Synthesis of N9- and N7-[2-hydroxy-3- (phosphonomethoxy)-propyl] derivatives of N6-substituted adenines, 2,6-diaminopurines and related compounds

Article abstract of DOI:10.1135/cccc20041889

Base-catalyzed reactions of diethyl [(oxiranylmethoxy)methyl]phosphonate (2) with purine bases (adenine, 2,6-diaminopurine, 6-chloropurine and 2-amino-6-chloropurine) gave corresponding 9- or 7-[2-hydroxy-3- (phosphonomethoxy)propyl] purines. The adenine

Full text of DOI:10.1135/cccc20041889

Acyclic Nucleotide Analogues
1889
SYNTHESIS OF N9- AND N7-[2-HYDROXY-3-(PHOSPHONOMETHOXY)-
PROPYL] DERIVATIVES OF N6-SUBSTITUTED ADENINES,
2,6-DIAMINOPURINES AND RELATED COMPOUNDS
1,
2
3
Marcela KREČMEROVÁ *, Milena MASOJÍDKOVÁ and Antonín HOLÝ
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
1
2
166 10 Prague 6, Czech Republic; e-mail: marcela@uochb.cas.cz, masojid@uochb.cas.cz,
holy@uochb.cas.cz
3
Received May 11, 2004
Accepted Jun e 10, 2004
Dedicated with due respect to Professor Miloslav Černý on the occasion of his 75th birthday in
recognition of his outstanding contributions to carbohydrate chemistry.
Base-catalyzed reaction s of dieth yl [(oxiran ylm eth oxy)m eth yl]ph osph on ate (2) with purin e
bases (aden in e, 2,6-diam in opurin e, 6-ch loropurin e an d 2-am in o-6-ch loropurin e) gave corre-
spon din g 9- or 7-[2-h ydroxy-3-(ph osph on om eth oxy)propyl] purin es. Th e aden in e an d
2,6-diam in opurin e derivatives cyclize to cyclic ph osph on ates 4 an d 6. Th e 9-[2-h ydroxy-
3-(ph osph on om eth oxy)propyl] derivatives of N6-substituted aden in e an d 2,6-diam in opurin e
(15–27) were prepared by th e treatm en t of dieth yl {[3-(6-ch loropurin -9-yl)-2-h ydroxy-
propoxy]m eth yl}ph osph on ate (11) or dieth yl {[3-(2-am in o-6-ch loropurin -9-yl)-2-h ydroxy-
propoxy]m eth yl}ph osph on ate (13) with prim ary or secon dary am in es. Th e reaction of
6-ch loro- or 2-am in o-6-ch loropurin e derivatives (11, 13) with th iourea gave th e correspon d-
in g dieth yl purin e-6-th iol or 2-am in opurin e-6-th iol ph osph on ates 47, 48. Th e guan in e de-
rivative 49 was prepared by th e treatm en t of com poun d 13 with 80% acetic acid. All dieth yl
ph osph on ates were tran sform ed to free ph osph on ic acids (31–43, 50–52) by th e action of
brom otrim eth ylsilan e an d subsequen t h ydrolysis.
Keyw ord s: Acyclic n ucleotide an alogues; Ph osph on ates; Purin es; Oxiran e rin g open in g;
Epoxides; Nucleotides; Nucleosides; An tivirals; HPMPA.
In th e course of system atic studies of various types of acyclic n ucleotide an a-
logues we paid con siderable atten tion to a group of th e so-called HPMP de-
rivatives, i.e. (S)-9-[3-h ydroxy-2-(ph osph on om eth oxy)propyl] derivatives of
purin e an d pyrim idin e bases1. Som e of th ese com poun ds exh ibit sign ifican t
an tiviral activity, in particular th e aden in e an d diam in opurin e derivatives
(HPMPA, HPMPDAP)1,2, th eir guan in e coun terpart HPMPG 3 an d th e cyto-
sin e derivative HPMPC (cidofovir4), an active con stituen t of Vistide , th e
Collect. Czech. Chem. Commun. (Vol. 69) (2004)
doi:10.1135/cccc20041889

1890
Krečmerová, Masojídková, Holý:
drug used for th e treatm en t of CMV retin itis in AIDS patien ts an d
papillom atosis caused by HPV an d oth er viral diseases5.
Several approach es to th e preparation of HPMP derivatives h ave been
elaborated, m ost of th em usin g attach m en t of th e ph osph on om eth oxy
group to th e 3′-protected 9-(2,3-dih ydroxypropyl) derivatives by th e action
of a [(tosyloxy)m eth yl]ph osph on ic acid diester in th e presen ce of alkali6–8
.
Th e startin g N-(2,3-dih ydroxypropyl) derivatives are easily accessible eith er
by reaction of an appropriate n ucleobase with (R)-glycidol butyrate (in th e
case of R en an tiom ers), or by reaction of a n ucleobase with (R)-2,2-di-
m eth yl-4-[(tosyloxy)m eth yl]-1,3-dioxolan e (in th e case of S en an tiom ers).
Both types of reaction s are catalyzed by cesium carbon ate4.
In spite of n um erous studies dealin g with HPMP derivatives, th ere is on ly
a lim ited kn owledge about th eir structural isom ers bearin g a ph osph on o-
m eth yl eth er group at th e 3′-position , i.e. 9-[2-h ydroxy-3-(ph osph on o-
m eth oxy)propyl] derivatives of n ucleobases. Several years ago, we described
a first represen tative of th is group, 9-[2-h ydroxy-3-(ph osph on om eth oxy)-
propyl]aden in e8 (1). Un fortun ately, it proved to be biologically in active1a.
Later, we also described m ixtures of 2-h ydroxy-3-(ph osph on om eth oxy)-
propyl an d 3-h ydroxy-2-(ph osph on om eth oxy)propyl an alogues derived
from cytosin e an d 5-m eth ylcytosin e, as well as th eir deam in ated products
uracil an d th ym in e9. Th e form ation of such com poun ds was observed in
th e course of studies of esterification reaction s of 9-(2,3-dih ydroxypropyl)-
purin e derivatives, e.g. 9-(2,3-dih ydroxypropyl)aden in e (DHPA) with
(ch lorom eth yl)ph osph on ic acid derivatives8–10. Such reaction s afford al-
ways a m ixture of 2′- an d 3′-(ch lorom eth yl)ph osph on ates, wh ich un dergo
in tram olecular cyclization in aqueous alkali to correspon din g cyclic ph os-
ph on ates an d subsequen t rin g open in g to give a m ixture of both n ucleotide
an alogues: HPMPA from 3′-(ch lorom eth yl)ph osph on ate an d 3′-ph osph on o-
m eth oxy derivative 1 from 2′-(ch lorom eth yl)ph osph on ate. Both free acids
(HPMPA an d 1) are separable on ly by HPLC tech n ique, so th is m eth od is
useful on ly for m icro an d sem im icro scale. In som e cases th e differen ce in
HPLC m obilities of 2′- an d 3′-ph osph on ates is so sm all th at th eir separation
is im possible9.
NH2
NH2
N
N
N
N
N
N
N
N
OH
P(O)(OH)2
O
P(O)(OH)2
O
OH
1
HPMPA
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1891
In order to verify th e gen eral validity of th e rule accordin g to wh ich th e
biological activity is associated with th e 2′-ph osph on om eth yl isom ers on ly,
we h ave perform ed th e presen t study; its aim is th e elaboration of prepara-
tive syn th esis of a large group of racem ic 9-[2-h ydroxy-3-(ph osph on o-
m eth oxy)propyl] derivatives of various purin e bases: 2,6-diam in opurin e,
guan in e, 2-am in o-6-ch loropurin e, N6-(alkylam in o)purin es, 2-am in o-N6-
(alkylam in o)purin es, purin e-6-th iol an d 2-am in opurin e-6-th iol. Special care
is paid to N6-substituted aden in e an d 2,6-diam in opurin e derivatives with
respect to th e fact th at th e N6-substitution can con siderably in crease an tivi-
ral an d cytostatic activity of som e acyclic n ucleoside ph osph on ates. Th is ef-
fect is m ost eviden t with 2,6-diam in o-9-[2-(ph osph on om eth oxy)eth yl]-
purin e (PMEDAP), wh ere th e N-substitution of 6-am in o group leads to n u-
m erous com poun ds with a h igh cytostatic an d an tiviral activity11. Th e best
results were obtain ed with 2,2,2-trifluoroeth yl, allyl, 2-(dim eth ylam in o)-
eth yl, cyclopropyl an d dim eth yl N6-derivatives.
Our syn th eses of 2-h ydroxy-3-(ph osph on om eth oxy)propyl derivatives are
based on th e oxiran e rin g open in g of racem ic dieth yl [(oxiran ylm eth oxy)-
m eth yl]ph osph on ate (2) with n ucleobases. Th e syn th on 2 was prepared ac-
cordin g to a literature procedure12 startin g from epich loroh ydrin an d a so-
dium salt of dieth yl (h ydroxym eth yl)ph osph on ate, accessible by th e
Arbuzov reaction of dieth yl ph osph ite an d paraform aldeh yde13 (Sch em e 1).
O
P
O
P
C2H5O
C2H5O
Na
OC2H5
OC2H5
CH2ONa
(CH2O)x
H
+
ether
CH2 CH CH2Cl
O
O
OC2H5
OC2H5
O
CH2
P
CH2 CH CH2
O
2
SCHEME 1
Reactivity of dieth yl [(oxiran ylm eth oxy)m eth yl]ph osph on ate (2) towards
various n ucleobases was studied. Th e reaction was carried out with aden in e,
2,6-diam in opurin e, 6-ch loropurin e an d 2-am in o-6-ch loropurin e un der
stan dard con dition s: DMF as a solven t, tem perature 120 °C an d cesium car-
bon ate as a base catalyst. As expected, th e n ucleoph ilic open in g of oxiran e
rin g in 2 took place always at th e term in al C-3 position givin g regio-
specifically 9-[2-h ydroxy-3-(ph osph on om eth oxy)propyl] derivatives. Som e
of th ese com poun ds un derwen t an in tram olecular n ucleoph ilic attack of
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1892
Krečmerová, Masojídková, Holý:
2-h ydroxy group at th e ph osph orus atom form in g six-m em bered cyclic
ph osph on ates. Such situation occurs especially wh en th e reaction is carried
out with aden in e an d 2,6-diam in opurin e.
Th us, th e reaction of oxiran e 2 with aden in e afforded a m ixture of di-
eth yl ester of 9-[2-h ydroxy-3-(ph osph on om eth oxy)propyl]aden in e (3) an d
th e cyclization product 4 in th e ratio of 1:1. Th e cyclic ph osph on ate 4 was
an equim olar m ixture of two ch rom atograph ically easily separable dia-
stereoisom ers (4a an d 4b); both of th em were fully ch aracterized. On th e
oth er h an d, th e ch rom atograph ic m obilities of th e un cyclized com poun d 3
an d cyclic ph osph on ate 4a are practically th e sam e an d th eir separation
was possible on ly by acetylation of th e m ixture tran sform in g com poun d 3
to th e faster m ovin g acetate 5 (Sch em e 2).
NH2
NH2
N
N
N
N
N
N
+
R1
N
O
P
i
N
R
O
O
OC2H5
OR2
O
P
OC2H5
O
3, R1 = R2 = H
OC2H5
5, R1 = H, R2 = COCH3
7, R1 = NH2, R2 = H
4, R = H
6, R = NH2
O
P(O)(OC2H5)2
O
2
R
N
N
N
Cl
N
N
+
O
ii
N
N
N
Cl
O
P
OC2H5
R
O
OH
OC2H5
O
P
OC2H5
OH
OC2H5
11, R = H
13, R = NH2
12, R = H
14, R = NH2
(i) adenine or 2,6-diaminopurine; (ii) 6-chloropurine or 2-amino-6-chloropurine
in both cases: DMF, 110 oC, Cs2CO3, 2 h
SCHEME 2
With 2,6-diam in opurin e, oxiran e 2 affords th e cyclic ph osph on ate 6
practically as a sin gle reaction product. Com poun d 7 was presen t on ly in
trace am oun ts. In order to prepare th e required 9-[2-h ydroxy-3-(ph os-
ph on om eth oxy)propyl]purin e-2,6-diam in e (8), cyclic ph osph on ate 6 was
utilized. It was treated first with sodium m eth oxide to give 9-{2-h ydroxy-
3-[(h ydroxym eth oxyph osph oryl)m eth oxy]propyl}purin e-2,6-diam in e (9).
Rem oval of th e m eth yl ester group in 9 with brom otrim eth ylsilan e gave
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1893
free acid 8 as fin al product. An oth er m eth od for th e preparation of 8 con -
sists in tran sform ation of cyclic ph osph on ate 6 to free cyclic ph osph on ic
acid 10 an d its treatm en t with aqueous sodium h ydroxide (Sch em e 3).
However, th is m eth od is n ot so advan tageous as th e above m en tion ed pro-
cedure because of partial deam in ation of th e product.
NH2
NH2
N
N
N
N
N
N
i
N
N
H2N
H2N
O
O
O
O
10
iii
6
P
P
O
O
OH
OC2H5
ii
NH2
NH2
i
N
N
N
N
N
N
H2N
N
O
H2N
N
O
O
P
OCH3
O
P
OH
OH
OH
OH
OH
9
8
(i) (CH3)3SiBr, CH3CN, r.t.; (ii) CH3ONa/CH3OH, reflux; (iii) 1 M NaOH, 80 oC, 3 h
SCHEME 3
Th e treatm en t of dieth yl [(oxiran ylm eth oxy)m eth yl]ph osph on ate (2)
with 6-ch loropurin e gave a m ixture of N9- an d N7-alkylated products:
6-ch loro-9-{2-h ydroxy-3-[(dieth oxyph osph oryl)m eth oxy]propyl}purin e (11)
an d 6-ch loro-7-{2-h ydroxy-3-[(dieth oxyph osph oryl)m eth oxy]propyl}purin e
(12) in th e ratio 3:1. Both isom ers can be separated ch rom atograph ically. In
th is case, n o form ation of cyclic ph osph on ate was observed. Sim ilarly, n o
cyclization product was form ed in th e treatm en t of oxiran e 2 with
2-am in o-6-ch loropurin e; also in th is case on ly th e N9- an d N7-alkylated
products 13 an d 14 were isolated (in th e ratio 7:2). It can be speculated th at
in both cases th e electron -with drawin g effect of ch lorin e in th e purin e
m oiety decreases th e n ucleoph ilic ch aracter of 2-h ydroxy group an d m akes
its attack on ph osph orus difficult.
9-[2-Hydroxy-3-(ph osph on om eth oxy)propyl] derivatives of N6-substituted
aden in e an d 2,6-diam in opurin e derivatives were syn th esized by th e reac-
tion of prim ary or secon dary am in es with 6-ch loropurin e derivative 11 or
2-am in o-6-ch loropurin e derivative 13 (Table I, Sch em e 4). Th e reaction was
perform ed in boilin g aceton itrile un der exclusion of CO2, except for
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1894
Krečmerová, Masojídková, Holý:
R1
R1
N
Cl
ii
i
N
N
N
N
N
N
N
N
R2
OC2H5
R2
N
O
O
P
N
R
N
O
P
O
P
OH
O
OC2H5
O
OH
OC2H5
OH
OH
OC2H5
OH
11, R = H
13, R = NH2
31-43
15-27
(i) primary and secondary amines or dimethylammonium N,N-dimethylcarbamate, CH3CN, reflux;
(ii) (CH3)3SiBr, CH3CN, r.t.
SCHEME 4
TABLE I
Tran sform ation of 6-ch loro- an d 2-am in o-6-ch loropurin e derivatives of N9-[2-h ydroxy-
3-(ph osph on om eth oxy)propyl] series (11, 13) to correspon din g N6-substituted an alogues
Yield of
ph osph on ic
acid, %
Startin g
com poun d
Ph osph on ate
ester
Yield
of ester, %
Ph osph on ic
acid
R1
R2
11
11
11
11
11
11
11
11
13
13
13
13
13
15
16
17
18
19
20
21
22
23
24
25
26
27
–N(CH3)2
H
H
88
98
58
91
86
96
87
57
95
95
93
94
97
31
32
33
34
35
36
37
38
39
40
41
42
43
77
77
42
76
72
52
65
74
77
52
62
52
69
N
H
N
H
–NHCH2CH2OCH3
–NHCH2CH2OH
–NHCH2CH=CH2
(CH3)2NCH2CH2NH–
–NHCH2CF3
H
H
H
H
H
–N(CH3)2
NH2
NH2
NH2
NH2
NH2
N
H
–NHCH2CF3
–NHCH2CH=CH2
(CH3)2NCH2CH2NH–
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1895
2,2,2-trifluoroeth ylam in e, wh ere it was perform ed in an autoclave, an d for
dim eth ylam in e wh ich was replaced by dim eth ylam m on ium N,N-dim eth yl-
carbam ate. Th e th us obtain ed in term ediate dieth yl ph osph on ates 15–27
were purified by colum n ch rom atograph y on silica gel with th e exeption of
com poun ds with a low ch rom atograph ic m obility, i.e. N6-[2-(dim eth yl-
am in o)eth yl] derivatives 21 an d 27, wh ich were desalted by ion exch an ge
ch rom atograph y. An alogously, we prepared also som e N6-substituted ade-
n in e derivatives of th e 7-[2-h ydroxy-3-(ph osph on om eth oxy)propyl] series
(28–30) by th e treatm en t of 6-ch loropurin e derivative 12 with various
am in es un der th e above described con dition s (Table II, Sch em e 5). All di-
eth yl ph osph on ates were subsequen tly con verted to free ph osph on ic acids
by th e action of brom otrim eth ylsilan e in aceton itrile followed by de-
ion ization an d ion exch an ge ch rom atograph y on Dowex 50 resin . Th us, we
obtain ed a series of free 2-h ydroxy-3-(ph osph on om eth oxy)propyl deriva-
tives of N6-substituted aden in e an d diam in opurin e 31–43 an d 44–46 as
fin al products.
N
N
R
N
R
N
N
N
N
N
N
i
ii
N
N
N
O
P
O
P
O
Cl
O
OC2H5
O
OC2H5
O
P
OH
OH
OH
28-30
OH
44-46
OC2H5
OH
OC2H5
12
(i) amines or dimethylammonium N,N-dimethylcarbamate, CH3CN, reflux; (ii) (CH3)3SiBr, CH3CN, r.t.
SCHEME 5
TABLE II
Preparation of som e N6-substituted aden in e derivatives of N7-[2-h ydroxy-3-(ph osph on o-
m eth oxy)propyl] series from com poun d 12
Yield of
ph osph on ic
acid, %
Ph osph on ate
ester
Yield
of ester, %
Ph osph on ic
acid
R
28
29
30
–N(CH3)2
63
65
87
44
45
46
55
49
53
N
H
–NHCH2CH=CH2
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1896
Krečmerová, Masojídková, Holý:
An oth er tran sform ation of ch lorin e atom in position 6 of purin e con sists
in its con version to th iol group. Th us, th e treatm en t of dieth yl ph os-
ph on ates 11 an d 13 with th iourea in eth an ol afforded th e correspon din g
th iol derivatives 47 an d 48. Tran sform ation of ch lorin e atom in 2-am in o-
6-ch loropurin e com poun d 13 to guan in e derivative 49 was perform ed by
h eatin g with 80% acetic acid. Also dieth yl ph osph on ates 47–49 were
treated with brom otrim eth ylsilan e to give a series of free ph osph on ic acids
50–52.
X
47, X = S, R1 = H, R2 = C2H5
48, X = S, R1 = NH2, R2 = C2H5
49, X = O, R1 = NH2, R2 = C2H5
50, X = S, R1 = R2 = H
51, X = S, R1 = NH2, R2 = H
52, X = O, R1 = NH2, R2 = H
N
HN
N
O
N
R1
O
P
OR2
OH
OR2
In con clusion , we developed a n ovel syn th etic approach leadin g selec-
tively to 9- or 7-[2-h ydroxy-3-(ph osph on om eth oxy)propyl] derivatives of
purin e n ucleobases, th e structural isom ers of biologically active HPMP com -
poun ds.Usin g th e m eth od based on utilization of dieth yl [(oxiran yl-
m eth oxy)m eth yl]ph osph on ate as an easily accessible syn th on bearin g a
ph osph on om eth yl group, we h ave prepared a large series of racem ic 9- or
7-[2-h ydroxy-3-(ph osph on om eth oxy)propyl]purin e derivatives for biologi-
cal screen in g. All fin al products were tested for cytostatic activity in vitro.
In con trast to N6-substituted aden in es an d 2,6-diam in opurin es of N-[2-
(ph osph on om eth oxy)eth yl] series prepared earlier11, n o cytostatic activity
was foun d for th ese com poun ds. Th e an tiviral screen in g is un der way.
EXPERIMENTAL
Un less stated oth erwise, solven ts were evaporated at 40 °C/2 kPa an d com poun ds were dried
at 13 Pa. Meltin g poin ts were determ in ed on a Kofler block an d are un corrected. An alytical
TLC were perform ed on Silufol UV254 plates (Kavalier, Votice, Czech Republic) in system s
eth yl acetate–aceton e–eth an ol–water (18:3:1:1, system S1; 18:3:2:2, system S2 or 15:3:4:3,
system S3). Oth er ch rom atograph ic system s are described in text. Colum n ch rom atograph y
was perform ed on silica gel 60 µm (Fluka). 1H NMR spectra were m easured on a Varian
Un ity 500 in strum en t (at 500 MHz) in DMSO-d6 solution s (referen ced to th e solven t sign al
at δ 2.50) or in D2O solution s with in tern al stan dard sodium 3-(trim eth ylsilyl)propan e-
1-sulfon ic acid (DSS). 1H NMR ch em ical sh ifts (δ, ppm ) an d couplin g con stan ts (J, Hz) were
obtain ed by first-order an alysis of th e spectra. 13C NMR spectra were recorded on th e sam e
in strum en t (at 125.7 MHz) usin g APT pulse sequen ce in DMSO-d6 (referen ced to th e solven t
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1897
sign al δ 39.70). Th e n um berin g system for assign m en t of NMR sign als is outlin ed in Fig. 1.
Mass spectra were m easured on a ZAB-EQ (VG An alytical) spectrom eter usin g FAB (ion iza-
tion with xen on , acceleratin g voltage 8 kV, glycerol m atrix).
Materials an d Solven ts
Most of ch em icals an d ion -exch an ge resin (Dowex 50W X8-200) were purch ased from
Sigm a–Aldrich (Czech Republic). 2,2,2-Trifluoroeth ylam in e was a product of Fluka. Di-
m eth ylform am ide an d aceton itrile were dried by distillation from CaH2 (DMF in vacuo) an d
stored over m olecular sieves (4 Å).
Dieth yl [(Oxiran ylm eth oxy)m eth yl]ph osph on ate (2)
Th e com poun d was prepared by publish ed procedure12 from dieth yl ph osph ite (27.6 g,
0.2 m ol). Yield 30 g (67%) of a colorless liquid, b.p. 130 °C/133 Pa. 1H NMR (DMSO-d6):
4.05 dq, 2 H an d 4.03 dq, 2 H, J(CH2,CH3) = 7.1, J(P,OCH) = 7.6 (P-OCH2); 3.85 d, 2 H,
J(P,CH) = 8.3 (PCH2); 3.82 dd, 1 H, J(3a,2) = 3.7, J(gem ) = 11.6 (H-3a); 3.35 dd, 1 H, J(3b,2) =
6.6 (H-3b); 3.12 m , 1 H (H-2); 2.73 dd, 1 H, J(1b,2) = 4.4, J(gem ) = 5.0 (H-1b); 2.55 dd, 1 H,
J(1a,2) = 2.7 (H-1a); 1.24 t, 6 H (CH3). 13C NMR ((DMSO-d6): 73.56 d, J(P,C) = 11.7 (C-3);
64.40 d, J(P,C) = 163.6 (PCH2); 61.92 d, 2 C, J(P,C) = 6.3 (P-OCH2); 50.12 (C-2); 43.45 (C-1);
16.98 d, 2 C, J(P,C) = 5.4 (CH3).
Reaction of Dieth yl [(Oxiran ylm eth oxy)m eth yl]ph osph on ate (2) with Aden in e
A suspen sion of aden in e (270 m g, 2 m m ol) an d Cs2CO3 (65 m g, 0.2 m m ol) in dry DMF (10 m l)
was h eated to 120 °C. Oxiran e 2 (448 m g, 2 m m ol) was added an d th e reaction m ixture was
stirred at 120 °C for 1 h . After coolin g to room tem perature, th e m ixture was n eutralized
with acetic acid to pH 7, taken down in vacuo, th e residue codistilled with xylen e (2 × 10 m l)
an d th en applied on to a colum n of silica gel (150 m l) in system S2. Th e ch rom atograph y
gave, in elution sequen ce, a n on -separable m ixture of 4b an d 3 as a wh ite solid (330 m g to-
geth er) an d pure diastereoisom er 4a.
9-[(2-Ethoxy-2-oxo-2λ5-1,4,2-dioxaphosphinan-6-yl)methyl]-9H-purin-6-amine (4a). Isolated as
wh ite crystals (113 m g, 18%), m .p. 172–174 °C (eth an ol). FAB MS, m/z (%): 314 (100) [M + H],
136 (11) [aden in e + H]. 1H NMR (DMSO-d6): 8.16 s, 1 H an d 8.10 s, 1 H (H-2, H-8); 7.30 br s,
2 H (NH2); 4.99 m , 1 H, ΣJ = 28.7, J(2′,P) = 8.0 (H-2′); 4.47 ddd, 1 H, J(1′a,P) = 0.9, J(1′a,2′) =
7.3, J(gem ) = 14.6 (H-1′a); 4.43 ddd, 1 H, J(1′b,P) = 1.6, J(1′b,2′) = 4.8 (H-1′b); 4.06 m , 2 H
1´´
3´´
R
N
6
Ha
Hb
2´´
Ha
O
N
N
2
1
3
8
O
P
OCH2CH3
OCH2CH3
Hb
N
O
X
N
2
H
O
3´
2´
1´
O
P
OR
OH
OR
FIG. 1
Gen eral n um berin g sch em e for assign m en t of NMR sign als
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1898
Krečmerová, Masojídková, Holý:
(P-OCH2); 4.01 d, 2 H, J(P,CH) = 5.0 (PCH2); 3.94 dd, 1 H, J(3′a,2′) = 1.8, J(gem ) = 12.8
(H-3′a); 3.56 dd, 1 H, J(3′b,2′) = 6.7 (H-3′b); 1.22 t, 3 H, J(CH3,CH2) = 7.1 (CH3). 13C NMR
(DMSO-d6): 156.20 (C-6); 152.75 (C-2); 149.875 (C-4); 141.72 (C-8); 118.72 (C-5); 78.33 d,
J(P,C) = 8.3 (C-2′); 67.67 d, J(P,C) = 5.4 (C-3′); 63.94 d, J(P,C) = 141.6 (P-C); 62.84 d, J(P,C) =
5.9 (P-OCH2); 43.84 d, J(P,C) = 2.9 (C-1′); 16.47 d, J(P,C) = 4.9 (CH3).
Diethyl {[3-(6-amino-9H-purin-9-yl)-2-hydroxypropyloxy]methyl}phosphonate (3) and 9-[(2-
ethoxy-2-oxo-2λ5-1,4,2-dioxaphosphinan-6-yl)methyl]-9H-purin-6-amine (4b). FAB MS, m/z (%):
360 (40) [M + H, 3], 314 (55) [M + H, 4b]. Separation an d ch aracterization of both com -
poun ds after acetylation .
Acetylation of a Mixture of Com poun ds 3 an d 4b. Separation of Products
A m ixture of 3 an d 4b (330 m g) in aceton itrile (20 m l) was stirred with acetic an h ydride
(178 m g, 1.8 m m ol) an d 4-(dim eth ylam in o)pyridin e (10 m g, 0.08 m m ol) at am bien t tem pera-
ture for 1 h , th en diluted with m eth an ol (5 m l) an d taken down . Th e residue was ch ro-
m atograph ed on a colum n of silica gel (100 m l) in system S3. Two products were obtain ed
in th e followin g elution sequen ce.
2-(6-Amino-9H-purin-9-yl)-1-{[(diethoxyphosphoryl)methoxy]methyl}ethyl acetate (5). Yield
250 m g (31%, based on aden in e), wh ite solid, m .p. 87–88 °C. For C15H24N5O6P (401.4) cal-
culated: 44.89% C, 6.03% H, 17.45% N, 7.72% P; foun d: 44.33% C, 6.30% H, 17.61% N,
7.33% P. FAB MS, m/z (%): 402 (100) [M + H], 136 (28) [aden in e + H]. 1H NMR (DMSO-d6):
8.16 s, 1 H an d 8.10 s, 1 H (H-2, H-8); 7.31 br s, 2 H (NH2); 5.29 m , 1 H (H-2′); 4.41 dd, 1 H,
J(1′a,2′) = 3.9, J(gem ) = 14.7 (H-1′a); 4.34 dd, 1 H, J(1′b,2′) = 7.2 (H-1′b); 4.04 m , 4 H
(P-OCH2); 3.89 dd, 1 H, J(P,CHa) = 7.9, J(gem ) = 14.2 (PCHa); 3.85 dd, 1 H, J(P,CHb) = 7.9
(PCHb); 3.72 dd, 1 H, J(3′a,2′) = 4.0, J(gem ) = 10.9 (H-3′a); 3.62 dd, 1 H, J(3′b,2′) = 6.1
(H-3′b); 1.90 s, 3 H (acetyl); 1.23 t, 3 H an d 1.225 t, 3 H, J(CH3,CH2) = 7.1 (CH3). 13C NMR
(DMSO-d6): 169.69 (COCH3); 155.97 (C-6); 152.49 (C-2); 149.97 (C-4); 141.395 (C-8); 118.63
(C-5); 71.36 d, J(P,C) = 10.7 (C-3′); 70.33 (C-2′); 64.58 d, J(P,C) = 162.1 (P-C); 62.00 d, 2 C,
J(P,C) = 6.3 (P-OCH2); 43.26 (C-1′); 20.77 (COCH3); 16.46 d, 2 C, J(P,C) = 5.4 (CH3).
9-[(2-Ethoxy-2-oxo-2λ5-1,4,2-dioxaphosphinan-6-yl)methyl]-9H-purin-6-amine (4b). Isolated as
wh ite crystals, m .p. 183–184 °C (eth an ol). Yield 107 m g (16%). For C11H16N5O4P·0.5C2H5OH
(336.3) calculated: 42.86% C, 5.69% H, 20.83% N, 9.21% P; foun d: 42.72% C, 5.90% H,
20.32% N, 8.82% P. FAB MS, m/z (%): 314 (100) [M + H], 136 (12) [aden in e + H]. 1H NMR
(DMSO-d6): 8.165 s, 1 H an d 8.11 s, 1 H (H-2, H-8); 7.33 br s, 2 H (NH2); 4.91 m , 1 H, ΣJ =
24.9, J(2′,P) = 2.0 (H-2′); 4.39 ddd, 1 H, J(1′a,2′) = 4.3, J(gem ) = 15.6, J(1′a,P) = 1.8 (H-1′a);
4.35 ddd, 1 H, J(1′b,P) = 1.4, J(1′b,2′) = 6.7 (H-1′b); 4.17 dd, 1 H, J(P,CHa) = 10.0, J(gem ) =
14.9 (PCHa); 3.99 dt, 1 H, J(3′a,P) = J(3′a,2′) = 1.8, J(gem ) = 12.8 (H-3′a); 3.90 m , 2 H
(P-OCH2); 3.75 d, 1 H, J(gem ) = 14.9 (PCHb); 3.40 dd, 1 H, J(3′b,2′) = 10.1 (H-3′b); 1.08 t,
3 H, J(CH3,CH2) = 7.1 (CH3). 13C NMR (DMSO-d6): 156.145 (C-6); 152.68 (C-2); 149.84
(C-4); 141.56 (C-8); 118.72 (C-5); 79.37 d, J(P,C) = 8.3 (C-2′); 67.92 d, J(P,C) = 4.4 (C-3′);
62.54 d, J(P,C) = 141.1 (P-C); 61.36 d, J(P,C) = 6.3 (P-OCH2); 43.69 d, J(P,C) = 5.4 (C-1′);
16.16 d, J(P,C) = 5.4 (CH3).
9-[(2-Eth oxy-2-oxo-2λ5-1,4,2-dioxaph osph in an -6-yl)m eth yl]-9H-purin e-2,6-diam in e (6)
A suspen sion of 2,6-diam in opurin e (500 m g, 3.33 m m ol) an d Cs2CO3 (130 m g, 0.4 m m ol)
in DMF (18 m l) was h eated to 120 °C. Oxiran e 2 (747 m g, 3.33 m m ol) was added an d th e
h eatin g con tin ued for 3 h . Th e reaction m ixture was worked up in th e sam e m an n er as de-
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1899
scribed for aden in e derivatives 3, 4. Th e crude m ixture of products was ch rom atograph ed on
a silica gel colum n (300 m l) in system ch loroform –m eth an ol (4:1) givin g 6 as a wh ite solid
(700 m g, 64%) con tam in ated by a ch rom atograph ically n on -separable com poun d 7 (ca. 10%
accordin g to NMR spectrum ). For ch aracterization of 6, acetylation of such obtain ed product
was perform ed: A m ixture of crude 6 (700 m g, 2.1 m m ol), aceton itrile (25 m l) an d
4-(dim eth ylam in o)pyridin e (20 m g, 0.16 m m ol) was stirred with acetic an h ydride (80 m g,
0.8 m m ol) at 25 °C for 2 h . After addition of m eth an ol (5 m l), th e m ixture was evaporated
an d th e residue ch rom atograph ed on silica gel (200 m l) in th e system ch loroform –m eth an ol
(4:1). A m ixture of ch rom atograph ically faster m ovin g acetates was eluted first, followed by
a pure com poun d 6 wh ich is a m ixture of two diastereoisom ers (6a an d 6b, 3:1), yield
500 m g (46%), wh ite crystals, m .p. 218–220 °C (CH3OH). HR MS (FAB): For C11H18N6O4P
[M + H] calculated: 329.1127; foun d: 329.1147. FAB MS, m/z (%): 329 (85) [M + H]. 1H NMR
(DMSO-d6), m ajor diastereoisom er 6a: 7.66 s, 1 H (H-8); 6.74 br s, 2 H (NH2); 5.85 br s,
2 H (NH2); 4.94 m , 1 H (OCH); 4.27 br dd, 1 H, J(1′a,P) ≤ 1.0, J(1′a,2′) = 7.2, J(gem ) = 14.6
(H-1′a); 4.22 ddd, 1 H, J(1′b,P) = 1.6, J(1′b,2′) = 5.1 (H-1′b); 4.07 m , 2 H (P-OCH2); 4.03 dd,
1 H, J(P,CH) = 5.5, J(gem ) = 14.3 an d 4.00 dd, 1 H, J(P,CH) = 4.5 (PCH2); 3.88 dd, 1 H,
J(3′a,2′) = 2.8, J(gem ) = 12.9 (H-3′a); 3.54 dd, 1 H, J(3′b,2′) = 6.8 (H-3′b); 1.24 t, 3 H,
J(CH3,CH2) = 7.1 (CH3). 13C NMR (DMSO-d6): 160.56 (C-2); 156.34 (C-6); 152.09 (C-4);
137.93 (C-8); 113.15 (C-5); 78.16 d, J(P,C) = 8.3 (C-2′); 67.72 d, J(P,C) = 4.9 (C-3′); 64.01 d,
J(P,C) = 142.1 (P-C); 62.81 d, J(P,C) = 5.9 (P-OCH2); 43.41 d, J(P,C) = 2.9 (C-1′); 16.49 d,
J(P,C) = 5.4 (CH3). 1H NMR (DMSO-d6), m in or diastereoisom er 6b: 7.67 s, 1 H (H-8); 6.74 br s,
2 H (NH2); 5.85 br s, 2 H (NH2); 4.88 m , 1 H (OCH); 4.18 dd, 1 H, J(P,CH) = 10.0, J(gem ) =
14.9 an d 3.76 d, 1 H, J(P,CH) = 0, J(gem ) = 14.9 (PCH2); 4.16 m , 2 H (H-1′); 3.95 m , 2 H
(P-OCH2); 3.93 dt, 1 H, J(3′a,P) = J(3′a,2′) = 1.8, J(gem ) = 12.9 (H-3′a); 3.40 dd, 1 H, J(3′b,2′) =
10.1 (H-3′b); 1.13 t, 3 H, J(CH3,CH2) = 7.1 (CH3). 13C NMR (DMSO-d6): 160.56 (C-2); 156.34
(C-6); 152.06 (C-4); 138.07 (C-8); 113.15 (C-5); 79.26 d, J(P,C) = 8.3 (C-2′); 68.02 d, J(P,C) =
4.2 (C-3′); 62.58 d, J(P,C) = 141.1 (P-C); 61.365 d, J(P,C) = 5.4 (P-OCH2); 43.25 d, J(P,C) = 5.4
(C-1′); 16.17 d, J(P,C) = 5.4 (CH3).
Meth yl {[3-(2,6-Diam in o-9H-purin -9-yl)-2-h ydroxypropoxy]m eth yl}ph osph on ate (9)
A solution of 6 (390 m g, 1.2 m m ol) in 1 M sodium m eth oxide in m eth an ol (25 m l) was
refluxed for 6 h , th en left to cool to room tem perature an d n eutralized with Dowex 50 (H+
form ). Hydroch loric acid (36%) was added un til th e precipitated product dissolved. Th e m ix-
ture was applied on to a colum n of Dowex 50 (H+ form , 100 m l). Elution was carried out
with water (1200 m l) an d th en con tin ued with 2.5% aqueous am m on ia. Th e UV absorbin g
am m on ia eluate was collected an d evaporated in vacuo. Yield 307 m g (69.5%) of wh ite crys-
tals, m .p. 235–238 °C (eth an ol). For C10H17N6O5P·2H2O (368.3) calculated: 32.61% C,
5.75% H, 22.82% N, 8.41% P; foun d: 32.75% C, 5.55% H, 23.30% N, 8.32% P. HR MS (FAB):
for C10H18N6O5P [M + H] calculated: 333.1076; foun d: 333.1066. FAB MS, m/z (%): 333 (40)
[M + H]. 1H NMR (D2O + NaOD): 7.81 s, 1 H (H-8); 4.19 dd, 1 H, J(1′a,2′) = 4.2, J(gem ) =
13.4 (H-1′a); 4.15 m , 1 H (H-2′); 4.06 dd, 1 H, J(1′b,2′) = 6.7 (H-1′b); 3.72 dd, 1 H an d
3.69 dd, 1 H, J(P,CH) = 8.4, J(gem ) = 10.4 (P-CH2); 3.62 d, 3 H, J(P,OCH) = 10.4 (P-OCH3);
3.61 dd, 1 H, J(3′a,2′) = 4.5, J(gem ) = 10.3 (H-3′a); 3.54 dd, 1 H, J(3′b,2′) = 5.2 (H-3′b).
13C NMR (D2O + NaOD): 159.96 (C-2); 156.04 (C-6); 151.28 (C-4); 140.75 (C-8); 112.82
(C-5); 74.60 d, J(P,C) = 11.2 (C-3′); 68.28 (C-2′); 65.56 d, J(P,C) = 157.72 (PCH2); 51.93 d,
J(P,C) = 5.9 (P-OCH3); 46.44 (C-1′).
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1900
Krečmerová, Masojídková, Holý:
{[3-(2,6-Diam in o-9H-purin -9-yl)-2-h ydroxypropoxy]m eth yl}ph osph on ic Acid (8)
Method A. From 2,6-diam in opurin e (550 m g, 3.66 m m ol) an d oxiran e 2 (830 m g, 3.7 m m ol)
in th e sam e m an n er as described for com poun d 6. Th e crude product (con tain in g sm all
am oun t of 7) was dried in vacuo at 60 °C an d treated with brom otrim eth ylsilan e (4.05 m l,
30 m m ol) in aceton itrile (30 m l) at room tem perature for 48 h . Th e reaction m ixture was
th en poured to 1 M trieth ylam m on ium h ydrogen carbon ate (100 m l), evaporated to dryn ess
an d th e residue coevaporated with water (3 × 100 m l). 1 M NaOH (25 m l) was added, th e so-
lution h eated to 80 °C for 4 h , th en cooled to room tem perature, n eutralized with acetic
acid to pH 6 an d evaporated to ca. quarter of its origin al volum e. Hydroch loric acid (36%)
was added un til th e product dissolved an d th e solution was applied on to a colum n of
Dowex 50 (H+ form , 300 m l). Elution was carried out first with water (3.5 l; th orough
wash in g with water is n ecessary to rem ove con tam in atin g guan in e derivatives) an d th en con -
tin ued with 1.5% am m on ia. Th e UV absorbin g am m on ia eluate was taken down to give a
crude 8 togeth er with a sm all am oun t of 10. Purification of th is product by HPLC was per-
form ed on C-18 reverse ph ase with elution with water. Th e ph osph on ate 8 was eluted first,
followed by com poun d 10. Fraction s of 8 were evaporated to dryn ess an d th e residue was
crystallized from 80% aqueous eth an ol. Yield 600 m g (48%) of wh ite crystals, m .p. 222–
225 °C (decom p.). For C9H15N6O5P·1.5H2O (345.3) calculated: 31.31% C, 5.25% H, 24.34% N,
8.97% P; foun d: 31.42% C, 4.97% H, 24.02% N, 9.02% P. FAB MS, m/z (%): 319 (10) [M + H].
1H NMR (D2O): 7.78 s, 1 H (H-8); 4.20 dd, 1 H, J(1′a,2′) = 3.7, J(gem ) = 14.2 (H-1′a); 4.18 m ,
1 H (H-2′); 4.10 dd, 1 H, J(1′b,2′) = 9.2 (H-1′b); 3.69 dd, 1 H, J(3′a,2′) = 3.7, J(gem ) = 10.6
(H-3′a); 3.57 dd, 1 H, J(3′b,2′) = 5.6 (H-3′b); 3.64 dd, 1 H, J(P,CH) = 8.8, J(gem ) = 12.7 an d
3.61 dd, 1 H, J(P,CH) = 8.7 (P-CH2). 13C NMR (D2O): 159.59 (C-2); 155.98 (C-6); 150.69
(C-4); 140.56 (C-8); 112.63 (C-5); 73.71 d, J(P,C) = 11.2 (C-3′); 68.56 d, J(P,C) = 154.8 (P-C);
68.29 (C-2′); 45.86 (C-1′).
Method B. A solution of 6 (390 m g, 1.2 m m ol) in 1 M sodium m ethoxide in m ethanol (25 m l)
was refluxed for 6 h , th en left stan din g at room tem perature overn igh t an d n eutralized with
Dowex 50 (H+ form ). Hydroch loric acid (36%) was added to dissolution of precipitated prod-
uct. Th e m ixture was applied on to a colum n of Dowex 50 (H+ form , 100 m l), th e elution
was perform ed with water (1.2 l) an d th en with 2.5% aqueous am m on ia. Th e UV absorbin g
eluate was evaporated in vacuo an d th e residue stirred with a m ixture of eth an ol an d ace-
ton e (1:1, 10 m l). Th e solid product was filtered off an d dried in vacuo. Yield 130 m g (68%)
of wh ite solid (an h ydrous).
6-[(2,6-Diam in o-9H-purin -9-yl)m eth yl]-2-h ydroxy-2λ5-1,4,2-dioxaph osph in an -2-on e (10)
Th e com poun d was isolated as a side product in preparation of 8. Yield 250 m g (22%) of
wh ite crystals, m .p. > 300 °C. For C9H13N6O4P·0.5H2O (309.2) calculated: 34.96% C, 4.56% H,
27.18% N, 10.02% P; foun d: 34.76% C, 4.39% H, 26.77% N, 9.79% P. FAB MS, m/z (%): 301
(70) [M + H]. 1H NMR (D2O + NaOD): 7.80 s, 1 H (H-8); 4.71 m , 1 H (H-2′); 4.16 m , 2 H
(H-1′); 3.98 br dt, 1 H, J(3′a,2′) ≈ J(3′a,P) ≤ 1.0, J(gem ) = 12.2 (H-3′a); 3.92 dd, 1 H, J(P,CH) =
8.7, J(gem ) = 13.9 an d 3.61 dd, 1 H, J(P,CH) = 1.8 (P-CH2); 3.32 dd, 1 H, J(3′b,2′) = 10.2,
J(gem ) = 12.2 (H-3′b). 13C NMR (D2O + NaOD): 159.86 (C-2); 155.83 (C-6); 150.94 (C-4);
140.51 (C-8); 112.51 (C-5); 76.34 d, J(P,C) = 6.4 (C-2′); 68.40 d, J(P,C) = 3.0 (C-3′); 65.51 d,
J(P,C) = 143.5 (P-C); 43.95 d, J(P,C) = 5.4 (C-1′).
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1901
Reaction of Dieth yl [(Oxiran ylm eth oxy)m eth yl]ph osph on ate (2) with 6-Ch loropurin e an d
2-Am in o-6-ch loropurin e. Gen eral Procedure
A suspen sion of an appropriate n ucleobase (21 m m ol) an d Cs2CO3 (652 m g, 2 m m ol) in dry
DMF (100 m l) was evaporated to h alf of its volum e an d th en h eated to 115 °C. Com poun d 2
(4.42 g, 19.7 m m ol) was added an d th e reaction m ixture stirred at 115 °C for 2 h . After cool-
in g to room tem perature th e m ixture was n eutralized with acetic acid to pH 7, evaporated in
vacuo an d th e residue codistilled with xylen e (2 × 50 m l). Th e crude m ixture of products
was ch rom atograph ed on a silica gel colum n (1000 m l) in system S1 in separation of 11 an d
12 or S2 in separation of 13 an d 14. In both cases th e N9-isom ers are ch rom atograph ically
faster th en th e N7-isom ers.
Diethyl {[3-(6-chloro-9H-purin-9-yl)-2-hydroxypropoxy]methyl}phosphonate (11). Yield 4.2 g
(56%) of a colorless syrup. For C13H20ClN4O5P (378.8) calculated: 41.23% C, 5.32% H,
9.36% Cl, 14.79% N, 8.18% P; foun d: 41.24% C, 5.63% H, 9.26% Cl, 14.89% N, 7.85% P.
FAB MS, m/z (%): 379 (100) [M + H], 155 (20) [6-ch loropurin e + H]. 1H NMR (DMSO-d6):
8.77 s, 1 H an d 8.62 s, 1 H (H-2, H-8); 5.36 d, 1 H, J(OH,2′) = 5.6 (OH); 4.40 dd, 1 H,
J(1′a,2′) = 3.6, J(gem ) = 13.9 (H-1′a); 4.21 dd, 1 H, J(1′b,2′) = 8.4 (H-1′b); 4.07 m , 1 H (H-2′);
4.06 br pen t, 4 H, J(CH2,CH3) ≈ J(P,CH2) = 7.1 (P-OCH2); 3.87 d, 2 H, J(P,CH) = 7.8 (P-CH2);
3.56 dd, 1 H, J(3′a,2′) = 5.2, J(gem ) = 10.0 (H-3′a); 3.52 dd, 1 H, J(3′b,2′) = 5.9 (H-3′b); 1.24 t,
6 H (CH3). 13C NMR (DMSO-d6): 152.41 (C-4); 151.56 (C-2); 148.99 (C-6); 148.36 (C-8);
130.94 (C-5); 74.66 d, J(P,C) = 10.3 (C-3′); 67.31 (C-2′); 64.70 d, J(P,C) = 162.1 (P-C); 61.97 d
an d 61.92 d, J(P,C) = 5.9 (P-OCH2); 47.31 (C-1′); 16.51 d, 2 C, J(P,C) = 5.4 (CH3).
Diethyl {[3-(6-chloro-7H-purin-7-yl)-2-hydroxypropoxy]methyl}phosphonate (12). Yield 1.4 g
(19%) of wh ite crystals, m .p. 98–100 °C (eth an ol–aceton e 1:1). For C13H20ClN4O5P (378.8)
calculated: 41.23% C, 5.32% H, 9.36% Cl, 14.79% N, 8.18% P; foun d: 41.53% C, 5.21% H,
9.30% Cl, 15.25% N, 7.82% P. FAB MS, m/z (%): 379 (100) [M + H], 155 (17) [6-chloropurine + H].
1H NMR (DMSO-d6): 8.80 s, 1 H an d 8.71 s, 1 H (H-2, H-8); 5.36 d, 1 H, J(OH,2′) = 5.6 (OH);
4.67 dd, 1 H, J(1′a,2′) = 3.1, J(gem ) = 14.3 (H-1′a); 4.31 dd, 1 H, J(1′b,2′) = 9.1 (H-1′b); 4.06
br pen t, 4 H, J(CH2,CH3) = J(P,CH2) = 7.1 (P-OCH2); 4.00 m , 1 H (H-2′); 3.62 dd, 1 H,
J(3′a,2′) = 5.0, J(gem ) = 10.0 (H-3′a); 3.57 dd, 1 H, J(3′b,2′) = 5.9 (H-3′b); 1.25 t, 6 H (CH3).
13C NMR (DMSO-d6): 161.84 (C-4); 151.97 (C-8); 151.54 (C-2); 142.25 (C-6); 122.445 (C-5);
74.57 d, J(P,C) = 10.7 (C-3′); 68.47 (C-2′); 64.75 d, J(P,C) = 162.5 (P-C); 61.91 d, 2 C, J(P,C) =
5.9 (P-OCH2); 50.04 (C-1′); 16.50 d, 2 C, J(P,C) = 5.4 (CH3).
Diethyl {[3-(2-amino-6-chloro-9H-purin-9-yl)-2-hydroxypropoxy]methyl}phosphonate (13). Yield
4.5 g (58%) of a colorless syrup. For C13H21ClN5O5P (393.8) calculated: 39.65% C, 5.38% H,
9.00% Cl, 17.79% N, 7.87% P; foun d: 39.66% C, 5.43% H, 9.06% Cl, 17.72% N, 7.78% P.
FAB MS, m/z (%): 394 (100) [M + H], 170 (50) [2-am in o-6-ch loropurin e + H]. 1H NMR
(DMSO-d6): 8.03 s, 1 H (H-8); 6.91 br s, 2 H (NH2); 5.33 d, 1 H, J(OH, 2′) = 5.3 (2′-OH);
4.05 dq, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 8.2 (P-OCH2); 4.13 dd, 1 H, J(1′a,2′) = 3.0,
J(gem ) = 13.2 (H-1′a); 3.95 dd, 1 H, J(1′b,2′) = 8.5 (H-1′b); 4.02 m , 1 H (H-2′); 3.86 d, 2 H,
J(P,CH) = 7.8 (PCH2); 3.53 dd, 1 H, J(3′a,2′) = 5.1, J(gem ) = 10.0 (H-3′a); 3.49 dd, 1 H,
J(3′b,2′) = 5.4 (H-3′b); 1.24 t, 6 H (CH3). 13C NMR (DMSO-d6): 159.91 (C-2); 154.50 (C-4);
149.37 (C-6); 144.20 (C-8); 123.46 (C-5); 74.97 d, J(P,C) = 9.8 (C-3′); 67.19 (C-2′); 64.75 d,
J(P,C) = 162.1 (P-C); 61.94 d, 2 C, J(P,C) = 6.4 (P-OCH2); 46.59 (C-1′); 16.52 d, 2 C, J(P,C) =
5.4 (CH3).
Diethyl {[3-(2-amino-6-chloro-7H-purin-7-yl)-2-hydroxypropoxy]methyl}phosphonate (14). Yield
1.3 g (17%) of a wh ite solid, m .p. 70–72 °C. For C13H21ClN5O5P·0.5H2O (402.8) calculated:
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1902
Krečmerová, Masojídková, Holý:
38.77% C, 5.51% H, 8.80% Cl, 17.38% N, 7.69% P; foun d: 38.94% C, 5.55% H, 8.86% Cl,
17.11% N, 7.30% P. FAB MS, m/z (%): 394 (100) [M + H], 170 (20) [2-am ino-6-chloropurine + H].
1H NMR (DMSO-d6): 8.24 s, 1 H (H-8); 6.16 br s, 2 H (NH2); 5.33 d, 1 H, J(OH,2′) = 5.4
(2′-OH); 4.47 dd, 1 H, J(1′a,2′) = 3.2, J(gem ) = 14.2 (H-1′a); 4.11 dd, 1 H, J(1′b,2′) = 9.0
(H-2′b); 4.05 br pen t, 4 H, J(CH2,CH3) ≈ J(P,OCH) ≈ 7.2 (P-OCH2); 3.93 m , 1 H (H-2′); 3.87 d,
2 H, J(P,CH) = 7.8 (PCH2); 3.57 dd, 1 H, J(3′a,2′) = 5.1, J(gem ) = 10.0 (H-3′a); 3.52 dd, 1 H,
J(3′b,2′) = 5.7 (H-3′b); 1.24 t, 6 H, J(CH3,CH2) = 7.1 (CH3). 13C NMR (DMSO-d6): 164.47
(C-4); 160.00 (C-2); 150.54 (C-8); 142.29 (C-6); 115.20 (C-5); 74.71 d, J(P,C) = 10.7 (C-3′);
68.31 (C-2′); 64.73 d, J(P,C) = 162.1 (P-C); 61.92 d, 2 C, J(P,C) = 6.4 (P-OCH2); 49.75 (C-1′);
16.51 d, 2 C, J(P,C) = 5.4 (CH3).
Dieth yl Esters of N6-Substituted 9- an d 7-[2-Hydroxy-3-(ph osph on om eth oxy)propyl]
Derivatives of Aden in e an d 2,6-Diam in opurin e. Gen eral Procedure
A m ixture of an appropriate 6-ch loro derivative 11, 12 or 13 (1 m m ol), prim ary or secon d-
ary am in e, or dim eth ylam m on ium N,N-dim eth ylcarbam ate (5 m m ol) in aceton itrile (10 m l)
was refluxed for 0.5–4 h . Th e reaction course was m on itored by TLC in system S1, S2 or S3.
Th e m ixture was evaporated to dryn ess in vacuo an d th e residue ch rom atograph ed on a
silica gel colum n (100 m l) in th e system described below.
Diethyl ({3-[6-(dimethylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonate (15).
Ch rom atograph ed in system S2. Yield 342 m g (88%) of a colorless syrup. For C15H26N5O5P
(387.4) calculated: 46.51% C, 6.76% H, 18.08% N, 8.00% P; foun d: 46.32% C, 6.98% H,
17.97% N, 7.66% P. FAB MS, m/z (%): 388 (100) [M + H], 164 (15) [6-(dimethylamino)purine + H].
1H NMR (DMSO-d6): 8.20 s, 1 H an d 8.06 s, 1 H (H-2, H-8); 5.33 d, 1 H, J(OH,2′) = 5.4
(2′-OH); 4.26 dd, 1 H, J(1′a,2′) = 3.4, J(gem ) = 13.4 (H-1′a); 4.04 dd, 1 H, J(1′b,2′) = 8.0
(H-1′b); 4.06 br pen t, 4 H, J(CH2,CH3) ≈ J(P,CH2) = 7.1 (P-OCH2); 4.00 m , 1 H (H-2′); 3.86 d,
2 H, J(P,CH) = 7.7 (PCH2); 3.50 d, 2 H, J(3′,2′) = 5.4 (H-3′); 3.45 br, 6 H (NCH3); 1.24 t, 6 H,
J(CH3,CH2) = 7.0 (CH3). 13C NMR (DMSO-d6): 154.405 (C-6); 151.77 (C-2); 150.59 (C-4);
140.56 (C-8); 119.25 (C-5); 74.86 d, J(P,C) = 10.2 (C-3′); 67.60 (C-2′); 64.72 d, J(P,C) = 162.1
(P-C); 61.91 d, J(P,C) = 5.9 (P-OCH2); 61.90 d, J(P,C) = 5.9 (P-OCH2); 46.42 (C-1′); 39.95, 2 C
(NCH3); 16.46 d, 2 C, J(P,C) = 5.4 (CH3).
Diethyl ({3-[6-(cyclopropylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonate (16).
Ch rom atograph ed in system S3. Yield 397 m g (98%) of a wh ite solid. For C16H26N5O5P·
0.25H2O (403.9) calculated: 47.58% C, 6.61% H, 17.34% N, 7.67% P; foun d: 47.58% C,
6.91% H, 17.18% N, 7.29% P. FAB MS, m/z (%): 400 (100) [M
+ H], 176 (12)
[6-(cyclopropylam in o)purin e + H]. 1H NMR (DMSO-d6): 8.23 s, 1 H an d 8.04 s, 1 H (H-2,
H-8); 7.83 br s, 1 H (NH); 5.32 d, 1 H, J(OH,2′) = 5.4 (2′-OH); 4.26 dd, 1 H, J(1′a,2′) = 2.8,
J(gem ) = 13.2 (H-1′a); 4.05 br pen t, 4 H, J(CH2,CH3) ≈ J(P,CH2) = 7.1 (P-OCH2); 4.03 overlay,
1 H (H-1′b); 4.01 m , 1 H (H-2′); 3.51 dd, 1 H, J(3′a,2′) = 5.1, J(gem ) = 11.1 (H-3′a); 3.48 dd, 1 H,
J(3′b,2′) = 5.4 (H-3′b); 3.05 m , 1 H (CH-cyclopropyl); 1.24 t, 6 H, J(CH3,CH2) = 7.0 (CH3);
0.72 m , 2 H an d 0.61 m , 2 H (CH2-cyclopropyl). 13C NMR (DMSO-d6): 155.66 (C-6); 153.94
(C-4); 152.31 (C-2); 141.515 (C-8); 119.10 (C-5); 74.84 d, J(P,C) = 10.2 (C-3′); 67.62 (C-2′);
64.70 d, J(P,C) = 161.6 (P-C); 61.89 d an d 61.87 d, J(P,C) = 6.4 (P-OCH2); 46.42 (C-1′); 29.80
(cyclopropyl); 16.46 d, 2 C, J(P,C) = 5.4 (CH3); 6.55 (cyclopropyl).
Diethyl {[2-hydroxy-3-(6-piperidino-9H-purin-9-yl)propoxy]methyl}phosphonate (17). Ch rom ato-
graph ed in system S1. Yield 249 m g (58%) of a wh ite solid. For C18H30N5O5P·0.25H2O
(431.9) calculated: 50.05% C, 7.12% H, 16.21% N, 7.17% P; foun d: 49.99% C, 7.30% H,
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1903
16.22% N, 7.28% P. FAB MS, m/z (%): 428 (100) [M + H], 204 (17) [6-piperidin opurin e + H].
1H NMR (DMSO-d6): 8.20 s, 1 H an d 8.06 s, 1 H (H-2, H-8); 5.32 d, 1 H, J(OH,2′) = 5.4
(2′-OH); 4.26 dd, 1 H, J(1′a,2′) = 3.3, J(gem ) = 13.4 (H-1′a); 4.20 m , 4 H (NCH2-piperidin yl);
4.06 d pen t, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 8.2 (P-OCH2); 4.03 dd, 1 H, J(1′b,2′) = 8.2
(H-1′b); 4.00 m , 1 H (H-2′); 3.87 d, 2 H, J(P,CH) = 7.8 (PCH2); 3.51 dd, 1 H, J(3′a,2′) = 5.2,
J(gem ) = 10.4 (H-3′a); 3.49 dd, 1 H, J(3′b,2′) = 5.4 (H-3′b); 1.67 m , 2 H an d 1.57 m , 4 H
(CH2-piperidin yl); 1.24 t, 6 H (CH3). 13C NMR (DMSO-d6): 153.31 (C-6); 151.89 (C-2);
150.87 (C-4); 140.56 (C-8); 118.97 (C-5); 74.89 d, J(P,C) = 9.8 (C-3′); 67.59 (C-2′); 64.72 d,
J(P,C) = 161.6 (P-C); 61.95 d an d 61.92 d, J(P,C) = 6.4 (P-OCH2); 47.20 br, 2 C (NCH2); 46.49
(C-1′); 25.88, 2 C (C-CH2); 24.49 (C-CH2); 16.51 d, 2 C, J(P,C) = 5.4 (CH3).
Diethyl [(2-hydroxy-3-{6-[(2-methoxyethyl)amino]-9H-purin-9-yl}propoxy)methyl]phosphonate
(18). Ch rom atograph ed in system S1. Yield 386 m g (91%) of a wh ite solid. For C16H28N5O6P·
0.25H2O (421.9) calculated: 45.55% C, 6.81% H, 16.60% N, 7.34% P; foun d: 45.65% C,
6.90% H, 16.65% N, 7.24% P. FAB MS, m/z (%): 418 (100) [M + H]. 1H NMR (DMSO-d6):
8.20 s, 1 H an d 8.05 s, 1 H (H-2, H-8); 7.60 br s, 1 H (NH); 5.33 d, 1 H, J(OH,2′) = 5.3
(2′-OH); 4.25 dd, 1 H, J(1′a,2′) = 3.0, J(gem ) = 13.2 (H-1′a); 4.06 dq, 4 H, J(CH2,CH3) = 7.0,
J(P,OCH) = 8.2 (P-OCH2); 4.04 dd, 1 H, J(1′b,2′) = 8.0 (H-1′b); 4.01 m , 1 H (H-2′); 3.86 d, 2 H,
J(P,CH) = 7.8 (PCH2); 3.64 m , 2 H (NCH2); 3.52 t, 2 H, J(CH2,CH2) = 5.9 (OCH2); 3.50 dd,
1 H an d 3.48 dd, 1 H, J(3′a,2′) = J(3′b,2′) = 5.0, J(gem ) = 10.5 (H-3′); 3.26 s, 3 H (OCH3);
1.24 t, 6 H, J(CH3,CH2) = 7.0 (CH3). 13C NMR (DMSO-d6): 154.60 (C-6); 152.13 (C-2); 149.50
(C-4); 141.34 (C-8); 118.88 (C-5); 74.36 d, J(P,C) = 9.8 (C-3′); 70.63 (OCH2); 67.58 (C-2′);
64.68 d, J(P,C) = 162.1 (P-C); 61.71 d, 2 C, J(P,C) = 6.4 (P-OCH2); 57.86 (OCH3); 46.31
(C-1′); 45.70 (NCH2); 16.24 d, 2 C, J(P,C) = 5.4 (CH3).
Diethyl [(2-hydroxy-3-{6-[(2-hydroxyethyl)amino]-9H-purin-9-yl}propoxy)methyl]phosphonate
(19). Ch rom atograph ed in system S3. Yield 348 m g (86%) of a colorless syrup. For
C
15H26N5O6P (403.4) calculated: 44.66% C, 6.50% H, 17.36% N, 7.68% P; foun d: 44.37% C,
6.51% H, 17.48% N, 7.49% P. FAB MS, m/z (%): 404 (100) [M + H]. 1H NMR (DMSO-d6):
8.20 br s, 1 H an d 8.05 s, 1 H (H-2, H-8); 7.54 br s, 1 H (NH); 5.34 d, 1 H, J(OH,2′) = 5.4
(2′-OH); 4.78 br t, 1 H, J(OH,CH2) = 5.0 (OH); 4.25 dd, 1 H, J(1′a,2′) = 3.2, J(gem ) = 13.2
(H-1′a); 4.06 dq, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 8.1 (P-OCH2); 4.03 overlay, 1 H (H-1′b);
4.01 m , 1 H (H-2′); 3.86 d, 2 H, J(P,CH) = 7.8 (PCH2); 3.57 m , 4 H (NCH2 an d OCH2);
3.51 dd, 1 H an d 3.48 dd, 1 H, J(3′a,2′) = J(3′b,2′) = 5.0, J(gem ) = 10.5 (H-3′); 1.24 t, 6 H
(CH3). 13C NMR (DMSO-d6): 154.72 (C-6); 152.14 (C-2); 150.0 (C-4); 141.325 (C-8); 118.89
(C-5); 74.73 d, J(P,C) = 10.3 (C-3′); 67.59 (C-2′); 64.75 d, J(P,C) = 162.1 (P-C); 61.72 d, 2 C,
J(P,C) = 6.3 (P-OCH2); 60.06 (OCH2); 46.32 (C-1′); 45.0 (NCH2); 16.25 d, 2 C, J(P,C) = 5.4
(CH3).
Diethyl ({3-[6-(allylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonate (20). Ch ro-
m atograph ed in system S1. Yield 385 m g (96%) of a yellow solid. For C16H26N5O5P (399.4)
calculated: 48.12% C, 6.56% H, 17.54% N, 7.76% P; foun d: 47.76% C, 6.65% H, 17.36% N,
7.57% P. FAB MS, m/z (%): 400 (100) [M + H]. 1H NMR (DMSO-d6): 8.19 br s, 1 H an d 8.05 s,
1 H (H-2, H-8); 7.89 br s, 1 H (NH); 5.95 ddt, 1 H, J(2′′,1′′) = 5.1, J(2′′,3′′) = 10.3 (cis) an d
17.3 (tran s) (H-2′′); 5.34 d, 1 H, J(OH,2′) = 5.3 (2′-OH); 5.15 br dq, 1 H, J(3′′,1′′) ≈ J(gem ) =
1.6, J(3′′,2′′) = 17.3 (H-3′′tran s); 5.04 dq, 1 H, J(3′′,1′′) ≈ J(gem ) = 1.7, J(3′′,2′′) = 10.3
(H-3′′cis); 4.26 dd, 1 H, J(1′a,2′) = 2.7, J(gem ) = 12.9 (H-1′a); 4.11 m , 2 H (H-1′′); 4.06 br
pen t, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 7.8 (P-OCH2); 4.02 m , 1 H (H-1′b); 4.02 m , 1 H
(H-2′); 3.87 d, 2 H, J(P,CH) = 7.8 (PCH2); 3.51 dd, 1 H, J(3′a,2′) = 5.0, J(gem ) = 10.0 (H-3′a);
3.49 dd, 1 H, J(3′b,2′) = 5.4 (H-3′b); 1.24 t, 6 H (CH3). 13C NMR (DMSO-d6): 154.50 (C-6);
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1904
Krečmerová, Masojídková, Holý:
152.37 (C-2); 149.23 (C-4); 141.59 (C-8); 136.02 (C-2′′); 119.05 (C-5); 115.095 (C-3′′); 74.86 d,
J(P,C) = 10.2 (C-3′); 67.64 (C-2′); 64.70 d, J(P,C) = 162.1 (P-C); 61.91 d, 2 C, J(P,C) = 6.4
(P-OCH2); 46.47 (C-1′); 42.16 (C-1′′); 16.49 d, 2 C, J(P,C) = 5.4 (CH3).
Diethyl {[3-(6-{[2-(dimethylamino)ethyl]amino}-9H-purin-9-yl)-2-hydroxypropoxy]methyl}phos-
phonate (21). In stead of ch rom atograph y, th e crude reaction product was dissolved in 20%
aqueous m eth an ol (30 m l) an d Dowex 50 (H+ form ) was added un til th e solution was acid.
Th e m ixture was applied on to a colum n of th e sam e ion exch an ger (60 m l), elution per-
form ed first with 20% m eth an ol (1.5 l) an d th en with 2.5% am m on ia in 20% m eth an ol. Th e
UV absorbin g fraction s were collected an d taken down to give 21 as a yellowish syrup. Yield
373 m g (87%). Th e product of ca. 90% purity was used for subsequen t preparation of 37
with out furth er purification . FAB MS, m/z (%): 431 (30) [M + H].
Diethyl [(2-hydroxy-3-{6-[(2,2,2-trifluoroethyl)amino]-9H-purin-9-yl}propoxy)methyl]phosphonate
(22). Prepared with 5% solution of 2,2,2-trifluoreth ylam in e in eth an ol (20 m l) in an auto-
clave at 100 °C for 6 h . Ch rom atograph y in system S1 gave 250 m g (57%) of 22 as a yellow-
ish syrup. For C15H23F3N5O5P (441.4) calculated: 40.82% C, 5.25% H, 12.91% F, 15.87% N,
7.02% P; foun d: 40.74% C, 5.45% H, 12.82% F, 15.85% N, 7.31% P. FAB MS, m/z (%): 442
(100) [M + H]. 1H NMR (DMSO-d6): 8.32 br s, 1 H (NH); 8.30 s, 1 H an d 8.16 s, 1 H (H-2,
H-8); 5.34 d, 1 H, J(OH,2′) = 5.5 (2′-OH); 4.35 m , 2 H (NCH2); 4.29 dd, 1 H, J(1′a,2′) = 3.4,
J(gem ) = 13.6 (H-1′a); 4.06 dq, 4 H, J(CH2,CH3) = 7.1, J(P,CH) = 8.2 (P-OCH2); 4.05 m , 1 H
(H-2′); 4.02 dd, 1 H, J(1′b,2′) = 7.0 (H-1′b); 3.87 d, 2 H, J(P,CH) = 7.8 (PCH2); 3.52 dd, 1 H,
J(3′a,2′) = 5.2, J(gem ) = 10.0 (H-3′a); 3.50 dd, 1 H, J(3′b,2′) = 5.4 (H-3′b); 1.24 t, 6 H (CH3).
13C NMR (DMSO-d6): 154.16 (C-6); 152.05 (C-2); 150.0 (C-4); 142.59 (C-8); 125.61 (C-5);
125.30 q, J(C,F) = 280.3 (CF3); 74.83 d, J(P,C) = 10.2 (C-3′); 67.565 (C-2′); 64.70 d, J(P,C) =
161.6 (P-C); 61.93 d, 2 C, J(P,C) = 6.4 (P-OCH2); 43.08 q, J(C,F) = 31.3 (NCH2); 16.48 d, 2 C,
J(P,C) = 5.4 (CH3).
Diethyl ({3-[2-amino-6-(dimethylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonate
(23). Ch rom atograph y in system S3. Yield 384 m g (95%) of
a colorless syrup. For
C
15H27N6O5P (402.4) calculated: 44.77% C, 6.76% H, 20.89% N, 7.70% P; foun d: 44.36% C,
6.83% H, 20.37% N, 7.68% P. FAB MS, m/z (%): 403 (100) [M + H]. 1H NMR (DMSO-d6): 7.64 s,
1 H (H-8); 5.83 br s, 2 H (NH2); 5.38 br, 1 H (OH); 4.06 dd, 1 H, J(1′a,2′) = 3.5, J(gem ) = 13.2
(H-1′a); 4.05 dq, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 8.2 (P-OCH2); 3.945 m , 1 H (H-2′);
3.86 dd, 1 H, J(1′b,2′) = 7.3 (H-1′b); 3.86 d, 2 H, J(P,CH) = 7.7 (PCH2); 3.47 d, 2 H, J(3′,2′) =
5.0 (H-3′); 3.35 br s, 6 H (N-CH3); 1.24 t, 6 H (CH3). 13C NMR (DMSO-d6): 159.42 (C-2);
154.83 (C-6); 152.85 (C-4); 137.55 (C-8); 113.65 (C-5); 75.035 d, J(P,C) = 9.8 (C-3′); 67.63
(C-2′); 64.74 d, J(P,C) = 162.1 (P-CH2); 61.945 d, 2 C, J(C,P) = 6.4 (P-OCH2); 46.08 (C-1′);
37.84, 2 C (NCH3); 16.54 d, 2 C, J(C,P) = 5.9 (CH3).
Diethyl ({3-[2-amino-6-(cyclopropylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonate
(24). Ch rom atograph ed in system S3. Yield 392 m g (95%) of
a yellow syrup. For
C
16H27N6O5P (414.4) calculated: 46.37% C, 6.57% H, 20.28% N, 7.47% P; foun d: 45.99% C,
6.55% H, 20.01% N, 7.25% P. FAB MS, m/z (rel.%): 415 (100) [M + H]. 1H NMR (DMSO-d6):
7.61 s, 1 H (H-8); 7.28 d, 1 H, J(NH,CH) = 4.5 (NH); 5.84 br s, 2 H (NH2); 5.38 d, 1 H,
J(OH,2′) = 5.0 (2′-OH); 4.05 dq, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 8.3 (P-OCH2); 4.02 m , 3 H
(H-1′, H-2′); 3.86 d, 2 H, J(P,CH) = 7.7 (PCH2); 3.50 dd, 1 H, J(3′a,2′) = 3.8, J(gem ) = 10.0
(H-3′a); 3.44 dd, 1 H, J(3′b,2′) = 5.0 (H-3′b); 3.02 m , 1 H (H-1′′); 1.23 t, 6 H (CH3); 0.62 m ,
2 H an d 0.58 m , 2 H (H-2′′). 13C NMR (DMSO-d6): 160.31 (C-2); 156.11 (C-6); 151.64 (C-4);
138.29 (C-8); 113.48 (C-5); 75.07 d, J(C,P) = 10.0 (C-3′); 67.715 (C-2′); 64.75 d, J(C,P) = 161.9
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Acyclic Nucleotide Analogues
1905
(P-CH2); 62.01 d, 2 C, J(C,P) = 6.7 (P-OCH2); 46.10 (C-1′); 24.06 (cyclopropyl); 16.575 d, 2 C,
J(C,P) = 5.6 (CH3); 6.69, 2 C (cyclopropyl).
Diethyl [(3-{2-amino-6-[(2,2,2-trifluoroethyl)amino]-9H-purin-9-yl}-2-hydroxypropoxy)methyl]-
phosphonate (25). Prepared with a 5% solution of 2,2,2-trifluoreth ylam in e in eth an ol (20 m l)
in an autoclave at 100 °C for 12 h . Ch rom atograph y in system S2 gave 423 m g (93%) of 25
as a wh ite foam . For C15H24F3N6O5P (456.4) calculated: 39.48% C, 5.30% H, 12.49% F,
18.42% N, 6.79% P; 39.31% C, 5.17% H, 12.59% F, 18.13% N, 6.67% P. FAB MS, m/z (%):
457 (100) [M + H], 233 (40) [M + H of 2-am in o-6-[(trifluoroeth yl)am in o]purin e]. 1H NMR
(DMSO-d6): 7.78 br, 1 H (NH); 7.70 s, 1 H (H-8); 6.02 br s, 2 H (NH2); 5.35 d, 1 H, J(OH,2′) =
4.8 (2′-OH); 4.29 m , 2 H (H-1′′); 4.05 dq, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 8.2 (P-OCH2);
4.03 overlay, 1 H (H-1′a); 3.97 m , 1 H (H-2′); 3.89 dd, 1 H, J(1′b,2′) = 7.6, J(gem ) = 13.2
(H-1′b); 3.86 d, 2 H, J(P,CH) = 7.7 (PCH2); 3.49 d, 2 H, J(3′,2′) = 4.6 (H-3′); 1.24 t, 6 H (CH3).
13C NMR (DMSO-d6): 160.07 (C-2); 154.51 (C-6); 152.36 (C-4); 139.12 (C-8); 125.40 q,
J(C,F) = 280.1 (CF3); 113.315 (C-5); 75.055 d, J(C,P) = 9.8 (C-3′); 67.57 (C-2′); 64.74 d,
J(C,P) = 161.6 (P-C); 61.95 d, 2 C, J(C,P) = 6.3 (P-OCH2); 46.37 q, J(C,F) = 29.0 (NCH2);
46.13 (C-1′); 16.53 d, 2 C, J(C,P) = 5.4 (CH3).
Diethyl ({3-[6-(allylamino)-2-amino-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonate (26).
Ch rom atograph ed in system S2. Yield 391 m g (94%) of a colorless syrup. For C16H27N6O5P
(414.4) calculated: 46.37% C, 6.57% H, 20.28% N, 7.47% P; foun d: 45.86% C, 6.65% H,
20.08% N, 7.26% P. FAB MS, m/z (%): 415 (100) [M + H]. 1H NMR (DMSO-d6): 7.63 s, 1 H
(H-8); 7.32 br s, 1 H (NH); 5.93 ddt, 1 H, J(2′′,1′′) = 5.1, J(2′′,3′′cis) = 10.4, J(2′′,3′′tran s) =
17.3 (H-2′′); 5.83 br s, 2 H (NH2); 5.37 d, 1 H, J(OH,2′) = 5.4 (2′-OH); 5.14 dq, 1 H, J(3′′,1′′) ≈
J(gem ) = 1.8 (H-3′′tran s); 5.03 dq, 1 H, J(3′′,1′′) = 1.8 (H-3′′cis); 4.06 dd, 1 H, J(1′a,2′) = 3.8,
J(gem ) = 13.8 (H-1′a); 4.05 dq, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 8.2 (P-OCH2); 4.05 m , 2 H
(H-1′′); 3.96 m , 1 H (H-2′); 3.87 dd, 1 H (1′b,2′) = 7.8 (H-1′b); 3.86 d, 2 H, J(P,CH) = 7.7
(PCH2); 3.49 dd, 1 H, J(3′a,2′) = 5.4, J(gem ) = 10.0 (H-3′a); 3.46 dd, 1 H, J(3′b,2′) = 5.2
(H-3′b); 1.24 t, 6 H (CH3). 13C NMR (DMSO-d6): 160.25 (C-2); 154.91 (C-6); 150.0 (C-4);
138.225 (C-8); 136.52 (C-2′′); 114.85 (C-3′′); 113.285 (C-5); 75.02 d, J(C,P) = 9.8 (C-3′); 67.68
(C-2′); 64.74 d, J(C,P) = 162.1 (P-C); 61.92 d an d 61.90 d, J(C,P) = 6.4 (P-OCH2); 46.07 (C-1′);
43.33 (C-1′′); 16.50 d, 2 C, J(C,P) = 5.4 (CH3).
Diethyl {[3-(2-amino-6-{[2-(dimethylamino)ethyl]amino}-9H-purin-9-yl)-2-hydroxypropoxy]methyl}-
phosphonate (27). Th e crude reaction product was desalted on Dowex 50 (H+ form ) in th e
sam e way as described for com poun d 21. Yield 434 m g (97%) of a foam . HR MS (FAB): For
C
17H33N7O5P [M + H] calculated: 446.2278; foun d: 446.2281. FAB MS, m/z (%): 446 (8) [M +
H]. 1H NMR (DMSO-d6): 7.64 s, 1 H (H-8); 6.85 br s, 1 H (NH); 5.86 br s, 2 H (NH2); 4.04 dd,
1 H, J(1′a,2′) = 3.2, J(gem ) = 12.8 (H-1′a); 3.90 m , 1 H (H-2′); 3.85 dd, 1 H, J(H-1′b,2′) = 8.1
(H-1′b); 3.76 pen t, 4 H, J(CH2,CH3) = J(P,OCH) = 7.1 (P-OCH2); 3.50 m , 2 H, J(CH2,CH2) =
6.4 (NCH2); 3.44 dd, 1 H, J(3′a,2′) = 5.4, J(gem ) = 10.4 (H-3′a); 3.43 dd, 1 H an d 3.40 dd,
1 H, J(P,CH) = 8.2, J(gem ) = 13.4 (PCH2); 3.39 dd, 1 H, J(3′b,2′) = 5.4 (H-3′b); 2.81 t, 2 H
(NCH2); 2.19 s, 6 H (NCH3); 1.05 t, 6 H (CH3). 13C NMR (DMSO-d6): 160.29 (C-2); 154.96
(C-6); 152.75 (C-4); 138.36 (C-8); 113.25 (C-5); 75.065 d, J(C,P) = 9.3 (C-3′); 68.42 d, J(C,P) =
153.3 (P-C); 68.02 (C-2′); 59.20 d, 2 C, J(C,P) = 6.4 (P-OCH2); 58.36 an d 47.02 (NCH2); 45.90
(C-1′); 45.27 an d 45.13 (NCH3); 17.20 d, 2 C, J(C,P) = 5.4 (CH3).
Diethyl ({3-[6-(dimethylamino)-7H-purin-7-yl]-2-hydroxypropoxy}methyl)phosphonate (28).
Ch rom atograph ed in system S3. Yield 244 m g (63%) of a colorless syrup. For C15H26N5O5P
(387.4) calculated: 46.51% C, 6.76% H, 18.08% N, 8.00% P; foun d: 46.66% C, 6.78% H,
17.87% N, 8.25% P. FAB MS, m/z (%): 388 (100) [M + H]. 1H NMR (DMSO-d6): 8.42 s, 1 H
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Krečmerová, Masojídková, Holý:
an d 8.37 s, 1 H (H-2, H-8); 5.25 d, 1 H, J(OH,2′) = 5.7 (2′-OH); 4.51 dd, 1 H, J(1′a,2′) = 3.2,
J(gem ) = 14.4 (H-1′a); 4.19 dd, 1 H, J(1′b,2′) = 9.2 (H-1′b); 4.06 br pen t, 4 H, J(P,OCH) =
J(CH2,CH3) = 7.1 (P-OCH2); 3.89 m , 1 H (H-2′); 3.85 d, 2 H, J(P,CH) = 8.1 (PCH2); 3.49 dd,
1 H, J(3′a,2′) = 5.0, J(gem ) = 10.9 (H-3′a); 3.43 dd, 1 H, J(3′b,2′) = 6.3 H-3′b); 3.01 s, 6 H
(NCH3); 1.25 t, 6 H, J(CH3,CH2) = 7.0 (CH3). 13C NMR (DMSO-d6): 161.53 (C-4); 155.64
(C-6); 150.975 (C-2); 148.69 (C-8); 114.66 (C-5); 74.79 d, J(C,P) = 10.7 (C-3′); 68.30 (C-2′);
64.63 d, J(C,P) = 162.6 (P-C); 61.86 d, 2 C, J(C,P) = 6.4 (P-OCH2); 50.94 (C-1′); 16.47 d, 2 C,
J(C,P) = 5.4 (CH3).
Diethyl ({3-[6-(cyclopropylamino)-7H-purin-7-yl]-2-hydroxypropoxy}methyl)phosphonate (29).
Ch rom atograph ed in system S3. Yield 264 m g (65%) of a wh ite solid. For C16H26N5O5P·
0.5H2O (408.4) calculated: 47.06% C, 6.66% H, 17.15% N, 7.58% P; foun d: 47.26% C, 6.74% H,
16.91% N, 7.45% P. FAB MS, m/z (%): 400 (100) [M + H]. 1H NMR (DMSO-d6): 8.33 s, 1 H
an d 8.14 s, 1 H (H-2, H-8); 7.31 br d, 1 H, J(NH,CH) = 2.8 (NH); 6.01 d, 1 H, J(OH,2′) = 4.9
(2′-OH); 4.48 dd, 1 H, J(1′a,2′) = 2.6, J(gem ) = 15.0 (H-1′a); 4.23 dd, 1 H, J(1′b,2′) = 7.7
(H-1′b); 4.07 dq, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) = 8.2 (P-OCH2); 3.92 m , 1 H (H-2′); 3.88 d,
2 H, J(P,CH) = 7.7 (PCH2); 3.53 dd, 1 H, J(3′a,2′) = 5.5, J(gem ) = 10.2 (H-3′a); 3.49 dd, 1 H,
J(3′b,2′) = 5.6 (H-3′b); 2.90 m , 1 H (CH-cyclopropyl); 1.25 t, 6 H (CH3); 0.77 m , 2 H an d
0.52 m , 2 H (CH2-cyclopropyl). 13C NMR (DMSO-d6): 159.00 (C-4); 152.04 (C-6); 151.84
(C-2); 146.23 (C-8); 112.64 (C-5); 73.96 d, J(C,P) = 9.8 (C-3′); 68.64 (C-2′); 64.815 d, J(C,P) =
162.1 (P-C); 61.75 d, 2 C, J(C,P) = 6.3 (P-OCH2); 49.38 (C-1′); 23.81 (cyclopropyl); 16.26 d,
2 C, J(C,P) = 5.4 (CH3); 6.59 an d 6.33 (cyclopropyl).
Diethyl ({3-[6-(allylamino)-7H-purin-7-yl]-2-hydroxypropoxy}methyl)phosphonate (30). Ch ro-
m atograph ed in system ch loroform –m eth an ol 8:1. Yield 347 m g (87%) of a wh ite solid. For
C
16H26N5O5P (399.4) calculated: 48.12% C, 6.56% H, 17.54% N, 7.76% P; foun d: 48.05% C,
6.76% H, 17.38% N, 7.78% P. FAB MS, m/z (%): 400 (100) [M + H]. 1H NMR (DMSO-d6):
8.26 s, 1 H an d 8.16 s, 1 H (H-2, H-8); 7.22 t, 1 H, J(NH,CH2) = 5.6 (NH); 5.97 ddt, 1 H,
J(2′′,1′′) = 5.1, J(2′′,3′′) = 10.4 (cis) an d 17.2 (tran s) (H-2′′); 5.90 d, 1 H, J(OH,2′) = 5.0
(2′-OH); 5.22 dq, 1 H, J(3′′tran s,1′′) ≈ J(gem ) = 1.7 (H-3′′tran s); 5.08 dq, 1 H, J(3′′cis,1′′) ≈
J(gem ) = 1.7 (H-3′′cis); 4.52 dd, 1 H, J(1′tran s,2′) = 2.8, J(gem ) = 15.0 (H-1′tran s); 4.28 dd,
1 H, J(1′cis,2′) = 7.9 (H-1′cis); 4.13 m , 2 H (H-1′′); 4.07 dq, 4 H, J(CH2,CH3) = 7.1, J(P,OCH) =
8.2 (P-OCH2); 3.95 dqd, 1 H (H-2′); 3.89 d, 2 H, J(P,CH) = 7.8 (PCH2); 3.55 dd, 1 H, J(3′a,2′) =
5.4, J(gem ) = 10.0 (H-3′a); 3.52 dd, 1 H, J(3′b,2′) = 5.5 (H-3′b); 1.25 t, 6 H (CH3). 13C NMR
(DMSO-d6): 159.24 (C-4); 151.92 (C-2); 150.89 (C-6); 146.45 (C-8); 135.545 (C-2′′); 115.30
(C-3′′); 112.43 (C-5); 74.115 d, J(C,P) = 9.8 (C-3′); 68.82 (C-2′); 64.805 d, J(C,P) = 162.6
(P-CH2); 61.94 d, 2 C, J(C,P) = 6.4 (P-OCH2); 49.47 (C-1′); 42.64 (C-1′′); 16.49 d, 2 C, J(C,P) =
5.4 (CH3).
Preparation of Free 9- an d 7-[2-Hydroxy-3-(ph osph on om eth oxy)propyl] Derivatives
of N6-Substituted Aden in es an d 2,6-Diam in opurin es 31–37, 39–43 an d 44–46.
Gen eral Procedure
Brom otrim eth ylsilan e (1.35 m l, 10 m m ol) was added to a solution (or suspen sion ) of an ap-
propriate dieth yl ph osph on ate 15–28 (1 m m ol) in aceton itrile (10 m l). Th e m ixture was
stirred at room tem perature in th e dark for 24 h , th e reaction was th en quen ch ed by addi-
tion of a m ixture of eth an ol, water an d (C2H5)3N (15:15:2; 32 m l), th e resultin g solution
evaporated to dryn ess an d codistilled with water (2 × 30 m l). Th e residue was dissolved in
water (5 m l) an d applied on to a colum n of Dowex 50X8 (H+ form , 50 m l) equilibrated in
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Acyclic Nucleotide Analogues
1907
water an d th e colum n was wash ed with water un til th e UV absorption of th e eluate (254 n m )
dropped to th e origin al value (1000 m l). Th e colum n was th en wash ed with 2.5% aqueous
am m on ia, th e UV absorbin g fraction s were collected an d evaporated to dryn ess. Th e am or-
ph ous residue was dissolved in water (3–5 m l), applied on to a colum n of Dowex 50X8 (Na+
form , 30 m l) an d th e colum n was wash ed with water. Th e UV absorbin g eluate was evapo-
rated to dryn ess, th e residue stirred with a m ixture of eth an ol (20 m l) an d petroleum eth er
(5 m l) for 1 h an d th en left stan din g in a refrigerator overn igh t. Th e obtain ed sodium salt of
th e product was filtered off, wash ed with eth an ol an d eth er an d dried in vacuo. Th e follow-
in g com poun ds were obtain ed as disodium salts.
({3-[6-(Dimethylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonic acid (31). Yield
303 m g (77%). For C11H16N5Na2O5P·H2O (393.2) calculated: 33.60% C, 4.61% H, 17.81% N,
7.88% P; foun d: 33.27% C, 4.78% H, 17.79% N, 7.66% P. FAB MS, m/z (%): 376 (9) [M + H,
sodium salt], 332 (14) [M + H, free acid]. 1H NMR (D2O): 7.98 s, 2 H (H-2, H-8); 4.30 m , 1 H
an d 4.21 m , 2 H (H-1′, H-2′); 3.72 dd, 1 H, J(3′a,2′) = 3.0, J(gem ) = 10.5 (H-3′a); 3.67 dd, 1 H,
J(P,CHa) = 9.0, J(gem ) = 12.9 (PCHa); 3.63 dd, 1 H, J(P,CHb) = 8.8 (PCHb); 3.59 dd, 1 H,
J(3′b,2′) = 5.2 (H-3′b); 3.26 br s, 6 H (NCH3). 13C NMR (D2O): 154.00 (C-6); 151.48 (C-2);
148.99 (C-4); 140.86 (C-8); 118.43 (C-5); 73.81 d, J((P,C) = 11.7 (C-3′); 68.39 d, J(P,C) =
154.3 (P-C); 68.28 (C-2′); 46.20 (C-1′); 38.81, 2 C (NCH3).
({3-[6-(Cyclopropylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonic acid (32). Yield
303 m g (77%). For C12H17N5NaO5P·1.5H2O (392.3) calculated: 36.74% C, 5.14% H, 17.85% N,
7.90% P; foun d: 36.66% C, 5.06% H, 18.32% N, 7.40% P. FAB MS, m/z (%): 366 (19) [M + H,
sodium salt], 344 (20) [M + H, free acid]. 1H NMR (D2O): 8.18 s, 1 H an d 8.07 s, 1 H (H-2,
H-8); 4.38 dd, 1 H, J(1′a,2′) = 3.3, J(gem ) = 13.7 (H-1′a); 4.28 dd, 1 H, J(1′b,2′) = 8.2 (H-1′b);
4.24 m , 1 H (H-2′); 3.72 dd, 1 H, J(3′a,2′) = 3.7, J(gem ) = 10.5 (H-3′a); 3.68 dd, 1 H an d 3.65 dd,
1 H, J(P,CH) = 8.8, J(gem ) = 12.9 (PCH2); 3.59 dd, 1 H, J(3′b,2′) = 5.2 (H-3′b); 2.80 m , 1 H,
0.92 m , 2 H an d 0.68 m , 2 H (cyclopropyl). 13C NMR (D2O): 155.37 (C-6); 152.14 (C-2);
148.03 (C-4); 142.48 (C-8); 118.53 (C-5); 73.73 d, J(P,C) = 10.7 (C-3′); 68.29 (C-2′); 68.01 d,
J(P,C) = 153.8 (P-C); 46.23 (C-1′); 23.26 an d 6.56, 2 C (cyclopropyl).
{[2-Hydroxy-3-(6-piperidino-9H-purin-9-yl)propoxy]methyl}phosphonic acid (33). Yield 181 m g
(42%). For C14H20N5Na2O5P·H2O (433.3) calculated: 38.81% C, 5.12% H, 16.16% N, 7.15% P;
foun d: 39.32% C, 5.32% H, 15.99% N, 6.89% P. FAB MS, m/z (%): 416 (60) [M + H, sodium
salt], 372 (5) [M + H, free acid]. 1H NMR (D2O): 8.03 s, 1 H an d 8.02 s, 1 H (H-2, H-8); 4.33 m ,
1 H an d 4.24 m , 2 H (H-1′, H-2′); 3.90 m , 4 H (NCH2-piperidin e); 3.73 dd, 1 H, J(3′a,2′) =
2.7, J(gem ) = 10.5 (H-3′a); 3.67 dd, 1 H an d 3.63 dd, 1 H, J(P,CH) = 8.8, J(gem ) = 12.9
(PCH2); 3.60 dd, 1 H, J(3′b,2′) = 5.1 (H-3′b); 1.70 m , 2 H an d 1.58 m , 4 H (CH2-piperidin e).
13C NMR (D2O): 153.02 (C-6); 151.67 (C-2); 149.36 (C-4); 140.71 (C-8); 118.30 (C-5); 73.75 d,
J(P,C) = 11.2 (C-3′); 68.59 d, J(P,C) = 153.3 (P-C); 68.25 (C-2′); 46.935, 2 C (piperidin e);
46.15 (C-1′); 25.55, 2 C an d 23.91 (piperidin e).
[(2-Hydroxy-3-{6-[(2-methoxyethyl)amino]-9H-purin-9-yl}propoxy)methyl]phosphonic acid (34).
Yield 310 m g (76%). FAB MS, m/z (%): 406 (15) [M + H, sodium salt], 362 (95) [M + H, free
acid]. 1H NMR (D2O): 8.12 s, 1 H an d 8.065 s, 1 H (H-2, H-8); 4.36 m , 1 H an d 4.26 m , 2 H
(H-1′, H-2′); 3.75 m , 4 H (NCH2, OCH2); 3.75 overlay (H-3′a); 3.73 dd, 1 H an d 3.70 dd, 1 H,
J(P,CH) = 8.8, J(gem ) = 13.1 (PCH2); 3.62 dd, 1 H, J(3′b,2′) = 5.4, J(gem ) = 10.6 (H-3′b);
3.45 s, 3 H (OCH3). 13C NMR (D2O): 154.51 (C-6); 152.33 (C-2); 148.24 (C-4); 142.52 (C-8);
118.62 (C-5); 73.98 d, J(P,C) = 11.7 (C-3′); 70.725 (OCH2); 68.49 (C-2′); 68.03 d, J(P,C) =
155.8 (P-C); 58.37 (OCH3); 46.45 (C-1′); 40.27 (NCH2).
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Krečmerová, Masojídková, Holý:
[(2-Hydroxy-3-{6-[(2-hydroxyethyl)amino]-9H-purin-9-yl}propoxy)methyl]phosphonic acid (35).
Yield 295 m g (72%). For C11H16N5Na2O6P·H2O (409.3) calculated: 32.28% C, 4.43% H,
17.11% N, 7.56% P; foun d: 32.36% C, 4.65% H, 16.86% N, 7.46% P. FAB MS, m/z (%): 392
(37) [M + H, sodium salt], 348 (5) [M + H, free acid]. 1H NMR (D2O): 8.08 s, 1 H an d 8.06 s,
1 H (H-2, H-8); 4.34 m , 1 H an d 4.25 m , 2 H (H-1′, H-2′); 3.86 t, 2 H, J(CH2,CH2) = 5.5 an d
3.67 m , 2 H (NCH2, OCH2); 3.74 dd, 1 H, J(3′a,2′) = 3.3, J(gem ) = 10.6 (H-3′a); 3.66 dd, 1 H
an d 3.64 dd, 1 H, J(P,CH) = 8.8, J(gem ) = 12.7 (PCH2); 3.61 dd, 1 H, J(3′b,2′) = 5.4 (H-3′b).
13C NMR (D2O): 154.29 (C-6); 152.025 (C-2); 148.00 (C-4); 142.25 (C-8); 118.35 (C-5); 73.69 d,
J(P,C) = 11.7 (C-3′); 68.72 d, J(P,C) = 152.8 (P-C); 68.31 (C-2′); 60.71 (OCH2); 46.22 (C-1′);
42.78 (NCH2).
({3-[6-(Allylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonic acid (36). Yield 200 m g
(52%). For C12H16N5Na2O5P (387.2) calculated: 37.22% C, 4.16% H, 18.09% N, 8.00% P;
foun d: 37.21% C, 4.58% H, 17.69% N, 8.10% P. FAB MS, m/z (%): 388 (16) [M + H]. 1H NMR
(D2O): 8.05 s, 2 H (H-2, H-8); 6.01 ddt, J(2′′,1′′) = 4.9, J(2′′,3′′cis) = 10.4, J(2′′,3′′tran s) = 17.3
(H-2′); 5.24 dq, 1 H, J(3′′tran s,1′′) = J(gem ) = 1.6 (H-3′′tran s); 5.19 dq, 1 H, J(3′′cis,1′′) = 1.6
(H-3′cis); 4.34 m , 1 H an d 4.25 m , 2 H (H-1′, H-2′); 4.09 m , 2 H (H-1′′); 3.74 dd, 1 H, J(3′a,2′) =
3.3, J(gem ) = 10.4 (H-3′a); 3.70 dd, 1 H an d 3.66 dd, 1 H, J(P,CH) = 8.8, J(gem ) = 12.8
(PCH2); 3.61 dd, 1 H, J(3′b,2′) = 5.3 (H-3′b). 13C NMR (D2O): 154.04 (C-6); 152.13 (C-2);
147.91 (C-4); 142.24 (C-8); 134.00 (C-2′′); 118.23 (C-5); 115.54 (C-3′′); 73.72 d, J(P,C) = 11.7
(C-3′); 68.28 (C-2′); 68.23 d, J(P,C) = 154.1 (P-C); 46.22 (C-1′); 42.77 (C-1′′).
{[3-(6-{[2-(Dimethylamino)ethyl]amino}-9H-purin-9-yl)-2-hydroxypropoxy]methyl}phosphonic
acid (37). Yield 273 m g (65%). FAB MS, m/z (%): 419 (1) [M + H, sodium salt], 375 (5) [M +
H, free acid]. 1H NMR (D2O): 8.20 s, 1 H an d 8.13 s, 1 H (H-2, H-8); 4.39 dd, 1 H, J(1′a,2′) =
3.6, J(gem ) = 14.2 (H-1′a); 4.31 dd, 1 H, J(1′b,2′) = 8.1 (H-1′b); 4.25 m , 1 H (H-2′); 3.97 t, 2 H,
J(CH2,CH2) = 6.1 (NCH2); 3.72 dd, 1 H, J(3′a,2′) = 3.5, J(gem ) = 10.5 (H-3′a); 3.60 dd, 1 H,
J(3′b,2′) = 6.0 (H-3′b); 3.60 dd, 1 H an d 3.56 dd, 1 H, J(P,CH) = 8.8, J(gem ) = 12.2 (PCH2);
3.41 t, 2 H, J(CH2,CH2) = 6.1 (NCH2); 2.92 s, 6 H (NCH3). 13C NMR (D2O): 154.20 (C-6);
152.095 (C-2); 148.42 (C-4); 142.83 (C-8); 118.61 (C-5); 73.65 d, J(P,C) = 11.7 (C-3′); 69.53 d,
J(P,C) = 150.9 (P-C); 68.375 (C-2′); 56.91 (NCH2); 50.51 (NCH3); 46.32 (C-1′); 43.225
(NCH3); 36.31 (NCH2).
({3-[2-Amino-6-(dimethylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonic acid (39).
Yield 290 m g (77%). FAB MS, m/z (%): 391 (45) [M + H]. 1H NMR (D2O): 7.68 s, 1 H (H-8);
4.18 dd, 1 H, J(1′a,2′) = 3.3, J(gem ) = 14.0 (H-1′a); 4.14 m , 1 H (H-2′); 4.06 dd, 1 H, J(1′b,2′) =
8.8 (H-1′b); 3.71 dd, 1 H, J(3′a,2′) = 3.0, J(gem ) = 10.6 (H-3′a); 3.55 d, 2 H, J(P,CH) = 8.6
(PCH2); 3.54 dd, 1 H, J(3′b,2′) = 5.5 (H-3′b); 3.21 s, 6 H (CH3). 13C NMR (D2O): 159.61 (C-2);
155.06 (C-6); 151.69 (C-4); 138.95 (C-8); 113.60 (C-5); 74.20 d, J(P,C) = 11.7 (C-3′); 70.02 d,
J(P,C) = 150.8 (P-C); 68.77 (C-2′); 46.275 (C-1′); 38.775, 2 C (CH3).
({3-[2-Amino-6-(cyclopropylamino)-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonic acid
(40). Yield 208 m g (52%). For C12H17N6Na2O5P (402.3) calculated: 35.83% C, 4.26% H,
20.89% N, 7.70% P; foun d: 35.80% C, 4.53% H, 20.62% N, 7.48% P. FAB MS, m/z (%): 403
(10) [M + H, sodium salt], 359 (17) [M + H, free acid]. 1H NMR (D2O): 7.76 s, 1 H (H-8);
4.20 m , 2 H (H-1′a, H-2′); 4.10 dd, 1 H, J(1′b,2′) = 8.0, J(gem ) = 14.2 (H-1′b); 3.69 dd, 1 H,
J(3′a,2′) = 3.4, J(gem ) = 10.6 (H-3′a); 3.56 d, 2 H, J(P,CH) = 8.8 (PCH2); 3.54 dd, 1 H, J(3′b,2′) =
5.6 (H-3′b); 2.81 m , 1 H, 0.88 m , 2 H an d 0.65 m , 2 H (cyclopropyl). 13C NMR (D2O): 160.35
(C-2); 156.475 (C-6); 150.51 (C-4); 140.385 (C-8); 113.42 (C-5); 74.25 d, J(P,C) = 11.6 (C-3′);
69.99 d, J(P,C) = 150.4 (P-C); 68.825 (C-2′); 46.35 (C-1′); 23.63 an d 7.155, 2 C (cyclopropyl).
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1909
[(3-{2-Amino-6-[(2,2,2-trifluoroethyl)amino]-9H-purin-9-yl}-2-hydroxypropoxy)methyl]phosphonic
acid (41). Yield 300 m g (62%). For C11H14F3N6Na2O5P·2H2O (480.2) calculated: 11.86% F,
17.50% N, 6.45% P; foun d: 12.16% F, 17.16% N, 6.33% P. FAB MS, m/z (%): 445 (80) [M + H,
sodium salt]. 1H NMR (D2O): 7.83 s (H-8); 4.34 br q, 2 H, J(H,F) = 9.5 (NCH2); 4.24 dd, 1 H,
J(1′a,2′) = 3.5, J(gem ) = 13.2 (H-1′a); 4.21 m , 1 H (H-2′); 4.14 dd, 1 H, J(1′b, 2′) = 7.1 (H-1′b);
3.70 dd, 1 H, J(3′a,2′) = 3.3, J(gem ) = 10.5 (H-3′a); 3.60 dd, 1 H an d 3.56 dd, 1 H, J(P,CH) =
8.7, J(gem ) = 12.4 (PCH2); 3.57 dd, 1 H, J(3′b,2′) = 4.9 (H-3′b). 13C NMR (D2O): 159.95 (C-2);
154.88 (C-6); 151.23 (C-4); 140.655 (C-8); 124.68 q, J(C,F) = 278.8 (CF3); 113.19 (C-5);
73.68 d, J(C,P) = 11.3 (C-3′); 69.24 d, J(C,P) = 151.4 (P-C); 68.39 (C-2′); 45.85 (C-1′); 41.37 q,
J(C,F) = 29.0 (NCH2).
({3-[6-(Allylamino)-2-amino-9H-purin-9-yl]-2-hydroxypropoxy}methyl)phosphonic acid (42).
Yield 220 m g (52%). For C12H17N6Na2O5P·H2O (420.3) calculated: 34.29% C, 4.56% H,
19.99% N, 7.37% P; foun d: 34.42% C, 4.38% H, 19.70% N, 7.37% P. FAB MS, m/z (%): 403
(30) [M + H]. 1H NMR (D2O): 7.79 s, 1 H (H-8); 6.02 ddt, 1 H, J(2′′,1′′) = 5.0, J(2′′,3′′cis) =
10.4, J(2′′,3′′tran s) = 17.3 (H-2′′); 5.26 dq, 1 H, J(3′′tran s,1′′) ≈ J(gem ) = 1.6 (H-3′′tran s);
5.19 dq, 1 H, J(3′′cis,1′′) = 1.6 (H-3′′cis); 4.23 dd, 1 H, J(1′a,2′) = 3.7, J(gem ) = 14.6 (H-1′a);
4.21 m , 1 H (H-2′); 4.14 m , 2 H (H-1′′); 4.13 dd, 1 H, J(1′b,2′) = 9.8 (H-1′b); 3.69 dd, 1 H,
J(3′a,2′) = 3.2, J(gem ) = 10.5 (H-3′a); 3.55 dd, 1 H, J(3′b,2′) = 5.0 (H-3′b); 3.57 dd, 1 H an d
3.53 dd, 1 H, J(P,CH) = 8.8, J(gem ) = 12.4 (PCH2). 13C NMR (D2O): 160.14 (C-2); 155.11
(C-6); 150.10 (C-4); 140.04 (C-8); 134.61 (C-2′′); 115.41 (C-3′′); 112.995 (C-5); 73.70 d,
J(P,C) = 11.7 (C-3′); 69.62 d, J(P,C) = 150.9 (P-C); 68.45 (C-2′); 45.84 (C-1′); 42.66 (C-1′′).
{[3-(2-Amino-6-{[2-(dimethylamino)ethyl]amino}-9H-purin-9-yl)-2-hydroxypropoxy]methyl}-
phosphonic acid (43). Yield 300 m g (69%), wh ite solid. HR MS (FAB): For C13H23N7Na2O5P
[M + H] calculated: 434.1294; foun d: 434.1267. FAB MS, m/z (%): 434 (12) [M + H]. 1H NMR
(D2O): 7.785 s, 1 H (H-8); 4.23 dd, 1 H, J(1′a,2′) = 3.8, J(gem ) = 14.8 (H-1′a); 4.21 m , 1 H
(H-2′); 4.13 dd, 1 H, J(1′b,2′) = 9.2 (H-1′b); 3.70 dd, 1 H, J(3′a,2′) = 3.3, J(gem ) = 10.7 (H-3′a);
3.66 m , 2 H (NCH2); 3.58 dd, 1 H an d 3.55 dd, 1 H, J(P,CH) = 8.7, J(gem ) = 12.4 (PCH2);
3.56 dd, 1 H, J(3′b,2′) = 5.3 (H-3′b); 2.70 t, 2 H, J(CH2,CH2) = 6.7 (NCH2); 2.33 s, 6 H (CH3).
13C NMR (D2O): 160.11 (C-2); 155.13 (C-6); 150.445 (C-4); 139.99 (C-8); 113.10 (C-5); 73.74 d,
J(P,C) = 11.2 (C-3′); 69.65 d, J(P,C) = 149.9 (P-C); 68.46 (C-2′); 57.36 (NCH2); 45.87 (C-1′);
44.045, 2 C (NCH3); 37.93 (NCH2).
({3-[6-(Dimethylamino)-7H-purin-7-yl]-2-hydroxypropoxy}methyl)phosphonic acid (44). Yield
206 m g (55%). Hygroscopic m aterial. FAB MS, m/z (%): 376 (10) [M + H, sodium salt], 332
(100) [M + H, free acid]. 1H NMR (D2O): 8.37 s, 1 H an d 8.30 s, 1 H (H-2, H-8); 4.69 dd, 1 H,
J(1′a,2′) = 3.7, J(gem ) = 14.9 (H-1′a); 4.47 dd, 1 H, J(1′b,2′) = 8.8 (H-1′b); 4.32 m , 1 H (H-2′);
3.64 d, 2 H, J(P,CH) = 8.8 (PCH2); 3.63 dd, 1 H, J(3′a,2′) = 4.1, J(gem ) = 10.5 (H-3′a); 3.52 dd,
1 H, J(3′b,2′) = 5.4 (H-3′b); 3.15 s, 6 H (NCH3). 13C NMR (D2O): 159.50 (C-4); 156.19 (C-6);
151.18 (C-2); 148.69 (C-8); 114.30 (C-5); 73.67 d, J(P,C) = 11.2 (C-3′); 69.01 (C-2′); 67.88 d,
J(P,C) = 155.8 (P-C); 51.07 (C-1′); 40.98, 2 C (NCH3).
({3-[6-(Cyclopropylamino)-7H-purin-7-yl]-2-hydroxypropoxy}methyl)phosphonic acid (45). Yield
200 m g (49%). For C12H16N5Na2O5P·H2O (405.3) calculated: 35.56% C, 4.48% H, 17.28% N,
7.64% P; foun d: 35.61% C, 4.70% H, 17.11% N, 7.49% P. FAB MS, m/z (%): 388 (12) [M + H].
1H NMR (D2O): 8.22 s, 1 H an d 8.18 s, 1 H (H-2, H-8); 4.55 dd, 1 H, J(1′a,2′) = 2.9, J(gem ) =
15.5 (H-1′a); 4.40 dd, 1 H, J(1′b,2′) = 5.3 (H-1′b); 4.19 m , 1 H (H-2′); 3.68 dd, 1 H an d
3.65 dd, 1 H, J(P,CH) = 8.2, J(gem ) = 12.8 (PCH2); 3.65 dd, 1 H, J(3′a,2′) = 5.5, J(gem ) = 10.4
(H-3′a); 3.60 dd, 1 H, J(3′b,2′) = 6.2 (H-3′b); 2.78 m , 1 H, 0.94 m , 2 H an d 0.67 m , 2 H
(cyclopropyl). 13C NMR (D2O): 156.675 (C-4); 152.36 (C-6); 151.93 (C-2); 146.64 (C-8);
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1910
Krečmerová, Masojídková, Holý:
112.09 (C-5); 73.11 d, J(P,C) = 10.7 (C-3′); 68.84 (C-2′); 68.37 d, J(P,C) = 153.3 (P-C); 49.74
(C-1′); 23.52, 6.62 an d 6.49 (cyclopropyl).
({3-[6-(Allylamino)-7H-purin-7-yl]-2-hydroxypropoxy}methyl)phosphonic acid (46). Yield 205 m g
(53%). FAB MS, m/z (%): 388 (72) [M + H]. 1H NMR (D2O): 8.28 s, 1 H an d 8.25 s, 1 H (H-2,
H-8); 6.07 ddt, 1 H, J(2′′,1′′) = 4.6, J(2′′,3′′cis) = 10.4, J(2′′,3′′tran s) = 17.3 (H-2′′); 5.26 dq, 1 H,
J(3′′tran s,1′′) ≈ J(gem ) = 1.6 (H-3′′tran s); 5.22 dq, 1 H, J(3′′cis,1′′) ≈ J(gem ) = 1.6 (H-3′′cis);
4.68 dd, 1 H, J(1′a,2′) = 3.1, J(gem ) = 15.4 (H-1′a); 4.53 dd, 1 H, J(1′b,2′) = 7.6 (H-1′b); 4.27 m ,
1 H (H-2′); 4.20 dt, 2 H (H-1′′); 3.72 dd, 1 H, J(3′a,2′) = 4.4, J(gem ) = 10.4 (H-3′a); 3.63 dd, 1 H,
J(3′b,2′) = 6.5 (H-3′b); 3.61 dd, 1 H an d 3.57 dd, 1 H, J(P,CH) = 8.7, J(gem ) = 12.4 (PCH2).
13C NMR (D2O): 157.02 (C-4); 152.21 (C-2); 151.40 (C-6); 146.69 (C-8); 133.92 (C-2′′);
115.295 (C-3′′); 112.16 (C-5); 73.13 d, J(P,C) = 11.2 (C-3′); 69.37 d, J(P,C) = 150.9 (P-C);
69.07 (C-2′); 49.75 (C-1′); 42.87 (C-1′′).
Preparation of 6-Th ioxo Derivatives 47 an d 48. Gen eral Procedure
Th iourea (152 m g, 2 m m ol) was added to a solution of an appropriate 6-ch loro derivative 11
or 13 (1.5 m m ol) in absolute eth an ol (15 m l). Th e reaction m ixture was refluxed for 3 h ,
th en cooled to room tem perature an d taken down . Th e residue was applied on to a colum n
of silica gel (150 m l) an d ch rom atograph ed in system ch loroform –m eth an ol 4:1 (com poun d
47) or in system S3 (com poun d 48). Product-con tain in g fraction s were evaporated an d th e
th us obtain ed solid residue furth er recrystallized. Th e followin g com poun ds were obtain ed.
Diethyl {[2-hydroxy-3-(6-thioxo-1,6-dihydro-9H-purin-9-yl)propoxy]methyl}phosphonate (47) was
prepared from com poun d 11. Yield 460 m g (81%), wh ite crystals, m .p.137–138 °C (eth yl
acetate). HR MS (FAB): For C13H22N4O5PS [M + H] calculated: 377.1049; foun d: 377.1032.
FAB MS, m/z (%): 377 (100) [M + H]. 1H NMR (DMSO-d6): 13.70 br s, 1 H (NH); 8.195 s, 1 H
an d 8.19 s, 1 H (H-2, H-8); 5.36 br s, 1 H (2′-OH); 4.26 dd, 1 H, J(1′a,2′) = 3.7, J(gem ) = 13.8
(H-1′a); 4.07 dd, 1 H, J(1′b,2′) = 7.2 (H-1′b); 4.05 br pen t, 4 H, J(CH2,CH3) ≈ J(P,OCH) = 7.2
(P-OCH2); 4.00 m , 1 H (H-2′); 3.86 d, 2 H, J(P,CH) = 7.8 (PCH2); 3.53 dd, 1 H, J(3′a,2′) = 5.2,
J(gem ) = 10.0 (H-3′a); 3.49 dd, 1 H, J(3′b,2′) = 5.7 (H-3′b); 1.24 t, 6 H (CH3). 13C NMR
(DMSO-d6): 175.86 (C-6); 144.94 (C-2); 144.57 (C-4); 143.93 (C-8); 135.03 (C-5); 74.69 d,
J(P,C) = 10.3 (C-3′); 67.62 (C-2′); 64.70 d, J(P,C) = 162.1 (P-C); 61.94 d, 2 C, J(P,C) = 6.4
(P-OCH2); 46.90 (C-1′); 16.51 d, 2 C, J(P,C) = 5.4 (CH3).
Diethyl {[3-(2-amino-6-thioxo-1,6-dihydro-9H-purin-9-yl)-2-hydroxypropoxy]methyl}phosphonate
(48) was prepared from com poun d 13. Yield 437 m g (74%), wh ite crystals, m .p. 146–148 °C
(eth an ol). For C13H22N5O5PS (391.4) calculated: 39.89% C, 5.66% H, 17.89% N, 7.91% P,
8.19% S; foun d: 39.52% C, 5.62% H, 17.77% N, 7.84% P, 8.37% S. FAB MS, m/z (%): 392
(100) [M + H]. 1H NMR (DMSO-d6): 11.85 br s, 1 H (NH); 7.80 s, 1 H (H-8); 6.78 br s, 2 H
(NH2); 5.31 d, 1 H, J(OH,2′) = 5.5 (2′-OH); 4.05 dq, 4 H, J(CH2,CH3) = 7.1, J(P,CH) = 8.3
(P-OCH2); 4.04 dd, 1 H, J(1′a,2′) = 3.4, J(gem ) = 13.6 (H-1′a); 3.95 m , 1 H (H-2′); 3.86 dd,
1 H, J(1′b,2′) = 8.4 (H-1′b); 3.85 d, 2 H, J(P,CH) = 7.6 (PCH2); 3.51 dd, 1 H, J(3′a,2′) = 5.4,
J(gem ) = 10.0 (H-3′a); 3.47 dd, 1 H, J(3′b,2′) = 5.4 (H-3′b); 1.24 t, 6 H (CH3). 13C NMR
(DMSO-d6): 174.88 (C-6); 153.065 (C-2); 148.22 (C-4); 141.50 (C-8); 128.32 (C-5); 74.97 d,
J(P,C) = 9.8 (C-3′); 67.42 (C-2′); 64.735 d, J(P,C) = 161.6 (P-C); 61.91 d, 2 C, J(P,C) = 6.4
(P-OCH2); 46.30 (C-1′); 16.50 d, 2 C, J(P,C) = 5.4 (CH3).
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1911
Dieth yl {[3-(2-Am in o-6-oxo-1,6-dih ydro-9H-purin -9-yl)-2-h ydroxypropoxy]m eth yl}-
ph osph on ate (49)
A m ixture of 2-am in o-6-ch loro derivative 13 (825 m g, 2.1 m m ol) an d 80% acetic acid (20 m l)
was h eated to reflux for 2 h , th en cooled to room tem perature an d taken down . Th e residue
was coevaporated with toluen e (3 × 20 m l) an d th en ch rom atograph ed on a silica gel col-
um n (100 m l) in system ch loroform –m eth an ol (4:1). Yield 580 m g (74%), wh ite crystals,
m .p. 139–142 °C (eth an ol). For C13H22N5O6P (375.3) calculated: 41.60% C, 5.91% H, 18.66% N,
8.25% P; foun d: 41.11% C, 5.97% H, 18.36% N, 8.25% P. FAB MS, m/z (%): 376 (100) [M + H],
152 (58) [guan in e + H]. 1H NMR (DMSO-d6): 10.75 br s, 1 H (NH); 7.60 s, 1 H (H-8);
6.44 br s, 2 H (NH2); 5.50 d, 1 H, J(OH,2′) = 5.4 (2′-OH); 4.03 br pen t, 4 H, J(CH2,CH3) = 7.1,
J(P,OCH) = 8.0 (P-OCH2); 3.99 dd, 1 H, J(1′a,2′) = 2.9, J(gem ) = 13.8 (H-1′a); 3.92 m , 1 H
(H-2′); 3.86 dd, 1 H, J(1′b,2′) = 8.2 (H-2′b); 3.82 d, 2 H, J(P,CH) = 7.8 (PCH2); 3.46 d, 2 H,
J(3′,2′) = 5.0 (H-3′); 1.21 t, 6 H (CH3). 13C NMR (DMSO-d6): 158.08 (C-6); 154.39 (C-2);
152.30 (C-4); 139.75 (C-8); 116.715 (C-5); 75.57 d, J(P,C) = 10.7 (C-3′); 68.44 (C-2′); 65.25 d,
J(P,C) = 162.6 (P-C); 63.26 d, 2 C, J(P,C) = 6.4 (P-OCH2); 46.90 (C-1′); 17.06 d, 2 C, J(P,C) =
5.4 (CH3).
{[2-Hydroxy-3-(6-th ioxo-1,6-dih ydro-9H-purin -9-yl)propoxy]m eth yl}-
ph osph on ic Acid (50)
Brom otrim eth ylsilan e (0.79 m l, 6 m m ol) was added to a suspen sion of th ioxo derivative 47
(346 m g, 0.92 m m ol) in aceton itrile (10 m l). Th e reaction m ixture was stirred in th e dark at
room tem perature for 20 h , taken down in vacuo at 30 °C an d th e residue coevaporated
with toluen e (2 × 20 m l). Th e residue was dissolved in water (10 m l) an d n eutralized
dropwise with 5% LiOH to pH 7. Th e n eutral solution was evaporated to a sm aller volum e
(ca. 2–3 m l), applied on to a colum n of Dowex 50 (Li+ form , 50 m l) an d eluted with water.
Com bin ed UV-absorbin g fraction s were evaporated, th e solid residue m ixed with aceton e
(20 m l) an d set aside. Th e aceton e solution (con tain in g LiBr) was rem oved, th e sedim en t
treated th ree tim es m ore with aceton e (20 m l) to rem ove all LiBr. Th e solid residue was th en
filtered with suction , wash ed th orough ly with aceton e an d fin ally with eth er an d dried in
vacuo. Yield 255 m g (83%) of dilith ium salt of 50. Yellowish solid. FAB MS, m/z (%): 333
(30) [M + H of lith ium salt]. 1H NMR (D2O): 8.30 s, 1 H an d 8.29 s, 1 H (H-2, H-8); 4.46 dd,
1 H, J(1′a,2′) = 3.5, J(gem ) = 14.4 (H-1′a); 4.35 dd, 1 H, J(1′b, 2′) = 8.3 (H-1′b); 4.28 m , 1 H
(H-2′); 3.73 dd, 1 H, J(3′a,2′) = 3.5, J(gem ) = 10.5 (H-3′a); 3.63 d, 2 H, J(P,CH) = 8.9 (PCH2);
3.62 dd, 1 H, J(3′b,2′) = 6.0 (H-3′b). 13C NMR (D2O): 174.685 (C-6); 145.57 (C-2); 145.05
(C-8); 144.53 (C-4); 134.51 (C-5); 73.62 d, J(P,C) = 11.2 (C-3′); 68.69 d, J(P,C) = 152.8 (P-C);
68.35 (C-2′); 46.74 (C-1′).
Con version of Esters 22, 48 an d 49 to Free Acids
A m ixture of an appropriate dieth yl ph osph on ate (1 m m ol), aceton itrile (10 m l) an d brom o-
trim eth ylsilan e (1.35 m l, 10 m m ol) was treated an d th en worked up an alogously as de-
scribed in th e preparation of com poun ds 31–37 or 39–43. Th e crude product was desalted
on a colum n of Dowex 50X8 (H+ form , 50 m l) by elution with water. Elution of th e product
began after ca. 700–1000 m l of eluate. UV-absorbin g fraction s con tain in g th e product were
evaporated to dryn ess an d th e residue was crystallized from aqueous eth an ol. Th e followin g
com poun ds were obtain ed as free acids.
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

1912
Krečmerová, Masojídková, Holý:
[(2-Hydroxy-3-{6-[(2,2,2-trifluoroethyl)amino]-9H-purin-9-yl}propoxy)methyl]phosphonic acid
(38). Yield 300 m g (74%), m .p. 99–101 °C. For C11H15F3N5PO5·H2O (403.3) calculated:
32.76% C, 4.25% H, 14.13% F, 17.36% N, 7.68% P; foun d: 32.74% C, 4.24% H, 14.07% F,
17.66% N, 7.89% P. FAB MS, m/z (%): 386 (100) [M + H]. 1H NMR (D2O): 8.48 s, 1 H an d
8.44 s, 1 H (H-2, H-8); 4.60 m , 2 H (NCH2); 4.54 dd, 1 H, J(1′a,2′) = 3.9, J(gem ) = 14.7
(H-1′a); 4.44 dd, 1 H, J(1′b,2′) = 7.8 (H-1′b); 4.28 m , 1 H (H-2′); 3.73 dd, 1 H an d 3.70 dd,
1 H, J(P,CH) = 8.7, J(gem ) = 12.4 (PCH2); 3.71 dd, 1 H, J(3′a,2′) = 3.4, J(gem ) = 10.5 (H-3′a);
3.66 dd, 1 H, J(3′b,2′) = 5.0 (H-3′b). 13C NMR (D2O): 153.20 (C-6); 151.25 (C-2); 148.30
(C-4); 144.67 (C-8); 123.99 q, J(C,F) = 278.8 (CF3); 117.92 (C-5); 73.59 d, J(P,C) = 11.7 (C-3′);
68.08 (C-2′); 67.25 d, J(P,C) = 157.2 (P-C); 46.935 (C-1′); 42.72 q, J(C,F) = 30 (NCH2).
{[3-(2-Amino-6-thioxo-1,6-dihydro-9H-purin-9-yl)-2-hydroxypropoxy]methyl}phosphonic acid
(51). Yield 230 m g (69%) of wh ite crystals, m .p. 232–234 °C. For C9H14N5O5PS (335.3) calcu-
lated: 32.24% C, 4.21% H, 20.89% N, 9.24% P, 9.56% S; foun d: 31.94% C, 4.23% H, 20.53% N,
8.90% P, 9.81% S. FAB MS, m/z (rel.%): 336 (6) [M + H]. 1H NMR (D2O + NaOD): 7.87 s, 1 H
(H-8); 4.21 dd, 1 H, J(1′a,2′) = 4.0, J(gem ) = 14.8 (H-1′a); 4.20 m , 1 H (H-2′); 4.09 dd, 1 H,
J(1′b,2′) = 9.2 (H-1′b); 3.66 dd, 1 H, J(3′a,2′) = 3.5, J(gem ) = 10.6 (H-3′a); 3.54 dd, 1 H,
J(3′b,2′) = 5.6 (H-3′b); 3.56 dd, 1 H an d 3.52 dd, 1 H, J(P,CH) = 8.8, J(gem ) = 12.4 (PCH2).
13C NMR (D2O + NaOD): 177.925 (C-6); 158.75 (C-2); 148.06 (C-4); 141.17 (C-8); 129.42
(C-5); 74.13 d, J(P,C) = 11.2 (C-3′); 69.66 d, J(P,C) = 150.4 (P-C); 68.46 (C-2′); 46.16 (C-1′).
{[3-(2-Amino-6-oxo-1,6-dihydro-9H-purin-9-yl)-2-hydroxypropoxy]methyl}phosphonic acid (52).
Yield 202 m g (55%). For C9H14N5O6P·2.5H2O (364.3) calculated: 29.68% C, 5.26% H,
19.23% N, 8.50% P; foun d: 29.61% C, 4.98% H, 19.23% N, 8.24% P. FAB MS, m/z (%): 320
(30) [M + H], 152 (5) [guan in e + H]. 1H NMR (D2O + NaOD): 7.75 s, 1 H (H-8); 4.16 m , 1 H
(H-2′); 4.16 dd, 1 H, J(1′a,2′) = 5.6, J(gem ) = 14.6 (H-1′a); 4.00 dd, 1 H, J(1′b,2′) = 8.8 (H-1′b);
3.58 dd, 1 H, J(3′a,2′) = 3.8, J(gem ) = 10.0 (H-3′a); 3.51 d, 2 H, J(P,CH) = 8.8 (PCH2); 3.47 dd,
1 H, J(3′b,2′) = 5.6 (H-3′b). 13C NMR (D2O + NaOD): 160.39 (C-6); 155.21 (C-2); 150.87
(C-4); 138.68 (C-8); 116.73 (C-5); 74.57 d, J(P,C) = 11.2 (C-3′); 68.78 d, J(P,C) = 150.4 (P-C);
67.73 (C-2′); 46.31 (C-1′).
As a research project of the Institute of Organic Chemistry and Biochemistry, Academy of Sciences
of the Czech Republic (No. 4055905), this study was supported by the Programme of Targeted Projects
of the Academy of Sciences of the Czech Republic (No. S4055109), by amfAR (grant No.
106456-34-RGNT), by the Descartes Prize 2001 of the European Union and by Gilead Sciences (Foster
City, CA, U.S.A.). The authors’ thanks are due to Prof. E. De Clercq and his group in the Rega Insti-
tute, Catholic University Leuven (Belgium) for antiviral screening and to Dr I. Votruba of the Institute
of Organic Chemistry and Biochemistry for the estimation of cytostatic activity.
REFERENCES
1. a) De Clercq E., Holý A., Rosenberg I., Sakuma T., Balzarini J., Maudgal P. C.: Nature
1986, 323, 464; b) De Clercq E., Sakuma T., Baba M., Pauwels R., Balzarini J., Rosenberg I.,
Holý A.: Antiviral Res. 1987, 8, 261; c) Holý A., Votruba I., Merta A., Černý J., Veselý J.,
Vlach J., Šedivá K., Rosenberg I., Otmar M., Hřebabecký H., Trávníček M., Vonka V.,
Snoeck R., De Clercq E.: Antiviral Res. 1990, 13, 295; d) Holý A., Dvořáková H., Jindřich J.
in: Antibiotics and Antiviral Compounds (K. Krohn, H. A. Kirst and H. Maag, Eds), p. 455.
Verlag Chemie, Weinheim–New York 1993; e) Holý A. in: Advances in Antiviral Drug
Collect. Czech. Chem. Commun. (Vol. 69) (2004)

Acyclic Nucleotide Analogues
1913
Design (De Clercq E., Ed.), p. 179. JAI Press, Inc., Greenwich (CT) 1994; f) Holý A. in:
Recent Advances in Nucleosides: Chemistry and Chemotherapy (C. K. Chu, Ed.), p. 167.
Elsevier, Amsterdam 2002; g) Holý A.: Curr. Pharm. Des. 2003, 9, 2567.
2. Baba M., Konno K., Shigeta S., De Clercq E.: Eur. J. Clin. Microbiol. Infect. Dis. 1987, 6,
158.
3. Terry B. J., Mazina K. E., Tuomari A. V., Haffey M. L., Hagen M., Feldman A., Slusarchyk
W. A., Young M. G., Zahler R., Field A. K.: Antiviral Res. 1988, 10, 235.
4. Holý A.: Collect. Czech. Chem. Commun. 1993, 58, 649.
5. a) Naesens L., De Clercq E.: Nucleosides Nucleotides 1997, 16, 983; b) Rahhal F. M.,
Arevalo J. F., Munguia D., Taskintuna I., Delapaz E. C., Azen S. P., Freeman W. R.:
Ophthalmology 1996, 103, 1078; c) De Clercq E.: Collect. Czech. Chem. Commun. 1998,
63, 480.
6. Rosenberg I., Holý A.: Collect. Czech. Chem. Commun. 1982, 47, 3447.
7. Webb R. R., Martin J. C.: Tetrahedron Lett. 1987, 28, 4963.
8. Holý A., Rosenberg I.: Collect. Czech. Chem. Commun. 1987, 52, 2775.
9. Holý A., Rosenberg I., Dvořáková H.: Collect. Czech. Chem. Commun. 1989, 54, 2470.
10. Rosenberg I., Holý A.: Collect. Czech. Chem. Commun. 1987, 52, 2792.
11. Holý A., Votruba I., Tloušꢀová E., Masojídková M.: Collect. Czech. Chem. Commun. 2001,
66, 1545.
12. Orlov N. F., Viktorov O. F., Elkina N. S.: Dokl. Akad. Nauk SSSR 1966, 166, 378.
13. Arbuzov B. A., Bogonostseva N. P.: Zh. Obshch. Khim. 1959, 29, 2617.
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