Syntheses of base and side-chain modified pyrimidine 1-[2- (phosphonomethoxy)propyl] derivatives as potent inhibitors of thymidine phosphorylase (PD-ECGF) from SD-lymphoma

Article abstract of DOI:10.1135/cccc20060595

In this study we synthesized a series of thymine and 5-ethyluracil acyclic nucleoside phosphonates bearing hydroxymethyl, methoxymethyl, azidomethyl, aminomethyl and (trimethylammonio)methyl group in side chain as potent inhibitors of thymidine phosphoryl

Full text of DOI:10.1135/cccc20060595

Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
595
SYNTHESES OF BASE AND SIDE-CHAIN MODIFIED PYRIMIDINE
1-[2-(PHOSPHONOMETHOXY)PROPYL] DERIVATIVES AS POTENT
INHIBITORS OF THYMIDINE PHOSPHORYLASE (PD-ECGF) FROM
SD-LYMPHOMA
1,
2
3
4
Karel POMEISL *, Ivan VOTRUBA , Antonín HOLÝ and Radek POHL
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
1
2
166 10 Prague 6, Czech Republic; e-mail: pomeislk@uochb.cas.cz, votruba@uochb.cas.cz,
holy@uochb.cas.cz, pohl@uochb.cas.cz
3
4
Received Jan uary 23, 2006
Accepted March 10, 2006
In th is study we syn th esized a series of th ym in e an d 5-eth yluracil acyclic n ucleoside
ph osph on ates bearin g h ydroxym eth yl, m eth oxym eth yl, azidom eth yl, am in om eth yl an d
(trim eth ylam m on io)m eth yl group in side ch ain as poten t in h ibitors of th ym idin e ph os-
ph orylase. In addition , we in vestigated in particular th e n ovel syn th eses of fluorin ated de-
rivatives con tain in g fluorom eth yl or trifluorom eth yl groups in side ch ain such as 5-eth yl-
1-[(S)-3-fluoro-2-(ph osph on om eth oxy)propyl]uracil (8) or 5-eth yl-1-[3,3,3-trifluoro-2-(ph os-
ph on om eth oxy)propyl]uracil an d th ym in e derivatives 27 an d 28. Uracil acyclic n ucleoside
ph osph on ates 1-{2-[(diisopropoxyph osph oryl)m eth oxy]eth yl}uracil (12) an d 1-{2-[(diiso-
propoxyph osph oryl)m eth oxy]-3,3,3-trifluoropropyl}uracil (19) were fluorin ated to corre-
spon din g 5-fluorouracil derivatives. Wh ile th e 5-fluorouracil derivatives exh ibit a m argin al
in h ibitory effect, th ym in e an d 5-eth yluracil com poun d with fluorin e in side ch ain s possess
con siderable in h ibitory poten cy toward th ym idin e ph osph orylase from rat spon tan eous
T-cell lym ph om a.
Keyw ord s: Acyclic n ucleoside ph osph on ates; Acyclic n ucleotide an alogues; Nucleotides;
Th ym idin e ph osph orylase; Fluorin ation ; Pyrim idin es; Uracil; Th ym in e.
Acyclic n ucleoside ph osph on ates (ANPs) exh ibit various kin ds of biological
activities such as an tiviral, cytostatic, an tiparasitic an d im m un om odulatory
effects1. At presen t th ese com poun ds are in vestigated in con n ection with
th eir possible use as poten t in h ibitors of th ym idin e ph osph orylase2. Th y-
m idin e ph osph orylase [dTh d, EC 2.4.2.4. platelet-derived en doth elial-cell
growth factor (PD-ECGF)], an im portan t salvage-path way en zym e, catalyzes
ph osph orolysis of th ym idin e to th ym in e an d 2′-deoxy-D-ribose 1-ph os-
ph ate. Th e deph osph orylated product of th e latter, 2′-deoxy-D-ribose, h as
a ch em otactic activity in vitro an d an giogen ic activity in vivo stim ulatin g
en doth elial-cell m igration . Th is process is crucial for th e form ation of n ew
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624
© 2006 Institute of Organic Chemistry and Biochemistry
doi:10.1135/cccc20060595

596
Pomeisl, Votruba, Holý, Pohl:
blood vessels in a tum or wh ich overexpress th ym idin e ph osph orylase. Th us
th e in h ibition of 2′-deoxy-D-ribose release from en doth elial cells is a poten -
tial an ti-an giogen ic target in can cer ch em oth erapy.
Th e aim of our work h as been th e developm en t of n ew in h ibitors of
th ym idin e ph osph orylase based on th e structure of specifically m odified
an d m etabolically stable pyrim idin e ANPs. In th is study we syn th esized a
series of th ym in e an d 5-eth yluracil ANPs bearin g h ydroxym eth yl, m eth oxy-
m eth yl, azidom eth yl, am in om eth yl an d (trim eth ylam m on io)m eth yl group
in side ch ain (com poun ds I, Fig. 1). In addition , we in vestigated in particu-
lar th e n ovel syn th eses of fluorin ated derivatives con tain in g fluorom eth yl
(FPMP com poun ds) or trifluorom eth yl groups in side ch ain because fluorin e-
con tain in g substituen ts are often powerful m odifiers of ch em ical an d bio-
logical properties. For th e sam e reason we fluorin ated 1-[2-(ph osph on o-
m eth oxy)eth yl]uracil an d 1-[3,3,3-trifluoro-2-(ph osph on om eth oxy)propyl]-
uracil in th e rin g position C-5 to com pare th e in h ibitory effect of th e corre-
spon din g 5-fluorouracil derivatives (com poun ds II, Fig. 1). Th e obtain ed
com poun ds were tested as poten t in h ibitors of th ym idin e ph osph orylase
(PD-ECGF) expressed in V79 ch in ese h am ster cells, as well as th ym idin e
ph osph orylase from SD-lym ph om a an d h um an placen ta.
O
O
R2
R1
F
compounds I
compounds II
HN
HN
O
N
O
N
R
O
P(O)(OH)2
O
P(O)(OH)2
R1 = CH2OH, CH2OMe, CH2N3, CH2NH2,
CH2N+(CH3)3, CH2F, CF3
R2 = Me, Et
R = H, CF3
O
Me
R
HN
O
N
O
P(O)(OH)2
PMPT, R = Me
HPMPT, R = CH2OH
FPMPT, R = CH2F
FIG. 1
Structures of com poun ds un der study
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
597
RESULTS AND DISCUSSION
It is well kn own th at a n um ber of side-ch ain m odified pyrim idin e ANPs
possess con siderable in h ibitory activity on th ym idin e ph osph orylase iso-
lated from rat livers3. Th is is docum en ted by a kin etic study of several th y-
m in e ANP derivatives4 wh ich sh owed th e efficien t in h ibitory effect of
th ym idin e ph osph orylase for 1-[2-(ph osph on om eth oxy)propyl]th ym in e
(PMPT), 1-[3-h ydroxy-2-(ph osph on om eth oxy)propyl]th ym in e (HPMPT) an d
1-[3-fluoro-2-(ph osph on om eth oxy)propyl]th ym in e (FPMPT) (see Fig. 1).
Sim ilarly to previously reported procedure4 developed for th e preparation
of HPMPT or FPMPT, we used th is sim ple m eth od for syn th esis of com -
poun ds 5a an d 5b to com pare th e in fluen ce of th eir bulkier alkyl at posi-
tion C-5 of th e uracil m oiety on th e bioactivity in n ew syn th esized com -
poun ds 5a, 5b an d 8 (Sch em e 1 an d 2). N1-Alkylation 5 of protected base 1
with ch iral oxiran es 2a an d 2b takes place in dim eth ylform am ide in th e
presen ce of cesium carbon ate as catalyst affordin g com poun ds 3a an d 3b in
good preparative yields. For th e preparation of 4a an d 4b we h ave used a
gen eral procedure3,5 with (diisopropoxyph osph oryl)m eth yl tosylate in tro-
ducin g a startin g ph osph on ate block. Th e rem oval of trityl group in 4a an d
th e followin g replacem en t of h ydroxy group with fluorin e in th e in ter-
m ediate 6 was perform ed usin g perfluorobutan e-1-sulfon yl fluoride3 in th e
O
CH2CH3
HN
OCH3
CH2CH3
O
N
N
OH
O
N
H
O
P(O)(OH)2
1
5a,5b
OTr
2a,2b
O
a, (R)-enantiomers
b, (S)-enantiomers
(i)
(iii)
OCH3
N
OCH3
CH2CH3
CH2CH3
N
N
(ii)
O
O
N
OTr
OTr
OH
3a,3b
O
4a,4b
P(O)(OiPr)2
(i) Cs2CO3, DMF, 80 °C; (ii) TsOCH2P(O)(OiPr)2, NaH, THF, -20 °C-40 °C;(iii) (CH3)3SiBr, CH3CN, r.t.
SCHEME 1
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598
Pomeisl, Votruba, Holý, Pohl:
presen ce of DBU to give 7 in a preparative yield (Sch em e 2). Fin ally, th e
obtain ed ph osph on ates 4a, 4b an d 7 were deprotected by treatm en t with
brom otrim eth ylsilan e followed by h ydrolysis to give 5a, 5b an d 8.
O
OCH3
OCH3
OCH3
CH2CH3
CH2CH3
(iii)
CH2CH3
CH2CH3
HN
N
N
N
(i)
(ii)
O
N
O
N
O
N
O
N
F
P(O)(OH)2
F
OH
OTr
O
O
P(O)(OiPr)2
O
P(O)(OiPr)2
O
P(O)(OiPr)2
4a
6
7
8
(i) H2, Pd/C, 99% AcOH, MeOH; (ii) CF3(CF2)3SO2F, DBU, toluene, rt→90 °C;
(iii) (CH3)3SiBr, CH3CN, r.t.
SCHEME 2
Accordin g to previous studies6, a h alogen atom in C-5 position of a uracil
m oiety can also sign ifican tly m odify th e in h ibitory activity of th ym idin e
ph osph orylase. For exam ple, th e in creased in h ibition effect2,7 of 6-am in o-
5-brom ouracil ANPs com pared with th ym in e derivatives can be m en tion ed.
In addition , 5-h alouracil derivatives also exh ibit various kin ds of biological
activities8,9. Our effort was directed to in vestigate in h ibitory poten cy to-
ward th ym idin e ph osph orylase of m etabolically stable 5-fluorouracil ANP
derivatives 15 an d 21.
Com poun d 15 was prepared by a m ultistep syn th esis usin g direct fluori-
n ation of th e uracil m oiety in easily avalaible ph osph on ate 12 with F2 di-
luted with n itrogen , in acetic acid solution (Sch em e 3). Th e reaction course
O
OCH3
OCH3
HN
N
Cl
P(O)(OiPr)2
O
10
N
(ii)
O
N
O
N
O
N
H
(i)
O
P(O)(OiPr)2
O
P(O)(OiPr)2
9
12
11
O
N
O
O
F
F
F
HN
O
HN
HN
OAc
(iv)
O
N
(v)
O
N
(iii)
O
P(O)(OiPr)2
O
P(O)(OiPr)2
O
P(O)(OH)2
14
15
13
(i) NaH, DMF, 90 °C; (ii) Dowex 50 (H+), 90% aq. MeOH, ∆;
(iii) 5-10% F2/N2, 99% AcOH; (iv) Et3N, EtOH, reflux; (v) (CH3)3SiBr, CH3CN, r.t.
SCHEME 3
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Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
599
m ay be assum ed to in clude th e prim ary addition of h ardly form ed
CH3COOF to 5,6-double boun d of base10 to form in term ediate adducts 13,
wh ich com pletely con verted to 14 by treatm en t with trieth ylam in e. Th e re-
quired com poun d 12 was obtain ed by reaction of 4-m eth oxypyrim idin -
2(1H)-on e11 9 with diisopropyl [(2-ch loroeth oxy)m eth yl]ph osph on ate (10)
in th e presen ce of sodium h ydride followed by h ydrolysis of com poun d 11
in aqueous m eth an ol usin g Dowex 50 (H+ form ).
Fluorin ation of a h eterocyclic base was furth er used in th e syn th esis of
com poun d 21 bearin g th ree addition al fluorin e atom s in th e side ch ain
(Sch em e 4). For prelim in ary biological studies we developed th e syn th esis
of on ly for racem ic tetrafluoroph osph on ate 21 startin g from com m ercially
available (trifluorom eth yl)oxiran e (16). Its reaction with 4-m eth oxy-
pyrim idin -2(1H)-on e (9) in th e presen ce of cesium carbon ate afforded
h ydroxyl derivative 17 wh ich was furth er alkylated with (diisopropoxy-
ph osph oryl)m eth yl tosylate in th e presen ce of sodium h ydride. Th e
m eth oxy group of th e obtain ed ph osph on ate 18 was rem oved by h ydrolysis
on a Dowex 50 (H+ form ) to give 19 wh ich was fluorin ated un der usual con -
dition s12.
OCH3
OCH3
CF3
OCH3
N
O
N
16
N
(ii)
O
N
O
N
CF3
(i)
CF3
O
N
H
O
P(O)(OiPr)2
OH
9
17
18
O
O
N
O
N
F
F
HN
HN
HN
(vi)
O
N
O
(iii)
O
(iv), (v)
CF3
P(O)(OH)2
CF3
CF3
P(O)(OiPr)2
19
O
O
P(O)(OiPr)2
O
21
20
(i) Cs2CO3, DMF, 80 °C; (ii) TsOCH2P(O)(OiPr)2, NaH, THF, -20 °C-r.t.; (iii) Dowex 50 (H+), 90% aq.
MeOH; (iv) 5-10% F2/N2, 99% AcOH; (v) Et3N, EtOH, reflux; (vi) (CH3)3SiBr, CH3CN, r.t.
SCHEME 4
A sim ilar procedure h as used to syn th esize ph osph on ates 27 an d 28 con -
tain in g th ym in e or 5-eth yluracil m oiety (Sch em e 5). Th e preparation of
bulky quatern ary (trim eth ylam m on io)m eth yl derivative 32 followed th e
sam e route13 usin g com m ercially avalaible glycidyltrim eth ylam m on ium
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

600
Pomeisl, Votruba, Holý, Pohl:
ch loride (29) as a startin g com poun d. Th e separation of th e product from
in term ediate 31 by ion exch an ge ch rom atograph y on a Dowex 50 (H+
form ) afforded com poun d 32.
O
OCH3
OCH3
R
OCH3
R
R
HN
N
N
R
N
(ii)
(iii)
O
N
(i)
O
N
O
N
CF3
O
N
H
CF3
P(O)(OiPr)2
CF3
OH
CF3
O
16
O
P(O)(OH)2
O
27; R = Et
28; R = Me
1; R = Et
25; R = Et
26; R = Me
23; R = Et
24; R = Me
22; R = Me
O
OCH3
CH3
OCH3
CH3
CH3
HN
N
OCH3
CH3
N
Cl-
N
O
(iii)
N
N
Cl-
N+(CH3)3
O
O
N
(ii)
N+(CH3)3
(i)
N+(CH3)3
O
OH
O
N
H
N+(CH3)3
Cl-
O
P
O
O
P(O)(OiPr)2
31
O-
OH
22
29
32
30
(i) Cs2CO3, DMF, 80 °C; (ii) TsOCH2P(O)(OiPr)2, NaH, THF, -20 °C-r.t.; (iii) (CH3)3SiBr, CH3CN, rt
SCHEME 5
For th e preparation of racem ic azido an d am in o derivatives 36 an d 37 we
used ph osph on ate 33, wh ich was easily obtain ed by a previously reported
m ultistep syn th esis13 startin g from oxiran e 2 (Sch em e 6). Th e alkylation of
4-m eth yl-5-m eth oxypyrim idin -2(1H)-on e (22) with 33 proceeded sm ooth ly
in th e presen ce of sodium h ydride. Separation of N1- an d O2-regioisom ers
by colum n ch rom atograph y gave a m ixture of 34a an d 34b in th e ratio
43:57. N-1-Alkylated regioisom er 34a was th en con verted to th e corre-
spon din g am in o derivative 35 by h ydrogen ation in m eth an ol over 10% pal-
ladium on ch arcoal followed by deprotection of th e 4-OH group in aqueous
m eth an ol usin g ion exch an ger resin Dowex 50 (H+ form ).
Likewise, m eth oxy derivatives 45a, 45b an d 46a, 46b were prepared from
ch iral precursors 40a an d 40b (Sch em e 7). Sim ilarly to th e previously re-
ported procedure14 usin g azido derivative 33 or th e oth er related com -
poun ds15, we syn th esized th e n ew ph osph on ate buildin g blocks from ch iral
oxiran es 2a an d 2b. Th eir reaction with a sodium m eth oxide in th e pres-
en ce of cesium carbon ate afforded quan titatively 38a an d 38b . Th e
alkylation of obtain ed derivatives proceeded by usual reaction with a
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Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
601
OCH3
CH3
OCH3
CH3
OCH3
CH3
N
O
N
(i)
N
N3
OTs
+
+
O
N
O
N
O
P(O)(OiPr)2
O
N
H
N3
N3
33
22
P(O)(OiPr)2
O
P(O)(OiPr)2
34a
34b
(iv)
O
(ii), (iii)
O
N
O
CH3
CH3
CH3
HN
HN
HN
(iv)
O
N
O
O
N
N3
P(O)(OH)2
NH2
P(O)(OiPr)2
NH2
O
O
O
P(O)(OH)2
35
37
36
(i) NaH, DMF, 90 °C; (ii) H2, Pd/C, MeOH, r.t.; (iii) Dowex 50 (H+), 90% aq. MeOH;
(iv) (CH3)3SiBr, CH3CN, r.t.
SCHEME 6
(iii), (iv)
(i)
(ii)
OTr
MeO
OTr
MeO
OTr
P(O)(OiPr)2
39a,39b
MeO
OTs
O
OH
O
O
P(O)(OiPr)2
2a,2b
38a,38b
40a,40b
OCH3
R
N
O
O
N
OCH3
R
N
OMe
P(O)(OiPr)2
a, (R)-enantiomers
b, (S)-enantiomers
O
N
H
43a,43b; R = Et
44a,44b; R = Me
1, 22
40a,40b
(v)
OCH3
O
R
R
N
HN
(vi)
O
N
O
N
OMe
P(O)(OiPr)2
OMe
P(O)(OH)2
O
O
45a,45b; R = Et
46a,46b; R = Me
41a,41b; R = Et
42a,42b; R = Me
(i) MeONa, Cs2CO3, DMF, 95 °C; (ii) NaH, TsOCH2P(O)(OiPr)2, THF, -20 °C-40 °C;
(iii) Dowex 50 (H+), MeOH, H2O, reflux; (iv) TsCl, Et3N, DMAP, CH2Cl2; 0°C-reflux;
(v) NaH, DMF, 100 °C; (vi) (CH3)3SiBr, CH3CN, r.t.
SCHEME 7
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602
Pomeisl, Votruba, Holý, Pohl:
ph osph on ate buildin g block to form 39a an d 39b. Desired ph osph on ates
40a an d 40b were obtain ed by rem oval of trityl group an d furth er
tosylation of HO in term ediates with tosyl ch loride an d trieth ylam in e in th e
presen ce of 4-(dim eth ylam in o)pyridin e in good preparative yields. Alkyl-
ation of protected pyrim idin -2(1H)-on es 1 an d 22 in N1- an d O2-position s
with syn th on s 40a an d 40b gave ch iral (R)- an d (S)-derivatives of 41, 42
an d 43, 44. After separation of both regioisom ers N1-derivatives were
deprotected as well as com poun ds 15, 21, 27, 28, 32, 36, 37, 41a, 41b an d
42a, 42b with brom otrim eth ylsilan e an d h ydrolysis.
Th e structures of all prepared com poun ds were con firm ed by NMR spec-
troscopy with com plete proton an d carbon assign m en t usin g H,C-h etero-
correlated experim en ts (HSQC an d HMBC). Th e N1- an d O2-regioisom ers
can be easily distin guish ed based on th eir 13C an d HMBC spectra. Table I
sh ows ch aracteristic ch em ical sh ifts for N1- an d O2-regioisom ers, wh erein 13C
ch em ical sh ift of CH2-1′ fragm en t is particularly n oteworth y. H,C-HMBC
spectra of N1-isom ers sh ow crosspeaks of H-1′ to C-6 an d C-2, wh ile spectra
of O2-isom ers sh ow crosspeaks of H-1′ to on ly C-2. Th e optical purity of
selected en an tiom eric products was ch ecked by 1H NMR m easurem en t of
dyn am ic com plexes with ch iral solvatin g agen t (–)-(R)-1-(9-an th ryl)-
2,2,2-trifluoroeth an -1-ol (TFAE)16 in CDCl3 an d com pared with th e dia-
stereom eric m ixtures of correspon din g racem ic com poun ds (Fig. 2). Th e
optical purity was >95% ee in all cases.
In con clusion , th e series of th ym in e an d 5-eth yluracil (ph osph on o-
m eth oxy)propyl derivatives substituted at th e C-3′ position of th e aliph atic
ch ain with several subtituen ts were evaluated for th eir ability to in h ibit
th ym idin e ph osph orylase (TP) from rat spon tan eous T-cell lym ph om a,
TABLE I
Ch aracteristic ch em ical sh ifts (in ppm ) for N1- an d O2-regioisom ers
Com poun d
(isom er)
H-1′
C-1′
C-2
C-6
34a (N1)
41 (N1)
42 (N1)
34b (O2)
43 (O2)
44 (O2)
3.86, 4.11
3.77, 4.19
3.73, 4.20
4.39, 4.48
4.41, 4.44
4.41, 4.44
50.97
50.96
50.84
65.66
65.50
66.56
156.74
156.67
156.77
163.03
163.21
163.25
145.64
145.06
145.92
156.92
156.03
156.99
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Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
603
h um an placen ta, an d com m ercial en zym es (h um an TP expressed in V79
cells, E. coli). Table II sh ows th at th e in h ibitory effect of th ese substituted
(ph osph on om eth oxy)alkyl derivatives on T-cell lym ph om a TP decreases ac-
cordin g to th e substitution in side ch ain of th e (ph osph on om eth oxy)alkyl
lin ker at th e position C-3′ in th e order: -CH2F > -CH2OCH3 > -CH2N3 > -CF3 >
-CH2N+(CH3)3 > -CH2OH > -CH2NH2 with on e exception to th is rule: Com -
poun d 5a is th e sam e or even stron ger in h ibitor of th e en zym e th an com -
poun d 8, wh ich in h ibits T-cell lym ph om a TP with h igh er efficien cy th an
(R)-FPMPT (Vi/V0 = 0.11)3 an d possesses also som e, th ough low, in h ibitory
activity towards h um an en zym es (Vi/V0 ~ 0.77). Very efficien t in h ibitors of
T-cell lym ph om a TP are also com poun ds 46a, 45a, 36, 45b, 46b an d 28.
A com parison of th e in h ibitory activity of com poun ds 46a, 45a, 45b, 46b
sh ows th e crucial effect of th e side-ch ain substitution with -CH2OCH3
group. Substitution of th e uracil rin g at position 5 with m eth yl an d/or eth yl
group in both (R)- an d (S)-con figuration s displays on ly a m argin al effect.
Th e in h ibitory poten cy of 5-fluorouracil derivatives 15 an d 21 is low com -
pared with th e syn th esized th ym in e an d 5-eth yluracil derivatives.
As in our previous study3,4, th e [(ph osph on om eth oxy)alkyl] th ym in es are
n ot efficien t in h ibitors of E. coli an d h um an TPs. Th ese differen ces in th e
recogn ition of active sites of rat T-cell lym ph om a by [(ph osph on om eth oxy)-
FIG. 2
Optical purity determ in ation of th e com poun ds 42a an d 42b. a Part of 1H NMR spectrum of
th e dyn am ic com plex 42a + TFAE; b part of 1H NMR spectrum of th e dyn am ic com plex 42b +
TFAE; c part of 1H NMR spectrum of th e dyn am ic com plex 42a + 42b + TFAE sh owin g
diastereom eric m ixture (racem ic m ixture was obtained by m ixing of 42a and 42b in ~2:1 ratio)
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

604
Pomeisl, Votruba, Holý, Pohl:
alkyl]th ym in es, com pared with h um an an d E. coli TP, could be th e result of
som e m utation or post-tran slation m odification .
Despite their TP inhibitory activity, any of the presented com pounds do not
possess at a con cen tration of 10 µm ol l–1, a sign ifican t cytostatic activity in
tissue cultures estimated in mouse lymphocytic leukemia L1210 cells (ATCC CCL
219), CCRF-CEM T lym ph oblastoid cells (h um an acute lym ph oblastic leuke-
m ia, ATCC CCL 119), hum an prom yelocytic leukem ia HL-60 cells (ATCC CCL
240) an d h um an cervix carcin om a HeLa S3 cells (ATCC CCL 2.2).
TABLE II
In h ibition of th ym idin e ph osph orylases by ANPs
In h ibition of th ym idin e ph osph orylasea, Vi/V0
Com poun d
Escherichia
coli
Hum an , V79
expressed
Hum an
placen ta
SD-Lym ph om a
5a
0.94
0.95
0.88
0.94
1.00
0.83
0.93
0.96
0.97
1.00
1.00
0.84
1.00
1.00
0.86
0.87
0.77
0.96
0.93
1.00
0.76
0.96
1.00
0.93
0.96
1.00
0.96
0.94
0.01
0.16
0.02
0.90
0.57
0.17
0.12
0.14
0.06
0.50
0.04
0.10
0.04
0.10
0.94
0.94
0.79
0.81
1.00
0.95
1.00
0.95
1.00
1.00
0.99
1.00
1.00
0.90
5b
8
15
21
27
28
32
36
37
45a
45b
46a
46b
a
100 µM [3H]-2′-deoxyth ym idin e, 250 µM Pi (pH 6.7), tested com poun d 10 µm ol l–1, an ap-
propriate am oun t of en zym e, 10 m in in cubation at 37 °C (refs4,17). Th e in h ibitory efficacy is
expressed by Vi/V0 (Vi, rate of ph osph orolysis in th e presen ce of in h ibitors; V0, rate of
ph osph orolysis in th e absen ce of in h ibitors). Pi, in organ ic ph osph ate
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
605
EXPERIMENTAL
Un less stated oth erwise, solven ts were evaporated at 40 °C/0.5–2 kPa an d com poun ds were
dried at 50 °C/13 Pa. Meltin g poin ts were determ in ed on a Kofler block an d are un corrected.
Optical rotation s were m easured on a Autopol IV polarim eter (Rudolph Research An alytical,
U.S.A.) at 25 °C; [α]D values are given in 10–1 deg cm 2 g–1. IR spectra were recorded on a
FTIR spectrom eter Bruker IFS 55 (Equin ox) in KBr, CCl4 an d CHCl3. An alytical TLC was car-
ried out on Silufol UV254 plates (Kavalier Votice, Czech Republic). Colum n ch rom atograph y
was perform ed on n eutral alum in um oxide 150 m esh 58 Å with 3% of water (Aldrich ) an d
silica gel 60 µm (Fluka). Preparative TLC was carried out on 45 × 18 × 0.4 cm loose-layer
silica gel con tain in g UV in dicator (system S1). Purification of ph osph on ates 5a, 5b, 15, 21,
27, 28, 32, 36, 37, 45a, 45b an d 46a, 46b was perform ed on Seph adex A-25-120 DEAE (Cl–
form , activated with 0.02 M trieth ylam m on ium h ydrogen carbon ate) an d eluted in 0–0.4
M
trieth ylam m on ium h ydrogen carbon ate buffer (TEAB) (system S2). Reversed-ph ase HPLC was
perform ed on a Waters Delta 600; Xterra , an alytical RP18 colum n 5 µm 3.9 × 150 m m ,
0.05 M TEAB (system S3) or 0.025 M TEAB (system S4) or 0.0025 M (system S5) an d Lun a
Ph en om en ex , preparative C18 colum n 21.20 × 250 m m , 0.025 M TEAB (system S6). NMR
spectra were recorded on Bruker Avan ce 400, Bruker Avan ce 500, Varian Un ity 200 or Varian
Un ity 500 spectrom eters (1H at 400 or 500, 13C at 100.6 or 125.8 , 31P at 162 an d 19F at
188.2 or 470.2 MHz) in CDCl3 with in tern al stan dard tetram eth ylsilan e , in DMSO-d6 solu-
tion s (referen ced to th e solven t sign al at δ 2.50) or in D2O solution s with dioxan e18 as an
in tern al stan dard (3.75 ppm for 1H an d an d 69.3 ppm for 13C). 31P an d 19F NMR spectra
were referen ced to th e sign al of H3PO4 (0 ppm ) an d h exafluoroben zen e (–163 ppm ), respec-
tively, wh ich were used as an in tern al stan dards or as an extern al stan dards in coaxial 2 m m
capillary tube. 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. Th e n um berin g system for assign m en t of NMR sign als is
outlin ed in Fig. 3. Mass spectra were m easured on a ZAB-EQ (VG An alytical) spectrom eter
usin g FAB (Xe ion ization , acceleratin g voltage 8 kV, glycerol m atrix) an d Q-Tof m icro
(Waters, Milford, MA, U.S.A.) equipped with an ion source for atm osph eric pressure ch em i-
cal ion ization (APCI probe tem perature 400 °C, sam ple con e voltage 30 V, coron a disch arge
curren t 5.0 µA).
O
4
OCH3
OCH3
4
R3
4
R3
R3
3
HN
5
5
5
N
N
6
6
6
2
2
O
N1
O
N1
2
O
N1
3'
3'
2'
2'
1'
2'
1'
R1
1'
R1
3' R1
P(O)(OR2)2
P(O)(OR2)2
O
P(O)(OR2)2
O
O
4'
4'
4'
FIG. 3
Gen eral n um berin g sch em e for assign m en t of NMR sign als
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

606
Pomeisl, Votruba, Holý, Pohl:
Materials an d Ch em icals
Stan dard ch em icals, ion -exch an ge resin (Dowex 50WX8-200) an d activated ch arcoal were
purch ased from Sigm a–Aldrich (Czech Republic). Ion -exch an ge resin (Seph adex A-25-120 DEAE)
was purch ased from Fluka. Perfluorobutan e-1-sulfon yl fluoride was obtain ed from
Fluoroch em (Wesley Street, Old Glossop). (Trifluorom eth yl)oxiran e was purch ased from
Tokyo Kasei Kogyo Co, Ltd. (Tosh im a, Kita-ku Tokyo, Japan ). Fluorin ation of uracil deriva-
tives 12 an d 19 was perform ed with 5–10% m ixture of fluorin e an d n itrogen (Messer
Tech n ogas). 4-Meth oxy-2-pyrim idin -2(1H)-on es 1, 9 an d 22 were prepared from relevan t
uracils as described in refs5,11. Oxiran es 2a an d 2b were obtain ed from Daiso Co, Ltd.
(Japan ). (Diisopropoxyph osph oryl)m eth yl tosylate was syn th esized as described in ref.19
diisopropyl [(2-ch loroeth oxy)m eth yl]ph osph on ate (10) was obtain ed accordin g to ref.20
,
.
Diisopropyl ({[1-azido-3-(tosyloxy)propan -2-yl]oxy}m eth yl)ph osph on ate (33) was prepared
from racem ic glycidyl trityl eth er accordin g to ref.14. Dim eth ylform am ide, tetrah ydrofuran ,
an d aceton itrile were dried by distillation from calcium h ydride an d stored over m olecular
sieves (4Å).
En zym e Assay
Th e stan dard reaction m ixture (50 µl) con tain ed 20 µM bis(Tris-HCl) pH 6.4, 1 m M EDTA an d
2 m M DTT, 100 µM [3H-m eth yl]th ym idin e, 250 µM potassium ph osph ate pH 6.7, tested com -
poun d 10 µm ol l–1 an d 25.5 pU of en zym e accordin g to ref.4. Th e reaction was carried out at
37 °C for 10 m in an d stopped by spottin g a 2 µl aliquot on to Silica gel 60 F254 plate th at
h ad been prespotted with 0.01 µm ol of each th ym in e an d th ym idin e. Th e plate was developed
in th e n on -aqueous ph ase of th e solven t system eth yl acetate–water–form ic acid (60:35:5).
Th e spots were visualised un der UV ligh t (254 n m ) an d cutted out for radioactivity determ i-
n ation in th e toluen e-based scin tillation cocktail.
Syn th esis of 5-Eth yl-4-m eth oxypyrim idin -2(1H)-on es 3a an d 3b. Gen eral Procedure
A m ixture of 5-eth yl-4-m eth oxypyrim idin -2(1H)-on e (1; 500 m g, 3.2 m m ol), [(trityloxy)-
m eth yl]oxiran e 2a or 2b (1.03 g, 3.2 m m ol) an d cesium carbon ate (106 m g, 032 m m ol) in
dim eth ylform am ide (40 m l) was h eated at 90 °C for 10 h . Th e m ixture was con cen trated in
vacuo to a m in im um volum e. Th e residue was codistilled with toluen e (2 × 10 m l) an d ch ro-
m atograph ed on n eutral alum in um oxide (toluen e followed by eth yl acetate an d eth yl ace-
tate–eth an ol 10:1). Th e fraction s con tain in g com poun ds 3a or 3b were evaporated to
dryn ess in vacuo.
5-Ethyl-1-[(R)-2-hydroxy-3-(trityloxy)propyl]-4-methoxypyrimidin-2(1H)-one (3a). Yield 871 m g
(59%) of a wh ite foam . IR (CCl4), νm ax: 3357, 1663, 1638, 1537, 1472, 1401, 1368, 1335,
1099, 633. [α]D +43.8 (c 0.195 g/100 m l, CHCl3). HR MS (FAB): for C29H31N2O4 calculated
471.2284, foun d 471.2270. For C29H30N2O4 (470.6) calculated: 74.02% C, 6.43% H, 5.95% N;
foun d: 74.07% C, 6.49% H, 5.63% N. FAB MS, m/z: 471 [MH]+ (4), 243 (100), 211 (15), 197
(15), 165 (38), 155 (11), 105 (10), 77 (9). 1H NMR (500 MHz, DMSO-d6): 1.00 (t, 3 H, Jvic
=
7.4, CH3CH2); 2.23 (q, 2 H, Jvic = 7.4, CH2CH3); 2.90 (dd, 1 H, Jgem = 9.4, J3b′,2′ = 5.4, H-3′b);
2.97 (dd, 1 H, Jgem = 9.4, J3′a,2′ = 5.0, H-3′a); 3.58 (dd, 1 H, Jgem = 12.9, J1′b,2′ = 8.3, H-1′b);
3.83 (s, 3 H, OCH3); 3.99 (m , 1 H, H-2′); 4.05 (dd, 1 H, Jgem = 12.9, J1′a,2′ = 4.0, H-1′a); 5.27
(d, 1 H, JOH,2′ = 5.9, OH-2′); 7.26 (m , 3 H, H-p-Ph ); 7.33 (m , 6 H, H-m-Ph ); 7.42 (m , 6 H,
H-o-Ph ); 7.60 (s, 1 H, H-6). 13C NMR (125.8 MHz, DMSO-d6): 13.28 (CH3CH2); 19.20
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
607
(CH2CH3); 53.16 (CH2-1′); 53.95 (OCH3); 66.25 (CH2-3′); 67.08 (CH-2′); 86.03 (C-Tr); 109.60
(C-5); 127.16 (CH-p-Ph ); 128.02 (CH-m-Ph ); 128.42 (CH-o-Ph ); 143.69 (C-i-Ph ); 146.82
(CH-6); 155.56 (C-2); 169.79 (C-4).
5-Ethyl-1-[(S)-2-hydroxy-3-(trityloxy)propyl]-4-methoxypyrimidin-2(1H)-one (3b). Yield 944 m g
(64%) of a wh ite foam . 1H, 13C NMR, [α]D an d IR data were iden tical with th ose of com -
poun d reported in ref.5.
Syn th esis of 5-Eth yl-4-m eth oxypyrim idin -2(1H)-on es 4a an d 4b. Gen eral Procedure
A m ixture of com poun d 3a or 3b (787 m g, 1.7 m m ol), (diisopropoxyph osph oryl)m eth yl
tosylate (667 m g, 1.9 m m ol) an d 60% sodium h ydride dispersion (103 m g, 2.6 m m ol) in
tetrah ydrofuran (25 m l) was stirred at –20 °C. Th e suspen sion was allowed to warm to 20–40 °C
for 2 h an d stirred at room tem perature overn igh t. Th e m ixture was con cen trated in vacuo
an d th e residue was diluted with ch loroform (10 m l). After coolin g th e m ixture was evapo-
rated to dryn ess. Th e residue was filtered on n eutral alum in um oxide (eth yl acetate an d fol-
lowed by eth yl acetate–ch loroform –m eth an ol 21:20:1). Th e crude product was purified by
ch rom atograph y on preparative TLC plate (S1, eth yl acetate–ch loroform –m eth an ol 21:20:1).
Th e fraction s con tain in g product 4a or 4b were evaporated to dryn ess in vacuo.
1-{(R)-[2-(Diisopropoxyphosphoryl)methoxy]-3-(trityloxy)propyl}-5-ethyl-4-methoxypyrimidin-
2(1H)-one (4a). Yield 363 g (32%) of a wh ite foam . IR (CCl4), νm ax: 1674, 1653, 1536, 1471,
1403, 1386, 1331, 1256, 1007, 706. [α]D +35.1 (c 0.362 g/100 m l, CHCl3). HR MS (ESI): for
C
36H46N2O7P calculated 649.3043, foun d 649.3059. ESI MS, m/z: 649 [MH]+ (7). 1H NMR
(400 MHz, CDCl3): 1.10 (t, 3 H, Jvic = 7.4, CH3CH2); 1.25, 1.26, 1.28 an d 1.30 (4 × d, 4 × 3 H,
Jvic = 6.2, (CH3)2CH); 2.31 (q, 2 H, Jvic = 7.4, CH2CH3); 3.10 (dd, 1 H, Jgem = 10.7, J3b′,2′
6.5, H-3′b); 3.40 (dd, 1 H, Jgem = 10.7, J3′a,2′ = 3.4, H-3′a); 3.55 (dd, 1 H, Jgem = 13.4, JH,P
10.2, H-4′b); 3.80 (dd, 1 H, Jgem = 13.6, J1′b,2′ = 8.2, H-1′b); 3.85 (dd, 1 H, Jgem = 13.4, JH,P
=
=
=
8.7, H-4′a); 3.91 (dq, 1 H, J2′,1′ = 8.2, 3.7, J2′,3′ = 4.0, 3.4, H-2′); 3.96 (s, 3 H, OCH3); 4.27 (dd,
1 H, Jgem = 13.6, J1′a,2′ = 3.7, H-1′a); 4.67 an d 4.70 (2 × dh , 2 × 1 H, JH,P = 7.7, Jvic = 6.2,
CH(CH3)2); 7.21–7.33 (m , 10 H, H-6 + H-m,p-Ph ); 7.44 (m , 6 H, H-o-Ph ). 13C NMR (100.6 MHz,
CDCl3): 12.96 (CH3CH2); 19.75 (CH2CH3); 23.93, 24.00 an d 24.04 (d, JC,P = 4, (CH3)2CH);
51.55 (CH2-1′); 54.38 (OCH3); 62.37 (CH2-3′); 64.90 (d, JC,P = 169, CH2-4′); 70.79 an d 70.97
(d, JC,P = 6, CH(CH3)2); 78.96 (d, JC,P = 13, CH-2′); 86.82 (C-Tr); 109.71 (C-5); 127.16
(CH-p-Ph ); 127.91 (CH-m-Ph ); 128.60 (CH-o-Ph ); 143.54 (C-i-Ph ); 145.17 (CH-6); 158.56
(C-2); 170.59 (C-4).
1-{(S)-[2-(Diisopropoxyphosphoryl)methoxy]-3-(trityloxy)propyl}-5-ethyl-4-methoxypyrimidin-
2(1H)-one (4b). Yield 363 m g (32%) of a wh ite foam . 1H, 13C NMR, [α]D an d IR data were
iden tical with th ose of com poun d reported in ref.5. Th e procedure was repeated to obtain
8.6 g of 4a wh ich was furth er utilized for th e preparation of 8.
Syn th esis of 5-Eth yluracils 5a an d 5b. Gen eral Procedure
A m ixture of com poun d 4a or 4b (302 m g, 0.5 m m ol), aceton itrile (10 m l) an d brom o-
trim eth ylsilan e (0.7 m l) was stirred overn igh t at room tem perature. Th e m ixture was con -
cen trated in vacuo an d th en codistilled with water (2 × 2 m l). Th e residue in water (10 m l)
was h eated with Dowex 50X8 (H+ form ; 2 m l) at 100 °C for 1 h . Th e m ixture was cooled to
room tem perature an d th en filtered. Th e filtrate was con cen trated in vacuo an d purified on
40 m l of DEAE-Seph adex (S2) with subsequen t deion ization on activated ch arcoal. Th e rele-
van t fraction s were com bin ed, evaporated in vacuo an d codistilled with water (3 × 5 m l).
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

608
Pomeisl, Votruba, Holý, Pohl:
Th e residue was dissolved in water (3 m l), applied on to a colum n of Dowex 50X8 (Li+ form ;
30 m l) an d th en th e colum n was wash ed with water. Th e appropriate UV absorbin g fraction
con tain in g product 5a or 5b was evaporated to dryn ess in vacuo. Th e residue was dissolved
in water an d lyoph ilized. Th e followin g com poun ds were obtain ed as dilith ium salts:
5-Ethyl-1-[(R)-3-hydroxy-2-(phosphonomethoxy)propyl]uracil (5a). Yield 129 g (87%) of a
wh ite solid, b.p. > 300 °C. IR (KBr), νm ax: 3418, 3260, 1684, 1471, 1462, 1440, 1076, 994,
918. [α]D +31.7 (c 0.365 g/100 m l, CHCl3). HPLC, 99% (S3). HR MS (FAB): for C10H16Li2N2O7P
calculated 321.1015, foun d 321.1011. FAB MS, m/z: 321 [MH]+ (9). 1H NMR (400 MHz, D2O,
ref(dioxan e) 3.75 ppm ): 1.09 (t, 3 H, Jvic = 7.5, CH3CH2); 2.31 (qd, 2 H, Jvic = 7.5, JCH2,6
1.0, CH3CH2); 3.56 (dd, 1 H, Jgem = 12.2, J3b′,2′ = 4.7, H-3′b); 3.60 (dd, 1 H, Jgem = 12.7, JH,P
=
=
9.4, H-4′b); 3.64 (dd, 1 H, Jgem = 12.7, JH,P = 9.1, H-4′a); 3.73 (m , 1 H, H-2′); 3.79 (dd, 1 H,
Jgem = 12.2, J3′a,2′ = 3.4, H-3′a); 3.87 (dd, 1 H, Jgem = 14.4, J1′b,2′ = 6.8, H-1′b); 3.94 (dd, 1 H,
Jgem = 14.4, J1′a,2′ = 4.9, H-1′a); 7.49 (t, 1 H, J6,CH2 = 1.0, H-6). 13C NMR (100.6 MHz, D2O,
ref(dioxan e) 69.3 ppm ): 12.66 (CH3CH2); 19.97 (CH3CH2); 49.19 (CH2-1′); 61.09 (CH2-3′);
67.80 (d, JC,P = 154, CH2-4′); 80.50 (d, JC,P = 11, CH-2′); 116.91 (C-5); 144.01 (CH-6); 153.00
(C-2); 167.29 (C-4). 31P NMR (162 MHz, D2O): 15.05 (t, J = 9.4, 9.1).
5-Ethyl-1-[(S)-3-hydroxy-2-(phosphonomethoxy)propyl]uracil (5b). Yield 100 m g (67%) of a
wh ite solid, m .p. > 300 °C. HPLC, 99% (S3). [α]D –30.5 (c 0.414 g/100 m l, H2O). FAB MS,
m/z: 321 [MH]+ (15). 1H, 13C NMR, HR MS an d IR data were iden tical with th ose of com -
poun d 5a.
1-{(S)-2-[(Diisoph osph oryl)m eth oxy]-3-h ydroxypropyl}-
5-eth yl-4-m eth oxypyrim idin -2(1H)-on e (6)
Com poun d 4a (8.6 g, 13.3 m m ol) in m eth an ol (125 m l) an d glacial acetic acid (4 m l) was
h ydrogen ated over 10% palladium on ch arcoal (0.7 g) at room tem perature for 32 h un til
th e con version of startin g ph osph on ate to 6 was com plete (TLC in ch loroform –m eth an ol
25:1). Th e m ixture was th en n eutralized with solid sodium h ydrogen carbon ate an d filtered
th rough a Celite pad. Th e filtrate was con cen trated to a m in im um volum e. Th e residue was
ch rom atograph ed on n eutral alum in um oxide (ch loroform followed by ch loroform –m eth an ol
25:1). Yield 2.9 g (54%) of 6 as a colorless oil. IR (CCl4), νm ax: 3406, 1674, 1663, 1641, 1536,
1472, 1403, 1386, 1375, 1375, 1254, 1011, 993. [α]D –7.3 (c 0.109 g/100 m l, CHCl3). HR MS
(FAB): for C17H32N2O7P calculated 407.1947, foun d 407.1933. FAB MS, m/z: 407 [MH]+
(100). 1H NMR (400 MHz, CDCl3): 1.14 (t, 3 H, Jvic = 7.5, CH3CH2); 1.31, 1.33 an d 1.34
(3 × d, 12 H, Jvic = 6.2, (CH3)2CH); 2.37 (qd, 2 H, Jvic = 7.5, JCH2,6 = 1.0, CH2CH3); 3.52–3.65
(m , 2 H, H-3′); 3.76 (dd, 1 H, Jgem = 14.0, JH,P = 8.8, H-4′b); 3.80 (dd, 1 H, Jgem = 14.0, JH,P
=
7.9, H-4′a); 3.84 (m , 1 H, H-2′); 3.99 (dd, 1 H, Jgem = 14.0, J1′b,2′ = 3.9, H-1′b); 4.00 (s, 3 H,
OCH3); 4.13 (dd, 1 H, Jgem = 14.0, J1′a,2′ = 5.7, H-1′a); 4.29 (dd, 1 H, JOH,3′ = 8.9, 5.7, OH-3′);
4.71 an d 4.77 (2 × dh , 2 × 1 H, JH,P = 7.6, Jvic = 6.2, CH(CH3)2); 7.35 (t, 1 H, J6,CH2 = 1.0,
H-6). 13C NMR (100.6 MHz, CDCl3): 13.07 (CH3CH2); 19.71 (CH2CH3); 23.99 an d 24.05 (d,
JC,P = 4, (CH3)2CH); 50.05 (CH2-1′); 54.56 (OCH3); 59.63 (CH2-3′); 64.88 (d, JC,P = 170,
CH2-4′); 71.43 (d, JC,P = 7, CH(CH3)2); 80.57 (d, JC,P = 8, CH-2′); 110.46 (C-5); 145.18 (CH-6);
157.51 (C-2); 170.82 (C-4).
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Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
609
1-{(S)-2-[(Diisopropoxyph osph oryl)m eth oxy]-3-fluoropropyl}-
5-eth yl-4-m eth oxypyrim idin -2(1H)-on e (7)
A m ixture of 1,8-diazabicyclo[5.4.0]un dec-7-en e (1.3 g, 8.5 m m ol) an d 92% perfluorobutan e-
1-sulfon yl fluoride (3 g, 9.1 m m ol) was stirred in toluen e (15 m l) at room tem perature for
10 m in . A solution of com poun d 6 (2.6 g, 6.4 m m ol) in toluen e (30 m l) was added an d th e
resultin g m ixture was stirred for 8 h at room tem perature an d th en h eated at 80 °C for 1 h
un til th e con version of startin g ph osph on ate to 7 was com plete (TLC in ch loroform –m eth an ol
25:1). Th e m ixture was con cen trated in vacuo an d filtered over n eutral alum in um oxide
(ch loroform followed by ch loroform –m eth an ol 25:1). After evaporation of solven ts th e resi-
due was ch rom atograph ed on preparative TLC plate (S1, ch loroform –m eth an ol 15:1). Yield
792 m g (30%) of a colorless oil. IR (CCl4), νm ax: 1674, 1651, 1536, 1471, 1403, 1386, 1376,
1255, 1106, 1007, 992. [α]D –45.0 (c 0.549 g/100 m l, CHCl3). For C17H30FN2O6P (408.4) cal-
culated: 50.00% C, 7.40% H, 4.65% F, 6.86% N, 7.58% P; foun d: 49.84% C, 7.49% H,
4.90% F, 6.69% N, 7.64% P. FAB MS, m/z: 409 [MH]+ (100). 1H NMR (500 MHz, CDCl3): 1.15
(t, 3 H, Jvic = 7.5, CH3CH2); 1.29, 1.31 an d 1.32 (4 × d, 4 × 3 H, Jvic = 6.2, (CH3)2CH); 2.37
(m , 2 H, CH2CH3); 3.71 (dd, 1 H, Jgem = 13.6, JH,P = 9.4, H-4′b); 3.81 (dd, 1 H, Jgem = 13.8,
J1′b,2′ = 7.5, H-1′b); 3.89 (dd, 1 H, Jgem = 13.6, JH,P = 8.8, H-4′a); 3.99 (s, 3 H, OCH3); 4.05 (m ,
1 H, H-2′); 4.20 (dd, 1 H, Jgem = 13.8, J1′a,2′ = 3.8, H-1′a); 4.45 (ddd, 1 H, JH,F = 47.3, Jgem
=
10.7, J3′b,2′ = 4.6, H-3′b); 4.71 (m , 2 H, CH(CH3)2); 4.72 (ddd, 1 H, JH,F = 47.3, Jgem = 10.7,
J3′a,2′ = 2.5, H-3′a); 7.32 (t, 1 H, J6,CH2 = 1.0, H-6). 13C NMR (125.8 MHz, CDCl3): 13.02
(CH3CH2); 19.73 (CH2CH3); 23.90, 23.94 an d 24.03 (d, JC,P = 4, (CH3)2CH); 50.19 (d, JC,F = 9,
CH2-1′); 54.52 (OCH3); 65.45 (d, JC,P = 169, CH2-4′); 71.08 an d 71.21 (d, JC,P = 7, CH(CH3)2);
78.42 (dd, JC,F = 18, JC,P = 11, CH-2′); 82.51 (d, JC,F = 173, CH2-3′); 110.31 (C-5); 144.79
(CH-6); 156.60 (C-2); 170.83 (C-4). 19F NMR (188.2 MHz, CDCl3): –232.48 (td, JF,H-3′ = 47.3,
JF,H-2′ = 25.1).
5-Eth yl-1-[(S)-3-fluoro-2-(ph osph on om eth oxy)propyl]uracil (8)
Com poun d 8 was obtain ed as dilith ium salt by th e sam e procedure reported for 5a an d 5b.
A m ixture of com poun d 7 (302 m g, 0.74 m m ol) in aceton itrile (10 m l) an d brom o-
trim eth ylsilan e (1 m l) afforded 150 m g (59%) of 8 as a wh ite solid, m .p. 249–250 °C. IR
(KBr), νm ax: 3065, 1700, 1670, 1627, 1375, 1465, 1458, 1118, 1079, 996, 924, 583, 546. [α]D
–12.3 (c 0.141 g/100 m l, H2O). HPLC, 99% (S3). HR MS (FAB): for C10H15FLi2N2O6P calcu-
lated 323.0971, foun d 323.0958. For C10H14FLi2N2O6P·1.3H2O (345.5) calculated: 34.76% C,
4.78% H, 5.50% F, 8.12% N, 8.96% P; foun d: 35.03% C, 4.59% H, 5.31% F, 7.59% N, 8.61% P.
FAB MS, m/z: 323 [MH]+ (41). 1H NMR (500 MHz, D2O, ref(dioxan e) 3.75 ppm ): 1.09 (t, 3 H,
Jvic = 7.5, CH3CH2); 2.31 (qd, 2 H, Jvic = 7.5, JCH2,6 = 1.1, CH2CH3); 3.64 an d 3.72 (2 × dd,
2 H, Jgem = 13.0, JH,P = 9.2, H-4′); 3.91-4.11 (m , 3 H, H-1′ an d H-2′); 4.53 (ddd, 1 H, JH,F
=
47.0, Jgem = 10.6, J3′b,2′ = 3.6, H-3′b); 4.70 (ddd, 1 H, JH,F = 47.0, Jgem = 10.6, J3′a,2′ = 3.2,
H-3′a); 7.50 (t, 1 H, J6,CH2 = 1.1, H-6). 13C NMR (125.8 MHz, D2O, ref(dioxan e) 69.3 ppm ):
12.83 (CH3CH2); 19.97 (CH2CH3); 48.49 (d, JC,F = 7, CH2-1′); 67.64 (d, JC,P = 155, CH2-4′);
78.31 (dd, JC,F = 18, JC,P = 11, CH-2′); 82.96 (d, JC,F = 168, CH2-3′); 117.00 (C-5); 143.91
(CH-6); 152.97 (C-2); 167.32 (C-4). 19F NMR (188.2 MHz, D2O): –230.65 (td, JF,H-3′ = 47.0,
JF,H-2′ = 26.6).
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610
Pomeisl, Votruba, Holý, Pohl:
1-{2-[(Diisopropoxyph osph oryl)m eth oxy]eth yl}-4-m eth oxypyrim idin -2(1H)-on e (11)
A m ixture of com pound 9 (4 g, 32 m m ol) and 60% sodium hydride suspension (1.3 g, 33 m m ol)
in dim eth ylform am ide (200 m l) was stirred at room tem perature for 1 h . Th e com poun d 10
(10 g, 39 m m ol) was added an d th e resultin g m ixture was stirred at room tem perature for
2 h an d th en h eated at 90 °C for 12 h . Th e m ixture was evaporated an d codistilled with
toluen e (50 m l). Th e residue was diluted in dich lorom eth an e (300 m l) an d filtered th rough a
Celite pad. Th e residue was ch rom atograph ed on n eutral alum in um oxide (eth yl acetate–
m eth an ol 20:1 followed by ch loroform –m eth an ol 25:1) an d appropriate fraction s were evap-
orated in vacuo to dryn ess. Yield 5.4 g (49%) of 11 as a wh ite solid. IR (CCl4), νm ax: 1678,
1638, 1543, 1483, 1415, 1386, 1369, 1257, 1011, 990. For C14H25N2O6P (348.3) calculated:
48.27% C, 7.23% H, 8.04% N, 8.89% P; foun d: 48.05% C, 7.25% H, 7.87 N, 8.70% P. MS
(FAB), m/z: 349 (100) [MH]+. 1H NMR (500 MHz, CDCl3): 1.29 an d 1.32 (2 × d, 2 × 6 H, Jvic
=
6.2, (CH3)2CH); 3.70 (d, 2 H, JH,P = 8.6, H-4′); 3.85 (dd, 2 H, J2′,1′ = 5.0 an d 3.8, H-2′); 3.95
(s, 3 H, OCH3); 4.05 (dd, 2 H, J1′,2′ = 5.0 an d 3.8, H-1′); 4.71 (dh , 2 H, JH,P = 7.7, Jvic = 6.2,
CH(CH3)2); 5.84 (d, 1 H, J5,6 = 7.2, H-5); 7.52 (d, 1 H, J6,5 = 7.2, H-6). 13C NMR (125.8 MHz,
CDCl3): 23.99 an d 24.04 (d, JC,P = 4, (CH3)2CH); 49.99 (CH2-1′); 54.35 (OCH3); 66.05 (d,
JC,P = 169, CH2-4′); 70.81 (d, JC,P = 12, CH2-2′); 71.07 (d, JC,P = 7, CH(CH3)2); 94.97 (CH-5);
148.36 (CH-6); 156.50 (C-2); 171.90 (C-4).
1-{2-[(Diisopropoxyph osph oryl)m eth oxy]eth yl}uracil (12)
Com poun d 11 (5 g, 14.4 m m ol) was dissolved in 90% aqueous m eth an ol (100 m l). Dowex 50
(H+ form ; 20 m l) was added an d th e suspen sion was refluxed for 3.5 h un til th e con version
of startin g ph osph on ate to 12 was com plete (TLC in ch loroform –m eth an ol 25:1). Th e m ix-
ture was con cen trated in vacuo, ch rom atograph ed on n eutral alum in um oxide (ch loro-
form –m eth an ol 15:1) an d appropriate fraction s were evaporated in vacuo. Th e product was
triturated with h exan e. Th e solven t was decan ted an d th e obtain ed product was dried in
vacuo. Yield 4.4 g (91%) of 12 as a wh ite solid. IR (CCl4), νm ax: 3410, 3169, 3056, 1725,
1709, 1692, 1634, 1386, 1376, 1348, 1255, 1243, 1179, 1142, 1106, 1010, 992. For
C
13H23N2O6P (334.1) calculated: 46.71% C, 6.93% H, 8.38% N, 9.27% P; foun d: 46.42% C,
7.00% H, 8.13% N, 9.34% P. FAB MS, m/z: 335 [MH]+ (57). 1H NMR (500 MHz, CDCl3): 1.31
an d 1.33 (2 × d, 2 × 6 H, Jvic = 6.2, (CH3)2CH); 3.73 (d, 2 H, JH,P = 8.4, H-4′); 3.82 (dd, 2 H,
J2′,1′ = 5.2 an d 3.9, H-2′); 3.95 (dd, 2 H, J1′,2′ = 5.2 an d 3.9, H-1′); 4.73 (dh , 2 H, JH,P = 7.7,
Jvic = 6.2, CH(CH3)2); 5.66 (d, 1 H, J5,6 = 7.9, H-5); 7.35 (d, 1 H, J6,5 = 7.9, H-6); 9.04 (bs,
1 H, NH). 13C NMR (125.8 MHz, CDCl3): 24.00 an d 24.03 (d, JC,P = 4, (CH3)2CH); 48.30
(CH2-1′); 66.07 (d, JC,P = 169, CH2-4′); 70.92 (d, JC,P = 11, CH2-2′); 71.16 (d, JCP = 7,
CH(CH3)2); 101.50 (CH-5); 145.84 (CH-6); 150.77 (C-2); 163.61 (C-4).
1-{2-[(Diisopropoxyph osph oryl)m eth oxy]eth yl}-5-fluorouracil (14)
Com poun d 12 (1.5 g, 4.5 m m ol) was dissolved in glacial acetic acid (200 m l). A m ixture of
5–10% fluorin e in n itrogen was in troduced to th e solution un til th e con version of startin g
ph osph on ate was com plete (UV absorption disappears on TLC). Excess fluorin e was rem oved
by a stream of n itrogen an d th e m ixture was con cen trated in vacuo an d codistilled with
eth an ol (2 × 50 m l). Th e residue was dissolved in eth an ol (100 m l) an d trieth ylam in e (2 m l).
Th e m ixture was vigorously refluxed for 40 h un til th e con version of in term ediate dih ydro
adducts to 14 was com plete (m on itorin g by 19F NMR). Th e m ixture was con cen trated in
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
611
vacuo, th e residue ch rom atograph ed on silica gel (ch loroform –m eth an ol 25:1 an d 10:1) an d
appropriate fraction s were evaporated in vacuo. Th e product was triturated with h exan e.
Th e solven t was decan ted an d th e product dried un der reduced pressure. For an alytical pur-
poses th e product was crystallized in h exan e–aceton e. Yield 1.1 g (68%) of 14 as a wh ite
solid, m .p. 110–112 °C of crystallin e product. IR (CCl4), νm ax: 3403, 3146, 3078, 1729, 1713,
1670, 1386, 1378, 1356, 1255, 1239, 1178, 1142, 1105, 1010, 992. For C13H22FN2O6P (352.3)
calculated: 44.32% C, 6.29% H, 5.39% F, 7.95% N, 8.79% P; foun d: 44.33% C, 6.33% H,
5.17% F, 7.81% N, 8.69% P. FAB MS, m/z: 353 [MH]+ (63). 1H NMR (400 MHz, CDCl3): 1.31
an d 1.33 (2 ( d, 2 ( 6 H, Jvic = 6.5, (CH3)2CH); 3.74 (d, 2 H, JH,P = 8.3, H-4′); 3.82 (dd, 2 H,
J2′,1′ = 5.1 an d 3.6, H-2′); 3.93 (dd, 2 H, J1′,2′ = 5.1 an d 3.6, H-1′); 4.74 (dh , 2 H, JH,P = 7.7,
Jvic = 6.5, CH(CH3)2); 7.52 (d, 1 H, JH,F = 5.8, H-6); 9.35 (bd, 1 H, JH,F = 3.4, NH). 13C NMR
(100.6 MHz, CDCl3): 23.96 an d 23.99 (d, JC,P = 4, (CH3)2CH); 48.40 (CH2-1′); 66.13 (d, JC,P
=
169, CH2-4′); 70.95 (d, JC,P = 11, CH2-2′); 71.32 (d, JC,P = 7, CH(CH3)2); 130.26 (d, JC,F = 33,
CH-6); 139.93 (d, JC,F = 236, C-5); 149.41 (C-2); 157.18 (d, JC,F = 27, C-4). 19F NMR (470.2 MHz,
CDCl3): –168.38 (t, JF,H = 5.8 an d 3.4).
5-Fluoro-1-[2-(ph osph on om eth oxy)eth yl]uracil (15)
Com poun d 15 was obtain ed as dilith ium salt by th e procedure reported for 5a an d 5b. A
m ixture of com poun d 14 (540 m g, 1.53 m m ol) in aceton itrile (15 m l) an d brom otrim eth yl-
silan e (2 m l) afforded 177 m g (38%) of 15 as a wh ite solid, m .p. > 300 °C. IR (KBr), νm ax
:
3242, 3172, 3080, 1691, 1475, 1444, 1383, 1101, 994, 925, 570, 467. HPLC, 99% (S3). For
C7H9FLi2N2O6P·1.2H2O (301.6) calculated: 27.88% C, 3.45% H, 6.30% F, 9.29% N, 10.27% P;
foun d: 27.62% C, 3.05% H, 6.36% F, 9.33% N, 10.55% P. FAB MS, m/z: 281 [MH]+ (9). 1H NMR
(500 MHz, D2O, ref(dioxan e) 3.75 ppm ): 3.58 (d, 2 H, JH,P = 8.6, H-4′); 3.81 (dd, 2 H, J2′,1′
=
5.3 an d 4.3, H-2′); 3.97 (dd, 2 H, J1′,2′ = 5.3 an d 4.3, H-1′); 7.92 (d, 1 H, JH,F = 6.1, H-6).
13C NMR (125.8 MHz, D2O, ref(dioxan e) 69.3 ppm ): 51.05 (CH2-1′); 70.45 (d, JC,P = 155,
CH2-4′); 72.60 (d, JC,P = 11, CH2-2′); 134.58 (d, JC,F = 33, CH-6); 143.04 (d, JC,F = 231, C-5);
154.59 (C-2); 162.92 (d, JC,F = 29, C-4). 19F NMR (470.2 MHz, D2O): –167.13 (d, JF,H = 6.1).
31P NMR (162 MHz, D2O): 15.18 (t, JP,H = 8.6).
4-Meth oxy-1-(3,3,3-trifluoro-2-h ydroxypropyl)pyrim idin -2(1H)-on e (17)
A m ixture of 4-m eth oxypyrim idin -2(1H)-on e (9; 2 g, 15.9 m m ol), trifluorom eth yloxiran e 16
(1.8 g, 16.0 m m ol) an d cesium carbon ate (543 m g, 1.7 m m ol) in dim eth ylform am ide (140 m l)
was h eated at 80 °C for 8 h . Th e m ixture was con cen trated in vacuo to a m in im um volum e.
Th e residue was codistilled with toluen e (2 × 20 m l) an d ch rom atograph ed on n eutral alu-
m in um oxide (toluen e followed by eth yl acetate an d eth yl acetate–m eth an ol 75:25). Th e
fraction s con tain in g com poun d 17 were evaporated in vacuo. Th e residue was triturated
with h exan e. Th e solven t was decan ted an d th e obtain ed product dried un der reduced pres-
sure. Yield 2.6 g (69%) of 17 as a wh ite solid. IR (CHCl3), νm ax: 3607, 3289, 1668, 1654,
1638, 1543, 1487, 1487, 1416, 1321, 1273, 1116. For C8H9F3N2O3 (238.2) calculated: 40.34%
C, 3.81% H, 23.93% F, 11.76% N; foun d: 40.19% C, 3.72% H, 23.63% F, 11.78% N. FAB MS,
m/z: 239 [MH]+ (100). 1H NMR (400 MHz, DMSO-d6): 3.72 (dd, 1 H, Jgem = 13.4, J1′b,2′ = 9.4,
H-1′b); 3.82 (s, 3 H, OCH3); 4.19 (dd, 1 H, Jgem = 13.4, J1′a,2′ = 3.2, H-1′a); 4.31 (m , 1 H,
H-2′); 6.02 (d, 1 H, J5,6 = 7.2, H-5); 6.66 (d, 1 H, JOH,2′ = 6.5, OH-2′); 7.92 (d, 1 H, J6,5 = 7.2,
H-6). 13C NMR (100.6 MHz, DMSO-d6): 45.59 (CH2-1′); 53.97 (OCH3); 65.99 (q, JC,F = 30,
CH-2′); 94.21 (CH-5); 125.08 (q, JC,F = 283, CF3); 150.64 (CH-6); 155.52 (C-2); 171.70 (C-4).
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612
Pomeisl, Votruba, Holý, Pohl:
5-Eth yl-1-{2-[(diisopropoxyph osph oryl)m eth oxy]-3,3,3-trifluoropropyl}-
4-m eth oxypyrim idin -2(1H)-on e (18)
A m ixture of com poun d 17 (2.6 g, 10.9 m m ol), (diisopropoxyph osph oryl)m eth yl tosylate
(4.6 g, 13.1 m m ol) an d 60% sodium h ydride dispersion (655 m g, 13.4 m m ol) in tetra-
h ydrofuran (60 m l) was stirred at –20 °C. Th e suspen sion was allowed to warm to room tem -
perature for 1 h an d stirred at th e sam e tem perature overn igh t. Th e m ixture was filtered
th rough a Celite pad an d con cen trated in vacuo. Th e residue was ch rom atograph ed on n eu-
tral alum in um oxide (eth yl acetate followed by eth yl acetate–ch loroform –m eth an ol 17:10:1)
an d appropriate fraction s con tain in g com poun d 18 were evaporated in vacuo. Yield 2.5 g
(55%) of crude product 18 as a sligh tly yellow oil. It was used for preparation of 19 with out
furth er purification . HR MS (FAB): for C15H25F3N2O6P calculated 417.3380, foun d 417.3383.
FAB MS, m/z: 417 [MH]+ (26). 1H NMR (400 MHz, CDCl3): 1.29, 1.30, 1.31 an d 1.32 (4 × d,
4 × 3 H, Jvic = 6.2, (CH3)2CH); 3.60 (dd, 1 H, Jgem = 13.9, J1′b,2′ = 8.9, H-1′b); 3.71 (dd, 1 H,
Jgem = 13.3, JH,P = 10.8, H-4′b); 4.03 (dd, 1 H, Jgem = 13.3, JH,P = 9.6, H-4′a); 4.27 (m , 1 H,
J2′,1′ = 8.9, 3.4, JH,F = 6.0, H-2′); 4.49 (dd, 1 H, Jgem = 13.9, J1′a,2′ = 3.4, H-1′a); 4.63–4.78 (m ,
2 H, CH(CH3)2); 5.90 (d, 1 H, J5,6 = 7.2, H-5); 7.49 (d, 1 H, J6,5 = 7.2, H-6).
1-{2-[(Diisopropoxyph osph oryl)m eth oxy]-3,3,3-trifluoropropyl}uracil (19)
Com poun d 18 (2.4 g, 5.8 m m ol) was dissolved in 90% aqueous m eth an ol (40 m l). Dowex 50
(H+ form ; 10 m l) was added to th e solution an d th e suspen sion was h eated at 100 °C for 4 h
un til th e con version of 18 to 19 was com plete (TLC in eth yl acetate–ch loroform –m eth an ol
17:10:1). Th e m ixture was con cen trated in vacuo. Th e residue was ch rom atograph ed on n eu-
tral alum in um oxide (eth yl acetate–ch loroform –m eth an ol 17:10:1) an d appropriate fraction s
con tain in g com poun d 19 were evaporated in vacuo. Yield 780 m g (34%) of 19 as a colorless
oil. IR (CCl4), νm ax: 3407, 3161, 1719, 1691, 1634, 1433, 1276, 1356, 1255, 1177, 1147,
1017, 996. HR MS (FAB): for C14H23F3N2O6P calculated 403.1246, foun d 403.1260. For
C
14H22F3N2O6P (402.3) calculated: 41.80% C, 5.51% H, 14.17% F, 6.96% N, 7.70% P; foun d:
41.90% C, 5.42% H, 13.83% F, 6.77% N, 7.56% P. FAB MS, m/z: 403 [MH]+ (36). 1H NMR
(400 MHz, CDCl3): 1.31, 1.32, 1.33 an d 1.34 (4 × d, 4 × 3 H, Jvic = 6.2, (CH3)2CH); 3.60 (dd,
1 H, Jgem = 14.4, J1′b,2′ = 8.5, H-1′b); 3.82 (dd, 1 H, Jgem = 13.7, JH,P = 9.0, H-4′b); 4.07 (dd, 1 H,
Jgem = 13.7, JH,P = 9.4, H-4′a); 4.23 (m , 1 H, H-2′); 4.35 (dd, 1 H, Jgem = 14.4, J1′a,2′ = 3.3,
H-1′a); 4.72 an d 4.74 (2 × dh , 2 × 1 H, JH,P = 7.6, Jvic = 6.2, CH(CH3)2); 5.71 (d, 1 H, J5,6
=
8.0, H-5); 7.29 (d, 1 H, J6,5 = 8.0, H-6); 8.92(bs, 1 H, NH). 13C NMR (100.6 MHz, CDCl3):
23.91 an d 24.02 (d, JC,P = 4, (CH3)2CH); 47.30 (CH2-1′); 67.18 (d, JC,P = 169, CH2-4′); 71.55
an d 71.71 (d, JC,P = 7, CH(CH3)2); 77.00 (qd, JC,F = 30, JC,P = 12, CH-2′); 102.40 (CH-5);
123.77 (q, JC,F = 284, CF3); 145.44 (CH-6); 150.59 (C-2); 163.22 (C-4).
1-{2-[(Diisopropoxyph osph oryl)m eth oxy]-3,3,3-trifluoropropyl}-5-fluorouracil (20)
Com poun d 19 (778 m g, 1.9 m m ol) was dissolved in glacial acetic acid (90 m l). A m ixture of
5–10% fluorin e in n itrogen was in troduced to th e solution un til th e con version of startin g
ph osph on ate was com plete (UV absorption disappears on TLC plate). Excess fluorin e was re-
m oved by a stream of n itrogen , th e m ixture was con cen trated in vacuo an d codistilled with
eth an ol (2 × 20 m l). Th e residue was dissolved in eth an ol (45 m l) an d trieth ylam in e (3 m l).
Th e m ixture was vigorously refluxed for 11 h un til th e con version of in term ediate dih ydro
adducts to 20 was com plete (m on itorin g by 19F NMR). Th e m ixture was con cen trated in
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Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
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vacuo an d th e residue was subjected to preparative TLC (S1, ch loroform –m eth an ol 20:1).
Yield 365 m g (45%) of 20 as a colorless oil. IR (CCl4), νm ax: 3405, 3168, 3065, 2983, 1723,
1703, 1670, 1387, 1377, 1248, 1274, 1181, 1158, 1105, 1013, 996. For C14H21F4N2O6P
(420.3) calculated: 40.01% C, 5.04% H, 6.67% N, 7.37% P; foun d: 40.05% C, 5.17% H,
6.34% N, 7.37% P. FAB MS, m/z: 421 [MH]+ (13). 1H NMR (400 MHz, CDCl3): 1.31, 1.32,
1.33 an d 1.34 (4 × d, 4 × 3 H, Jvic = 6.2, (CH3)2CH); 3.51 (dd, 1 H, Jgem = 14.4, J1′b,2′ = 8.6,
H-1′b); 3.85 (dd, 1 H, Jgem = 14.1, JH,P = 5.7, H-4′b); 4.10 (dd, 1 H, Jgem = 14.1, JH,P = 8.8,
H-4′a); 4.29 (m , 1 H, H-2′); 4.37 (dd, 1 H, Jgem = 14.4, J1′a,2′ = 3.0, H-1′a); 4.73 (dh , 2 H,
JH,P = 7.6, Jvic = 6.2, CH(CH3)2); 7.39 (d, 1 H, JH,F = 5.5, H-6). 13C NMR (100.6 MHz, CDCl3):
23.84, 23.90 an d 23.99 (d, JC,P = 4, (CH3)2CH); 47.45 (CH2-1′); 67.10 (d, JC,P = 168, CH2-4′);
71.76 an d 71.86 (d, JC,P = 7, CH(CH3)2); 76.75 (qd, JC,F = 30, JC,P = 14, CH-2′); 123.75 (q,
JC,F = 284, CF3); 129.86 (d, JC,F = 33, CH-6); 140.28 (d, JC,F = 237, C-5); 149.35 (C-2); 157.13
(d, JC,F = 27, C-4). 19F NMR (188.2 MHz, CDCl3): –166.04 (d, JF,H-6 = 5.5, F-5); –75.92 (dd,
JF,H-2′ = 6.0, JF,H-1′a = 2.0, CF3).
5-Fluoro-1-[3,3,3-trifluoro-2-(ph osph on om eth oxy)propyl]uracil (21)
Com poun d 21 was obtain ed as dilith ium salt usin g th e sam e procedure as reported for 5a
an d 5b. A m ixture of com poun d 20 (248 m g, 0.59 m m ol) in aceton itrile (5 m l) an d brom o-
trim eth ylsilan e (0.8 m l) afforded 124 m g (59%) of 21 as a wh ite solid, m .p. > 300 °C. IR
(KBr), νm ax: 3250, 3080, 1701, 1669, 1482, 1442, 1378, 1252, 1179, 1154, 1136, 1079, 991,
921, 789, 759, 726, 685, 537, 469. HPLC, 99% (S3). HR MS (FAB): for C8H8F4Li2N2O6P calcu-
lated 349.0376, foun d 349.0386. For C8H7F4Li2N2O6P·0.5H2O (357.0) calculated: 26.91% C,
2.26% H, 22.28% F, 7.84% N, 8.68% P; foun d: 26.79% C, 2.42% H, 22.46% F, 7.43% N,
8.62% P. FAB MS, m/z: 349 [MH]+ (36). 1H NMR (500 MHz, D2O, ref(dioxan e) 3.75 ppm ):
3.69 an d 3.90 (2 × dd, 2 H, Jgem = 12.6, JH,P = 9.3, H-4′); 3.96 (dd, 1 H, Jgem = 14.8, J1′b,2′
=
7.4, H-1′b); 4.25 (dd, 1 H, Jgem = 14.8, J1′a,2′ = 3.7, H-1′a); 4.32 (m , 1 H, H-2′); 7.97 (d, 1 H,
JH,F = 6.0, H-6). 13C NMR (125.8 MHz, D2O, ref(dioxan e) 69.3 ppm ): 49.66 (CH2-1′); 72.61
(d, JC,P = 153, CH2-4′); 79.28 (qd, JC,F = 30, JC,P = 12, CH-2′); 126.84 (q, JC,F = 284, CF3);
142.18 (d, JC,F = 34, CH-6); 143.17 (d, JC,F = 232, C-5); 152.30 (d, JC,F = 30, C-4); 154.04
(C-2). 19F NMR (188.2 MHz, D2O): –165.22 (d, JF,H-6 = 6.0, F-5); –73.63 (dd, JF,H-2′ = 6.5, CF3).
31P NMR (162 MHz, D2O): 13.36 (t, JP,H-4′ = 9.3).
Syn th esis of 1-(3,3,3-Trifluoro-2-h ydroxypropyl)pyrim idin -2(1H)-on e
Derivatives 23 an d 24. Gen eral Procedure
A m ixture of pyrim idin -2(1H)-on e 1 or 22 (6.5 m m ol), trifluorom eth yloxiran e 16 (6.5 m m ol)
an d cesium carbon ate (0.7 m m ol) in dim eth ylform am ide (70 m l) was h eated at 80 °C for
15 h . Th e m ixture was con cen trated in vacuo to a m in im um volum e. Th e residue was
codistilled with toluen e (2 × 10 m l) an d ch rom atograph ed on n eutral alum in um oxide
(toluen e followed by eth yl acetate an d eth yl acetate–m eth an ol 60:40). Th e fraction s con tain -
in g com poun d 23 or 24 were evaporated to dryn ess in vacuo. Th e product was triturated
with h exan e an d th e solven t was decan tated. Th e product was dried un der reduced pressure.
5-Ethyl-4-methoxy-1-(3,3,3-trifluoro-2-hydroxypropyl)pyrimidin-2(1H)-one (23). Yield 951 m g
(55%) of a wh ite solid. IR (CCl4), νm ax: 3606, 3280, 1663, 1630, 1537, 1475, 1402, 1271. For
C
10H13F3N2O3 (266.2) calculated: 45.12% C, 4.92% H, 21.41% F, 10.52% N; foun d: 45.10% C,
4.87% H, 21.76% F, 10.47% N. FAB MS, m/z: 267 [MH]+ (100). 1H NMR (400 MHz,
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

614
Pomeisl, Votruba, Holý, Pohl:
DMSO-d6): 1.06 (t, 3 H, Jvic = 7.4, CH3CH2); 2.30 (qd, 2 H, Jvic = 7.4, JCH2,6 = 1.0, CH2CH3);
3.71 (dd, 1 H, Jgem = 13.4, J1′b,2′ = 9.4, H-1′b); 3.86 (s, 3 H, OCH3); 4.17 (dd, 1 H, Jgem = 13.4,
J1′a,2′ = 3.1, H-1′a); 4.32 (m , 1 H, H-2′); 6.62 (d, 1 H, JOH,2′ = 6.7, OH-2′); 7.74 (t, 1 H, J6,CH2
=
1.0, H-6). 13C NMR (100.6 MHz, DMSO-d6): 13.35 (CH3CH2); 19.20 (CH2CH3); 49.47
(CH2-1′); 54.20 (OCH3); 66.07 (q, JC,F = 30, CH-2′); 108.30 (C-5); 125.12 (q, JC,F = 283, CF3);
146.92 (CH-6); 155.39 (C-2); 170.25 (C-4).
4-Methoxy-5-methyl-1-(3,3,3-trifluoro-2-hydroxypropyl)pyrimidin-2(1H)-one (24). Yield 934 m g
(57%) of a wh ite solid. IR (CCl4), νm ax: 3623, 3273, 1671, 1650, 1542, 1478, 1399, 1339,
1270. For C9H11F3N2O3 (252.2) calculated: 42.86% C, 4.40% H, 22.60% F, 11.11% N; foun d:
42.47% C, 4.33% H, 22.10% F, 10.66% N. FAB MS, m/z: 253 [MH]+ (100). 1H NMR (400 MHz,
DMSO-d6): 1.87 (d, 3 H, JCH3,6 = 1.1, CH3-5); 3.66 (dd, 1 H, Jgem = 13.4, J1′b,2′ = 9.4, H-1′b);
3.85 (s, 3 H, OCH3); 4.16 (dd, 1 H, Jgem = 13.4, J1′a,2′ = 3.2, H-1′a); 4.31 (bm , 1 H, H-2′); 6.64
(bs, 1 H, OH-2′); 7.78 (q, 1 H, J6,CH3 = 1.1, H-6). 13C NMR (100.6 MHz, DMSO-d6): 11.65
(CH3-5); 49.45 (CH2-1′); 54.22 (OCH3); 65.74 (q, JC,F = 29, CH-2′); 102.40 (C-5); 125.09 (q,
JC,F = 283, CF3); 147.56 (CH-6); 155.49 (C-2); 170.54 (C-4).
Syn th esis of 1-{2-[(Diisopropoxyph osph oryl)m eth oxy]-3,3,3-trifluoropropyl}-
pyrim idin -2(1H)-on e Derivatives 25 an d 26. Gen eral Procedure
A m ixture of com poun d 23 or 24 (3.7 m m ol), (diisopropoxyph osph oryl)m eth yl tosylate
(4.1 m m ol) an d 60% sodium h ydride dispersion (5.5 m m ol) in tetrah ydrofuran (20 m l) was
stirred at –20 °C. Th e suspen sion was allowed to warm to room tem perature for 1 h an d
stirred at th e sam e tem perature overn igh t. Th e m ixture was filtered th rough a Celite pad un der
pressure an d con cen trated in vacuo to a m in im um volum e. Th e residue was ch rom atograph ed
on n eutral alum in um oxide (eth yl acetate followed by eth yl acetate–ch loroform –m eth an ol
17:10:1). Th e crude product was purified by preparative TLC (S1, eth yl acetate–ch loroform –
m eth an ol 17:10:1). Th e fraction s con tain in g product 25 or 26 were evaporated to dryn ess in
vacuo.
1-{2-[(Diisopropoxyphosphoryl)methoxy]-3,3,3-trifluoropropyl}-5-ethyl-4-methoxy-pyrimidin-
2(1H)-one (25). Yield 563 m g (35%) of a wh ite solid, m .p. > 300 °C. IR (CCl4), νm ax: 2981,
2959, 2937, 2900, 1678, 1653, 1537, 1537, 1471, 1403, 1366, 1268, 1255, 1179, 1149, 1130,
1111, 1006, 992. For C17H28F3N2O6P (444.4) calculated: 45.95% C, 6.35% H, 12.83% F,
6.30% N, 6.97% P; foun d: 46.08% C, 6.43% H, 12.73% F, 6.24% N, 7.06% P. FAB MS, m/z:
445 [MH]+ (100). 1H NMR (500 MHz, CDCl3): 1.16 (t, 3 H, Jvic = 7.5, CH3CH2); 1.27, 1.29,
1.31 an d 1.32 (4 × d, 4 × 3 H, Jvic = 6.2, (CH3)2CH); 2.36 an d 2.40 (2 × m , 2 H, Jgem = 15.1,
Jvic = 7.5, JCH2,6 = 1.1, CH2CH3); 3.64 (dd, 1 H, Jgem = 13.9, J1′b,2′ = 8.6, H-1′b); 3.72 (dd, 1 H,
Jgem = 13.3, JH,P = 10.3, H-4′b); 4.00 (s, 3 H, OCH3); 4.01 (dd, 1 H, Jgem = 13.3, JH,P = 9.6,
H-4′a); 4.29 (m , 1 H, H-2′); 4.43 (dd, 1 H, Jgem = 13.9, J1′a,2′ = 3.7, H-1′a); 4.67 an d 4.73 (2 ×
dh , 2 H, JH,P = 7.6, Jvic = 6.2, CH(CH3)2); 7.23 (t, 1 H, J6,CH2 = 1.1, H-6). 13C NMR (125.8 MHz,
CDCl3): 12.93 (CH3CH2); 19.75 (CH2CH3); 23.77, 23.85 an d 23.99 (d, JC,P = 4, (CH3)2CH);
49.18 (CH2-1′); 54.62 (OCH3); 67.39 (d, JC,P = 170, CH2-4′); 71.24 an d 71.54 (d, JC,P = 7,
CH(CH3)2); 76.54 (qd, JC,F = 30, JC,P = 14, CH-2′); 110.78 (C-5); 123.93 (q, JC,F = 284, CF3);
144.51 (CH-6); 156.25 (C-2); 171.08 (C-4).
1-{2-[(Diisopropoxyphosphoryl)methoxy]-3,3,3-trifluoropropyl}-4-methoxy-5-methyl-pyrimidin-
2(1H)-one (26). Yield 382 m g (24%) of a wh ite solid, m .p. > 300 °C. IR (CCl4), νm ax: 2982,
1679, 1654, 1541, 1476, 1398, 1386, 1376, 1366, 1337, 1267, 1255, 1013, 992. For C16H26F3N2O6P
(430.4) calculated: 44.65% C, 6.09% H, 13.24% F, 6.51% N, 7.20% P; foun d: 44.40% C,
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
615
5.86% H, 13.06% F, 6.48% N, 7.33% P. FAB MS, m/z: 431 [MH]+ (82). 1H NMR (400 MHz,
CDCl3): 1.29, 1.30 an d 1.31 (3 × d, 12 H, Jvic = 6.2, (CH3)2CH); 1.96 (d, 3 H, JCH3,6 = 1.1,
CH3-5); 3.60 (dd, 1 H, Jgem = 13.9, J1′b,2′ = 8.8, H-1′b); 3.72 (dd, 1 H, Jgem = 13.3, JH,P = 10.4,
H-4′b); 4.00 (s, 3 H, OCH3); 4.02 (dd, 1 H, Jgem = 13.3, JH,P = 9.5, H-4′a); 4.29 (m , 1 H, J2′,1′
=
8.8, 3.5, JH,F = 6.2, H-2′); 4.44 (dd, 1 H, Jgem = 13.9, J1′a,2′ = 3.5, H-1′a); 4.68 an d 4.73 (2 ×
dh , 2 H, JH,P = 7.6, Jvic = 6.2, CH(CH3)2); 7.27 (q, 1 H, J6,CH3 = 1.1, H-6). 13C NMR (100.6 MHz,
CDCl3): 12.04 (CH3-5); 23.78, 23.85 an d 23.99 (d, JC,P = 5, (CH3)2CH); 49.04 (CH2-1′); 54.69
(OCH3); 67.36 (d, JC,P = 169, CH2-4′); 71.27 an d 71.54 (d, JC,P = 7, CH(CH3)2); 76.68 (qd,
JC,F = 30, JC,P = 14, CH-2′); 104.80 (C-5); 123.94 (q, JC,F = 285, CF3); 145.36 (CH-6); 156.35
(C-2); 171.32 (C-4).
Syn th esis of 5-Eth yl-1-[3,3,3-trifluoro-2-(ph osph on om eth oxy)propyl]uracil an d
Th ym in e Derivatives 27 an d 28. Gen eral Procedure
Com poun ds 27 an d 28 were obtain ed as dilith ium salt by th e sam e procedure reported for
5a an d 5b.
5-Ethyl-1-[3,3,3-trifluoro-2-(phosphonomethoxy)propyl]uracil (27). A m ixture of com poun d 25
(432 m g, 0.97 m m ol) in aceton itrile (10 m l) an d brom otrim eth ylsilan e (1.3 m l) afforded
after work-up 173 m g (48%) of 27 as a wh ite solid, m .p. > 300 °C. IR (KBr), νm ax: 3254,
1685, 1474, 1459, 1440, 1375, 1354, 1266, 1167, 1137, 1223, 1087, 989, 916, 568, 542, 444.
HPLC, 99% (S5). For C10H12F3Li2N2O6P·0.5H2O (367.1) calculated: 32.72% C, 3.54% H,
15.52% F, 7.63% N, 8.43% P; foun d: 32.58% C, 3.64% H, 15.51% F, 7.34% N, 8.27% P. FAB
MS, m/z: 359 [MH]+ (4). 1H NMR (400 MHz, D2O, ref(dioxan e) 3.75 ppm ): 1.09 (t, 3 H, Jvic
=
7.5, CH3CH2); 2.32 (qd, 2 H, Jvic = 7.5, JCH2,6 = 1.1, CH2CH3); 3.69 (dd, 1 H, Jgem = 12.5,
JH,P = 9.2, H-4′b); 3.84 (dd, 1 H, Jgem = 12.5, JH,P = 9.5, H-4′a); 4.05 (dd, 1 H, Jgem = 14.8,
J1′b,2′ = 6.8, H-1′b); 4.21 (dd, 1 H, Jgem = 14.8, J1′a,2′ = 4.5, H-1′a); 4.32 (m , 1 H, J2′,1′ = 6.8,
4.5, JH,F = 6.5, H-2′); 7.56 (t, 1 H, J6,CH2 = 1.1, H-6). 13C NMR (125.8 MHz, D2O, ref(dioxan e)
69.3 ppm ): 14.94 (CH3CH2); 22.10 (CH2CH3); 49.25 (CH2-1′); 72.90 (d, JC,P = 152, CH2-4′);
79.15 (qd, JC,F = 30, JC,P = 12, CH-2′); 119.29 (C-5); 126.97 (q, JC,F = 284, CF3); 145.84
(CH-6); 154.90 (C-2); 169.36 (C-4). 19F NMR (188.2 MHz, D2O): –73.63 (dd, JF,H-2′ = 6.5,
CF3). 31P NMR (162 MHz, D2O): 12.94 (t, JP,H-4′ = 9.5, 9.2).
1-[3,3,3-Trifluoro-2-(phosphonomethoxy)propyl]thymine (28). A m ixture of com poun d 26
(418 m g, 0.97 m m ol) in aceton itrile (10 m l) an d brom otrim eth ylsilan e (1.3 m l) afforded
after work-up 207 m g (60%) of 28 as a wh ite solid, m .p. > 300 °C. IR (KBr), νm ax: 3243,
1690, 1477, 1439, 1374, 1352, 1230, 1171, 1152, 1131, 1087, 992, 916, 563, 541, 473, 455.
HPLC, 99% (S3). For C9H10F3Li2N2O6P·0.5H2O (353.0) calculated: 30.61% C, 3.14% H,
16.14% F, 7.93% N, 8.77% P; foun d: 30.49% C, 3.18% H, 16.07% F, 7.79% N, 9.00% P. FAB
MS, m/z: 345 [MH]+ (17). 1H NMR (400 MHz, D2O, ref(dioxan e) 3.75 ppm ): 1.88 (d, 3 H,
JCH3,6 = 1.2, CH3-5); 3.68 an d 3.86 (2 × dd, 2 H, Jgem = 12.5, JH,P = 9.1, H-4′); 4.01 (dd, 1 H,
Jgem = 14.8, J1′b,2′ = 7.0, H-1′b); 4.22 (dd, 1 H, Jgem = 14.8, J1′a,2′ = 4.2, H-1′a); 4.30 (m , 1 H,
J2′,1′ = 7.0, 4.2, JH,F = 6.0, H-2′); 7.56 (q, 1 H, J6,CH3 = 1.2, H-6). 13C NMR (125.8 MHz, D2O,
ref(dioxan e) 69.3 ppm ): 14.05 (CH3-5); 49.15 (CH2-1′); 72.84 (d, JC,P = 153, CH2-4′); 79.27
(qd, JC,F = 30, JC,P = 12, CH-2′); 113.52 (C-5); 126.93 (q, JC,F = 284, CF3); 146.43 (CH-6);
154.98 (C-2); 169.80 (C-4). 19F NMR (188.2 MHz, D2O): –73.58 (dd, JF,H-2′ = 6.0, CF3). 31P NMR
(162 MHz, D2O): 13.02 (t, JP,H-4′ = 9.1).
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

616
Pomeisl, Votruba, Holý, Pohl:
1-[2-Hydroxy-3-(trim eth ylam m on io)propyl]-4-m eth oxy-5-m eth ylpyrim idin -2(1H)-on e
Ch loride (30)
A m ixture of 4-m eth oxy-5-m eth ylpyrim idin -2(1H)-on e (22) (1 g, 7.1 m m ol), trim eth yl-
glycidylam m on ium ch loride (29; 1.01 g, 7.1 m m ol) an d cesium carbon ate (273 m g, 0.8
m m ol) in dim eth ylform am ide (50 m l) was h eated at 80 °C for 7.5 h . Th e m ixture was con -
cen trated in vacuo to a m in im um volum e. Th e residue was codistilled with toluen e (20 m l)
an d th e residue was treated with aceton e (50 m l) an d eth an ol (5 m l). Th e resultin g suspen -
sion was h eated un til a wh ite precipitate was obtain ed. After coolin g th e precipitate was fil-
tered off an d dried un der reduced pressure. Yield 661 m g (32%) of crude product 30 as a
wh ite solid wh ich was used for preparation of 32 with out furth er purification . HR MS (FAB):
for C12H23ClN3O3 calculated 292.7820, foun d 292.7828. FAB MS, m/z: 292 [MH]+ (71). 1H
NMR (500 MHz, DMSO-d6): 1.88 (d, 3 H, JCH3,6 = 1.0, CH3-5); 3.13 (s, 9 H, (CH3)3N); 3.30
(dd, 1 H, Jgem = 13.6, J3′b,2′ = 11.2, H-3′b); 3.45 (dd, 1 H, Jgem = 13.6, J3′a,2′ = 1.9, H-3′a); 3.65
(dd, 1 H, Jgem = 13.4, J1′b,2′ = 7.4, H-1′b); 3.85 (s, 3 H, OCH3); 3.88 (dd, 1 H, Jgem = 13.4, J1′a,2′
= 4.1, H-1′a); 4.38 (m , 1 H, H-2′); 6.01 (bs, 1 H, OH-2′); 7.73 (q, 1 H, J6,CH3 = 1.0, H-6).
13C NMR (100.6 MHz, DMSO-d6): 11.77 (CH3-5); 53.11 (CH2-1′); 53.58 ((CH3)3N); 54.13
(OCH3); 63.52 (CH-2′); 67.51 (CH-3′); 102.20 (C-5); 147.43 (CH-6); 155.83 (C-2); 170.31
(C-4).
1-[2-(Ph osph on om eth oxy)-3-(trim eth ylam m on io)propyl]th ym in e (32)
A m ixture of com poun d 30 (661 m g, 2.3 m m ol), (diisopropoxyph osph oryl)m eth yl tosylate
(873 m g, 2.5 m m ol) an d 60% sodium h ydride dispersion (136 m g, 3.4 m m ol) in dim eth yl-
form am ide (15 m l) was stirred at –20 °C. Th e suspen sion was allowed to warm to room tem -
perature for 1 h an d stirred at th e sam e tem perature overn igh t. Th e m ixture was con cen -
trated in vacuo to a m in im um volum e an d th e residue was dissolved in water (3 m l). Th e
m ixture was applied on to a Dowex 50X8 (H+ form ; 30 m l) an d th e colum n was wash ed with
water un til th e UV absoption of th e eluate dropped to th e origin al value. Th e product was
th en eluted with 2.5% aqueous am m on ia. Th e fraction s con tain in g com poun d 31 were
evaporated to dryn ess in vacuo. Th e residue was dissolved in aceton itrile (5 m l) an d
brom otrim eth ylsilan e (0.6 m l) was added. Th e resultin g m ixture was stirred overn igh t at
room tem perature. Th e m ixture was con cen trated in vacuo an d th en codistilled with water
(2 × 2 m l). Th e residue in water (20 m l) was h eated with Dowex 50X8 (H+ form ; 2 m l) at
100 °C for 2 h . Th e suspen sion was applied on to a colum n wh ich was th en wash ed with
2.5% aqueous am m on ia. Th e filtrate was evaporated to dryn ess in vacuo. Th e residue was
purified on 30 m l of Seph adex (S2) with subsequen t deion ization on Dowex 50X8 (H+ form ;
30 m l). Th e relevan t fraction s were com bin ed an d evaporated in vacuo. Th e residue was dis-
solved in water an d lyoph ilized. Yield 96 m g (12%) of 32 as a sligh tly yellow solid. IR (KBr),
νm ax: 1685, 1605, 1524, 1175, 1477, 1436, 968, 1069, 1100, 935, 902, 540, 451. HPLC, 99%
(S3). HR MS (FAB): for
C12H23N3O6P calculated 336.1324, foun d 336.1338. For
C
12H22N3O6P·1.5H2O (362.3) calculated: 39.78% C, 6.96% H, 11.59% N, 8.56% P; foun d:
39.65% C, 6.58% H, 11.76% N, 8.17% P. FAB MS, m/z: 336 [MH]+ (26). 1H NMR (400 MHz,
D2O, ref(dioxan e) 3.75 ppm ): 1.89 (d, 3 H, JCH3,6 = 1.2, CH3-5); 3.23 (s, 9 H, (CH3)3N); 3.46
(dd, 1 H, Jgem = 14.2, J3′b,2′ = 2.4, H-3′b); 3.54 (dd, 1 H, Jgem = 14.2, J3′a,2′ = 9.4, H-3′a); 3.61
(dd, 1 H, Jgem = 12.4, JH,P = 10.2, H-4′b); 3.84 (dd, 1 H, Jgem = 12.4, JH,P = 9.6, H-4′a); 4.00
(dd, 1 H, Jgem = 15.1, J1′b,2′ = 5.1, H-1′b); 4.12 (dd, 1 H, Jgem = 15.1, J1′a,2′ = 3.6, H-1′a); 4.34
(m , 1 H, H-2′); 7.54 (q, 1 H, J6,CH3 = 1.2, H-6). 13C NMR (100.6 MHz, D2O, ref(dioxan e)
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Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
617
69.3 ppm ): 14.05 (CH3-5); 50.10 (CH2-1′); 57.16 ((CH3)3N); 69.26 (d, JC,P = 153, CH2-4′);
69.54 (CH-3′); 77.31 (d, JC,P = 13, CH-2′); 113.95 (C-5); 146.09 (CH-6); 155.32 (C-2); 169.63
(C-4). 31P NMR (162 MHz, D2O): 13.14 (t, JP,H-4′ = 10.2, 9.6).
Syn th esis of 1-{3-Azido-2-[(diisopropoxyph osph oryl)m eth oxy)]propyl}
Derivatives 34a an d 34b
A m ixture of 4-m eth oxy-5-m eth ylpyrim idin -2(1H)-on e (22) (616 m g, 4.4 m m ol) an d 60%
sodium h ydride dispersion (179 m g, 4.5 m m ol) in dim eth ylform am ide (40 m l) was stirred at
room tem perature for 1 h . Com poun d 33 (1.8 g, 4.4 m m ol) in dim eth ylform am ide (25 m l)
was added an d th e resultin g m ixture was h eated at 90 °C for 6.5 h . Th e m ixture was con cen -
trated in vacuo to a m in im um volum e. Th e residue was codistilled with toluen e (2 × 10 m l)
Th e residue was ch rom atograph ed on n eutral alum in um oxide (eth yl acetate followed by
eth yl acetate–ch loroform –m eth an ol 11:10:1). Th e relevan t fraction s were com bin ed an d
evaporated in vacuo.
1-{3-Azido-2-[(diisopropoxyphosphoryl)methoxy]propyl}-4-methoxy-5-methylpyrimidin-2(1H)-one
(34a). Yield 662 m g (36%) of a sligh tly yellow oil. RF 0.30 in eth yl acetate–ch loroform –
m eth an ol 11:10:1. IR (CCl4), νm ax: 2104, 1675, 1654, 1540, 1476, 1400, 1386, 1376, 1257,
1106, 1012, 992. HR MS (FAB): for C16H29N5O6P calculated 418.1855, foun d 418.1862. FAB
MS, m/z: 418 [MH]+ (100). 1H NMR (400 MHz, CDCl3): 1.31, 1.32, 1.33 an d 1.34 (4 × d, 4 ×
3 H, Jvic = 6.2, (CH3)2CH); 1.96 (d, 3 H, JCH3,6 = 1.1, CH3-5); 3.28 (dd, 1 H, Jgem = 13.5, J3′b,2′
5.3, H-3′b); 3.67 (dd, 1 H, Jgem = 13.5, J3′a,2′ = 3.2, H-3′a); 3.70 (dd, 1 H, Jgem = 13.7, JH,P
9.4, H-4′b); 3.86 (dd, 1 H, Jgem = 13.6, J1′b,2′ = 6.9, H-1′b); 3.90 (dd, 1 H, Jgem = 13.7, JH,P
=
=
=
8.5, H-4′a); 3.95 (m , 1 H, J2′,1′ = 6.9, 3.6, J2′,3′ = 5.3, 3.2, H-2′); 3.99 (s, 3 H, OCH3); 4.11 (dd,
1 H, Jgem = 13.6, J1′a,2′ = 3.6, H-1′a); 4.72 an d 4.73 (2 ( dh , 2 H, JH,P = 7.6, Jvic = 6.2,
CH(CH3)2); 7.42 (q, 1 H, J6,CH3 = 1.1, H-6). 13C NMR (100.6 MHz, CDCl3): 11.97 (CH3-5);
23.92, 23.96, 24.00 an d 24.03 (d, JC,P = 4, (CH3)2CH); 50.97 (CH2-1′); 51.69 (CH-3′); 54.56
(OCH3); 65.64 (d, JC,P = 169, CH2-4′); 71.14 an d 71.30 (d, JC,P = 7, CH(CH3)2); 79.14 (d, JC,P
10, CH-2′); 104.34 (C-5); 145.64 (CH-6); 156.74 (C-2); 171.04 (C-4).
=
2-{3-Azido-2-[(diisopropoxyphosphoryl)methoxy)]propoxy}-4-methoxy-5-methylpyrimidine (34b).
Yield 879 m g (48%) of a sligh tly yellow oil. RF 0.50 in eth yl acetate–ch loroform –m eth an ol
11:10:1. IR (CCl4), νm ax: 2104, 1608, 1576, 1423, 1386, 1375, 1295, 1261, 1106, 1085, 1077,
1009, 991. HR MS (FAB): for C16H29N5O6P calculated 418.1855, foun d 418.1845. FAB MS,
m/z: 418 [MH]+ (100). 1H NMR (400 MHz, CDCl3): 1.33, 1.34, 1.35 an d 1.36 (4 × d, 4 × 3 H,
Jvic = 6.2, (CH3)2CH); 2.06 (d, 3 H, JCH3,6 = 0.9, CH3-5); 3.50 (dd, 1 H, Jgem = 13.1, J3′b,2′
6.1, H-3′b); 3.59 (dd, 1 H, Jgem = 13.1, J3′a,2′ = 3.9, H-3′a); 3.94 (dd, 1 H, Jgem = 13.7, JH,P
=
=
8.7, H-4′b); 3.98 (m , 1 H, J2′,1′ = 6.0, 5.2, J2′,3′ = 6.1, 3.9, H-2′); 3.99 (s, 3 H, OCH3); 4.03 (dd,
1 H, Jgem = 13.7, JH,P = 8.9, H-4′a); 4.39 (dd, 1 H, Jgem = 11.4, J1′b,2′ = 6.0, H-1′b); 4.48 (dd, 1 H,
Jgem = 11.4, J1′a,2′ = 5.2, H-1′a); 4.76 an d 4.77 (2 × dh , 2 H, JH,P = 7.6, Jvic = 6.2, CH(CH3)2);
7.97 (q, 1 H, J6,CH3 = 0.9, H-6). 13C NMR (100.6 MHz, CDCl3): 11.82 (CH3-5); 23.92, 24.03
an d 24.05 (d, JC,P = 4, (CH3)2CH); 51.95 (CH-3′); 53.90 (OCH3); 65.22 (d, JC,P = 168, CH2-4′);
65.66 (CH2-1′); 71.18 (d, JC,P = 7, CH(CH3)2); 78.84 (d, JC,P = 11, CH-2′); 111.58 (C-5); 156.92
(CH-6); 163.03 (C-2); 169.65 (C-4).
1-{3-Am in o-2-[(diisopropoxyph osph oryl)m eth oxy]propyl}th ym in e (35)
Com poun d 34a (724 m g, 1.7 m m ol) in m eth an ol (25 m l) was h ydrogen ated over 10% palla-
dium on ch arcoal (733 m g) at room tem perature for 13 h . Th e m ixture was filtered th rough
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618
Pomeisl, Votruba, Holý, Pohl:
a Celite pad. Water (2.5 m l) an d Dowex 50X8 (H+ form ; 6 m l) were added to th e filtrate an d
th e resultin g suspen sion was refluxed for 2 h . Th e suspen sion was applied on to a Dowex
50X8 (H+ form ; 30 m l) colum n an d wash ed with 90% aqueous m eth an ol un til th e UV
absoption of th e eluate dropped to th e origin al value. Th e colum n was th en eluted with
2.5% am m on ia in 90% aqueous m eth an ol an d th e fraction con tain in g th e product was
evaporated in vacuo. Th e crude product was purified by ch rom atograph y on preparative TLC
plate (S1, m eth an ol). Yield 159 m g (24%) of 35 as a sligh tly yellow oil. IR (CCl4), νm ax
:
3411, 3172, 3060, 2981, 1693, 1608, 1424, 1255, 1010, 991. HR MS (FAB): for C15H29N3O6P
calculated 378.1794, foun d 378.1779. FAB MS, m/z: 378 [MH]+ (58). 1H NMR (400 MHz,
CDCl3): 1.32, 1.33 an d 1.34 (3 × d, 12 H, Jvic = 6.2, (CH3)2CH); 1.92 (d, 3 H, JCH3,6 = 1.2,
CH3-5); 2.75 (dd, 1 H, Jgem = 13.6, J3′b,2′ = 4.9, H-3′b); 2.93 (dd, 1 H, Jgem = 13.6, J3′a,2′ = 5.1,
H-3′a); 3.67 (m , 1 H, H-2′); 3.74 (dd, 1 H, Jgem = 13.6, JH,P = 8.8, H-4′b); 3.85 (dd, 1 H, Jgem
13.6, JH,P = 9.5, H-4′a); 3.86 (dd, 1 H, Jgem = 14.3, J1′b,2′ = 7.6, H-1′b); 3.96 (dd, 1 H, Jgem
=
=
14.3, J1′a,2′ = 4.1, H-1′a); 4.74 (m , 2 H, CH(CH3)2); 7.25 (q, 1 H, J6,CH3 = 1.2, H-6). 13C NMR
(100.6 MHz, CDCl3): 12.24 (CH3-5); 23.96, 24.00 an d 24.07 (d, JC,P = 4, (CH3)2CH); 41.54
(CH-3′); 48.84 (CH2-1′); 64.96 (d, JC,P = 170, CH2-4′); 71.14 an d 71.22 (d, JC,P = 7,
CH(CH3)2); 81.60 (d, JC,P = 11, CH-2′); 110.06 (C-5); 142.06 (CH-6); 151.07 (C-2); 163.89
(C-4).
Syn th esis of 1-[3-Azido-2-(ph osph on om eth oxy)propyl]th ym in e (36) an d
1-[3-Am in o-2-(ph osph on om eth oxy)propyl]th ym in e (37). Gen eral Procedure
Com poun ds 36 an d 37 were obtain ed by a procedure sim ilar to th at reported for 5a an d 5b.
1-[3-Azido-2-(phosphonomethoxy)propyl]thymine (36). A m ixture of com poun d 34a (532 m g,
1.27 m m ol) in aceton itrile (10 m l) an d brom otrim eth ylsilan e (1.8 m l) afforded 131 m g
(31%) of dilith ium salt of 36 as a wh ite solid, m .p. > 300 °C. After deion ization of product
on activated ch arcoal th e purification of 36 was perform ed by preparative HPLC (S6). IR
(KBr), νm ax: 3242, 2106, 1684, 1475, 1437, 1386, 1351, 1225, 1177, 1139, 1077, 989, 921,
556, 470. HPLC, 99% (S4). For C9H12Li2N5O6P·1.25H2O (335.6) calculated: 32.21% C,
3.75% H, 20.86% N, 9.23% P; foun d: 32.33% C, 3.82% H, 20.53% N, 11.26% P. FAB MS,
m/z: 332 [MH]+ (5). 1H NMR (400 MHz, D2O, ref(dioxan e) 3.75 ppm ): 1.88 (d, 3 H, JCH3,6
1.2, CH3-5); 3.42 (dd, 1 H, Jgem = 13.5, J3′b,2′ = 4.4, H-3′b); 3.59 (dd, 1 H, Jgem = 12.8, JH,P
9.4, H-4′b); 3.67 (dd, 1 H, Jgem = 13.5, J3′a,2′ = 4.2, H-3′a); 3.71 (dd, 1 H, Jgem = 12.8, JH,P
=
=
=
9.1, H-4′a); 3.88 (m , 1 H, H-2′); 3.91 (dd, 1 H, Jgem = 14.0, J1′b,2′ = 6.1, H-1′b); 4.00 (dd, 1 H,
Jgem = 14.0, J1′a,2′ = 4.5, H-1′a); 7.57 (q, 1 H, J6,CH3 = 1.2, H-6). 13C NMR (125.8 MHz, D2O,
ref(dioxan e) 69.3 ppm ): 14.04 (CH3-5); 51.63 (CH2-1′); 53.39 (CH-3′); 69.74 (d, JC,P = 154,
CH2-4′); 80.64 (d, JC,P = 11, CH-2′); 113.29 (C-5); 146.68 (CH-6); 155.20 (C-2); 169.83 (C-4).
31P NMR (162 MHz, D2O): 14.53 (t, JP,H-4′ = 9.4, 9.1).
1-[3-Amino-2-(phosphonomethoxy)propyl]thymine (37). A m ixture of com poun d 35 (125 m g,
0.33 m m ol) in aceton itrile (5 m l) an d brom otrim eth ylsilan e (0.5 m l) afforded 54 m g (56%)
of ph osph on ic acid 37 as a wh ite solid, m .p. 203–205 °C. Th e product was separated by
ch rom atograph y on 40 m l of Seph adex (S2) an d fin ally purified on Dowex 50X8 (H+ form )
colum n by wash in g with water. IR (KBr), νm ax: 3251, 3000, 1627, 1689, 1472, 1435, 1386,
1102, 1051, 914, 544, 459. HR MS (FAB): for C9H17N3O6P calculated 294.0855, foun d
294.0841. FAB MS, m/z: 294 [MH]+ (84). 1H NMR (400 MHz, D2O, ref(dioxan e) 3.75 ppm ):
1.88 (d, 3 H, JCH3,6 = 1.1, CH3-5); 3.02 (dd, 1 H, Jgem = 13.7, J3′b,2′ = 8.9, H-3′b); 3.26 (dd, 1 H,
Jgem = 13.7, J3′a,2′ = 2.6, H-3′b); 3.64 an d 3.86 (2 × dd, 2 H, Jgem = 12.8, JH,P = 9.6, H-4′b);
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Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
619
3.97–4.05 (m , 3 H, H-1′ an d H-2′); 7.49 (q, 1 H, J6,CH3 = 1.1, H-6). 13C NMR (125.8 MHz,
D2O, ref(dioxan e) 69.3 ppm ): 14.01 (CH3-5); 43.23 (CH-3′); 50.47 (CH2-1′); 69.27 (d, JC,P
=
157, CH2-4′); 79.33 (d, JC,P = 12, CH-2′); 113.74 (C-5); 146.20 (CH-6); 155.25 (C-2); 169.69
(C-4). 31P NMR (162 MHz, D2O): 15.60 (t, JP,H-4′ = 9.6).
Syn th esis of 1-Meth oxy-3-(trityloxy)propan -2-ols (38a, 38b). Gen eral Procedure
A m ixture of [(trityloxy)m eth yl]oxiran e 2a or 2b (12.6 m m ol) an d sodium m eth oxide
(12.7 m m ol) in m eth an ol (40 m l) was h eated in an autoclave at 95 °C for 5 h . Th e m ixture
was con cen trated in vacuo to a m in im um volum e. Th e residue was ch rom atograph ed on
silica gel (ch loroform followed by ch loroform –m eth an ol 20:1). Th e fraction s con tain in g
product 38a or 38b were evaporated to dryn ess in vacuo.
(R)-1-Methoxy-3-(trityloxy)propan-2-ol (38a). Yield 3.2 g (74%) of a wh ite solid. [α]D +18.4
(c 0.328 g/100 m l, CHCl3). HR MS (ESI): for C23H24O3Na calculated 371.1623, foun d
371.1605. For C23H24O3 (348.4) calculated: 79.28% C, 6.94% H; foun d: 79.11% C, 6.68% H.
ESI MS, m/z: 371 [MNa]+ (100). 1H NMR (400 MHz, CDCl3): 2.42 (bd, 1 H, JOH,CH = 4.4, OH);
3.18 (dd, 1 H, Jgem = 9.4, Jvic = 5.3, bCH2OTr); 3.20 (dd, 1 H, Jgem = 9.4, Jvic = 5.7, aCH2OTr);
3.36 (s, 3 H, OCH3); 3.43 (dd, 1 H, Jgem = 9.7, Jvic = 6.5, bCH2OMe); 3.49 (dd, 1 H, Jgem
=
9.7, Jvic = 4.1, aCH2OMe); 3.95 (m , 1 H, Jvic = 6.5, 5.7, 5.3, 4.1, CH); 7.20–7.33 (m , 9 H,
H-m,p-Tr); 7.41–7.46 (m , 6 H, H-o-Tr). 13C NMR (100.6 MHz, CDCl3): 59.12 (OCH3); 64.65
(CH2OTr); 69.81 (CH); 74.07 (CH2OMe); 86.67 (C-Tr); 127.06 (CH-p-Tr); 127.83 (CH-m-Tr);
128.64 (CH-o-Tr); 143.80 (C-i-Tr).
(S)-1-Methoxy-3-(trityloxy)propan-2-ol (38b). Yield 4.1 g (93%) of a wh ite solid. [α]D –17.5
(c 0.192 g/100 m l, CHCl3). For C23H24O3 (348.4) calculated: 79.28% C, 6.94% H; foun d:
78.71% C, 6.87% H. ESI MS, m/z: 371 [MNa]+ (100). 1H, 13C NMR an d HR MS data were
iden tical with com poun d 38a.
Syn th esis of Diisopropyl ({[1-Meth oxy-3-(trityloxy)-2-propan -2-yl]oxy}m eth yl)-
ph osph on ates (39a, 39b). Gen eral Procedure
A m ixture of com poun d 38a or 38b (11.4 m m ol), (diisopropoxyph osph oryl)m eth yl tosylate
(12.5 m m ol) an d 60% sodium h ydride dispersion (17.1 m m ol) in tetrah ydrofuran (60 m l)
was stirred at –20 °C. Th e suspen sion was allowed to warm to room tem perature for 1 h an d
th en stirred at th e sam e tem perature overn igh t. Th e suspen sion was h eated at 40 °C for 8 h
an d th e m ixture was th en filtered th rough a Celite pad. Th e m ixture was con cen trated in
vacuo to a m in im um volum e an d ch rom atograph ed on n eutral alum in um oxide (ch loro-
form ). Th e fraction s con tain in g product 39a or 39b were evaporated to dryn ess in vacuo.
Diisopropyl ({[(R)-1-methoxy-3-(trityloxy)-2-propan-2-yl]oxy}methyl)phosphonate (39a). Yield
5.3 g (88%) of a sligh tly yellow oil. IR (CCl4), νm ax: 3088, 3062, 3035, 2981, 2932, 2895,
2836, 1386, 1375, 1259, 1179, 1142, 1107, 1010, 991. HR MS (ESI): for C30H39O6NaP calcu-
lated 549.2382, foun d 549.2375. FAB MS, m/z: 549 [MNa]+ (35). 1H NMR (400 MHz, CDCl3):
1.30, 1.31, 1.32 an d 1.33 (4 × d, 4 × 3 H, Jvic = 6.2, (CH3)2CH); 3.22 (d, 2 H, Jvic = 5.1,
CH2OTr); 3.31 (s, 3 H, OCH3); 3.50 (dd, 1 H, Jgem = 10.4, Jvic = 6.0, bCH2OMe); 3.54 (dd, 1 H,
Jgem = 10.4, Jvic = 4.2, aCH2OMe); 3.73 (m , 1 H, Jvic = 6.0, 5.1, 4.2, CH); 3.91 (d, 2 H, JH,P
8.8, CH2P); 4.74 (dh , 2 H, JH,P = 7.5, Jvic = 6.2, CH(CH3)2); 7.20–7.33 (m , 9 H, H-m,p-Tr);
7.41–7.46 (m , 6 H, H-o-Tr). 13C NMR (100.6 MHz, CDCl3): 23.95, 23.98 an d 24.11 (d, JC,P
=
=
4, (CH3)2CH); 59.11 (OCH3); 63.39 (CH2OTr); 65.21 (d, JC,P = 167, CH2P); 70.92 an d 70.98
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620
Pomeisl, Votruba, Holý, Pohl:
(d, JC,P = 6, CH(CH3)2); 73.11 (CH2OMe); 80.33 (d, JC,P = 12, CH); 88.26 (C-Tr); 126.98
(CH-p-Tr); 127.77 (CH-m-Tr); 128.71 (CH-o-Tr); 143.90 (C-i-Tr).
Diisopropyl ({[(S)-1-methoxy-3-(trityloxy)-2-propan-2-yl]oxy}methyl)phosphonate (39b ). Yield
4.9 g (82%) of a sligh tly yellow oil. FAB MS, m/z: 549 [MNa]+ (100). 1H, 13C NMR, HR MS
an d IR data were iden tical with com poun d 39a.
Syn th esis of Diisopropyl ({[1-Meth oxy-3-(tosyloxy)-2-propan -2-yl]oxy}m eth yl)-
ph osph on ates (40a, 40b). Gen eral Procedure
A m ixture of com poun d 39a or 39b (6.6 m m ol) an d Dowex 50X8 (H+ form ; 15 m l) in 90%
aqueous m eth an ol (25 m l) was vigorously refluxed for 5 h un til th e trityl group of startin g
ph osph on ate was com pletely rem oved (TLC in ch loroform ). After coolin g th e m ixture was
filtered th rough a Celite pad an d con cen trated in vacuo to a m in im um volum e. Th e residue
was dissolved in dich lorom eth an e (20 m l) an d trieth ylam in e (5 m l). Th e solution of tosyl
ch loride (7.8 m m ol) in dich lorom eth an e (10 m l) was added at 0 °C an d th e resultin g m ix-
ture was stirred at room tem perarure for 2 h . 4-(Dim eth ylam in o)pyridin e (0.1 m m ol) was
added an d th e m ixture was th en refluxed for 10 h . Th e m ixture was con cen trated in vacuo
to a m in im um volum e, th e residue was ch rom atograph ed on silica gel (ch loroform followed
by ch loroform –m eth an ol 20:1) an d appropriate fraction s con tain in g 40a or 40b were evapo-
rated to dryn ess in vacuo.
Diisopropyl ({[(R)-1-Methoxy-3-(tosyloxy)-2-propan-2-yl]oxy}methyl)phosphonate (40a). Yield
1.3 g (46%) of a sligh tly yellow oil. IR (CCl4), νm ax: 2981, 2880, 1386, 1375, 1258, 1190,
1179, 1142, 1121, 1106, 1100, 1020, 815, 555. [α]D +3.6 (c 0.414 g/100 m l, CHCl3). HR MS
(FAB): for C18H32O8PS calculated 439.1556, foun d 439.1570. FAB MS, m/z: 439 [MH]+ (13).
1H NMR (500 MHz, CDCl3): 1.31, 1.32 an d 1.33 (3 × d, 12 H, Jvic = 6.2, (CH3)2CH); 2.46 (s,
3 H, CH3-Ts); 3.29 (s, 3 H, OCH3); 3.44 an d 3.47 (2 × dd, 2 H, Jgem = 10.3, Jvic = 5.1,
CH2OMe); 3.82 (dd, 1 H, Jgem = 13.8, JH,P = 8.6, bCH2P); 3.83 (m , 1 H, CH); 3.84 (dd, 1 H,
Jgem = 13.8, JH,P = 8.6, aCH2P); 4.06 (dd, 1 H, Jgem = 10.5, Jvic = 6.0, bCH2OTs); 4.15 (dd, 1 H,
Jgem = 10.5, Jvic = 4.2, aCH2OTs); 4.72 (m , 2 H, CH(CH3)2); 7.36 (m , 2 H, H-m-Ts); 7.80 (m , 2 H,
H-m-Ts). 13C NMR (125.8 MHz, CDCl3): 21.64 (CH3-Ts); 23.90, 23.93 an d 24.06 (d, JC,P = 4,
(CH3)2CH); 59.22 (OCH3); 65.10 (d, JC,P = 168, CH2P); 69.06 (CH2OTs); 70.98 (CH2OMe);
71.16 an d 71.18 (d, JC,P = 6, CH(CH3)2); 77.93 (d, JC,P = 10, CH); 127.96 (CH-o-Ts); 129.87
(CH-m-Ts); 132.48 (C-i-Ts); 144.93 (C-p-Tr).
Diisopropyl ({[(S)-1-methoxy-3-(tosyloxy)-2-propan-2-yl]oxy}methyl)phosphonate (40b ). Yield
2.7 g (91%) of a sligh tly yellow oil. [α]D –0.6 (c 0.490 g/100 m l, CHCl3). HR MS (FAB): for
C
18H32O8PS calculated 439.1556, foun d 439.1558. FAB MS, m/z: 439 [MH]+ (22). 1H, 13C NMR
an d IR data were iden tical with th ose of com poun d 40a.
Syn th esis of 2-[(Diisopropoxyph osph oryl)m eth oxy]-3-m eth oxypropyl Derivatives of
5-Alkyl-4-m eth oxypyrim idin -2(1H)-on e an d Pyrim idin e 41–44. Gen eral Procedure
A m ixture of pyrim idin -2(1H)-on e 1 or 22 (3 m m ol) an d 60% sodium h ydride dispersion
(3 m m ol) in dim eth ylform am ide (40 m l) was stirred at room tem perature for 1 h . Com -
poun d 40a or 40b (3 m m ol) in dim eth ylform am ide (25 m l) was added at 80 °C an d th e re-
sultin g m ixture was h eated at 100 °C for 9 h . Th e m ixture was con cen trated in vacuo to a
m in im um volum e. Th e residue was codistilled with toluen e (2 × 20 m l). Th e residue was
ch rom atograph ed on n eutral alum in um oxide (eth yl acetate followed by eth yl acetate–ch lo-
roform –m eth an ol 26:25:1). Th e crude product was purified by preparative TLC (S1, eth yl
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Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
621
acetate–ch loroform –m eth an ol 26:25:1). Th e relevan t fraction s were com bin ed an d evapo-
rated in vacuo.
1-{(R)-2-[(Diisopropoxyphosphoryl)methoxy]-3-methoxypropyl}-5-ethyl-4-methoxypyrimidin-2(1H)-
one (41a). Yield 441 m g (35%) of a sligh tly yellow oil. RF 0.25 (eth yl acetate–ch loroform –
m eth an ol 26:25:1). IR (CCl4), νm ax: 2998, 2937, 2980, 1673, 1652, 1536, 1471, 1403, 1255,
1107, 1007, 991. [α]D +49.0 (c 0.539 g/100 m l, CHCl3). HR MS (FAB): for C18H34N2O7P
calculated 421.2104, foun d 421.2120. FAB MS, m/z: 421 [MH]+ (100). 1H NMR (500 MHz,
CDCl3): 1.16 (t, 3 H, Jvic = 7.5, CH3CH2); 1.28, 1.29, 1.30 an d 1.31 (4 × d, 4 × 3 H, Jvic = 6.2,
(CH3)2CH); 2.34 an d 2.38 (2 × dqd, 2 H, Jgem = 17.3, Jvic = 7.5, JCH2,6 = 1.0, CH2CH3); 3.36
(s, 3 H, OCH3-3′); 3.44 (dd, 1 H, Jgem = 10.8, J3′b,2′ = 4.7, H-3′b); 3.63 (dd, 1 H, Jgem = 10.8,
J3′a,2′ = 3.3, H-3′a); 3.70 (dd, 1 H, Jgem = 13.7, JH,P = 9.2, H-4′b); 3.77 (dd, 1 H, Jgem = 13.8,
J1′b,2′ = 7.7, H-1′b); 3.91 (dd, 1 H, Jgem = 13.7, JH,P = 8.6, H-4′a); 3.95 (m , 1 H, H-2′); 3.98 (s, 3 H,
OCH3-4); 4.19 (dd, 1 H, Jgem = 13.8, J1′a,2′ = 3.7, H-1′a); 4.69 an d 4.71 (2 × dh , 2 H, JH,P
=
7.8, Jvic = 6.2, CH(CH3)2); 7.34 (t, 1 H, J6,CH2 = 1.0, H-6). 13C NMR (100.6 MHz, CDCl3):
13.03 (CH3CH2); 19.70 (CH2CH3); 23.90, 23.93, 24.02 an d 24.04 (d, JC,P = 4, (CH3)2CH);
50.96 (CH2-1′); 54.42 (OCH3-4); 59.27 (OCH3-3′); 65.04 (d, JC,P = 168, CH2-4′); 70.93 an d
71.02 (d, JC,P = 7, CH(CH3)2); 71.90 (CH-3′); 78.57 (d, JC,P = 11, CH-2′); 109.89 (C-5); 145.06
(CH-6); 156.67 (C-2); 170.64 (C-4).
1-{(S)-2-[(Diisopropoxyphosphoryl)methoxy]-3-methoxypropyl}-5-ethyl-4-methoxypyrimidin-2(1H)-
one (41b). Yield 440 m g (35%) of a sligh tly yellow oil. RF 0.25 in eth yl acetate–ch loroform –
m eth an ol 26:25:1. [α]D –45.3 (c 0.651 g/100 m l, CHCl3). HR MS (FAB): for C18H34N2O7P
calculated 421.2104, foun d 421.2083. FAB MS, m/z: 421 [MH]+ (100). 1H, 13C NMR an d IR
data were iden tical with com poun d 41a.
1-{(R)-2-[(Diisopropoxyphosphoryl)methoxy]-3-methoxypropyl}-4-methoxy-5-methylpyrimidin-
2(1H)-one (42a). Yield 304 m g (25%) of a sligh tly yellow oil. RF 0.25 in eth yl acetate–ch loro-
form –m eth an ol 26:25:1. IR (CCl4), νm ax: 2981, 2899, 2836, 1676, 1655, 1541, 1475, 1255,
1141, 1107, 1011, 991. [α]D +88.8 (c 0.526 g/100 m l, CHCl3). HR MS (FAB): for C17H32N2O7P
calculated 407.1947, foun d 407.1961. FAB MS, m/z: 407 [MH]+ (100). 1H NMR (400 MHz,
CDCl3): 1.29, 1.30, 1.31 an d 1.32 (4 × d, 4 × 3 H, Jvic = 6.2, (CH3)2CH); 1.94 (d, 3 H, JCH3,6
=
1.1, CH3-5); 3.36 (s, 3 H, OCH3-3′); 3.45 (dd, 1 H, Jgem = 10.8, J3′b,2′ = 4.9, H-3′b); 3.63 (dd,
1 H, Jgem = 10.8, J3′a,2′ = 3.3, H-3′a); 3.69 (dd, 1 H, Jgem = 13.7, JH,P = 9.2, H-4′b); 3.73 (dd,
1 H, Jgem = 13.8, J1′b,2′ = 7.7, H-1′b); 3.93 (dd, 1 H, Jgem = 13.7, JH,P = 8.8, H-4′a); 3.94 (m ,
1 H, H-2′); 3.98 (s, 3 H, OCH3-4); 4.20 (dd, 1 H, Jgem = 13.8, J1′a,2′ = 3.5, H-1′a); 4.70 (dh ,
2 H, JH,P = 7.7, Jvic = 6.2, CH(CH3)2); 7.38 (q, 1 H, J6,CH3 = 1.1, H-6). 13C NMR (100.6 MHz,
CDCl3): 12.02 (CH3-5); 23.91, 23.93 an d 24.04 (d, JC,P = 4, (CH3)2CH); 50.84 (CH2-1′); 54.49
(OCH3-4); 59.28 (OCH3-3′); 65.05 (d, JC,P = 168, CH2-4′); 70.90 an d 71.01 (d, JC,P = 7,
CH(CH3)2); 72.04 (CH-3′); 78.72 (d, JC,P = 11, CH-2′); 103.85 (C-5); 145.92 (CH-6); 156.77
(C-2); 170.88 (C-4).
1-{(S)-2-[(Diisopropoxyphosphoryl)methoxy]-3-methoxypropyl}-4-methoxy-5-methylpyrimidin-
2(1H)-one (42b). Yield 366 m g (30%) of a sligh tly yellow oil. RF 0.25 in eth yl acetate–ch loro-
form –m eth an ol 26:25:1. [α]D –79.4 (c 0.487 g/100 m l, CHCl3). HR MS (FAB): for C17H32N2O7P
calculated 407.1947, foun d 407.1934. FAB MS, m/z: 407 [MH]+ (100). 1H, 13C NMR an d IR
data were iden tical with th ose of com poun d 42a.
2-{(R)-2-[(Diisopropoxyphosphoryl)methoxy]-3-methoxypropoxy}-5-ethyl-4-methoxypyrimidine
(43a). Yield 252 m g (20%) of a sligh tly yellow oil. RF 0.50, eth yl acetate–ch loroform –m eth an ol
26:25:1). IR (CCl4), νm ax: 2980, 1603, 1571, 1424, 1399, 1386, 1375, 1352, 1258, 1107, 1011,
991. [α]D +12.8 (c 0.392 g/100 m l, CHCl3). HR MS (FAB): for C18H34N2O7P calculated
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622
Pomeisl, Votruba, Holý, Pohl:
421.2104, foun d 421.2124. FAB MS, m/z: 421 [MH]+ (100). 1H NMR (400 MHz, CDCl3): 1.16
(t, 3 H, Jvic = 7.5, CH3CH2); 1.31, 1.32 an d 1.33 (3 × d, 12 H, Jvic = 6.2, (CH3)2CH); 2.48 (qd,
2 H, Jvic = 7.5, JCH2,6 = 0.7, CH2CH3); 3.38 (s, 3 H, OCH3-3′); 3.60 (dd, 1 H, Jgem = 10.4, J3′b,2′
=
5.5, H-3′b); 3.64 (dd, 1 H, Jgem = 10.4, J3′a,2′ = 4.5, H-3′a); 3.98 (s, 3 H, OCH3-4); 3.99 (dd,
1 H, Jgem = 13.7, JH,P = 8.8, H-4′b); 4.01 (m , 1 H, H-2′); 4.03 (dd, 1 H, Jgem = 13.7, JH,P = 8.8,
H-4′a); 4.41 (dd, 1 H, Jgem = 11.4, J1′b,2′ = 5.2, H-1′b); 4.44 (dd, 1 H, Jgem = 11.4, J1′a,2′ = 5.8,
H-1′a); 4.75 (m , 2 H, CH(CH3)2); 7.97 (t, 1 H, J6,CH2 = 0.7, H-6). 13C NMR (100.6 MHz,
CDCl3): 13.43 (CH3CH2); 19.79 (CH2CH3); 23.90 an d 24.08 (d, JC,P = 4, (CH3)2CH); 53.80
(OCH3-4); 59.23 (OCH3-3′); 65.20 (d, JC,P = 167, CH2-4′); 66.50 (CH2-1′); 71.03 an d 71.08 (d,
JC,P = 6, CH(CH3)2); 72.48 (CH-3′); 78.83 (d, JC,P = 12, CH-2′); 117.07 (C-5); 156.03 (CH-6);
163.21 (C-2); 169.32 (C-4).
2-{(S)-2-[(Diisopropoxyphosphoryl)methoxy]-3-methoxypropoxy}-5-ethyl-4-methoxypyrimidine
(43b). Yield 530 m g (42%) of a sligh tly yellow oil. RF 0.50 in eth yl acetate–ch loroform –
m eth an ol 26:25:1. [α]D –10.2 (c 0.720 g/100 m l, CHCl3). HR MS (FAB): for C18H34N2O7P
calculated 421.2104, foun d 421.2124. FAB MS, m/z: 421 [MH]+ (100). 1H, 13C NMR an d IR
data were iden tical with th ose of com poun d 43a.
2-{(R)-2-[(Diisopropoxyphosphoryl)methoxy]-3-methoxypropoxy}-4-methoxy-5-methylpyrimidine
(44a). Yield 207 m g (17%) of a sligh tly yellow oil. RF 0.50 in eth yl acetate–ch loroform –
m eth an ol 26:25:1. IR (CCl4), νm ax: 2981, 1608, 1575, 1424, 1397, 1386, 1374, 1258, 1107,
1010, 992. [α]D +20.0 (c 0.231 g/100 m l, CHCl3). HR MS (FAB): for C17H32N2O7P calculated
407.1947, foun d 407.1936. FAB MS, m/z: 407 [MH]+ (100). 1H NMR (400 MHz, CDCl3): 1.31,
1.32, 1.33 an d 1.34 (4 × d, 4 × 3 H, Jvic = 6.2, (CH3)2CH); 2.05 (d, 2 H, JCH3,6 = 1.0, CH3-5);
3.37 (s, 3 H, OCH3-3′); 3.60 (dd, 1 H, Jgem = 10.4, J3′b,2′ = 5.5, H-3′b); 3.63 (dd, 1 H, Jgem
=
10.4, J3′a,2′ = 4.6, H-3′a); 3.98 (s, 3 H, OCH3-4); 3.99 (dd, 1 H, Jgem = 13.7, JH,P = 8.8, H-4′b);
4.01 (m , 1 H, H-2′); 4.03 (dd, 1 H, Jgem = 13.7, JH,P = 8.8, H-4′a); 4.41 (dd, 1 H, Jgem = 11.4,
J1′b,2′ = 5.2, H-1′b); 4.44 (dd, 1 H, Jgem = 11.4, J1′a,2′ = 5.8, H-1′a); 4.75 (m , 2 H, CH(CH3)2);
7.96 (q, 1 H, J6,CH3 = 1.1, H-6). 13C NMR (100.6 MHz, CDCl3): 11.84 (CH3-5); 23.93 an d
24.10 (d, JC,P = 4, (CH3)2CH); 53.87 (OCH3-4); 59.24 (OCH3-3′); 65.25 (d, JC,P = 167, CH2-4′);
66.56 (CH2-1′); 71.01 an d 71.06 (d, JC,P = 7, CH(CH3)2); 72.50 (CH-3′); 78.84 (d, JC,P = 12,
CH-2′); 111.23 (C-5); 156.99 (CH-6); 163.24 (C-2); 169.41 (C-4).
2-{(S)-2-[(Diisopropoxyphosphoryl)methoxy]-3-methoxypropoxy}-4-methoxy-5-methylpyrimidine
(44b). Yield 524 m g (43%) of a sligh tly yellow oil. RF 0.50 (TLC, eth yl acetate–ch loroform –
m eth an ol 26:25:1). [α]D –16.4 (c 0.268 g/100 m l, CHCl3). HR MS (FAB): for C17H32N2O7P
calculated 407.1947, foun d 407.1957. FAB MS, m/z: 407 [MH]+ (100). 1H, 13C NMR an d IR
data were iden tical with th ose of com poun d 44a.
Syn th esis of 1-[2-(Ph osph on om eth oxy)-3-m eth oxypropyl] Derivatives of
5-Eth yluracil an d Th ym in e 45 an d 46. Gen eral Procedure
A m ixture of com poun ds 41 or 42 (1 m m ol) in aceton itrile (10 m l) an d brom otrim eth yl-
silan e (10 m m ol) afforded correspon din g derivatives 45, 46 wh ich were obtain ed as
dilith ium salts by th e sam e procedure as described for 5a or 5b.
5-Ethyl-1-[(R)-2-(Phosphonomethoxy)-3-methoxypropyl]uracil (45a). Yield 243 m g (69%) of a
wh ite solid, m .p. > 300 °C. [α]D +22.1 (c 0.261 g/100 m l, H2O). IR (KBr), νm ax: 3188, 3035,
2894, 2831, 1689, 1460, 1107, 1090, 991. HPLC, 99% (S3). HR MS (FAB): for
C
11H18Li2N2O7P calculated 335.1171, foun d 335.1167. For C11H17Li2N2O7P·1H2O (352.1)
calculated: 37.57% C, 5.43% H, 7.96% N, 8.80% P; foun d: 37.57% C, 5.54% H, 7.79% N,
Collect. Czech. Chem. Commun. 2006, Vol. 71, No. 4, pp. 595–624

Pyrimidine 1-[2-(Phosphonomethoxy)propyl] Derivatives
623
8.44% P. FAB MS, m/z: 335 [MH]+ (100). 1H NMR (400 MHz, D2O, ref(dioxan e) 3.75 ppm ):
1.09 (t, 3 H, Jvic = 7.5, CH3CH2); 2.31 (qd, 2 H, Jvic = 7.5, JCH2,6 = 1.0, CH2CH3); 3.36
(OCH3); 3.47 (dd, 1 H, Jgem = 11.0, J3′b,2′ = 4.3, H-3′b); 3.58 (dd, 1 H, Jgem = 12.4, JH,P = 9.5,
H-4′b); 3.62 (dd, 1 H, Jgem = 12.4, JH,P = 9.4, H-4′a); 3.67 (dd, 1 H, Jgem = 11.0, J3′a,2′ = 3.8,
H-3′a); 3.83 (tt, 1 H, J2′,1′ = 5.8, J2′,3′ = 4.3, 3.8, H-2′); 3.92 (dd, 1 H, Jgem = 14.3, J1′b,2′ = 5.8,
H-1′b); 4.01 (dd, 1 H, Jgem = 14.3, J1′a,2′ = 5.8, H-1′a); 7.50 (t, 1 H, J6,CH2 = 1.0, H-6).
13C NMR (125.8 MHz, D2O, ref(dioxan e) 69.3 ppm ): 15.03 (CH3CH2); 22.08 (CH2CH3); 51.15
(CH2-1′); 61.21 (OCH3); 70.07 (d, JC,P = 154, CH2-4′); 73.48 (CH-3′); 80.23 (d, JC,P = 12,
CH-2′); 118.96 (C-5); 146.17 (CH-6); 155.10 (C-2); 169.43 (C-4). 31P NMR (162 MHz, D2O):
14.36 (t, JP,H-4′ = 9.5, 9.4).
5-Ethyl-1-[(S)-2-(phosphonomethoxy)-3-methoxypropyl]uracil (45b). Yield 239 m g (68%) of a
wh ite solid, m .p. > 300 °C. [α]D –20.1 (c 0.250 g/100 m l, H2O). HPLC, 99% (S3). HR MS
(FAB): for C11H18Li2N2O7P calculated 335.1171, foun d 335.1166. For C11H17Li2N2O7P·1H2O
(352.1) calculated: 37.57% C, 5.43% H, 7.96% N, 8.80% P; foun d: 37.56% C, 5.36% H,
7.72% N, 8.54% P. FAB MS, m/z: 335 [MH]+ (51). 1H, 13C NMR an d IR data were iden tical
with com poun d 45a.
1-[(R)-2-(Phosphonomethoxy)-3-methoxypropyl]thymine (46a). Yield 252 m g (71%) of a wh ite
solid, m .p. > 300 °C. IR (KBr), νm ax: 3260, 2902, 2837, 1690, 1474, 1439, 1105, 1081, 1073,
992. [α]D +27.9 (c 0.330 g/100 m l, H2O). HPLC, 99% (S3). HR MS (FAB): for C10H16Li2N2O7P
calculated 321.1015, foun d 321.1022. For
C10H15Li2N2O7P·2H2O (356.1) calculated:
33.73% C, 5.38% H, 7.86% N, 8.69% P; foun d: 34.07% C, 5.54% H, 7.88% N, 8.35% P. FAB
MS, m/z: 321 [MH]+ (37). 1H NMR (400 MHz, D2O, ref(dioxan e) 3.75 ppm ): 1.88 (d, 3 H,
JCH3,6 = 1.2, CH3-5); 3.36 (OCH3); 3.48 (dd, 1 H, Jgem = 11.0, J3′b,2′ = 4.5, H-3′b); 3.57 (dd, 1 H,
Jgem = 12.5, JH,P = 9.6, H-4′b); 3.64 (dd, 1 H, Jgem = 12.5, JH,P = 9.4, H-4′a); 3.67 (dd, 1 H,
Jgem = 11.0, J3′a,2′ = 3.9, H-3′a); 3.81 (bp, 1 H, J2′,1′ = 5.9, 5.1, J2′,3′ = 4.5, 3.9, H-2′); 3.93 (dd,
1 H, Jgem = 14.4, J1′b,2′ = 5.1, H-1′b); 3.97 (dd, 1 H, Jgem = 14.4, J1′a,2′ = 5.9, H-1′a); 7.55 (q, 1 H,
J6,CH3 = 1.2, H-6). 13C NMR (125.8 MHz, D2O, ref(dioxan e) 69.3 ppm ): 14.02 (CH3-5); 51.14
(CH2-1′); 61.23 (OCH3); 69.87 (d, JC,P = 154, CH2-4′); 73.56 (CH-3′); 80.46 (d, JC,P = 12,
CH-2′); 113.14 (C-5); 146.78 (CH-6); 155.20 (C-2); 169.86 (C-4). 31P NMR (162 MHz, D2O):
14.67 (t, JP,H-4′ = 9.6, 9.4).
1-[(S)-2-(Phosphonomethoxy)-3-methoxypropyl]thymine (46b). Yield 228 m g (64%) of a wh ite
solid, m .p. > 300 °C. [α]D –23.4 (c 0.280 g/100 m l, H2O). For C10H15Li2N2O7P·2H2O (356.1)
calculated: 33.73% C, 5.38% H, 7.86% N, 8.69% P; foun d: 34.88% C, 4.94% H, 7.52% N,
8.33% P. FAB MS, m/z: 321 [MH]+ (27). 1H, 13C NMR, HR MS an d IR data were iden tical with
com poun d 46a.
The study has been supported by the Grant Agency of the Czech Republic (grant No. 203/03/0089)
as a part of research project Z 40550506 of the Institute of Organic Chemistry and Biochemistry,
Prague and by the Descartes Prize HPAW-CT-2002-9001. The authors thank the Analytical
Department of the Institute (Dr K. Ubik, Head), and Dr O. Špalek and Dr V. Jirásek (Institute of
Physics, Academy of Sciences of the Czech Republic, Prague) for assistance. Excellent technical
assistance of Ms J. Křelinová is also acknowledged.
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