SYNTHESIS OF ACYCLIC NUCLEOTIDE ANALOGUES DERIVED FROM 2-(AMINOMETHYL)ADENINE AND 2-(AMINOMETHYL)HYPOXANTHINE

Article abstract of DOI:10.1135/cccc19950875

Synthesis of a series of 2-(aminomethyl)-9-(2-phosphonomethoxyalkyl)adenins VI and hypoxanthines VII is reported.The protected 2-(aminomethyl)adenine I was selectively alkylated with alkyl chlorides or tosylates II and the

Full text of DOI:10.1135/cccc19950875

Synthesis of Acyclic Nucleotide Analogues
875
SYNTHESIS OF ACYCLIC NUCLEOTIDE ANALOGUES DERIVED FROM
2-(AMINOMETHYL)ADENINE AND 2-(AMINOMETHYL)HYPOXANTHINE
Michal HOCEK, Milena MASOJIDKOVA and Antonin HOLY
Institute of Organic Chemistry and Biochemistry,
Academy of Sciences of the Czech Republic, 166 10 Prague 6, The Czech Republic
Received March 10, 1995
Accepted April 26, 1995
Synthesis of a series of 2-(aminomethyl)-9-(2-phosphonomethoxyalkyl)adenines VI and hypo-
xanthines VII is reported. The protected 2-(aminomethyl)adenine I was selectively alkylated with
[bis(2-propoxy)phosphonylmethoxy]alkyl chlorides or tosylates II and the obtained 9-[bis(2-propoxy)-
phosphonylmethoxy]alkyl-2-(benzyloxycarbonylaminomethyl)adenines IV were oxodeaminated to give
the corresponding hypoxanthine derivatives V. The intermediates IV and V were completely deprotected
by treatment with iodotrimethylsilane under formation of the title compounds VI and VII.
Phosphonomethoxyalkyl derivatives of pyrimidine and purine bases are potent
antivirals1. The most promising of this group are the 2-phosphonomethoxyethyl (PME),
(R)-2-phosphonomethoxypropyl (PMP) and (S)-3-hydroxy-2-phosphonomethoxypropyl
(HPMP) derivatives. In our recent structure–activity relationship study of base-
modified analogues of these compounds we have observed that the antiviral activity is
connected with the presence of amino groups at the heterocyclic moiety. Thus, adenine,
guanine, 2-aminopurine, 2,6-diaminopurine derivatives2 and their aza and deaza
analogues3–6, as well as cytosine derivatives2, exhibit potent antiviral effects while
uracil, thymine, hypoxanthine, xanthine and other derivatives lacking the amino
function are generally inactive2.
The role of the amino group may consist just in its basicity or in the formation of
hydrogen bonds with some (enzyme) counterparts. Several N(6)-monosubstituted and
N(6)-disubstituted derivatives of adenine and 2,6-diaminopurine, however, possess
some antiviral activity7 as well. Therefore, we decided to investigate the synthesis and
properties of acyclic nucleotide analogues bearing 2-aminomethyl function on the
purine ring.
In general, (aminomethyl)purines are known8–10 to be quite unstable due to their
strongly basic benzyl-type amino group and are usually isolated as stable N-acyl derivatives9,10
.
Recently we have published11 a facile synthesis of N-protected 2-(aminomethyl)purines;
now we report on alkylation of these bases leading to acyclic nucleotide analogues
(see Scheme 1).
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SCHEME 1
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Benzyloxycarbonyl-protected 2-(aminomethyl)adenine I (Cb = benzyloxycarbonyl)
was alkylated using the following synthons: bis(2-propyl) 2-chloroethoxymethane-
phosphonate3 (IIa), (R)-2-[bis(2-propoxy)phosphonylmethoxy]propyl tosylate12 (IIb)
and (R)-3-benzoyloxy-2-[bis(2-propoxy)phosphonylmethoxy]propyl tosylate13 (IIc).
Sodium salt of I was formed at room temperature by treatment with sodium hydride in
dimethylformamide. After addition of the alkylating agent, the mixture was heated for
2 – 3 h at 100 °C until most of the starting material disappeared (TLC). Since slow
decomposition of the product was observed after several hours of heating, the reaction
was stopped before reaching complete conversion and the products were isolated by
preparative TLC. After alkylation with the tosylate IIc the crude product was treated
with methanolic ammonia prior to chromatography. The 9-substituted 2-(aminomethyl)-
adenine derivatives IV were obtained in moderate yields and no substantial amount of
other regioisomers was observed. Attempts to crystallize these products failed and the
TLC-pure oily compounds were used in the further steps.
Attempted alkylation of the guanine analogue III by the same method was
unsuccessful due to its low stability under the alkylation conditions. The 9-substituted
2-(aminomethyl)hypoxanthine derivatives V were, however, prepared in moderate
yields by oxodeamination of IV using a mixture of 3-methylbutyl nitrite and dilute
sulfuric acid. This procedure avoids formation of acetylated byproducts observed
during usual oxodeamination with 3-methylbutyl nitrite in acetic acid and gives better
yields than the use of sodium nitrite in sulfuric acid.
Treatment of the protected intermediates IV and V with iodotrimethylsilane14 cleaved
simultaneously both the Cb and the 2-propyl protecting groups, while analogous
reaction with bromotrimethylsilane removed the 2-propyl functions only (a microscale
experiment with MS detection). Pure 2-(aminomethyl)-9-(phosphonomethoxyalkyl)-
adenines VI and hypoxanthines VII were obtained after ion exchange purification.
These were hygroscopic but otherwise stable in the presence of air, representing thus
the first example of stable N-unsubstituted aminomethylpurine derivatives. The
stability could be explained by formation of zwitterionic structures.
1
Structure of the compounds was proved by H and 13C NMR spectra. The observed
chemical shifts and interaction constants (see Experimental and Table I) are in good
accord with the reported15 values of 9-substituted adenine and hypoxanthine
derivatives. The downfield shifts of C-2 (δ 165) and C-6 (δ 167) carbon signals of
hypoxanthine derivatives VII in basic medium (NaOD) can be explained16 by anion
formation at N-1. This was also proved by 13C NMR spectrum of derivative VIIa in
D2O which exhibited the expected shifts at 149.87 and 158.62, respectively.
The UV spectra of 9-substituted 2-(aminomethyl)adenines VI and 2-(aminomethyl)-
hypoxanthines VII displayed maxima at 260 nm or 250 nm, respectively. The presence
of the strongly basic amino group manifests itself in the electrophoretic mobility of the
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Synthesis of Acyclic Nucleotide Analogues
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compounds VI and VII at slightly alkaline pH which is substantially lower compared to
that of phosphonomethoxyalkyl derivatives of adenine and guanine.
The 2-(aminomethyl)-9-(phosphonomethoxyalkyl)purines VI and VII obtained by the
above-described methods were tested17 for their cytostatic (L-1210, P-388 mouse
leukemia) and antiviral (selected RNA and DNA viruses and HIV-1, HIV-2) activity.
None of these compounds showed any significant antiviral or cytostatic activity, nor
did it exhibit any considerable cell toxicity.
EXPERIMENTAL
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. TLC was
performed on Silufol UV254 plates (Kavalier Votice, The Czech Republic) in a CHCl3–CH3OH (80 : 20)
mixture. Preparative TLC was carried out on 40 × 17 × 0.4 cm plates of silica gel containing UV
indicator. Paper electrophoresis was performed on a paper Whatman No. 3 MM at 40 V/cm for 1 h
in 0.05 M triethylammonium hydrogen carbonate, pH 7.5. The electrophoretical mobilities (EUp) are
referenced to uridine 3′-phosphate. NMR spectra were measured on a Varian Unity 500 spectrometer
1
(500 MHz for H and 125.7 MHz for 13C NMR) in hexadeuteriodimethyl sulfoxide referenced to the
solvent signals (2.5 ppm for 1H and 39.7 ppm for 13C NMR), or in deuterium oxide containing
sodium deuteroxide with sodium 3-(trimethylsilyl)propane sulfonate (DSS) as internal standard for
1H and dioxane as external standard for 13C NMR (δ(dioxane) 66.86). Mass spectra were measured
on a ZAB-EQ (VG Analytical) instrument using FAB technique (ionization by Xe, accelerating
voltage 8 kV, glycerol matrix). UV absorption spectra were measured on a Beckman DU-65
spectrometer in aqueous HCl (0.01 M) solutions. CD spectra were recorded on Jobin Yvon Mark V
spectrometer in aqueous HCl (0.01 M) solutions (c 1 . 10−3 mol/l). Dimethylformamide was distilled
from P2O5 and stored over molecular sieves (4A). Acetonitrile was refluxed with CaH2 and distilled.
Iodotrimethylsilane was purchased from Aldrich (U.S.A.) and 3-methylbutyl nitrite from Janssen
(Belgium). The starting 2-(aminomethyl)purines11 I and III as well as the alkylation synthons3,12,13 II
were prepared according to previously reported methods.
Alkylation of Compound I – General Procedure
Method A: Sodium hydride (60% dispersion in mineral oil, 80 mg, 2 mmol) was added to a stirred
solution of compound I (400 mg, 1.34 mmol) in DMF (20 ml) and stirring at room temperature was
continued for 2 h. The alkylating agent II (2 mmol) was added and the mixture was stirred at 100 °C
for 2 – 3 h. The solvent was evaporated, the residue was treated with water (50 ml) and extracted
with ethyl acetate (3 × 50 ml). The combined organic layers were evaporated and the residue was
chromatographed on a preparative layer of silica. The obtained chromatographically pure (TLC)
yellowish oils were used in the further steps.
Method B: The procedure was identical with method A except that after evaporation of the extracts
the residue was allowed to stand overnight with saturated methanolic ammonia (50 ml) and
evaporated before the chromatography.
Bis(2-propyl) 2-(benzyloxycarbonylaminomethyl)-9-(2-phosphonomethoxyethyl)adenine (IVa),
method A, yield 74%, RF 0.60. FAB MS, m/z (rel.%): 521 (18) [M + H]. 1H NMR spectrum
((CD3)2SO): 1.13 d, 6 H, J(CH3,CH) = 6.1 (2 × CH3); 1.17 d, 6 H, J(CH3,CH) = 6.1 (2 × CH3); 3.78 d,
2 H, J(P,CH2) = 8.2 (PCH2); 3.89 t, 2 H, J(2′,1′) = 4.7 (H-2′); 4.24 d, 2 H, J(CH2,NH) = 5.8
(CH2N); 4.31 t, 2 H, J(1′,2′) = 4.7 (H-1′); 4.49 m, 2 H (POCH); 5.06 s, 2 H (CH2Ph); 7.18 bs, 1 H
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and 7.58 bs, 1 H (NH2); 7.25 – 7.40 m, 5 H (arom. H); 7.55 bt, 1 H, J(NH,CH2) = 5.8 (NHCb); 8.14 s,
1 H (H-8).
Bis(2-propyl) (R)-2-(benzyloxycarbonylaminomethyl)-9-(2-phosphonomethoxypropyl)adenine (IVb),
method A, yield 56%, RF 0.65. FAB MS m/z (rel.%): 535 (100) [M + H]. 1H NMR spectrum
((CD3)2SO): 1.05 d, 3 H, J(3′,2′) = 6.3 (H-3′); 1.13 – 1.24 m, 12 H (4 × CH3-iPr); 3.77 m, 2 H
(PCH2); 3.96 m, 1 H (H-2′); 4.12 dd, 1 H, J(1′b,2′) = 6.6, J(gem) = 14.4 (Hb-1′); 4.20 d, 2 H,
J(CH2,NH) = 5.9 (CH2N); 4.22 dd, 1 H, J(1′a,2′) = 3.7, J(gem) = 14.4 (Ha-1′); 4.51 m, 2 H (2 ×
CH-iPr); 5.05 s, 2 H (CH2Ph); 7.30 – 7.40 m, 5 H (arom. H); 7.19 bs, 2 H (NH2); 7.42 t, 1 H,
J(NH,CH2) = 6.0 (NHCb); 8.01 s, 1 H (H-8).
Bis(2-propyl) (S)-2-(benzyloxycarbonylaminomethyl)-9-(3-hydroxy-2-phosphonomethoxypropyl)-
adenine (IVc), method B, yield 42%, RF 0.63. FAB MS, m/z (rel.%): 551 (100) [M + H]. 1H NMR
spectrum ((CD3)2SO): 1.155, 1.16, 1.24, 1.245 4 × d, 4 × 3 H, J(CH3,CH) = 6.1 (CH3-iPr); 3.42 m,
2 H (H-3′); 3.81 dd, 1 H, J(PCH) = 9.5, J(gem) = 13.7 (PC-Hb); 3.85 m, 1 H (H-2′); 3.88 dd, 1 H,
J(P,CH) = 8.8, J(gem) = 13.7 (PC-Ha); 4.19 dd, 1 H, J(1′b,2′) = 6.8, J(gem) = 14.4 (Hb-1′); 4.20 d,
2 H, J(CH2,NH) = 5.9 (CH2N); 4.33 dd, 1 H, J(1′a,2′) = 3.6, J(gem) = 14.4 (Ha-1′); 4.47 and 4.51 2 × dsept,
2 × 1 H, J(CH,CH3) = 6.1, J(P,OCH) = 7.8 (CH-iPr); 4.98 bs, 1 H (OH); 5.05 s, 2 H (CH2Ph); 7.20 – 7.40 m,
5 H (arom. H); 7.25 bs, 2 H (NH2); 7.44 t, 1 H, J(NH,CH2) = 5.9 (NHCb); 8.02 s, 1 H (H-8).
Oxodeamination of Adenines IV to Hypoxanthines V – General Procedure
A mixture of the adenine derivative IV (1.33 mmol), water (50 ml), 98% sulfuric acid (1.5 ml) and
3-methylbutyl nitrite (3 g, 25.6 mmol) was stirred for 7 days at room temperature. The resulting
yellow solution was neutralized with NaOH to pH 7 and extracted with ethyl acetate (4 × 100 ml).
The combined organic layers were evaporated and the residue was chromatographed on a preparative
layer of silica. The resulting (TLC pure) yellow oils were used in the further step.
Bis(2-propyl) 2-(benzyloxycarbonylaminomethyl)-9-(2-phosphonomethoxyethyl)hypoxanthine (Va),
yield 47%, RF 0.52. FAB MS, m/z (rel.%): 522 (100) [M + H]. 1H NMR spectrum ((CD3)2SO): 1.14
and 1.18 2 × d, 2 × 6 H, J(CH3,CH) = 6.1 (CH3-iPr); 3.77 d, 2 H, J(P,CH) = 8.5 (PCH2); 4.00 t, 2 H,
J(2′,1′) = 5.1 (H-2′); 4.20 d, 2 H, J(CH2,NH) = 6.1 (CH2N); 4.26 t, 2 H, J(1′,2′) = 5.1 (H-1′); 4.50 dsept,
2 H, J(CH,CH3) = 6.1, J(P,OCH) = 7.8 (CH-iPr); 5.06 s, 2 H (CH2Ph); 7.10 – 7.40 m, 5 H (arom. H);
7.70 t, 1 H, J(NH,CH2) = 6.0 (NHCb); 7.99 s, 1 H (H-8); 12.10 bs, 1 H (NH-1).
Bis(2-propyl) (R)-2-(benzyloxycarbonylaminomethyl)-9-(2-phosphonomethoxypropyl)hypoxanthine
1
(Vb), yield 52%, RF 0.59. FAB MS, m/z (rel.%): 536 (100) [M + H]. H NMR spectrum ((CD3)2SO):
1.02 d, 3 H, J(3′,2′) = 6.3 (H-3′); 1.14, 1.17, 1.18 and 1.21 4 × d, 4 × 3 H, J(CH3,CH) = 6.1
(CH3-iPr); 3.72 dd, 1 H, J(P,CH) = 9.5, J(gem) = 13.7 (PC-Hb); 3.78 dd, 1 H, J(P,CH) = 9.3, J(gem)
= 13.7 (PC-Ha); 3.93 m, 1 H (H-2′); 4.10 dd, 1 H, J(1′b,2′) = 6.3, J(gem) = 14.4 (Hb-1′); 4.19 d, 2 H,
J(CH2,NH) = 5.9 (CH2N); 4.21 dd, 1 H, J(1′a, 2′) = 3.9, J(gem) = 14.4 (Ha-1′); 4.51 and 4.52 2 × dsept,
2 × 1 H, J(CH,CH3) = 6.1, J(P,OCH) = 7.1 (CH-iPr); 5.06 s, 2 H (CH2Ph); 7.10 – 7.40 m, 5 H
(arom. H); 7.70 t, 1 H, J(NH,CH2) = 5.9 (NHCb); 7.96 s, 1 H (H-8); 12.15 bs, 1 H (NH-1).
Bis(2-propyl) (S)-2-(benzyloxycarbonylaminomethyl)-9-(3-hydroxy-2-phosphonomethoxypropyl)-
hypoxanthine (Vc), yield 46%, RF 0.58. FAB MS, m/z (rel.%): 552 (100) [M + H]. 1H NMR
spectrum ((CD3)2SO): 1.16, 1.165, 1.23, 1.24 4 × d, 4 × 3 H, J(CH3,CH) = 6.1 (CH3-iPr); 3.45 m, 2 H
(H-3′); 3.76 dd, 1 H, J(P,CH) = 9.5, J(gem) = 13.7 (PC-Hb); 3.85 m, 1 H (H-2′); 3.88 dd, 1 H,
J(P,CH) = 8.8, J(gem) = 13.7 (PC-Ha); 4.15 dd, 1 H, J(1′b,2′) = 7.6, J(gem) = 14.4 (Hb-1′); 4.21 d,
2 H, J(CH2,NH) = 6.0 (CH2N); 4.30 dd, 1 H, J(1′a,2′) = 3.5, J(gem) = 14.4 (Ha-1′); 4.49 m, 2 H
(CH-iPr); 5.00 bs, 1 H (OH); 5.06 s, 2 H (CH2Ph); 7.10 – 7.40 m, 5 H (arom. H); 7.75 t, 1 H,
J(NH,CH2) = 6.0 (NHCb); 7.98 s, 1 H (H-8); 12.10 bs, 1 H (NH-1).
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Deprotection of Intermediates IV and V – General Procedure
Iodotrimethylsilane (1.07 ml, 7.5 mmol) was added under argon to a stirred solution of protected
intermediates IV or V (0.75 mmol) in acetonitrile (25 ml). The mixture was stirred overnight at room
temperature, the solvent was evaporated in vacuo and the residue was codistilled with acetonitrile
and toluene (2 × 20 ml each) to remove excess iodotrimethylsilane. Water (50 ml) and triethylamine
(1 ml) were added and the mixture was washed with ether (3 × 50 ml). The aqueous layer was
applied on a Dowex D1 X2 (200 – 400 mesh, acetate form) column (50 ml). The column was washed
with water and the products were eluted with a gradient of acetic acid (0.01 – 0.1 mol/l). The
resulting white solid was recrystallized from water–ethanol.
2-(Aminomethyl)-9-(2-phosphonomethoxyethyl)adenine (VIa), yield 57%, colourless needles, m.p.
264 – 266 °C (dec.), EUp 0.56. For C9H15N6O4P (302.2) calculated: 35.76% C, 5.00% H, 27.81% N,
10.25% P; found: 35.57% C, 4.95% H, 27.52% N, 10.26% P. FAB MS, m/z (rel.%): 303 (80) [M + H].
1H NMR spectrum (D2O + NaOD): 3.52 d, 2 H, J(P, CH2) = 8.8 (PCH2); 3.96 t, 2 H, J(2′,1′) = 5.0
(H-2′); 4.02 s, 2 H (NCH2); 4.41 t, 2 H, J(1′,2′) = 5.0 (H-1′); 8.21 s, 1 H (H-8). UV spectrum (0.01 M HCl):
λmax 262 nm (log ε 4.03).
(R)-2-(Aminomethyl)-9-(2-phosphonomethoxypropyl)adenine(VIb), yield 51%, hygroscopic colourless
needles, m.p. 192 – 195 °C (dec.), EUp 0.53. CD spectrum (0.01 M HCl): λ 274 nm (∆ε 0.2). For
C10H17N6O4P . 2 H2O (352.3) calculated: 34.09% C, 6.01% H, 23.85% N, 8.85% P; found: 33.95% C,
5.79% H, 23.68% N, 8.85% P. FAB MS, m/z (rel.%): 317 (100) [M + H]. 1H NMR spectrum (D2O
+ NaOD): 1.14 d, 3 H, J(3′,2′) = 6.3 (H-3′); 3.54 dd, 1 H, J(P,CH) = 9.8, J(gem) = 12.2 (PC-Hb);
3.62 dd, 1 H, J(P,CH) = 9.5, J(gem) = 12.2 (PC-Ha); 3.98 m, 1 H (H-2′); 4.22 s, 2 H (NCH2); 4.22 dd,
1 H, J(1′b,2′) = 5.6, J(gem) = 14.7 (Hb-1′); 4.45 dd, 1 H, J(1′a,2′) = 3.6, J(gem) = 14.7 (Ha-1′); 8.19 s,
1 H (H-8). UV spectrum (0.01 M HCl): λmax 260 nm (log ε 4.06).
(S)-2-(Aminomethyl)-9-(3-hydroxy-2-phosphonomethoxypropyl)adenine (VIc), yield 68%, white
powder, m.p. 210 – 213 °C (dec.), EUp 0.49. CD spectrum (0.01 M HCl): λ 273 nm (∆ε 0.26). For
C10H17N6O5P . H2O (350.3) calculated: 34.29% C, 5.46% H, 23.99% N, 8.84% P; found: 33.90% C,
5.31% H, 23.60% N, 8.47% P. FAB MS, m/z (rel.%): 333 (100) [M + H]. 1H NMR spectrum (D2O
+ NaOD): 3.48 dd, 1 H, J(3′b,2′) = 4.6, J(gem) = 12.4 (Hb-3′); 3.49 dd, 1 H, J(P,CH) = 9.0, J(gem)
= 12.2 (PC-Hb); 3.54 dd, 1 H, J(P,CH) = 8.8, J(gem) = 12.2 (PC-Ha); 3.76 dd, 1 H, J(3′a,2′) = 3.7,
J(gem) = 12.4 (Ha-3′); 3.83 s, 2 H (NCH2); 3.86 m, 1 H (H-2′); 4.35 dd, 1 H, J(1′b,2′) = 6.3, J(gem)
= 14.6 (Hb-1′); 4.42 dd, 1 H, J(1′a,2′) = 4.4, J(gem) = 14.6 (Ha-1′); 8.21 s, 1 H (H-8). UV spectrum
(0.01 M HCl): λmax 262 nm (log ε 4.06).
2-(Aminomethyl)-9-(2-phosphonomethoxyethyl)hypoxanthine (VIIa), yield 66%, colourless needles,
m.p. >300 °C , EUp 0.83. For C9H14N5O5P (303.2) calculated: 35.65% C, 4.65% H, 23.10% N, 10.22% P;
1
found: 35.35% C, 4.69% H, 22.96% N, 9.88% P. FAB MS, m/z (rel.%): 304 (54) [M + H]. H NMR
spectrum (D2O + NaOD): 3.50 d, 2 H, J(P,CH2) = 8.3 (PCH2); 3.74 s, 2 H (NCH2); 3.96 t, 2 H,
J(2′,1′) = 5.1 (H-2′); 4.39 t, 2 H, J(1′,2′) = 5.1 (H-1′); 8.07 s, 1 H (H-8). UV spectrum (0.01 M HCl):
λmax 250 nm (log ε 3.95).
(R)-2-(Aminomethyl)-9-(2-phosphonomethoxypropyl)hypoxanthine (VIIb), yield 63%, white
hygroscopic powder, m.p. 204 – 207 °C (dec.), EUp 0.78. CD spectrum (0.01 M HCl): λ 266 nm (∆ε
0.08), λ 248 nm (∆ε −0.14), λ 223 nm (∆ε 0.13). For C10H16N5O5P . 3 H2O (371.3) calculated:
32.35% C, 5.97% H, 18.86% N, 8.34% P; found: 32.52% C, 5.80% H, 18.92% N, 8.16% P. FAB
MS, m/z (rel.%): 318 (100) [M + H]. 1H NMR spectrum (D2O + NaOD): 1.12 d, 3 H, J(3′,2′) = 6.3
(H-3′); 3.46 dd, 1 H, J(P,CH) = 9.0, J(gem) = 12.4 (PC-Hb); 3.55 dd, 1 H, J(P,CH) = 8.3, J(gem) = 12.4
(PC-Ha); 3.74 s, 2 H (NCH2); 4.01 m, 1 H (H-2′); 4.26 dd, 1 H, J(1′b,2′) = 5.6, J(gem) = 14.4
(Hb-1′); 4.35 dd, 1 H, J(1′a,2′) = 4.4, J(gem) = 14.4 (Ha-1′); 8.10 s, 1 H (H-8). UV spectrum (0.01 M
HCl): λmax 250 nm (log ε 4.00).
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(S)-2-(Aminomethyl)-9-(3-hydroxy-2-phosphonomethoxypropyl)hypoxanthine (VIIc), yield 66%,
colourless needles, m.p. 224 – 226 °C, EUp 0.80. CD spectrum (0.01 M HCl): λ 269 nm (∆ε 0.14),
λ 248 nm (∆ε −0.23), λ 223 (∆ε 0.27). For C10H16N5O6P . 2.5 H2O (378.3) calculated: 31.74% C,
5.59% H, 18.51% N, 8.19% P; found: 31.61% C, 5.18% H, 18.43% N, 8.78% P. FAB MS, m/z
1
(rel.%): 334 (60) [M + H]. H NMR spectrum (D2O + NaOD): 3.47 dd, 1 H, J(3′b,2′) = 4.4, J(gem)
= 12.4 (Hb-3′); 3.49 dd, 1 H, J(P,CH) = 9.5, J(gem) = 12.2 (PC-Hb); 3.57 dd, 1 H, J(P,CH) = 8.8,
J(gem) = 12.2 (PC-Ha); 3.75 dd, 1 H, J(3′a,2′) = 3.4, J(gem) = 12.4 (Ha-3′); 3.86 s, 2 H (NCH2);
3.88 m, 1 H (H-2′); 4.34 dd, 1 H, J(1′b,2′) = 6.1, J(gem) = 14.6 (Hb-1′); 4.41 dd, 1 H, J(1′a,2′) = 4.9,
J(gem) = 14.6 (Ha-1′); 8.08 s, 1 H (H-8). UV spectrum (0.01 M HCl): λmax 250 nm (log ε 3.94).
This work was supported by the Grant Agency of the Academy of Sciences of the Czech Republic
(Grants No. 455402 and No. 455407), by the State Grant Agency of the Czech Republic (Grant No.
203/93/0118) and by Gilead Sciences (Foster City, CA, U.S.A.). The antiviral activities were studied by
Professor E. De Clercq, Dr J. Balzarini and Dr R. Snoeck, Rega Institute, Catholic University, Leuven,
Belgium, the cytostatic activity by Dr I. Votruba of this Institute. The authors’ thanks are due to Dr H.
Votavova for measurement of CD spectra.
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1. Holy A., Dvorakova H., Jindrich J. in: Antiviral Acyclic Nucleotide Analogues in Antibiotics and
Antiviral Compounds (K. Krohn, H. A. Kirst and H. Maag, Eds), p. 455. VCH, Weinheim 1993.
2. Holy A., Votruba I., Merta A., Cerny J., Vesely J., Vlach J., Sediva K., Rosenberg I., Otmar M.,
Hrebabecky H., Travnicek M., Vonka V., Snoeck R., De Clercq E.: Antiviral Res. 13, 295
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Collect. Czech. Chem. Commun. (Vol. 60) (1995)