corticotropin-releasing
hormone antagonist activity of some 2,8,9-trisubsti-
tuted-6-arylpurines has been also reported.23
* To whom correspondence should be addressed. Fax: 420
3111271. E-mail: hocek@uochb.cas.cz.
2
C-Nucleosites bearing a 2,4-difluoro-5-methylphenyl
moiety24 are recognized by DNA polymerases as ana-
logues of dTMP and, if incorporated into single-stranded
† Academy of Sciences of the Czech Republic.
‡ Prague Institute of Chemical Technology.
10.1021/jm991167+ CCC: $19.00 © 2000 American Chemical Society
Published on Web 04/14/2000

1818 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 9
Hocek et al.
Sch em e 1a
a
Reagents and conditions: (i) R1,R2,R3PhB(OH)2 (5), K2CO3, Pd(PPh3)4, toluene; (ii) Dowex 50×8 (H+), MeOH, H2O; (iii) NaOMe (0.1
equiv), MeOH.
DNA, encode selectively for adenine in DNA replication.
halopurines with aryl- or alkenylboronic acid leading
to 6-aryl- and 6-alkenylpurines in good yields.31 Two
different optimized procedures have been elaborated.
Anhydrous conditions using K2CO3 in toluene were
favorable for the coupling of phenylboronic acids bearing
an electroneutral or electron-donor substituent, while
aqueous conditions using dimethoxyethane and satu-
rated aqueous Na2CO3 were successfully used for the
coupling reactions of alkenylboronic acids and phenyl-
boronic acids bearing an electron-withdrawing group.
Advantages of this reaction compared to the above-
mentioned methods are high stability of boronic acids,
tolerance toward the presence of diverse functions
(including amino and hydroxy groups), low toxicity of
boron compounds, easy workup and isolation of the
product, and the availability of starting boronic acids
(a number of boronic acids, in particular aromatic ones,
are commercially available). It is the methodology of
choice for the synthesis of a series of substituted
6-phenylpurines and their ribonucleosides.
Considering all the above-mentioned findings and sur-
prisingly high biocompatibility of the fluorophenyl group
in C-nucleosides, we have focused on the introduction
of phenyl and diverse fluorophenyl groups into position
6 of the purine ring. Here we report on the synthesis
and in vitro antineoplastic activity of 6-phenylpurine
bases and nucleosides against selected transformed cell
lines.
Ch em istr y
With the development of the cross-coupling methodol-
ogy, many 6-C-substituted purines have been prepared
in the past decade. Thus, 6-halopurine derivatives react
with arylmagnesium halides,25 alkyl(aryl)zinc or tin
reagents,26 trialkylaluminum,27 or alkylcuprates28 to
give the 6-alkylpurine derivatives. Also a reverse ap-
proach based on the reaction of purine-6-zinc iodide with
aryl or vinyl halides has recently been described.29 For
the synthesis of 6-arylpurines, an alternative approach
makes use of radical photochemical reactions of adenine
derivatives with aromatic compounds,30 but this method
is very unselective and for substituted benzenes, mix-
tures of ortho-, meta-, and para-substituted derivatives
were obtained.
Tetrahydropyran-2-yl (THP)-protected 6-chloropurine
132 and bis(THP)-protected 2-amino-6-chloropurine 2,33
as well as 2′,3′,5′-O-triacetyl 6-chloropurine riboside 334
and its 2-amino-6-chloropurine analogue 435 were cho-
sen as suitable starting compounds for the cross-
coupling reactions with unsubstituted and fluorinated
phenylboronic acids 5a -d . Due to the lability of the
Recently we have reported a preliminary communica-
tion on Suzuki-Miyaura type of cross-coupling of 6-

Substituted 6-Phenylpurine Bases and Nucleosides
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 9 1819
Ta ble 1. Cytostatic Activity of 6-Phenylpurine Nucleosides 12
and Their Triacetates 8
acetyl protection of starting nucleosides 3 and 4, only
the anhydrous conditions could be used. The reactions
were performed in toluene in the presence of potassium
carbonate (2 equiv) and Pd(PPh3)4 catalyst (5%) under
Ar atmosphere. The corresponding protected 6-phen-
ylpurines 6-9(a -d ) were obtained in good yields of 60-
95% after column chromatography. On the other hand,
the attempted reactions of 6-chloropurines 1-4 with an
extremely electron-poor pentafluorophenylboronic acid
were unsuccessful. The THP derivatives 6 and 7 were
deprotected by the use33 of wet Dowex 50×8 (H+) in
methanol to afford the 6-phenylpurine and 2-amino-6-
phenylpurine bases 10a -d and 11a -d , respectively.
The acetyl functions in compounds 8a -d and 9a -d
were cleaved by methanolysis to afford quantitatively
the free nucleosides 12a -d and 13a -d that were easily
purified by crystallization.
IC50 (µmol/L)a
compd
R1
R2
R3
L1210
HeLa
CCRF-CEM
12a
12b
12c
12d
12e
12f
12g
12h
12i
12j
8a
8b
8c
8d
8e
8f
8g
8h
8i
H
F
F
F
H
H
H
F
H
H
H
MeO
H
H
H
H
H
F
H
H
H
H
H
F
9.0
4.5
2.7
2.5
NA
2.5
NA
NA
4.3
NA
5.0
0.7
0.75
NA
1.4
NA
1.5
0.25
4.8
0.6
0.9
10.5
4.0
NA
8.3
NA
NA
4.3
NA
20.0
NA
NA
1.5
H
Me
H
H
H
H
H
H
H
F
H
Me
H
H
H
H
H
H
Me
MeO
H
EtO
Cl
H
F
F
F
H
Me
MeO
H
EtO
Cl
NA
2.5
NA
NA
NA
NA
NA
NA
NA
17.0
NA
NA
NA
5.0
NA
15.0
NA
15.0
NA
NA
19.0
NA
NA
NA
Since in our preliminary cytostatic activity screening
the promising cytostatic activity of 6-phenylpurine
ribonucleosides 12a -d had been found (vide infra),
further attempts focused on the synthesis of other
substituted 6-phenylpurine ribonucleosides bearing di-
verse types of substituents on the benzene moiety. Thus
the 6-chloropurine nucleoside 3 has been submitted in
a series of reactions with a variety of substituted
phenylboronic acids. In accord with our previous find-
ings,31 phenylboronic acids bearing electroneutral and
electron-donor substituents (methyl, alkoxy, and chloro
- compounds 5e-j) reacted smoothly under analogous
conditions giving the corresponding 6-arylpurines 8e-j
in high yields, while phenylboronic acids bearing electron-
withdrawing groups (e.g. 3-nitrophenyl, 4-formylphenyl,
and 4-acetylphenyl derivatives) did not give cross-
coupled products in satisfactory yields. Methanolysis of
acetates 8e-j gave the desired free nucleosides 12e-j.
In conclusion, the application of the Suzuki-Miyaura
reaction of 6-chloropurine derivatives with substituted
phenylboronic acids is a facile and effective approach
for the synthesis of a series of specifically substituted
6-phenylpurine bases and nucleosides. In comparison
with the previously known methods25-30 using other
types of organometallic reagents or photochemistry, this
method is more effective and selective, and therefore,
further applications in the synthesis of 6-C-substituted
purine derivatives may be expected.
MeO
H
H
NA
3.8
4.3
8j
a
NA, not active (inhibition of the cell growth at c ) 10 µmol/L
was lower than 20%).
inverse techniques HMQC and HMBC spectra, respec-
tively. The bis(THP)-protected 2-aminopurines 7 were
isolated, characterized, and used as diastereomeric
mixtures. They were chromatographically homogeneous,
but splitting of some carbon signals was observed in 13C
NMR spectra.
Cytosta tic Activity Eva lu a tion
The title substituted 6-phenylpurine bases and nu-
cleosides 10-13, as well as some acetyl derivatives 8,
were tested on their in vitro inhibition of the cell growth
in the following cell cultures: mouse leukemia L1210
cells (ATCC CCL 219); murine L929 cells (ATCC CCL
1); human cervix carcinoma HeLa S3 cells (ATCC CCL
2.2); and human T-lymphoblastoid CCRF-CEM cell line
(ATCC CCL 119). Only substituted 6-phenylpurine
ribonucleosides 12 and their triacetates 8 exhibited
significant activity in these assays (Table 1), while the
bases 10 and 11, as well as the 2-amino-6-phenylpurine
ribonucleosides 13, were entirely inactive.
The results (Table 1) show that the 6-phenylpurine
ribonucleosides 12 possess powerful cytostatic potency
against T-lymphoblastoids (CCRF-CEM: IC50 values
from 0.25 to 8.3 µmol/L) while HeLa S3 cells are less
sensitive (IC50 values from 2.5 to 19 µmol/L). Markedly
reduced cytostatic activity against L1210 cells and
marginal effects toward L929 cell line (data not shown)
might reflect a difference in the transport mechanism
of mentioned compounds into the different cell lines and/
or a putative process of the metabolic activation.
The structure-activity relationship of the series of
compounds shows that the most active compounds are
6-phenylpurine nucleoside 12a and its congeners bear-
ing a functionality in position 4 of the benzene ring
(compounds 12b, 12g, 12i, and 12j). Introduction of a
substituent into position 3 of the benzene moiety causes
a substantial decrease of activity (compounds 12d and
12h ), while the presence of a substituent in position 2
leads to inactive compounds (12c and 12e). Also the
presence of an amino function in position 2 of the purine
ring causes loss of activity. The acetyl derivatives 8 of
All compounds were fully characterized by MS and
1H and 19F NMR; 13C NMR was recorded for at least
one example of each class of compounds. UV spectra of
the parent unsubstituted 6-phenylpurine bases and
ribosides 10a , 11a , 12a , and 13a were recorded, as well
as of the whole series of substituted nucleosides 12a -j
showing expectable substituent effects: bathochromic
shifts in compounds bearing electron-donating substit-
uents in the para- or meta-positions of the phenyl ring
or hypsochromic shifts in compounds bearing ortho-
substituents. All target 6-phenylpurine bases and nu-
cleosides 10-13 were crystallized and characterized by
microanalysis. Some oily protected intermediates did not
give satisfactory analysis and were used in the depro-
tection step as TLC homogeneous material. For the
assignment of the NMR signals, standard 2D techniques
were used. 1H-1H spin network was determined by
COSY spectra. Carbon connectivities based on one-bond
1
and three-bond H-13C correlations were established by

1820 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 9
Hocek et al.
(400 MHz, CDCl3): 1.6-1.9 and 2.0-2.2 (2 × m, 6 H, CH2);
3.82 (dt, 1 H, J ) 2.6 and 11.6, H-5′a); 4.21 (m, 1 H, H-5′b);
5.85 (dd, 1 H, J ) 10.3 and 2.5, H-1′); 7.33 (m, 1 H, H-Ar);
8.34 (s, 1 H, H-8); 8.66-8.78 (m, 2 H, H-Ar); 8.99 (s, 1 H,
active compounds are considerably less active than the
parent free nucleosides 12.
In conclusion, a novel type of antineoplastic com-
pounds, substituted 6-phenylpurine ribonucleosides, has
been discovered. In our view, the cytostatic activity of
this class of compounds cannot be deduced from any
known cytostatic nucleoside or purine derivatives, and
therefore, it might be considered a new structural lead
in the search for antitumor compounds.
H-2). 19F NMR (376.5 MHz, CDCl3): -134.26 (dd, J 1 ) J 2
)
9.4) and -137.52 (pent, J ) 10.8, F2Ph). Anal. (C16H14F2N4O)
C, H, N.
6-P h en yl-9-(tetr ah ydr opyr an -2-yl)-2-[(tetr ah ydr opyr an -
2-yl)a m in o]p u r in e (7a ). Yellow amorphous solid, yield 95%.
FAB MS m/z (rel. %) 380 (25) [M + H], 212 (100) [M + 2H -
1
2THP]. H NMR (400 MHz, CDCl3): 1.5-2.1 (m, 12 H, CH2);
3.66-3.78 (m, 2 H, H-5′ and H-5"a); 4.05 (d, 1 H, J ) 12.8)
and 4.15 (d, 1 H, J ) 12.1, H-5′ and H-5"b); 5.53 (t, 1 H, J )
9.2) and 5.64-5.82 (m, 2 H, H-1′, H-1" and NH); 7.46-7.53
(m, 3 H, H-Ar); 8.03 (s, 1 H, H-8); 8.66-8.70 (m, 2 H, H-Ar).
13C NMR (100 MHz, CDCl3): 22.97, 23.03, 24.96, 25.35, 31.33,
31.61 and 31.78 (CH2); 66.49, 66.57 and 68.60 (CH2-5′ and CH2-
5′′); 80.32, 80.47, 81.18 and 81.62 (CH-1′ and CH-1′′); 125.96
and 126.03 (C-5); 128.73, 129.61 and 130.55 (CH-Ph); 136.04
and 136.10 (i-C-Ph); 139.27 and 139.41 (C-8); 153.51, 153.64,
155.54 and 155.67 (C-4 and C-6); 157.75 and 157.84 (C-2).
Exp er im en ta l Section
Unless otherwise stated, solvents were evaporated at 40 °C/2
kPa and compounds were dried at 60 °C/2 kPa over P2O5.
Melting points were determined on a Kofler block and are
uncorrected. NMR spectra were measured on Bruker AMX-3
400 (400 MHz for 1H, 100.6 MHz for 13C and 376.5 MHz for
19F nuclei), Bruker DRX 500 (500 MHz for 1H, 125.7 MHz for
13C and 470.59 MHz for 19F) and Varian Gemini 300HC
(300.075 MHz for 1H and 75.462 MHz for 13C). TMS was used
as internal standard for the H and 13C NMR spectra; CFCl3
1
6-(4-F lu or op h en yl)-9-(t et r a h yd r op yr a n -2-yl)-2-[(t et -
r a h yd r op yr a n -2-yl)a m in o]p u r in e (7b). Colorless amor-
phous solid, yield 81%. FAB MS m/z (rel. %): 398 (33) [M +
was an internal standard for 19F spectra; values are given in
δ (ppm) and J values are in Hz. Mass spectra were measured
on a ZAB-EQ (VG Analytical) spectrometer using FAB (ioniza-
tion by Xe, accelerating voltage 8 kV, glycerol matrix) or EI
(electron energy 70 eV) techniques. Toluene was degassed in
vacuo and stored over molecular sieves under Ar. Substituted
phenylboronic acids 5 were supplied by Aldrich.
1
H], 230 (100) [M + 2H - 2THP]. H NMR (400 MHz, CDCl3):
1.5-2.2 (m, 12 H, CH2); 3.68-3.78 (m, 2 H, H-5′ and H-5"a);
4.04 (d, 1 H, J ) 12.2) and 4.16 (d, 1 H, J ) 11.8, H-5′ and
H-5"b); 5.50 (t, 1 H) and 5.66 (t, 1 H, J ) 10.1, H-1′ and H-1");
7.18 (t, 2 H, J ) 8.6, H-o-Ar); 8.00 (s, 1 H, H-8); 8.71-8.75
(m, 2 H, H-m-Ar). 13C NMR (100 MHz, CDCl3): 22.97, 23.02,
24.95, 25.33, 31.35, 31.61 and 31.78 (CH2); 66.52, 66.58 and
68.62 (CH2-5′ and CH2-5′′); 80.31, 80.45, 81.22 and 81.62 (CH-
1′ and CH-1′′); 115.36 (d, J ) 21.3, m-CH-FPh); 125.69 and
125.76 (C-5); 131.79 (d, J ) 8.3, o-CH-FPh); 132.26 (i-C-FPh);
139.31 and 139.44 (C-8); 153.54, 153.67, 154.26 and 154.38
(C-4 and C-6); 157.70 and 157.78 (C-2); 164.42 (d, 1J (CF) )
251.0, CF). 19F NMR (376.5 MHz, CDCl3): -110.37 (s, F-Ph).
Anal. (C21H24FN5O2) C, H; N: calcd, 17.62; found, 17.12.
Su zu k i Cou p lin g of th e 6-Ch lor op u r in es a n d Su bsti-
tu ted P h en ylbor on ic Acid s: Gen er a l P r oced u r e. Toluene
(10 mL) was added to an argon-purged flask containing a
6-chloropurine 1-4 (1 mmol), K2CO3 (200 mg, 1.5 mmol),
substituted phenylboronic acid 5 (1.5 mmol) and Pd(PPh3)4 (59
mg, 0.05 mmol) and the mixture was stirred under argon at
100 °C for 8 h. After cooling to ambient temperature the
mixture was evaporated in vacuo and the residue was chro-
matographed on a silica gel column (50 g, ethyl acetate-light
petroleum 1:2 to 9:1). Evaporation and drying of the product
containing fractions afforded the 6-phenylpurines 6-9 as
foams or amorphous solids.
6-P h en yl-9-(tetr a h yd r op yr a n -2-yl)p u r in e (6a ). Color-
less amorphous solid, yield 95%. FAB MS m/z (rel. %): 281
(40) [M + H], 197 (100) [M + H - THP], 85 (14) [THP]. 1H
NMR (400 MHz, CDCl3): 1.6-1.9 and 2.0-2.2 (2 × m, 6 H,
CH2); 3.82 (dt, 1 H, J ) 2.5, 11.6, H-5′a); 4.21 (m, 1 H, H-5′b);
5.86 (dd, 1 H, J ) 10.4 and 2.7, H-1′); 7.5-7.6 (m, 3 H, H-Ar);
8.34 (s, 1 H, H-8); 8.77-8.80 (m, 2 H, H-Ar); 9.03 (s, 1 H,
H-2). Anal. (C16H16N4O) C, H, N.
6-(2,4-Diflu or oph en yl)-9-(tetr ah ydr opyr an -2-yl)-2-[(tet-
r a h yd r op yr a n -2-yl)a m in o]p u r in e (7c). Colorless amor-
phous solid, yield 60%. FAB MS m/z (rel. %): 416 (76) [M +
1
H], 332 (100) [M + H - THP], 248 (66) [M + 2H - 2THP]. H
NMR (400 MHz, CDCl3): 1.5-2.2 (m, 12 H, CH2); 3.65-3.82
(m, 2 H, H-5′and H-5"a); 4.03 (d, 1 H, J ) 12.0) and 4.17 (d, 1
H, J ) 11.8, H-5′ and H-5"b); 5.43-5.50 (m, 1 H) and 5.65-
5.77 (m, 2 H, NH, H-1′ and H-1"); 6.93-7.04 (m, 2 H, H-Ar);
7.91-8.00 (m, 1 H, H-Ar); 8.01 (s, 1 H, H-8). 19F NMR (376.5
MHz, CDCl3): -107.83 (brs, F-Ph). Anal. (C21H23F2N5O2) C,
H, N.
6-(4-Flu or oph en yl)-9-(tetr ah ydr opyr an -2-yl)pu r in e (6b).
Colorless amorphous solid, yield 84%. FAB MS m/z (rel. %):
299 (7) [M + H], 215 (100) [M + H - THP]. 1H NMR (400
MHz, CDCl3): 1.6-1.9 and 2.0-2.2 (2 × m, 6 H, CH2); 3.81
(dt, 1 H, J ) 2.2 and 11.5, H-5′a); 4.20 (brd, 1 H, J ) 11.5,
H-5′b); 5.84 (dd, 1 H, J ) 10.3 and 2.3, H-1′); 7.22 (t, 2 H, J )
8.7, H-o-Ar); 8.31 (s, 1 H, H-8); 8.84 (dd, 2 H, J ) 8.7 and 5.7,
H-m-Ar); 8.99 (s, 1 H, H-2). 13C NMR (100 MHz, CDCl3):
22.78, 24.87 and 31.86 (CH2); 68.87 (CH2-5′); 82.02 (CH-1′);
115.67 (d, J ) 21.4, m-CH-FPh); 130.84 (C-5); 131.73 (i-C-
FPh); 132.01 (d, J ) 8.5, o-CH-FPh); 142.04 (C-8); 152.36 (C-
2); 151.71 and 153.71 (C-4 and C-6); 164.64 (d, 1J (C,F) ) 252.0,
CF). 19F NMR (376.5 MHz, CDCl3): -109.56 (s, F-Ph). Anal.
(C16H15FN4O) C, H, N.
6-(3,4-Diflu or oph en yl)-9-(tetr ah ydr opyr an -2-yl)-2-[(tet-
r a h yd r op yr a n -2-yl)a m in o]p u r in e (7d ). Yellow amorphous
solid, yield 60%. FAB MS m/z (rel. %): 416 (88) [M + H], 332
1
(100) [M + H - THP], 248 (93) [M + 2H - 2THP]. H NMR
(400 MHz, CDCl3): 1.5-2.2 (m, 12 H, CH2); 3.65-3.80 (m, 2
H, H-5′and H-5"a); 4.04 (d, 1 H, J ) 11.5) and 4.15 (d, 1 H, J
) 10.9, H-5′ and H-5"b); 5.50 (brt, 1 H, J ) 9.6) and 5.62-
5.77 (m, 2 H, NH, H-1′ and H-1"); 7.23-7.31 (m, 1 H, H-Ar);
8.03 (s, 1 H, H-8); 8.55-8.62 (m, 2 H, H-Ar). 19F NMR (376.5
MHz, CDCl3): -134.93 and -137.95 (2 × brs, F2Ph).
9-(2,3,5-Tr i-O-a cet yl-â-D-r ib ofu r a n osyl)-6-p h en ylp u -
r in e (8a ). Colorless amorphous solid, yield 79%. FAB MS m/z
(rel. %): 455 (22) [M + H], 197 (100) [M + H - AcRf]. 1H NMR
(400 MHz, CDCl3): 2.09, 2.13 and 2.16 (3 × s, 3 × 3 H, CH3);
4.37-4.50 (m, 3 H, H-4′ and 2 × H-5′); 5.71 (dd, 1 H, J ) 3.6
and 5.4, H-3′); 6.02 (t, 1 H, J ) 5.4, H-2′); 6.30 (d, 1 H, J )
5.3, H-1′); 7.52-7.58 (m, 3 H, H-Ph); 8.28 (s, 1 H, H-8); 8.76
(dd, 2 H, J ) 1.6 and 8.0, H-Ph); 9.03 (s, 1 H, H-2). Anal.
(C22H22N4O7) C, H, N.
6-(2,4-Diflu or op h e n yl)-9-(t e t r a h yd r op yr a n -2-yl)p u -
r in e (6c). Colorless amorphous solid, yield 66%. FAB MS m/z
(rel. %): 317 (19) [M + H], 233 (100) [M + H - THP], 85 (30)
[THP]. 1H NMR (400 MHz, CDCl3): 1.60-1.85 and 2.0-2.2 (2
× m, 6 H, CH2); 3.75-3.85 (m, 1 H, H-5′a); 4.16-4.23 (m, 1 H,
H-5′b); 5.76-5.88 (m, 1 H, H-1′); 6.97-7.09 (m, 2 H, H-Ar);
8.04 (q, 1 H, J ) 6.7, H-Ar); 8.34 (s, 1 H, H-8); 9.07 (s, 1 H,
H-2). 19F NMR (376.5 MHz, CDCl3): -106.90 and -108.35 (2
× m, F2Ph).
9-(2,3,5-Tr i-O-a ce t yl-â-D-r ib ofu r a n osyl)-6-(4-flu or o-
p h en yl)p u r in e (8b). Colorless amorphous solid, yield 87%.
FAB MS m/z (rel. %): 473 (8) [M + H], 215 (100) [M + H -
AcRf]. 1H NMR (400 MHz, CDCl3): 2.07, 2.12 and 2.15 (3 × s,
3 × 3 H, CH3); 4.35-4.47 (m, 3 H, H-4 and 2 × H-5); 5.69 (dd,
1 H, J ) 4.1 and 5.2, H-3′); 6.00 (t, 1 H, J ) 5.2, H-2′); 6.27 (d,
6-(3,4-Diflu or op h e n yl)-9-(t e t r a h yd r op yr a n -2-yl)p u -
r in e (6d ). Colorless amorphous solid, yield 70%. FAB MS m/z
(rel. %): 317 (24) [M + H], 233 (100) [M + H - THP]. 1H NMR

Substituted 6-Phenylpurine Bases and Nucleosides
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 9 1821
1 H, J ) 5.1, H-1′); 7.22 (t, 2 H, J ) 8.4, H-o-Ar); 8.26 (s, 1 H,
H-8); 8.82 (m, 2 H, H-m-Ar); 8.98 (s 1 H, H-2). 13C NMR (100
MHz, CDCl3): 21.05, 21.19 and 21.42 (CH3); 63.75 (CH2-5′);
71.37 (CH-3′); 73.82 (CH-2′); 81.13 (CH-4′); 87.17 (CH-1′);
116.46 (d, J ) 21.6, m-CH-FPh); 132.07 (i-C-FPh); 132.29
(C-5); 132.81 (d, J ) 8.3, o-CH-FPh); 143.23 (C-8); 152.76 (C-
NMR (500 MHz, CDCl3): 1.46 (t, 3 H, J ) 6.9, CH3CH2); 2.09,
2.14 and 2.16 (3 × s, 3 × 3 H, CH3); 4.14 (q, 2 H, J ) 6.9,
CH2CH3); 4.38-4.49 (m, 3 H, H-4′ and 2 × H-5′); 5.71 (dd, 1
H, J ) 4.3 and 5.4, H-3′); 6.01 (t, 1 H, J ) 5.4, H-2′); 6.29 (d,
1 H, J ) 5.3, H-1′); 7.06 (d, 2 H, J ) 8.8, H-Ph); 8.24 (s, 1 H,
H-8); 8.78 (d, 2 H, J ) 8.8, H-Ph); 8.96 (s, 1 H, H-2). Anal.
(C24H26N4O8) C, H, N.
1
4); 153.34 (C-2); 154.93 (C-6), 165.48 (d, J (C,F) ) 251.0, CF);
170.03, 170.24 and 170.96 (3 × CO). 19F NMR (376.5 MHz,
CDCl3): -109.12 (s, F-Ph). Anal. (C22H21FN4O7) C, H, N.
9-(2,3,5-Tr i-O-a cet yl-â-D-r ib ofu r a n osyl)-6-(2,4-d iflu o-
r op h en yl)p u r in e (8c). Colorless amorphous solid, yield 65%.
FAB MS m/z (rel. %): 491 (19) [M + H], 233 (100) [M + H -
AcRf]. 1H NMR (400 MHz, CDCl3): 2.11, 2.13 and 2.17 (3 × s,
3 × 3 H, CH3); 4.38-4.50 (m, 3 H, H-4 and 2 × H-5); 5.71 (dd,
1 H, J ) 4.6 and 5.4, H-3′); 6.03 (t, 1 H, J ) 5.4, H-2′); 6.30 (d,
1 H, J ) 5.3, H-1′); 7.00-7.12 (m, 2 H, H-Ar); 8.03 (m, 1 H,
H-Ar); 8.28 (s, 1 H, H-8); 9.09 (s, 1 H, H-2). 19F NMR (376.5
MHz, CDCl3): -106.45 (t, J ) 7.3, FPh); -108.18 (q, J ) 8.8,
FPh). Anal. (C22H20F2N4O7) C, H, N.
9-(2,3,5-Tr i-O-a cet yl-â-D-r ib ofu r a n osyl)-6-(3,4-d iflu o-
r op h en yl)p u r in e (8d ). Colorless amorphous solid, yield 81%.
FAB MS m/z (rel. %): 491 (100) [M + H], 233 (66) [M + H -
AcRf]. 1H NMR (400 MHz, CDCl3): 2.09, 2.14 and 2.17 (3 × s,
9 H, CH3); 4.38-4.50 (m, 3 H, H-4 and 2 × H-5); 5.70 (m, 1 H,
H-3′); 6.00 (m, 1 H, H-2′); 6.28 (d, 1 H, J ) 5.2, H-1′); 7.29-
7.38 (m, 1 H, H-Ar); 8.28 (s, 1 H, H-8); 8.64-8.77 (m, 2 H,
H-Ar); 9.01 (s, 1 H, H-2). 19F NMR (376.5 MHz, CDCl3):
-133.76 and -137.32 (2 × m, F2Ph). Anal. (C22H20F2N4O7) C,
H, N.
9-(2,3,5-Tr i-O-a cet yl-â-D-r ib ofu r a n osyl)-6-(2-t olyl)p u -
r in e (8e). Colorless amorphous solid, yield 89%. FAB MS m/z
(rel. %): 469 (18) [M + H], 211 (100) [M + H - AcRf]. 1H NMR
(400 MHz, CDCl3): 2.10, 2.11 and 2.15 (3 × s, 3 × 3 H, CH3);
2.43 (s, 3 H, CH3Ph); 4.37-4.49 (m, 3 H, H-4′ and 2 × H-5′);
5.72 (dd, 1 H, J ) 5.0 and 5.3, H-3′); 6.02 (t, 1 H, J ) 5.3,
H-2′); 6.27 (d, 1 H, J ) 5.1, H-1′); 7.7.3-7.4 (m, 3 H, H-Ar);
7.66-7.69 (m, 1 H, H-Ar); 8.21 (s, 1 H, H-8); 9.05 (s 1 H, H-2).
13C NMR (100 MHz, CDCl3): 21.05-21.27 (m, 4 × CH3); 63.59
(C-5′); 71.20 (C-3′); 73.71 (C-2′); 80.98 (C-4′); 87.22 (C-1′);
126.34, 130.38, 131.34, 131.73 (CH-arom); 135.25, 137.75 (C-
arom); ∼143 (very weak, C-8); 151.93 (C-4); 153.07 (C-2);
160.17 (C-6), 169.89-170.80 (m, 3 × CO). Anal. (C23H24N4O7)
C, H, N.
9-(2,3,5-Tr i-O-a cet yl-â-D-r ib ofu r a n osyl)-6-(4-t olyl)p u -
r in e (8f). Colorless amorphous solid, yield 79%. FAB MS m/z
(rel. %): 469 (22) [M + H], 211 (100) [M + H - AcRf]. 1H NMR
(400 MHz, CDCl3): 2.07, 2.12 and 2.15 (3 × s, 3 × 3 H, CH3);
2.44 (s, 3 H, CH3Ph); 4.36-4.48 (m, 3 H, H-4′ and 2 × H-5′);
5.70 (dd, 1 H, J ) 4.5 and 5.4, H-3′); 6.00 (t, 1 H, J ) 5.4,
H-2′); 6.28 (d, 1 H, J ) 5.3, H-1′); 7.36 (d, 2 H, J ) 8.1, H-Ph);
8.24 (s, 1 H, H-8); 8.67 (d, 2 H, J ) 8.1, H-Ph); 8.99 (s, 1 H,
H-2). Anal. (C23H24N4O7) C, H, N.
9-(2,3,5-Tr i-O-a ce t yl-â-D-r ib ofu r a n osyl)-6-(4-ch lor o-
p h en yl)p u r in e (8j). Colorless amorphous solid, yield 65%.
FAB MS m/z (rel. %): 489 (19) [M + H], 231 (100) [M + H -
AcRf]. 1H NMR (500 MHz, CDCl3): 2.09, 2.14 and 2.17 (3 × s,
3 × 3 H, CH3); 4.39-4.51 (m, 3 H, H-4′ and 2 × H-5′); 5.71
(dd, 1 H, J ) 4.7 and 5.4, H-3′); 6.01 (t, 1 H, J ) 5.4, H-2′);
6.30 (d, 1 H, J ) 5.3, H-1′); 7.53 (d, 2 H, J ) 8.6, H-Ph); 8.29
(s, 1 H, H-8); 8.77 (d, 2 H, J ) 8.6, H-Ph); 9.02 (s, 1 H, H-2).
Anal. (C22H21ClN4O7) C, H, N.
9-(2,3,5-Tr i-O-a ce t yl-â-D-r ib ofu r a n osyl)-2-a m in o-6-
p h en ylp u r in e (9a ). Yellowish amorphous solid, yield 83%.
FAB MS m/z (rel. %): 470 (30) [M + H], 212 (100) [M + H -
AcRf]. 1H NMR (400 MHz, CDCl3): 2.09, 2.10 and 2.15 (3 × s,
3 × 3 H, CH3); 4.35-4.50 (m, 3 H, H-4 and 2 × H-5); 5.13 (s,
2 H, NH2); 5.84 (dd, 1 H, J ) 4.5 and 5.3, H-3′); 6.04 (dd, 1 H,
J ) 4.9 and 5.3, H-2′); 6.08 (d, 1 H, J ) 4.9, H-1′); 7.49-7.54
(m, 3 H, H-Ph); 7.90 (s, 1 H, H-8); 8.60-8.63 (m, 2 H, H-Ph).
Anal. (C22H23N5O7) C, H, N.
9-(2,3,5-Tr i-O-a cet yl-â-D-r ib ofu r a n osyl)-2-a m in o-6-(4-
flu or op h en yl)p u r in e (9b). Colorless amorphous solid, yield
80%. FAB MS m/z (rel. %): 488 (14) [M + H], 230 (100) [M +
H - AcRf]. 1H NMR (400 MHz, CDCl3): 2.08, 2.09 and 2.14 (3
× s, 3 × 3 H, CH3); 4.35-4.50 (m, 3 H, H-4 and 2 × H-5); 5.10
(s, 2 H, NH2); 5.82 (t, 1 H, J ) 4.8, H-3′); 6.02 (t, 1 H, J ) 4.9,
H-2′); 6.07 (d, 1 H, J ) 4.8, H-1′); 7.20 (m, 2 H, H-o-Ar); 7.89
(s, 1 H, H-8); 8.69 (m, 2 H, H-m-Ar). 13C NMR (100 MHz,
CDCl3): 21.41, 20.52 and 20.68 (CH3); 63.04 (CH2-5′); 70.61
(CH-3′); 72.79 (CH-2′); 79.89 (CH-4′); 86.32 (CH-1′); 115.49 (d,
J ) 21.1, m-CH-FPh); 125.80 (C-5); 131.78 (d, J ) 8.7, o-CH-
FPh); 139.91 (C-8); 153.80 (C-4); 155.21 (C-6); 159.52 (C-2),
1
164.55 (d, J (C,F) ) 251.6, CF); 169.35, 169.56 and 170.47 (3
× CO). 19F NMR (376.5 MHz, CDCl3): -109.81 (s, F-Ph). Anal.
(C22H22FN5O7) C, H, N.
9-(2,3,5-Tr i-O-a cetyl-â-D-r ibofu r a n osyl)-2-a m in o-6-(2,4-
d iflu or op h en yl)p u r in e (9c). Colorless amorphous solid,
yield 75%. FAB MS m/z (rel. %): 506 (27) [M + H], 248 (100)
1
[M + H - AcRf]. H NMR (400 MHz, CDCl3): 2.08, 2.11 and
2.16 (3 × s, 3 × 3 H, CH3); 4.35-4.50 (m, 3 H, H-4 and 2 ×
H-5); 5.20 (s, 2 H, NH2); 5.83 (dd, 1 H, J ) 4.3 and 5.2, H-3′);
6.04 (dd, 1 H, J ) 4.9 and 5.2, H-2′); 6.07 (d, 1 H, J ) 4.9,
H-1′); 6.95-7.06 (m, 2 H, H-Ar); 7.85-7.91 (m, 1 H, H-Ar);
7.89 (s, 1 H, H-8). 19F NMR (376.5 MHz, CDCl3): -108.20 and
-107.22 (2 × m, F2Ph). Anal. (C22H21F2N5O7) C, H, N.
9-(2,3,5-Tr i-O-a cetyl-â-D-r ibofu r a n osyl)-2-a m in o-6-(3,4-
d iflu or op h en yl)p u r in e (9d ). Colorless amorphous solid,
yield 81%. FAB MS m/z (rel. %): 506 (35) [M + H], 248 (100)
9-(2,3,5-Tr i-O-a cetyl-â-D-r ibofu r a n osyl)-6-(4-m eth oxy-
p h en yl)p u r in e (8g). Colorless amorphous solid, yield 84%.
FAB MS m/z (rel. %): 485 (24) [M + H], 227 (100) [M + H -
AcRf]. 1H NMR (400 MHz, CDCl3): 2.07, 2.12 and 2.14 (3 × s,
3 × 3 H, CH3); 3.88 (s, 3 H, OCH3); 4.36-4.48 (m, 3 H, H-4′
and 2 × H-5′); 5.69 (dd, 1 H, J ) 4.5 and 5.4, H-3′); 5.99 (t, 1
H, J ) 5.4, H-2′); 6.28 (d, 1 H, J ) 5.4, H-1′); 7.06 (d, 2 H, J
) 9.0, H-Ph); 8.22 (s, 1 H, H-8); 8.79 (d, 2 H, J ) 9.0, H-Ph);
8.95 (s, 1 H, H-2). Anal. (C23H24N4O8) C, H, N.
1
[M + H - AcRf]. H NMR (400 MHz, CDCl3): 2.10, 2.11 and
2.16 (3 × s, 3 × 3 H, CH3); 4.35-4.50 (m, 3 H, H-4 and 2 ×
H-5); 5.12 (s, 2 H, NH2); 5.83 (t, 1 H, J ) 5.0, H-3′); 6.03 (t, 1
H, J ) 5.0, H-2′); 6.08 (d, 1 H, J ) 4.9, H-1′); 7.25-7.33 (m, 1
H, H-Ar); 7.91 (s, 1 H, H-8); 8.52-8.62 (m, 2 H, H-Ar). 19F
NMR (376.5 MHz, CDCl3): -137.75 and -134.48 (2 × m, F2-
Ph). Anal. (C22H21F2N5O7) C, H, N.
Clea va ge of th e THP -P r otected P u r in es 6 a n d 7:
Gen er a l P r oced u r e. A mixture of a THP-protected base 6
or 7 (0.6-0.8 mmol), Dowex 50×8 (H+) (ca. 300 mg), methanol
(10 mL) and water (1 mL) was refluxed for 1 h, then filtered
while hot and the resin was washed with saturated methanolic
ammonia (5 mL) followed by methanol (20 mL). The combined
filtrates were evaporated and the residue was codistilled with
toluene. Crystallization of the residue from methanol/toluene
with an addition of heptane afforded the free bases 10 or 11.
6-P h en ylp u r in e (10a ). Colorless crystals, yield 92%, mp
280-282 °C (lit.36 243 °C). EI MS m/z (rel. %): 196 (67) [M],
169 (54) [M - HCN], 141 (20) [M - 2 HCN], 41 (100). 1H NMR
(400 MHz, DMSO-d6): 7.54-7.62 (m, 3 H, H-Ar); 8.64 (s, 1
H, H-8); 8.83 (brm, 2 H, H-Ar); 8.96 (s, 1 H, H-2). UV (λmax
9-(2,3,5-Tr i-O-a cetyl-â-D-r ibofu r a n osyl)-6-(3-m eth oxy-
p h en yl)p u r in e (8h ). Colorless amorphous solid, yield 74%.
FAB MS m/z (rel. %): 485 (13) [M + H], 227 (100) [M + H -
AcRf]. 1H NMR (400 MHz, CDCl3): 2.12, 2.17 and 2.19 (3 × s,
3 × 3 H, CH3); 3.96 (s, 3 H, CH3O); 4.41-4.52 (m, 3 H, H-4′
and 2 × H-5′); 5.73 (dd, 1 H, J ) 4.5 and 5.4, H-3′); 6.04 (t, 1
H, J ) 5.4, H-2′); 6.33 (d, 1 H, J ) 5.3, H-1′); 7.12 (dd, 1 H, J
) 8.0, 2.6, H-Ph); 7.50 (t, 1 H, J ) 8.0, H-Ph); 8.30 (s, 1 H,
H-8); 8.36 (dd, 1 H, J ) 1.9, 2.4, H-Ph); 8.44 (d, 1 H, J ) 7.8,
H-Ph); 9.05 (s 1 H, H-2). Anal. (C23H24N4O8) C, H, N.
9-(2,3,5-Tr i-O-acetyl-â-D-r ibofu r an osyl)-6-(4-eth oxyph en -
yl)p u r in e (8i). Colorless amorphous solid, yield 80%. FAB MS
1
m/z (rel. %): 499 (30) [M + H], 241 (100) [M + H - AcRf]. H

1822 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 9
Hocek et al.
(ꢀ)) methanol: 289 (12900); water: pH 7, 288 (8600); pH 2,
298 (42100); pH 11, 298 (34700). Anal. (C11H8N4) C, H, N.
6-(4-F lu or op h en yl)p u r in e (10b). Colorless crystals, yield
83%, mp 299-302 °C. EI MS m/z (rel. %): 214 (100) [M], 187
FAB MS m/z (rel. %): 329 (35) [M + H], 197 (75) [M + H -
1
Rf]. H NMR (400 MHz, DMSO-d6): 3.31 (s, 3 H, CH3); 3.59-
3.65 and 3.70-3.76 (2 × m, 2 H, CH2-5′); 4.02 (m, 1 H, H-4′);
4.23 (m, 1 H, H-3′); 4.67 (ddd, 1 H, J ) 4.8, 5.5 and 5.7, H-2′);
5.13 (t, 1 H, J ) 5.4, 5′-OH); 5.25 (d, 1 H, J ) 5.0, 3′-OH); 5.56
(d, 1 H, J ) 5.7, 2′-OH); 6.11 (d, 1 H, J ) 5.5, H-1′); 7.55-7.65
(m, 3 H, H-Ph); 8.84 (d, 2 H, J ) 8.0, H-Ph); 8.93 (s, 1 H,
H-8); 9.02 (s, 1 H, H-2). UV (λmax (ꢀ)) methanol: 290 (18300);
water: pH 7, 289 (16800); pH 2, 298 (15700); pH 11, 289
(17100). Anal. (C16H16N4O4) C, H, N.
6-(4-F lu or op h en yl)-9-(â-D-r ibofu r a n osyl)p u r in e (12b).
Colorless crystals, yield 83%, mp 207-210 °C; [R]D -51.9 (c
0.5 DMF). FAB MS m/z (rel. %): 347 (6) [M + H], 215 (7) [M
+ H - Rf], 185 (40), 93 (100). 1H NMR (400 MHz, CDCl3):
3.58-3.64 and 3.70-3.75 (2 × m, 2 H, CH2-5′); 4.01 (m, 1 H,
H-4′); 4.23 (m, 1 H, H-3′); 4.66 (ddd, 1 H, J ) 4.9, 5.5 and 5.9,
H-2′); 5.13 (t, 1 H, J ) 5.5, 5′-OH); 5.24 (d, 1 H, J ) 5.1, 3′-
OH); 5.55 (d, 1 H, J ) 5.9, 2′-OH); 6.10 (d, 1 H, J ) 5.5, H-1′);
7.46 (t, 2 H, J ) 8.9, H-o-Ar); 8.89-8.94 (m, 2 H, H-m-Ar);
8.94 (s, 1 H, H-8); 9.01 (s, 1 H, H-2). 13C NMR (100 MHz,
DMSO-d6): 63.75 (CH2-5′); 71.33 (CH-3′); 73.88 (CH-2′); 85.78
(CH-4′); 87.86 (CH-1′); 115.89 (d, J ) 21.4, m-CH-FPh); 130.71
and 131.77 (i-C-FPh and C-5); 131.97 (d, J ) 8.7, o-CH-FPh);
145.11 (C-8); 151.96 (C-2); 152.01 and 152.31 (C-4 and C-6);
164.06 (d, 1J (C,F) ) 250.0, CF). 19F NMR (376.5 MHz, CDCl3):
-108.65 (m, FPh). UV (λmax (ꢀ)) methanol: 294 (25400). Anal.
(C16H15FN4O4) C, H, N.
1
(46) [M - HCN]. H NMR (400 MHz, CDCl3): 7.44 (t, 2 H, J
) 8.9, H-o-Ar); 8.67 (s, 1 H, H-8); 8.90-9.00 (brm, 2 H, H-m-
Ar); 8.96 (s, 1 H, H-2); 13.65 (brs, 1 H, NH). 13C NMR (100
MHz, CDCl3): 115.49 and 115.70 (CH-arom); 131.60 and
131.68 (CH-arom); 132.13 (C-5); 144.95 (C-8); ∼151 (very weak,
C-4); 151.77 (C-2); ∼154 (very weak, C-6); 163.71 (d, J (C,F) )
247.4, CF). 19F NMR (376.5 MHz, CDCl3): -109.24 (s, FPh).
Anal. (C11H7FN4) C, H. N.
6-(2,4-Diflu or op h en yl)p u r in e (10c). Yellowish crystals,
yield 81%, mp 304-307 °C. EI MS m/z (rel. %): 232 (100) [M],
205 (58) [M - HCN]. 1H NMR (400 MHz, DMSO-d6): 7.32 (m,
1 H, H-Ar); 7.48 (m, 1 H, H-Ar); 8.04 (br, 1 H, H-Ar); 8.65
(s, 1 H, H-8); 9.00 (s, 1 H, H-2); 13.56 (brs, 1 H, NH). 19F NMR
(376.5 MHz, DMSO-d6): -106.71 and -107.35 (2 × brs, F2-
Ph). Anal. (C11H6F2N4) C, H, N.
6-(3,4-Diflu or op h en yl)p u r in e (10d ). Colorless crystals,
yield 85%, mp 283-285 °C. EI MS m/z (rel. %): 232 (100) [M],
205 (52) [M - HCN]. 1H NMR (400 MHz, DMSO-d6): 7.67 (m,
1 H, H-Ar); 8.69 (s, 1 H, H-8); 8.73-8.77 (brm, 1 H, H-Ar);
8.80-8.87 (m, 1 H, H-Ar); 8.96 (s, 1 H, H-2). 19F NMR (376.5
MHz, DMSO-d6): -134.62 and -137.36 (2 × m, F2Ph). Anal.
(C11H6F2N4) C, H, N.
2-Am in o-6-p h en ylp u r in e (11a ). Yellowish crystals, yield
88%, mp 111-114 °C (loss of H2O), 249-252 °C (lit.37 257-
259 °C). EI MS m/z (rel. %): 211 (30) [M], 91 (90), 43 (100). 1H
NMR (400 MHz, DMSO-d6): 6.41 (brs, 2 H, NH2); 7.50-59 (m,
3 H, H-Ar); 8.14 (s, 1 H, H-8); 8.68-8.72 (brm, 2 H, H-Ar).
UV (λmax (ꢀ)) methanol: 334 (7900), 260 sh (11500), 243
(14300); water: pH 7, 327 (8400), 258 sh (10800), 244 (12600);
pH 2, 340 (10200), 259 (9100); pH 11, 331 (8100). Anal.
(C11H9N5‚1/2H2O) C, H, N.
2-Am in o-6-(4-flu or op h en yl)p u r in e (11b). Yellowish crys-
tals, yield 78%, mp 261-264 °C (H2O). EI MS m/z (rel. %):
229 (100) [M], 202 (6) [M - HCN], 149 (37). 1H NMR (400 MHz,
DMSO-d6): 6.40 (brs, 2 H, NH2); 7.38 (dt, 1 H, J ) 8.9 and
2.0, H-o-Ar); 8.14 (s, 1 H, H-8); 8.82 (m, 1 H, H-Ar); 12.70
(brs, 1 H, NH). 13C NMR (100 MHz, DMSO-d6): 115.09 and
115.30 (CH-arom); 132.40 (C-Ph); 131.29 and 131.37 (CH-
arom); 132.64 (C-5); 140.85 (C-8); 151.63 (C-4); 155.63 (C-6);
160.05 (C-2); 163.42 (d, J ) 246.6, CF). 19F NMR (376.5 MHz,
DMSO-d6): -110.10 (s, FPh). Anal. (C11H8FN5‚H2O) C, H, N.
2-Am in o-6-(2,4-d iflu or op h en yl)p u r in e (11c). Yellowish
crystals, yield 78%, mp 252-254 °C. EI MS m/z (rel. %): 247
(100) [M], 228 (29) [M - F], 220 (10) [M - HCN]. 1H NMR
(400 MHz, DMSO-d6): 6.49 (brs, 2 H, NH2); 7.25 (dt, 1 H, J )
8.5 and 2.3, H-Ar); 7.40 (dt, 1 H, J ) 10.4 and 2.4, H-Ar);
7.89 (q, 1 H, J ) 8.5, H-Ar); 8.12 (s, 1 H, H-8). 19F NMR (376.5
MHz, DMSO-d6): -107.43 (q, J ) 9.0) and -107.60 (pent, J )
7.9, F2Ph). Anal. (C11H7F2N5‚1/2H2O) C, H, N.
2-Am in o-6-(3,4-d iflu or op h en yl)p u r in e (11d ). Yellowish
crystals, yield 80%, mp 295-298 °C. EI MS m/z (rel. %): 247
(100) [M], 220 (5) [M - HCN]. 1H NMR (400 MHz, DMSO-d6):
6.37 (s, 2 H, NH2); 7.62 (m, 1 H, H-Ar); 8.14 (s, 1 H, H-8);
8.65-8.69 (brm, 1 H, H-Ar); 8.76 (ddd, 1 H, J ) 1.9, 8.2 and
10.2, H-Ar). 19F NMR (376.5 MHz, DMSO-d6): -135.70 and
-138.01 (2 × m, F2Ph). Anal. (C11H7F2N5) C, H, N.
6-(2,4-Diflu or oph en yl)-9-(â-D-r ibofu r an osyl)pu r in e (12c).
Colorless crystals, yield 91%, mp 89-92 °C; [R]D -49.0 (c 0.5
DMF). FAB MS m/z (rel. %): 365 (20) [M + H], 233 (24) [M +
1
H - Rf], 93 (100). H NMR (400 MHz, DMSO-d6): 3.57-3.63
and 3.68-3.75 (2 × m, 2 H, CH2-5′); 4.01 (m, 1 H, H-4′); 4.23
(m, 1 H, H-3′); 4.69 (ddd, 1 H, J ) 5.3, 5.7 and 5.9, H-2′); 5.10
(t, 1 H, J ) 5.5, 5′-OH); 5.25 (d, 1 H, J ) 5.0, 3′-OH); 5.56 (d,
1 H, J ) 5.9, 2′-OH); 6.10 (d, 1 H, J ) 5.7, H-1′); 7.0-7.35 (m,
1 H, H-5-Ph); 7.48 (ddd, 1 H, J ) 2.3, 9.7 and 10.7, H-3-Ph);
8.06 (dt, 1 H, J ) 8.5 and 6.7, H-6-Ar); 8.90 (s, 1 H, H-8); 9.05
(s, 1 H, H-2). 13C NMR (100 MHz, DMSO-d6): 61.21 (CH2-5′);
70.27 (CH-3′); 73.69 (CH-2′); 85.72 (CH-4′); 87.67 (CH-1′);
104.83 (t, J ) 25.9, CH-3-Ph); 111.91 (d, J ) 21.7, CH-5-Ph);
120.12 (C-1-Ph); 131.96 (C-5); 133.65 (d, J ) 6.7, CH-6-Ph);
145.25 (C-8); 151.22 (C-6); 151.62 (C-4); 151.92 (C-2); 160.35
(dd, J ) 12.8 and 256.0, CF-2-Ph); 163.51 (dd, J ) 11.8 and
250.3, CF-4-Ph). 19F NMR (376.5 MHz, DMSO-d6): -106.30
(pent, J ) 7.8) and -107.30 (q, J ) 9.6, F2Ph). UV (λmax (ꢀ))
methanol: 280 (26300). Anal. (C16H14F2N4O4) C, H, N.
6-(3,4-Diflu or op h e n yl)-9-(â-D-r ib ofu r a n osyl)p u r in e
(12d ).Colorless crystals, yield 93%, mp 189-191 °C; [R]D -48.0
(c 0.5 DMF). FAB MS m/z (rel. %): 365 (47) [M + H], 233 (100)
[M + H - Rf]. 1H NMR (400 MHz, DMSO-d6): 3.58-3.64 and
3.70-3.76 (2 × m, 2 H, CH2-5′); 4.02 (m, 1 H, H-4′); 4.23 (m,
1 H, H-3′); 4.66 (ddd, 1 H, J ) 4.7, 5.4 and 5.9, H-2′); 5.13 (t,
1 H, J ) 5.4, 5′-OH); 5.25 (d, 1 H, J ) 5.0, 3′-OH); 5.56 (d, 1
H, J ) 5.9, 2′-OH); 6.11 (d, 1 H, J ) 5.4, H-1′); 7.68 (q, 1 H, J
) 8.6, H-arom); 7.71-8.83 (m, 2 H, H-arom); 8.97 (s, 1 H, H-8);
9.02 (s, 1 H, H-2). 13C NMR (100 MHz, DMSO-d6): 61.11 (CH2-
5′); 70.15 (CH-3′); 73.79 (CH-2′); 85.64 (CH-4′); 87.76 (CH-1′);
117.87 (d, J ) 6.0, CH-5-Ph); 118.03 (CH-2-Ph); 126.57 (CH-
6-Ph); 130.67 (C-1-Ph); 132.71 (C-5); 145.32 (C-8); 149.48 (dd,
J ) 12.8 and 244.2, CF-Ph); 150.30 (C-6); 151.2248 (dd, J )
12.2 and 250.0, CF-Ph); 151.80 (C-2); 152.32 (C-4). 19F NMR
(376.5 MHz, DMSO-d6): -133.97 and -137.13 (2 × m, F2Ph).
UV (λmax (ꢀ)) methanol: 298 (36700). Anal. (C16H14F2N4O4) C,
H, N.
9-(â-D-Ribofu r a n osyl)-6-(2-tolyl)p u r in e (12e). Colorless
crystals, yield 91%, mp 79-81 °C; [R]D -51.6 (c 0.5 DMF). FAB
MS m/z (rel. %): 343 (12) [M + H], 211 (100) [M + H - Rf].
1H NMR (400 MHz, DMSO-d6): 2.37 (s, 3 H, CH3); 3.58-3.64
and 3.70-3.76 (2 × m, 2 H, CH2-5′); 4.02 (m, 1 H, H-4′); 4.23
(m, 1 H, H-3′); 4.71 (m, 1 H, H-2′); 5.14 (brs, 1 H, 5′-OH); 5.28
(brs, 1 H, 3′-OH); 5.58 (d, 1 H, J ) 5.6, 2′-OH); 6.11 (d, 1 H, J
) 5.8, H-1′); 7.12-7.68 (m, 4 H, H-Ph); 8.86 (s, 1 H, H-8);
9.04 (s, 1 H, H-2). UV (λmax (ꢀ)) methanol: 274 (23000). Anal.
(C17H18N4O4) C, H, N.
Dea cetyla tion of th e P r otected Nu cleosid es 8 a n d 9:
Gen er a l P r oced u r e. A 1 M solution of NaOMe (200 µL, 0.2
mmol) was added to the solution of the protected nucleoside 8
or 9 (0.5-0.8 mmol) in MeOH (20 mL) and the mixture was
stirred at ambient temperature overnight. The crystals (if
formed) were filtered off. Then the solution was neutralized
by addition of Dowex 50×8 (H+) (ca. 100 mg) and filtered. The
ion-exchanger was washed with saturated methanolic am-
monia (5 mL) followed by methanol (20 mL) and the combined
filtrates were evaporated to dryness. The collected crystals and
residue was recrystallized from EtOH/toluene to give the
nucleosides 12 or 13.
6-P h en yl-9-(â-D-r ibofu r a n osyl)p u r in e (12a ). Colorless
crystals, yield 66%, mp 228-230 °C; [R]D -56.1 (c 0.5 DMF).

Substituted 6-Phenylpurine Bases and Nucleosides
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 9 1823
9-(â-D-Ribofu r a n osyl)-6-(4-tolyl)p u r in e (12f). Colorless
crystals, yield 76%, mp 226-229 °C; [R]D -59.0 (c 0.5 DMF).
FAB MS m/z (rel. %): 343 (57) [M + H], 211 (72) [M + H -
(10100), 227 (23400); pH 11, 327 (9100), 247 (13800). Anal.
(C16H17N5O4) C, H, N.
2-Am in o-6-(4-flu or op h en yl)-9-(â-D-r ib ofu r a n osyl)p u -
r in e (13b). Yellowish crystals, yield 81%, mp 199-202 °C; [R]D
-29.1 (c 0.4 DMF). FAB MS m/z (rel. %): 362 (20) [M + H],
1
Rf], 201 (10), 185 (34), 93 (100). H NMR (400 MHz, DMSO-
d6): 3.31 (s, 3 H, CH3); 3.57-3.64 and 3.70-3.76 (2 × m, 2 H,
CH2-5′); 4.01 (m, 1 H, H-4′); 4.23 (m, 1 H, H-3′); 4.66 (ddd, 1
H, J ) 5.0, 5.5 and 5.9, H-2′); 5.12 (t, 1 H, J ) 5.6, 5′-OH);
5.22 (d, 1 H, J ) 5.1, 3′-OH); 5.53 (d, 1 H, J ) 5.9, 2′-OH);
6.10 (d, 1 H, J ) 5.5, H-1′); 7.42 (d, 2 H, J ) 8.2, H-Ph); 8.76
(d, 2 H, J ) 8.2, H-Ph); 8.90 (s, 1 H, H-8); 8.98 (s, 1 H, H-2).
UV (λmax (ꢀ)) methanol: 301 (22700). Anal. (C17H18N4O4) C, H,
N.
6-(4-Meth oxyph en yl)-9-(â-D-r ibofu r an osyl)pu r in e (12g).
Colorless crystals, yield 91%, mp 173-175 °C; [R]D -61.5 (c
0.5 DMF). FAB MS m/z (rel. %): 359 (9) [M + H], 227 (8) [M
+ H - Rf], 201 (16), 185 (40), 93 (100). 1H NMR (400 MHz,
DMSO-d6): 3.61 (dd, 1 H, J ) 3.8 and 12.0) and 3.73 (dd, 1 H,
J ) 3.8 and 12.0, CH2-5′); 3.87 (s, 3 H, OCH3); 4.01 (m, 1 H,
H-4′); 4.23 (dd, 1 H, J ) 3.6 and 4.9, H-3′); 4.66 (dd, 1 H, J )
4.9 and 5.5, H-2′); OH signals were exchanged; 6.09 (d, 1 H, J
) 5.5, H-1′); 7.16 (d, 2 H, J ) 8.9, H-Ph); 8.85 (d, 2 H, J )
8.9, H-Ph); 8.88 (s, 1 H, H-8); 8.94 (s, 1 H, H-2). 13C NMR
(100 MHz, DMSO-d6): 55.51 (CH3); 61.40 (CH2-5′); 70.43 (CH-
3′); 73.89 (CH-2′); 85.81 (CH-4′); 87.83 (CH-1′); 114.28 (d,
m-CH-OCH3Ph); 127.85 (i-C-OCH3Ph); 130.36 (C-5); 131.32
(o-CH-OCH3Ph); 144.51 (C-8); 151.98 (C-2); 152.07 (C-4);
152.96 (C-6); 161.88 (C-OCH3). UV (λmax (ꢀ)) methanol: 314
(21500). Anal. (C17H18N4O5) C, H, N.
6-(3-Meth oxyph en yl)-9-(â-D-r ibofu r an osyl)pu r in e (12h ).
Colorless crystals, yield 71%, mp 141-143 °C; [R]D -49.2 (c
0.5 DMF). FAB MS m/z (rel. %): 359 (93) [M + H], 227 (100)
[M + H - Rf], 201 (75). 1H NMR (400 MHz, DMSO-d6): 3.58-
3.64 and 3.70-3.75 (2 × m, 2 H, CH2-5′); 3.88 (s, 3 H, OCH3);
4.01 (m, 1 H, H-4′); 4.23 (m, 1 H, H-3′); 4.66 (ddd, 1 H, J )
5.1, 5.5 and 5.9, H-2′); 5.11 (t, 1 H, J ) 5.5, 5′-OH); 5.22 (d, 1
H, J ) 5.0, 3′-OH); 5.54 (d, 1 H, J ) 5.9, 2′-OH); 6.11 (d, 1 H,
J ) 5.5, H-1′); 7.17 (dd, 1 H, J ) 2.1 and 7.7, H-Ph); 7.53 (t,
1 H, J ) 8.0, H-Ph); 8.42 (brs, 1 H, H-Ph); 8.47 (d, 1 H, J )
7.9, H-Ph); 8.93 (s, 1 H, H-8); 9.01 (s, 1 H, H-2). UV (λmax (ꢀ))
methanol: 291 (29700). Anal. (C17H18N4O5) C, H, N.
1
230 (22) [M + H - Rf], 93 (100). H NMR (400 MHz, DMSO-
d6): 3.55-3.70 (m, 2 H, CH2-5′); 3.93 (m, 1 H, H-4′); 4.16 (dd,
1 H, J ) 3.6 and 4.7, H-3′); 4.55 (dd, 1 H, J ) 4.7 and 5.9,
H-2′); 5.13 (t, 1 H, J ) 5.5, 5′-OH); 5.08, 5.14 and 5.44 (3 ×
brs, 3 × 1 H, 3 × OH); 5.90 (d, 1 H, J ) 5.9, H-1′); 6.58 (s, 2
H, NH2); 7.39 (t, 2 H, J ) 8.9, H-o-Ar); 8.41 (s, 1 H, H-8); 8.79
(dd, 2 H, J ) 8.9 and 5.8, H-m-Ar). 19F NMR (376.5 MHz,
DMSO-d6): -109.62 (m, FPh). Anal. (C16H16FN5O4) C, H, N.
2-Am in o-6-(2,4-d iflu or op h en yl)-9-(â-D-r ibofu r a n osyl)-
p u r in e (13c). Yellowish crystals, yield 86%, mp 119-121 °C;
[R]D -24.6 (c 0.5 DMF). FAB MS m/z (rel. %): 380 (43) [M +
H], 248 (60) [M + H - Rf], 93 (100). 1H NMR (400 MHz,
DMSO-d6): 3.53-3.60 and 3.63-3.69 (2 × m, 2 H, CH2-5′);
3.93 (m, 1 H, H-4′); 4.15 (m, 1 H, H-3′); 4.55 (ddd, 1 H, J )
4.9, 6.0 and 6.0, H-2′); 5.05 (t, 1 H, J ) 5.4, 5′-OH); 5.16 (d, 1
H, J ) 4.7, 3′-OH); 5.46 (d, 1 H, J ) 6.0, 2′-OH); 5.89 (d, 1 H,
J ) 6.0, H-1′); 6.66 (s, 2 H, NH2); 7.26 (dt, 1 H, J ) 2.5 and
8.4, H-arom); 7.40 (dt, 1 H, J ) 2.4 and 10.0, H-arom); 7.87
(q, 1 H, J ) 7.1, H-arom); 8.34 (s, 1 H, H-8). 13C NMR (100
MHz, DMSO-d6): 61.38 (CH2-5′); 70.41 (CH-3′); 73.39 (CH-
2′); 85.37 (CH-4′); 86.39 (CH-1′); 104.57 (t, J ) 25.9, CH-3-
Ph); 111.91 (d, J ) 21.1, CH-5-Ph); 120.71 (C-1-Ph); 125.26
(C-5); 133.12 (d, J ) 9.5, CH-6-Ph); 140.82 (C-8); 152.29 (C-
6); 153.82 (C-4); 160.27 (C-2); 160.11 (dd, J ) 13.0 and 254.9,
CF-2-Ph); 163.16 (dd, J ) 12.4 and 249.6, CF-4-Ph). 19F NMR
(376.5 MHz, DMSO-d6): -107.33 - -107.45 (m, F2Ph). Anal.
(C16H15F2N5O4) C, H, N.
2-Am in o-6-(3,4-d iflu or op h en yl)-9-(â-D-r ibofu r a n osyl)-
p u r in e (13d ). Hygroscopic yellowish solid, yield 83%, mp
181-185 °C; [R]D -19.9 (c 0.4 DMF). FAB MS m/z (rel. %):
1
380 (20) [M + H], 248 (100) [M + H - Rf], 93 (77). H NMR
(400 MHz, DMSO-d6): 3.55-3.70 (m, 2 H, CH2-5′); 3.94 (m, 1
H, H-4′); 4.16 (dd, 1 H, J ) 3.3 and 4.9, H-3′); 4.54 (dd, 1 H, J
) 4.9 and 5.8, H-2′); signals of OH groups were exchanged;
5.90 (d, 1 H, J ) 5.8, H-1′); 6.65 (brs, 2 H, NH2); 7.64 (q, 1 H,
J ) 8.8, H-Ar); 8.45 (s, 1 H, H-8); 8.61-8.70 (m, 2 H, H-m-
Ar). 19F NMR (376.5 MHz, DMSO-d6): -134.96 and -137.72
(2 × m, F2Ph). Anal. (C16H15F2N5O4‚H2O) C, H. N.
6-(4-Eth oxyp h en yl)-9-(â-D-r ibofu r a n osyl)p u r in e (12i).
Colorless crystals, yield 89%, mp 173-176 °C; [R]D -57.5 (c
0.5 DMF). FAB MS m/z (rel. %): 373 (50) [M + H], 279 (63),
1
In h ibition of Cell Gr ow th . Mouse leukemia L1210 cells
(ATCC CCL 219) were cultivated in RPMI 1640 medium
containing 15% bovine serum38 using 24-well tissue culture
plates. The cells were seeded at 5 × 104 mL-1 and after a 24-h
incubation period (CO2 atmosphere, 37 °C) tested compounds
were added at five different concentrations. The endpoint of
the cell growth was 72 h following the addition. An appropriate
aliquot from every dish was then counted (cell counter Serono
150+). The inhibitory potency of the compounds tested was
expressed as IC50 values.
241 (100) [M + H - Rf]. H NMR (500 MHz, DMSO-d6): 1.38
(t, 3 H, J ) 6.9, CH3CH2); 3.61 (dd, 1 H, J ) 3.8 and 12.0) and
3.73 (dd, 1 H, J ) 3.8 and 12.0, CH2-5′); 4.01 (m, 1 H, H-4′);
4.15 (q, 2 H, J ) 6.9, CH2CH3); 4.22 (dd, 1 H, J ) 3.6 and 4.8,
H-3′); 4.66 (dd, 1 H, J ) 4.8 and 5.6, H-2′); OH signals were
exchanged; 6.09 (d, 1 H, J ) 5.6, H-1′); 7.15 (d, 2 H, J ) 8.9,
H-Ph); 8.84 (d, 2 H, J ) 8.9, H-Ph); 8.88 (s, 1 H, H-8); 8.94
(s, 1 H, H-2). UV (λmax (ꢀ)) methanol: 315 (24100). Anal.
(C18H20N4O5) C, H, N.
6-(4-Ch lor op h en yl)-9-(â-D-r ibofu r a n osyl)p u r in e (12j).
Colorless crystals, yield 82%, mp 206-208 °C; [R]D -55.5 (c
0.5 DMF). FAB MS m/z (rel. %): 363 (32) [M + H], 279 (100),
CCRF-CEM T-lymphoblastoid cells (ATCC CCL 119) were
cultivated in RPMI 1640 medium supplemented with L-
glutamine (0.3 g/L) containing 10% bovine serum using 24-
well tissue culture plates. The cells were seeded at 105 mL-1
and after a 24-h incubation period (CO2 atmosphere, 37 °C)
tested compounds were added at five different concentrations.
The endpoint of the cell growth was 72 h following the drug
addition. An appropriate aliquot from every dish was then
counted (cell counter Serono 150+). The inhibitory potency of
the compounds tested was expressed as IC50 values.
1
231 (81) [M + H - Rf]. H NMR (400 MHz, DMSO-d6): 3.58
(dd, 1 H, J ) 3.7 and 12.0) and 3.71 (dd, 1 H, J ) 3.8 and
12.0, CH2-5′); 3.99 (m, 1 H, H-4′); 4.20 (dd, 1 H, J ) 3.6 and
4.8, H-3′); 4.64 (dd, 1 H, J ) 4.8 and 5.4, H-2′); ca. 5.1, ca. 5.2
and ca. 5.55 (3 × brs, 3 × OH); 6.08 (d, 1 H, J ) 5.4, H-1′);
7.67 (d, 2 H, J ) 8.5, H-Ph); 8.84 (d, 2 H, J ) 8.5, H-Ph);
8.94 (s, 1 H, H-8); 9.00 (s, 1 H, H-2). UV (λmax (ꢀ)) methanol:
295 (26000). Anal. (C16H15ClN4O4) C, H, N, Cl.
Murine L929 cells (ATCC CCL 1) were placed in 24-well
tissue dishes and grown in Waymouth’s MB 752/1 medium
containing 5% fetal calf serum (105 cells/dish). After 24 h cell
monolayers were overlaid with fresh medium supplemented
with tested compounds at five different concentrations. After
the additional 48-h incubation at 37 °C (CO2 atmosphere)
cultures were stained with methylene blue (Sigma)39 and the
absorbance at 600 nm was then measured in 1% sarkosyl
(Sigma) extracts. The number of cells was calculated from the
calibration curve and results were expressed as IC50 values.
Human cervix carcinoma HeLa S3 cells (ATCC CCL 2.2)
2-Am in o-6-p h en yl-9-(â-D-r ib ofu r a n osyl)p u r in e (13a ).
Colorless crystals, yield 64%, mp 184-186 °C; [R]D -35.3 (c
0.3 DMF). FAB MS m/z (rel. %): 344 (11) [M + H], 212 (37)
[M + H - Rf], 115 (100). 1H NMR (400 MHz, DMSO-d6): 3.57
(dd, 1 H, J ) 4.0 and 11.9) and 3.67 (dd, 1 H, J ) 4.0 and
11.9, CH2-5′); 3.93 (m, 1 H, H-4′); 4.17 (dd, 1 H, J ) 3.7 and
5.0, H-3′); 4.51 (dd, 1 H, J ) 5.0 and 5.7, H-2′); OH signals
were exchanged; 5.90 (d, 1 H, J ) 5.7, H-1′); 6.55 (s, 2 H, NH2);
7.51-7.58 (m, 3 H, H-Ph); 8.41 (s, 1 H, H-8); 8.68-8.74 (m, 2
H, H-Ph). UV (λmax (ꢀ)) methanol: 335 (9600), 248 (16700);
water: pH 7, 327 (9200), 247 (13800); pH 2, 342 (11200), 259

1824 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 9
Hocek et al.
(9) Elion, G. B.; Hitchings, G. H. Metabolic basis for the actions of
analogues of purines and pyrimidines. Adv. Chemother. 1965,
2, 91-177.
(10) For review: Benezra, S. A.; Foss P. R. B. In Analytical Profiles
of Drug Substances;Florey, K., Ed.; Academic Press: New York,
1978; pp 343-357.
(11) (a) Avery, T. L.; Finch, R. A.; Vasquez, K. M.; Radparvar, S.;
Hanna, N. B.; Revankar, G. R.; Robins, R. K. Chemotherapeutic
characterization in mice of 2-amino-9-beta-D-ribofuranosylpu-
rine-6-sulfinamide (sulfinosine), a novel purine nucleoside with
unique antitumor properties. Cancer Res. 1990, 50, 2625-2630.
(b) Revankar, G. R.; Hanna, N. B.; Imamura, N.; Lewis, A. F.;
Larson, S. B.; Finch, R. A.; Avery, T. L.; Robins, R. K. Synthesis
and in vivo antitumor activity of 2-amino-9H-purine-6-sulfen-
amide, -sulfinamide, and -sulfonamide and related purine ribo-
nucleosides. J . Med. Chem. 1990, 33, 121-128. (c) Hanna, N.
B.; Bhattacharya, B. K.; Robins, R. K.; Avery, T. L.; Revankar,
G. R. Sulfinosine Congeners - Synthesis and Antitumor Activity
in Mice of Certain N9-Alkylpurines and Purine Ribonucleosides.
J . Med. Chem. 1994, 37, 177-183.
were seeded in 24-well dishes (RPMI 1640 HEPES modifica-
tion containing 5% fetal bovine serum) (7 × 104 cells/mL) and
incubated 24 h at 37 °C (CO2 atmosphere). Then the culture
medium was removed and fresh medium with test compound
was added at five different concentrations; 48 h following drug
addition cultivation was stopped and the cell growth was
evaluated after staining with methylene blue (Sigma)39 (see
above).
Cell Via bility. In parallel, the number of viable cells
(viability in cell population) was quantified using MTT stan-
dard spectrophotometric assay.40
Ack n ow led gm en t. This work was supported by the
Grant Agency of the Czech Republic (Grants 203/98/
P027 and 203/96/K001). The excellent technical as-
sistance of Ms. Kamila Havl´ıcˇkova´ is gratefully acknowl-
edged. The authors also thank the staff of the Mass
Spectrometry and Analytical Departments of this In-
stitute.
(12) Williams, B. A.; Blay, J .; Hoskin, D. W. 2-chloroadenosine
stimulates granule exocytosis from mouse natural killer cells:
Evidence for signal transduction through a novel extracellular
receptor. Exp. Cell Res. 1997, 233, 187-197.
(13) (a) Wataya, Y.; Hirota, Y.; Hiramoto-Yoshioka, A.; Tanaka, S.;
Otani, T.; Minowada, J .; Matsuda, A.; Ueda, T. The mechanism
of 2-chlorodeoxyadenosine-induced cell death. Adv. Exp. Med.
Biol. 1989, 253B, 227-234. (b) Carson, D. A.; Wasson, D. B.;
Taetle, R.; Yu, A. Specific toxicity of 2-chlorodeoxyadenosine
toward resting and proliferating human lymphocytes. Blood
1983, 62, 737-743. (c) Carson, D. A.; Wasson, D. B.; Beutler, E.
Antileukemic and immunosuppressive activity of 2-chloro-2′-
deoxyadenosine. Proc. Natl. Acad. Sci. U.S.A. 1984, 81, 2232-
2236.
Refer en ces
(1) (a) Robins, R. K.; Revankar, G. R. Purine analogues and related
nucleosides and nucleotides as antitumor agents. Med. Res. Rev.
1985, 5, 273-296. (b) Plunkett, W.; Saunders, P. P. Metabolism
and action of purine nucleoside analogues. Pharmacol. Ther.
1991, 49, 239-268. (c) Cheson, B. D. Perspectives on purine
analogues. Hematol. Cell Ther. 1996, 38 (Suppl. 2), S109-S116.
(d) Bergmann, L. Present status of purine analogues in the
therapy of chronic lymphocytic leukemias. Leukemia 1997, 11
(Suppl. 2), S29-S34.
(2) (a) Vesely´, J .; Havl´ıcˇek, L.; Strnad, M.; Blow, J . J .; Donella-
Deana, A.; Pinna, L.; Letham, D. S.; Kato, J .; Detivaud, L.;
Leclerc, S. Inhibition of cyclin-dependent kinases by purine
analogues. Eur. J . Biochem. 1994, 224, 771-786. (b) Havl´ıcˇek,
L.; Hanus, J .; Vesely´, J .; Leclerc, S.; Meijer, L.; Shaw, G.; Strnad,
M. Cytokinin-derived cyclin-dependent kinase inhibitors: syn-
thesis and cdc2 inhibitory activity of olomoucine and related
compounds. J . Med. Chem. 1997, 40, 408-412. (c) Legraverend,
M.; Ludwig, O.; Bisagni, E.; Leclerc, S.; Meijer, L. Synthesis of
C2 alkynylated purines, a new family of potent inhibitors of
cyclin-dependent kinases. Bioorg. Med. Chem. Lett. 1998, 8,
793-798. (d) Legraverend, M.; Ludwig, O.; Bisagni, E.; Leclerc,
S.; Meijer, L.; Giocanti, N.; Sadri, R.; Favaudon, V. Synthesis
and in vitro evaluation of novel 2,6,9-trisubstituted purines
acting as cyclin-dependent kinase inhibitors. Bioorg. Med. Chem.
1999, 7, 1281-1293. (e) N.Oh, C. H.; Lee, S. C.; Lee, K. S.; Woo,
E. R.; Hong, C. Y.; Yang, B. S.; Baek, D. J .; Cho, J . H. Synthesis
and biological activities of C-2, N-9 substituted 6-benzylamino-
purine derivatives as cyclin-dependent kinase inhibitors. Arch.
Pharm. (Weinheim) 1999, 332, 187-190.
(3) Revankar, G. R.; Robins, R. K. Heterocyclic Analogues of Purine
Nucleosides and Nucleotides. In Chemistry of Nucleosides and
Nucleotides; Townsend, L. B., Ed.; Plenum Press: New York/
London, 1988; Vol. 2, pp 200-246.
(4) (a) Cristalli, G.; Franchetti, P.; Grifantini, M.; Vittori, S.;
Bordoni, T.; Geroni, C. Improved synthesis and antitumor
activity of 1-deazaadenosine. J . Med. Chem. 1987, 30, 1686-
1688. (b) Cristalli, G.; Vittori, S.; Eleuteri, A.; Grifantini, M.;
Volpini, R.; Lupidi, G.; Capolongo, L.; Pesenti, E. Purine and
1-deazapurine ribonucleosides and deoxyribonucleosides: syn-
thesis and biological activity. J . Med. Chem. 1991, 34, 2226-
2230.
(14) (a) Reiter, Z.; Tomson, S.; Ozes, O. N.; Taylor, M. W. Combination
treatment of 2-chlorodeoxyadenosine and type I interferon on
hairy cell leukemia-like cells: cytotoxic effect and MHC-
unrestricted killer cell regulation. Blood 1993, 81, 1699-1708.
(b) Saven, A.; Carrera, C. J .; Carson, D. A.; Beutler, E.; Piro, L.
D. 2-Chlorodeoxyadenosine: an active agent in the treatment
of cutaneous T-cell lymphoma. Blood 1992, 80, 587-592. (c)
Arner, E. S. J . On the phosphorylation of 2-chlorodeoxyadenosine
(CdA) and its correlation with clinical response in leukemia
treatment. Leuk. Lymphoma 1996, 21, 225-231.
(15) (a) Keating, M. J .; Kantarian, H.; Talpaz, M. Fludarabine:
A
new agent with major activity against chronic lymphocytic
leukemia. Blood 1989, 74, 19-25. (b) Pawelski, S. Fludarabine
in the treatment of chronic lymphocytic leukemia and other
lymphoproliferative disorders. Acta Haematol. Pol. 1995, 26,
263-268. (c) Fludarabine phosphate: an effective therapy for
lymphoid malignancies. Semin. Oncol. 1990, 17, 1-78.
(16) Matsuda, A.; Shinozaki, M.; Yamaguchi, T.; Homma, H.; Nomoto,
R.; Miyasaka, T.; Watanabe, Y.; Abiru, T. Nucleosides and
Nucleotides. 103. 2-Alkynyladenosines - A Novel Class of
Selective Adenosine-A2 Receptor Agonists with Potent Antihy-
pertensive Effects. J . Med. Chem. 1992, 35, 241-252.
(17) Lambe, C. U.; Averett, D. R.; Paff, M. T.; Reardon, J . E.; Wilson,
J . G.; Krenitsky, T. A. 2-Amino-6-methoxypurine arabinoside:
an agent for T-cell malignancies. Cancer Res. 1995, 55, 3352-
3356.
(18) (a) Gibboney, D. S.; French, B. T.; Patrick, D. E.; Trewyn, R. W.
6-ethylmercaptopurine-mediated growth inhibition of HL-60
cells in vitro irrespective of purine salvage. Cancer Chemother.
Pharmacol. 1989, 25, 189-194. (b) Akiyama, S. I.; Kuwano, M.;
Komiyama, S.; Saneyoshi, M. Antitumor effect of a combination
of 6-methylthioinosine and amphotericin B on mouse leukemia
L1210. Cancer Lett. 1980, 9, 305-311.
(19) (a) McGarrity, G. J .; Carson, D. A. Adenosine phosphorylase-
mediated nucleoside toxicity. Application towards the detection
of mycoplasmal infection in mammalian cell cultures. Exp. Cell
Res. 1982, 139, 199-205. (b) Whitaker, A. M.; Windsor, G. D.;
Burnett, C. M.; Taylor, C. H. A rapid and sensitive method for
the detection of mycoplasmas in infected cell cultures using
6-methylpurine deoxyriboside. Dev. Biol. Stand. 1987, 66, 503-
509. (c) Ishiguro, K.; Taira, S.; Sasaki, T.; Nariuchi, H. Depletion
of mycoplasma from infected cell lines by limiting dilution in
6-methylpurine deoxyriboside. J . Immunol. Methods 1988, 108,
39-43.
(20) (a) Parker, W. B.; King, S. A.; Allan, P. W.; Bennett, L. L., J r.;
Secrist, J . A.; Montgomery, J . A.; Gilbert, K. S.; Waud, W. R.;
Wells, A. H.; Gillespie, G. Y.; Sorscher, E. J . In vivo gene therapy
of cancer with E. coli purine nucleoside phosphorylase. Hum.
Gene Ther. 1997, 8, 1637-1644. (b) Parker, W. B.; Allan, P. W.;
Shaddix, S. C.; Rose, L. M.; Speegle, H. F.; Gillespie, G. Y.;
Bennett, L. L., J r. Metabolism and metabolic actions of 6-meth-
ylpurine and 2-fluoroadenine in human cells. Biochem. Phar-
macol. 1998, 55, 1673-1681.
(5) Franchetti, P.; Cappellacci, L.; Grifantini, M.; Lupidi, G.; No-
centini, G.; Barzi, A. 8-Aza Analogues of Deaza Purine Nucleo-
sides
- Synthesis and Biological Evaluation of 8-Aza-1-
deazaadenosine and 2′-Deoxy-8-aza-1-deazaadenosine. Nucleosides
Nucleotides 1992, 11, 1059-1076.
(6) (a) Leopold, W. R.; Fry, D. W.; Boritzki, T. J .; Besserer, J . A.;
Pattison, I. C.; J ackson, R. C. Deazaguanine mesylate: a new
antipurine antimetabolite. Invest. New Drugs 1985, 3, 223-231.
(7) (a) Page, T.; J acobsen, S. J .; Smejkal, R. M.; Scheele, J .; Nyhan,
W. L.; Mangum, J . H.; Robins, R. K. Studies on the mechanism
of cytotoxicity of 3-deazaguanosine in human cancer cells. Cancer
Chemother. Pharmacol. 1985, 15, 59-62. (b) J acobsen, S. J .;
Page, T.; Diala, E. S.; Nyhan, W. L.; Robins, R. K.; Mangum, J .
H. Synergistic activity of purine metabolism inhibitors in
cultured human tumor cells. Cancer Lett. 1987, 35, 97-104.
(8) Pudlo, J . S.; Nassiri, M. R.; Kern, E. R.; Wotring, L. L.; Drach,
J . C.; Townsend, L. B. Synthesis, antiproliferative, and antiviral
activity of certain 4-substituted and 4,5-disubstituted 7-[(1,3-
dihydroxy-2-propoxy)methyl]pyrrolo[2,3-d]pyrimidines. J . Med.
Chem. 1990, 33, 1984-1992.

Substituted 6-Phenylpurine Bases and Nucleosides
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 9 1825
(21) Brathe, A.; Gundersen, L. L.; Rise, E.; Eriksen, A. B.; Vollsnes,
A. V.; Wang, L. N. Synthesis of 6-alkenyl- and 6-alkynylpurines
with cytokinin activity. Tetrahedron 1999, 55, 211-228.
(22) (a) Kobayashi, Y.; Yamamoto, K.; Asai, T.; Nakano, M.; Ku-
madaki, I. Studies on Organic Fluorine Compounds. Part 35.
Trifluoromethylation of Pyrimidine- and Purine-nucleosides with
Trifluoromethyl-Copper Complex. J . Chem. Soc., Perkin Trans.
1 1980, 2755-2761. (b) Hockova´, D.; Hocek, M.; Dvorˇa´kova´, H.;
Votruba, I. Synthesis and Cytostatic Activity of Nucleosides and
Acyclic Nucleoside Analogues Derived form 6-(Trifluoromethyl)-
purines. Tetrahedron 1999, 55, 11109-11118.
(23) Cocuzza, A. J .; Chidester, D. R.; Culp, S.; Fitzgerald, L.; Gilligan,
P. Use of Suzuki Reaction for the Synthesis of Aryl-substituted
Heterocycles as Corticotropin-Releasing Hormone (CHR) An-
tagonists. Bioorg. Med. Chem. Lett. 1999, 9, 1063-1066.
(24) (a) Moran, S.; Ren, R. X. F.; Rumney, S., IV; Kool, E. T.
Difluorotoluene, A Nonpolar Isostere for Thymine, Codes Specif-
ically and Efficiently for Adenine in DNA Replication. J . Am.
Chem. Soc. 1997, 119, 2056-2057. (b) Liu, D.; Moran, S.; Kool,
E. T. Bi-stranded, multisite replication of a base pair between
difluorotoluene and adenine: confirmation by ′inverse′ sequenc-
ing. Chem. Biol. 1997, 4, 919-926 (c) Guckian, K. M.; Krugh,
T. R.; Kool, E. T. Solution structure of a DNA duplex containing
a replicable difluorotoluene-adenine pair. Nat. Struct. Biol. 1998,
5, 954-959.
(25) (a) Bergstrom, D. E.; Reddy, P. A. Synthesis of 6-Alkyl and 6-Aryl
Substituted 9-â-D-ribofuranosyl Purines via the Nickel Cata-
lyzed Coupling of Grignard Reagents to 2′,3′,5′-Tris-O-(tert-
butyldimethylsilyl)-9-â-D-ribofuranosyl-6-chloropurine. Tetra-
hedron Lett. 1982, 23, 4191-4194. (b) Estep, K. G.; J osef, K. A.;
Bacon, E. R.; Carabates, P. M.; Rumney, S., IV; Pilling, G. M.;
Krafte, D. S.; Volberg, W. A.; Dillon, K.; Dugrenier, N.; Briggs,
G. M.; Canniff, P. C.; Gorczyca, W. P.; Stankus, G. P.; Ezrin, A.
M. Synthesis and Structure-Activity Relationship of 6-Hetero-
cyclic-Substituted Purines as Inactivation Modifiers of Cardiac
Sodium Channels. J . Med. Chem. 1995, 38, 2582-2595.
(26) (a) Gundersen, L. L. 6-Chloropurines and Organostannanes in
Paladium Catalyzed Cross Coupling Reactions. Tetrahedron Lett.
1994, 35, 3155-3158. (b) Gundersen, L. L.; Bakkestuen, A. K.;
Aasen, A. J .; Lveras, H.; Rise, F. 6-Halopurines in Palladium-
Catalyzed Coupling with Organotin and Organozinc Reagents.
Tetrahedron 1994, 50, 9743-9756. (c) Hocek, M.; Masoj´ıdkova´,
M.; Holy´, A. Synthesis of Acyclic Nucleotide Analogues Derived
from N-substituted 6-(1-Aminomethyl)purines via 6-Acetylpu-
rine Derivatives. Tetrahedron 1997, 53, 2291-2302. (d) Hocek,
M.; Masoj´ıdkova´, M.; Holy´, A. Synthesis of Acyclic Nucleotide
Analogues Derived from 6-Hetarylpurines via Cross-Coupling
Reactions of 9-[2-(Diethoxyphosphonylmethoxy)ethyl]-6-iodopu-
rine with Hetaryl Organometallic Reagents. Collect. Czech.
Chem. Commun. 1997, 62, 136-146. (e) Van Aerschot, A. A.;
Mamos, P.; Weyns, N. J .; Ikeda, S.; De Clercq, E.; Herdewijn,
P. Antiviral Activity of C-Alkylated Purine Nucleosides Obtained
by Cross-Coupling with Tetraalkyltin Reagents. J . Med. Chem.
1995, 36, 2938-2942.
Reagents: A Convenient Route to sec- and tert- 6-Alkylpurines.
Tetrahedron Lett. 1996, 37, 1285-1288. (c) Dvorˇa´kova´, H.;
Dvorˇa´k, D.; Holy´, A. Synthesis of Acyclic Nucleotide Analogues
Derived from 6-(sec- or tert-Alkyl)purines via Coupling of
6-Chloropurine Derivatives with Organocuprates. Collect. Czech.
Chem. Commun. 1998, 63, 2065-2074. (d) Hocek, M.; Holy´, A.
Perfluoroalkylation of 6-Iodopurines by Trimethyl(perfluoro-
alkyl)silanes. Synthesis of 6-(Perfluoroalkyl)purine Bases, Nu-
cleosides and Acyclic Nucleotide Analogues. Collect. Czech.
Chem. Commun. 1999, 64, 229-241.
(29) (a) Stevenson, T. M.; Prasad, A. S. B.; Citineni, J . B.; Knochel,
P. Preparation of Zinc Organometallics Derived from Nucleosides
and Nucleic Acid Bases and Pd(0) Catalyzed Coupling with Aryl
Iodides. Tetrahedron Lett. 1996, 37, 8375-8378. (b) Prasad, A.
S. B.; Stevenson, T. M.; Citineni, J . B.; Nyzam, V.; Knochel, P.
Preparation and Reactions of New Zincated Nitrogen-Containing
Heterocycles. Tetrahedron 1997, 53, 7237-7254.
(30) (a) Nair, V.; Richardson, S. G.; Coffman, R. E. Arylation and
Heteroarylation of Photochemically Generated Purinyl Radicals.
J . Org. Chem. 1982, 47, 4520-4524. (b) McKenzie, T. C.; Epstein,
J . W. Coupling of Diazopurines, a Curious Steric Effect in a Free
Radical Reaction. J . Org. Chem. 1982, 47, 4881-4884. (c) Nair,
V.; Young, D. A. Synthetic Transformations of Transient Purinyl
Radicals: Formation of Mono- and Diarylated and Heteroary-
lated Nucleosides. J . Org. Chem. 1984, 49, 4340-4344.
(31) Havelkova´, M.; Hocek, M.; Cˇ esnek, M.; Dvorˇa´k, D. The Suzuki-
Miyaura Cross-Coupling Reactions of 6-Halopurines with Bo-
ronic Acids Leading to 6-Aryl- and 6-Alkenylpurines. Synlett
1999, 1145-1147.
(32) Robins, R. K.; Godefroi, E. F.; Taylor, E. C.; Lewis, L. R.; J ackson,
A. Purine Nucleosides. I. The Synthesis of Certain 6-Substituted-
9-(tetrahydro-2-pyranyl)-purines as Model of Purine Deoxy-
nucleotides. J . Am. Chem. Soc. 1961, 83, 2574-2579.
(33) Hocek, M.; Holy´, A. A Facile Synthesis of 6-Cyanopurine Bases.
Collect. Czech. Chem. Commun. 1995, 60, 1386-1389.
(34) Buck, I. M.; Reese, C. B. An Unambiguous Synthesis of Ade-
nylsuccinic Acid and its Constituent Nucleoside. J . Chem. Soc.,
Perkin Trans. 1 1990, 2937-2942.
(35) Robins, M. J .; Uznanski, B. Nucleic Acids Related Compounds.
33. Conversions of Adenosine and Guanosine to 2,6-Dichloro,
2-Amino-6-chloro, and Derived Purine Nucleosides. Can. J .
Chem. 1981, 59, 2601-2607.
(36) Sugimura, H.; Takei, H. Synthesis of 6-Alkylpurine Derivatives
by Nickel-Complex-Catalyzed Coupling Reaction of 6-(Meth-
ylthio)purine derivatives with Grignard Reagents. Bull. Chem.
Soc. J pn. 1985, 58, 664-666.
(37) Kos, N. J .; van der Plas, H. C. Occurrence of the SNANRORC
Mechanism in the Amination of 2-Substituted Purines with
Potassium Amide in Liquid Ammonia. J . Org. Chem. 1980, 45,
2942-2945.
(38) Vesely´, J . Synergistic effect of cis-dichlorodiammineplatinum and
5-aza-2′-deoxycytidine on mouse leukemic cells in vivo and in
vitro. Int. J . Cancer 1982, 29, 81-85.
(39) Lillie, R. D. H. J . Conn’s Biological Stains, 9th ed.; Williams &
Wilkins: Baltimore, 1977.
(27) Hirota, K.; Kitade, Y.; Kanbe, Y.; Maki, Y. Convenient Method
for the Synthesis of C-Alkylated Purine Nucleosides: Palladium-
Catalyzed Cross-Coupling Reaction of Halopurine Nucleosides
with Trialkylaluminums. J . Org. Chem. 1992, 57, 5268-5270.
(28) (a) McKenzie, T. C.; Glass, D. The Reaction of 6-Halopurines
with Phenyl Metal Complexes. J . Heterocycl. Chem. 1987, 24,
1551-1553. (b) Dvorˇa´kova´, H.; Dvorˇa´k, D.; Holy´, A. Coupling
of 6-Chloropurines with Organocuprates Derived from Grignard
(40) Carmichael, J .; DeGraff, W. G.; Gazdar, A. F.; Minna, J . D.;
Mitchell J . B. Evaluation of a tetrazolium-based semiautomated
colorimetric assay: assessment of chemosensitivity testing.
Cancer Res. 1987, 47, 936-942.
J M991167+