Synthesis and fungicidal activity of O-alkyl O-aryl O-2-(stearamido)ethyl phosphates

Article abstract of DOI:10.1002/hc.20485

N-stearoylethanolamine (NAE18:0) was reacted with O-alkyl O-aryl chlorophosphate and a series of O-alkyl O-aryl O-2-(stearamido)ethyl phosphates were synthesized to explore their antifungal activity. Compared with parent NAE18:0, title compounds without substitution or with methyl substitution on a benzene ring exhibited improved antifungal activity.

Full text of DOI:10.1002/hc.20485

Heteroatom Chemistry
Volume 19, Number 6, 2008
Synthesis and Fungicidal Activity of O-Alkyl
O-Aryl O-2-(Stearamido)ethyl Phosphates
Liang Han,1 Jian-Rong Gao,1 and Zheng-Ming Li2
1State Key Laboratory Breeding Base of Green Chemistry-Synthesis Technology,
Zhejiang University of Technology, Hangzhou 310014, People’s Republic of China
2National Key Laboratory of Elemento-Organic Chemistry, Tianjin 300071,
People’s Republic of China
Received 11 May 2007; revised 28 November 2007, 25 March 2008
ity [2,3], and at elevated levels, interfered with nor-
ABSTRACT: N-stearoylethanolamine (NAE18:0)
mal seedling root development [4–6]. The applica-
was reacted with O-alkyl O-aryl chlorophosphate and
tion of exogenous NAEs further confirmed the recog-
a series of O-alkyl O-aryl O-2-(stearamido)ethyl phos-
nition of their important role in plant [2f,7,8]. Al-
phates were synthesized to explore their antifungal
though the above reports are exciting, NAEs have
activity. Compared with parent NAE18:0, title com-
very low solubility in water and limited solubility
pounds without substitution or with methyl substitu-
in most common organic solvents, which discour-
tion on a benzene ring exhibited improved antifungal
ages their further study and application. In our
ꢀ
C
activity. 2008 Wiley Periodicals, Inc. Heteroatom Chem
previous work, we found that NAE derivatives ex-
hibit improved bioactivity and solubility. For ex-
ample, after introduction of N-stearoylethanolamine
19:602–608, 2008; Published online in Wiley InterScience
(www.interscience.wiley.com). DOI 10.1002/hc.20485
into nitrogen mustard phosphorate, a series of
O-substituted phenyl O-2-stearamidoethyl Nꢁ N-
INTRODUCTION
bis(2-chloroethy1)phosphoramidate with improved
solubility and antifungal activity were obtained [9].
Moreover, the compounds, synthesized through the
acylation of NAEs with aryloxyacetic acids exhibited
better activity than 2,4-dichlorophenyloxyacetic acid
(a traditional plant growth regulator) in stimulating
hypocotyls elongation of rape [10].
In organisms, NAEs are formed from N-acylated
phosphatidylethanolamine (PE) molecular species
by the action of a phosphodiesterase, phospholipase
D (PLD). This reaction produces a corresponding
NAE and a molecule of phosphatidic acid (PA). NAEs
are further hydrolyzed by an enzyme designated
fatty acid amide hydrolase (FAAH), producing free
fatty acid (FFA) and ethanolamine. Free fatty acids
are utilized by the enzyme designated NAPE (N-
acylphosphatidylethanolamine) synthase to acylate
the amino head group of PE and produce NAPE, the
precursor of NAEs, thus realizing the metabolism
N-acylethanolamines (NAEs), a kind of minor mem-
brane lipid constituent, exhibit important biologi-
cal and pharmacological activities in organisms [1].
More investigation supported the fact that NAEs play
a lipid mediator role in plants. NAEs were implicated
as transducers in plant defense signaling, identified
as inhibitors of phospholipase D (α isoform) activ-
Correspondence to: Liang Han; e-mail: hanliang814@163.com.
Contract grant sponsor: National Basic Research Program of
China.
Contract grant number: 2003CB114400.
Contract grant sponsor: National Science Foundation of China-
Key Project.
Contract grant number: 20432010.
Contract grant sponsor: Scientific Research Fund of Zhejiang
Education Department.
Contract grant number: 20070310.
2008 Wiley Periodicals, Inc.
c
ꢀ
602

Synthesis and Fungicidal Activity of O-Alkyl O-Aryl O-2-(Stearamido)ethyl Phosphates 603
O
OH
P
O
O
O
O
O
HN
O
O
OH
O
P
O
O
-
OH
O
P
O
O
O
O
O
O
O
O
H2
N
O
O
PA
PE
NAPE
NAPE synthase
PLD
O
O
FAAH
N
HO
H
OH
Free fatty acid
NAE18:0
H2
N
OH
SCHEME 1 The metabolism mechanism of N-stearoylethanolamine.
cycle of NAEs [3a]. The metabolism mechanism
CH3(CH2)16COOH
NH2CH2CH2OH
+
of N-stearoylethanolamine (NAE18:0) is shown in
Scheme 1. This metabolism was stimulated for sev-
eral hours following treatment with a pathogen elici-
tor and believed to be involved in signaling pathogen
perception. New bioactive compounds with good
solubility may be obtained through the prepara-
tion of compounds similar with the precursor of
NAEs. Therefore, a series of O-alkyl O-aryl O-2-
(stearamido)ethyl phosphates were synthesized and
their antifungal activity were evaluated.
O
HN
HO
NAE(18)
O
O
Et3N/CHCl3
Cl
NAE(18)
Cl
+
ArOH
EtO
P
EtO P
Cl
OAr
Et3N/CHCl3
O
O
HN
O
EtO P
OAr
1
SCHEME 2
RESULTS AND DISCUSSION
Synthesis of O-Ethyl O-Aryl
O-2-(Stearamido)ethyl Phosphates
from the solution of trichloromethane and acetone
instead of 95% ethanol as mentioned in Ref. [8].
First, NAE18:0 was reacted with O-ethyl O-aryl
chlorophosphate in the presence of Et3N, but a poor
yield was obtained. The well-known acylating cata-
lyst 4-dimethylaminopyridine (DMAP) was used to
improve the reaction yield, and the preparative re-
action of compound 1o was carried out under the
optimum conditions. The reaction yield improved to
38% from 26% with 0.1 equiv DMAP as the cata-
lyst, whereas the yield increased to 48% with 1 equiv
DMAP as both the catalyst and the acid receptor.
Therefore, NAE18:0 and 1 equiv DMAP were added,
respectively, into the CHCl3 solution of O-ethyl
O-aryl chlorophosphate and the suspension was ob-
tained owing to the poor solubility of NAE18:0.
With the progress of reaction, NAE18:0 dissolved
Several NAE types have been identified in a variety
of plant species. These NAE types identified contain
acyl chains of 12–18 C in length and up to three dou-
ble bonds, reflecting the typical acyl moieties preva-
lent in higher plants [3a]. In terms of bioactivity,
plants are particularly responsive to low concentra-
tion of medium chain saturated NAEs, whereas ani-
mal physiology is largely regulated by low concentra-
tion of long chain polyunsaturated NAEs (e.g., NAE
20:4) [5]. Therefore, in our primary work NAE18:0
was selected to be modified and a series of O-alkyl
O-aryl O-2-(stearamido)ethyl phosphates were pre-
pared as shown in Scheme 2.
NAE18:0 was prepared by refluxing 1 mol stearic
acid with 1.5 mol ethanolamine for 6 h in 98% yield
[11]. The product was purified by recrystallization
Heteroatom Chemistry DOI 10.1002/hc

604 Han, Gao, and Li
gradually and the reactant solution turned clear,
which indicated the reaction happened and the prod-
ucts had better solubility than NAE18:0 in CHCl3.
After the usual workup, title compounds 1 were pu-
rified by column chromatography. Most products
were solids with low melting points and the yields
were still somewhat lower due to the existence of
long fatty chain in NAEs.
1a show improved inhibition activity against all the
tested fungi.
The bioactivities of title compounds 1b–1p with
different substitutions on a benzene ring were inves-
tigated. Compounds 1l, 1m, and 1n with a methyl
group on the benzene ring exhibit improved ac-
tivities against all the tested fungi compared with
NAE18:0. For example, at the concentration of
50 mg/L, the inhibition percent of compound 1m
against Phoma asparagi, is 52.8%, whereas NAE18:0
shows no activity. However, the presence of a halo-
gen atom on the benzene ring was unfavorable and
compounds 1c–1e showed no fungicidal activity.
The bioactivity seemed to be associated with the
electron density on the benzene ring. Compounds
1m, 1n with high electron density on the benzene
ring exhibit better antifungal activities than parent
NAE18:0, whereas those with low electron density on
the benzene ring such as 1b–1e and 1i have inferior
inhibition activities to NAE18:0. Surprisedly, com-
pound 1g, bearing a fluorine atom on the benzene
ring, exhibited greater increased fungicidal activities
against Gillerella zeae and Alternaria solani compared
with NAE18:0. Another exception is compound 1k
Biological Activities
Since NAEs were implicated as transducers in plant
defense signaling and they could activate the expres-
sion of PAL2 (phenylalanine-ammonia lyase) [1],
we primarily studied the antifungal activities of the
derivatives in vitro against Gibberella zeae, Alternaria
solani, Phoma asparagi, Physalospora piricola, and
Cercospora arachi dicola. The inhibition percent in
these assays over control samples without fungi-
cides is listed in Table 1 and compared with that
of NAE18:0. The data indicate that NAE18:0 only
exhibits certain inhibition activity against Alternaria
solani and Physalospora piricola, whereas compound
TABLE 1 Antifungal Activity of Title Compounds and NAE(18:0) In Vitro (50 mg/L, Relative Inhibitory Ratio %)
O
O
R2
HN
OR1
O
O P
Ratio (%)
Phoma
asparagi
Gibberella
Alternaria
solani
Physalospora
piricola
Cercospora
arachi dicola
Compound
R1
R2
zeae
1a
1b
1c
1d
1e
1f
1g
1h
1i
1j
1k
1l
1m
1n
1o
1p
1q
1r
Et
Et
Et
Et
Et
Et
Et
Et
Et
Et
Et
Et
Et
Et
Et
Et
Pr
ClEt
H
3-Cl
4-Cl
2,4-Cl
2-Br
4-Br
4-F
2-NO2
4-NO2
3-CH3O
10
0
0
0
0
0
16.0
0
0
0
42.9
0
0
0
0
0
40.5
0
0
0
33.3
0
0
0
0
0
0
22.2
0
44.4
0
22.2
52.8
30.6
0
0
0
0
19.4
0
33.8
23.1
0
0
0
30.8
20
23.1
12.3
30.8
27.7
12.3
38.5
30.8
12.3
0
12.3
12.3
0
12.3
24.4
20.6
0
0
0
0
0
0
11.8
0
20.6
0
14.7
14.7
14.7
0
11.8
0
0
0
0
0
2-CH3
3-CH3
3,4-CH3
0
0
16
40
14
0
0
0
0
0
0
0
23.8
42.9
23.8
0
0
0
0
19.4
0
2,6-CH3
4-tBu
2-CH3-4-NO2
H
H
H
H
–
1s
1t
NAE18
BnOEt
a
–
14.3
0
a(Tetrahydrofuran-2-yl)methyl substituted.
Heteroatom Chemistry DOI 10.1002/hc

Synthesis and Fungicidal Activity of O-Alkyl O-Aryl O-2-(Stearamido)ethyl Phosphates 605
with ortho methyl substitution on the benzene ring
50 ppm tested compound. A bacterium tray of 4-mm
diameter cut along the external edge of the hypase
was removed to the flat containing tested compound
and put in equilateral triangular style. Later, the cul-
ture plate was cultured at 24 1◦C and the expanded
diameter of bacterium tray was measured after 48 h
and compared with that treated with sterilization
water to estimate the activity. Two replicates were
included in the evaluation.
that only exhibits comparable fungicidal activity
against Physalospora piricola with NAE18:0 and fails
to act against other fungi.
In addition, we replaced an ethyl group with
other alkyl groups and prepared compounds 1q–1t
to investigate the fungicidal activity variety along
with different alkyl groups. Unfortunately, the
change of the alkyl group does not contribute to the
improvement of inhibition activity and the bioactiv-
ities of compounds 1q–1t are inferior to those of 1a
and even parent NAE18:0.
General Synthetic Procedure for Title
Compounds
Ethyl or phenyl phosphorodichloride (3 mmol) was
dissolved in CHCl3 (5 mL), and the solution was
cooled to −10◦C. Phenol or alcohol (3 mmol) and
Et3N (3 mmol) in CHCl3 (5 mL) were dripped into the
stirred solution. The reaction mixture was continued
to be stirred in ice bath for 1 h. The solution of DMAP
(2.1 mmol) in CHCl3 (5 mL) was dripped into the
reaction solution after NAE18:0 (0.7 g) was added.
Then the reaction solution was allowed to rise to
room temperature, and during this period NAE18:0
dissolved gradually and the solution turned clear and
light yellow. After stirring for 12 h, the reaction so-
lution was washed with water and then dried over
anhydrous magnesium sulfate. The solvent was re-
moved, and column chromatography of the residue
with PE/EtOAc (2:1, v/v) gave the product as a solid.
CONCLUSION
In conclusion, a series of the phosphate deriva-
tives of NAE18:0 were synthesized through reac-
tion of NAE18:0 with O-alkyl O-aryl chlorophos-
phates. All compounds have better solubility than
NAE18:0 and are soluble in CHCl3. The bioactivity
data indicate that title compounds without substitu-
tion or with methyl substitution on benzene ring ex-
hibit improved fungicidal activities compared with
NAE18:0. Certain relationship is observed between
the inhibition activity and the electron density on
the benzene ring and those with high electron den-
sity on the benzene ring show better bioactivities and
broader antibacterial spectrum than NAE18:0. The
further study for the bioactivity evaluation of these
compounds and NAE18:0 is under consideration.
O-Ethyl O-phenyl O-2-(stearamido)ethyl phos-
phate (1a). Yellowy solid, mp 44–46◦C, yield 44%.
1H NMR (CDCl3, 300 MHz) δ: 7.38–7.33 (t, 2H, ArH),
7.22–7.20 (d, 3H, ArH), 6.13 (bs, 1H, NH), 4.29–4.19
(m, 4H, NHCH2CH2O, CH3CH2OP), 3.58–3.53 (dd,
2H, NHCH2CH2O), 2.15–2.10 (t, 2H, CH2CONH),
1.60–1.56 (t, 2H, CH2CH2CONH), 1.38–1.34 (t, 3H,
CH3CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–0.86 (t, 3H,
CH3(CH2)14). 31P NMR (CDCl3, 121.5 MHz): −4.84.
Anal Calcd for C28H50NO5P: C 65.73, H 9.85, N 2.74;
found: C 65.60, H 10.01, N 2.86.
EXPERIMENTAL
General
Melting points were determined on a Yanaco melt-
ing point apparatus and uncorrected. 1H NMR spec-
tra were recorded on a Bruker Avance 300 M NMR
spectrometer in CDCl3 with TMS as internal stan-
dard, and coupling constants (J) are expressed in
hertz. Elemental analyses were done on a Yanaco
CHN CORDER MT-3. The reaction progress is fol-
lowed with TLC plates run in PE-EtOAc solvent sys-
tems. Spots were visualized by exposure to UV light
(254 nm) followed by I2 vapor. Flash column chro-
matographies were carried out on silica gel with
PE-EtOAc mixtures under positive pressure.
O-Ethyl O-(3-chlorophenyl) O-2-(stearamido)-
ethyl phosphate (1b). Yellowy solid, mp <30◦C,
yield 40%. 1H NMR (CDCl3, 300 MHz) δ: 7.28–
7.24 (d, 1H, ArH), 7.22–7.19 (t, 1H, ArH), 7.17–
7.09 (dd, 2H, ArH), 6.20 (bs, 1H, NH), 4.26–4.18
(t, 4H, NHCH2CH2O, CH3CH2OP), 3.55–3.51 (dd,
2H, NHCH2-CH2O), 2.15–2.11 (t, 2H, CH2CONH),
1.58–1.55 (t, 2H, CH2CH2CONH), 1.36–1.33 (t, 3H,
CH3CH2OP), 1.23 (s, 28H, (CH2)14), 0.87–0.84 (t, 3H,
CH3(CH2)14). 31P NMR (CDCl3, 121.5 MHz): −5.21.
Anal Calcd for C28H49ClNO5P: C 61.58, H 9.04, N
2.56; found: C 61.29, H 9.43, N 2.39.
Biological Assay
The fungicidal activities of title compounds were
evaluated using mycelium growth rate test. One-
milliliter solution of tested compound in acetone was
added into a culture plate, and then 9 mL PDA cul-
ture medium was added to obtain the flat containing
Heteroatom Chemistry DOI 10.1002/hc

606 Han, Gao, and Li
O-Ethyl O-(4-chlorophenyl) O-2-(stearamido)-
3.59–3.54 (dd, 2H, NHCH2CH2O), 2.17–2.12 (t, 2H,
CH2CONH), 1.62–1.57 (t, 2H, CH2CH2CONH), 1.38–
1.33 (t, 3H, CH3CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–
0.86 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5
MHz): −4.64. Anal Calcd for C28H49FN O5P: C 63.49,
H 9.32, N 2.64; found: C 63.58, H 9.43, N 2.37.
ethyl phosphate (1c). ivory solid, mp 33–34◦C, yield
1
51%. H NMR (CDCl3, 300 MHz) δ: 7.33–7.30 (d,
2H, ArH), 7.17–7.15 (d, 2H, ArH), 6.08 (bs, 1H,
NH), 4.28–4.18 (m, 4H, NHCH2CH2O, CH3CH2OP),
3.58–3.53 (dd, 2H, NHCH2CH2O), 2.16–2.11 (t, 2H,
CH2CONH), 1.61–1.57 (t, 2H, CH2CH2CONH), 1.38–
1.33 (t, 3H, CH3-CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–
0.86 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5
MHz): −5.03. Anal Calcd for C28H49ClNO5P: C 61.58,
H 9.04, N 2.56; found: C 61.59, H 9.03, N 2.68.
O-Ethyl O-(2-nitrophenyl) O-2-(stearamido)ethyl
phosphate (1h). Yellow solid, mp 34–36◦C, yield
33%. 1H NMR (CDCl3, 300 MHz) δ: 7.96–7.93
(d, 1H, ArH), 7.66–7.48 (m, 2H, ArH), 7.37–7.32
(t, 1H, ArH), 6.16 (bs, 1H, NH), 4.36–4.24 (m,
4H, NHCH2CH2O, CH3CH2OP), 3.59–3.54 (dd, 2H,
NHCH2CH2O), 2.20–2.15 (t, 2H, CH2CONH), 1.63–
1.58 (t, 2H, CH2CH2CONH), 1.40–1.35 (t, 3H,
CH3CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–0.86 (t, 3H,
CH3(CH2)14). 31P NMR (CDCl3, 121.5 MHz): −5.94.
Anal Calcd for C28H49N2O7P: C 60.41, H 8.87, N 5.03;
found: C 60.11, H 8.36, N 4.87.
O-Ethyl O-(2,4-dichlorophenyl) O-2-(stearamido)-
ethyl phosphate (1d). yellowy solid, mp <30◦C,
yield 35%. 1H NMR (CDCl3, 300 MHz) δ: 7.44–
7.43 (d, 1H, ArH), 7.39–7.36 (d, 1H, ArH), 7.25–
7.21 (dd, 1H, ArH), 6.25 (bs, 1H, NH), 4.32–4.23
(m, 4H, NHCH2CH2O, CH3CH2OP), 3.60–3.55 (dd,
2H, NHCH2CH2O), 2.18–2.13 (t, 2H, CH2CONH),
1.62–1.57 (t, 2H, CH2CH2CONH), 1.40–1.35 (t, 3H,
CH3CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–0.86 (t, 3H,
CH3(CH2)14). 31P NMR (CDCl3, 121.5 MHz): −5.47.
Anal Calcd for C28H48Cl2 N O5P: C 57.93, H 8.33, N
2.41; found: C 57.82, H 8.25, N 2.41.
O-Ethyl O-(4-nitrophenyl) O-2-(stearamido)ethyl
phosphate (1i). Yellow solid, mp 75–77◦C, yield
1
41%. H NMR (CDCl3, 300 MHz) δ: 8.27–8.24 (d,
2H, ArH), 7.40–7.37 (d, 2H, ArH), 6.24 (bs, 1H,
NH), 4.32–4.23 (m, 4H, NHCH2CH2O, CH3CH2OP),
3.61–3.56 (dd, 2H, NHCH2CH2O), 2.19–2.14 (t, 2H,
CH2CONH), 1.62–1.58 (t, 2H, CH2CH2CONH), 1.41–
1.36 (t, 3H, CH3CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–
0.85 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5
MHz): −5.85. Anal Calcd for C28H49N2O7P: C 60.41,
H 8.87, N 5.03; found: C60.39, H 8.85, N 5.03.
O-Ethyl O-(2-bromophenyl) O-2-(stearamido)-
ethyl phosphate (1e). Yellowy solid, mp <30◦C, yield
1
45%. H NMR (CDCl3, 300 MHz) δ: 7.59–7.57 (d,
1H, ArH), 7.44–7.42 (d, 1H, ArH), 7.31–7.26 (dd,
1H, ArH), 7.08–7.04 (t, 1H, ArH), 6.16 (bs, 1H,
NH), 4.32–4.24 (m, 4H, NHCH2CH2O, CH3CH2OP),
3.58–3.54 (dd, 2H, NHCH2CH2O), 2.16–2.12 (t, 2H,
CH2CONH), 1.59–1.56 (t, 2H, CH2CH2CONH), 1.38–
1.35 (t, 3H, CH3CH2OP), 1.24 (s, 28H, (CH2)14), 0.88–
0.85 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5
MHz): −5.46. Anal Calcd for C28H49BrNO5P: C 56.94,
H 8.36, N 2.37; found: C 56.98, H 8.44, N 2.42.
O-Ethyl O-(3-methyloxyphenyl) O-2-(stearamido)-
ethyl phosphate (1j). Ivory solid, mp 41–43◦C, yield
53%.1H NMR (CDCl3, 300 MHz) δ: 7.27–7.24 (d,
1H, ArH), 7.18–7.13 (t, 1H, ArH), 6.98–6.89 (dd,
2H, ArH), 6.42 (bs, 1H, NH), 4.29–4.24 (t, 4H,
NHCH2CH2O, CH3CH2OP), 3.87 (s, 3H, CH3– OAr),
3.59–3.54 (dd, 2H, NHCH2CH2O), 2.16–2.11 (t, 2H,
CH2CONH), 1.61–1.57 (t, 2H, CH2CH2CONH), 1.39–
1.35 (t, 3H, CH3CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–
0.86 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5
MHz): −4.57. Anal Calcd for C29H52N O6P: C 64.30,
H 9.68, N 2.59; found: C 64.18, H 9.66, N 2.59.
O-Ethyl O-(4-bromophenyl) O-2-(stearamido)-
ethyl phosphate (1f). Off-white solid, mp 38–40◦C,
1
yield 49%. H NMR (CDCl3, 300 MHz) δ: 7.45–7.43
(d, 2H, ArH), 7.10–7.07 (d, 2H, ArH), 6.19 (bs, 1H,
NH), 4.24–4.14 (m, 4H, NHCH2CH2O, CH3CH2OP),
3.55–3.51 (dd, 2H, NHCH2CH2O), 2.13–2.09 (t, 2H,
CH2CONH), 1.58–1.55 (t, 2H, CH2CH2CONH), 1.35–
1.32 (t, 3H, CH3-CH2OP), 1.23 (s, 28H, (CH2)14), 0.87–
0.84 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5
MHz): −5.11. Anal Calcd for C28H49BrNO5P: C 56.94,
H 8.36, N 2.37; found: C 57.10, H 8.38, N 2.30.
O-Ethyl O-(2-methylphenyl) O-2-(stearamido)-
ethyl phosphate (1k). Off-white solid, mp <30◦C,
yield 43%. 1H NMR (CDCl3, 300 MHz) δ: 7.27–
7.06 (m, 4H, ArH), 6.19 (bs, 1H, NH), 4.28–4.18
(m, 4H, NHCH2CH2O, CH3CH2OP), 3.56–3.52 (dd,
2H, NHCH2CH2O), 2.31 (s, 3H, ArCH3), 2.14–2.09
(t, 2H, CH2CONH), 1.60–1.56 (t, 2H, CH2CH2CONH),
1.38–1.33 (t, 3H, CH3CH2OP), 1.25 (s, 28H, (CH2)14),
0.90–0.85 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3,
121.5 MHz): −4.57. Anal Calcd for C29H52NO5P: C
O-Ethyl O-(4-fluorophenyl) O-2-(stearamido)-
ethyl phosphate (1g). Yellowy solid, mp 38–39◦C,
1
yield 40%. H NMR (CDCl3, 300 MHz) δ: 7.20–7.16
(dd, 2H, ArH), 7.06–7.01 (t, 2H, ArH), 6.10 (bs, 1H,
NH), 4.28–4.18 (m, 4H, NHCH2CH2O, CH3CH2OP),
Heteroatom Chemistry DOI 10.1002/hc

Synthesis and Fungicidal Activity of O-Alkyl O-Aryl O-2-(Stearamido)ethyl Phosphates 607
66.26, H 9.97, N 2.66; found: C 66.37, H 10.02, N
2.68.
O-Ethyl O-(2-nitro-4-methylphenyl) O-2-(stear-
amido)ethyl phosphate (1p). Yellow solid, mp 40–
1
42◦C, yield 42%. H NMR (CDCl3, 300 MHz) δ: 7.74
(s, 1H, ArH), 7.44–7.37 (dd, 2H, ArH), 6.12 (bs, 1H,
NH), 4.32–4.22 (m, 4H, NHCH2CH2O, CH3CH2OP),
3.59–3.53 (dd, 2H, NHCH2CH2O), 2.42 (s, 3H,
ArCH3), 2.20–2.15 (t, 2H, CH2CONH), 1.63–1.58 (t,
2H, CH2CH2CONH), 1.39–1.34 (t, 3H, CH3CH2OP),
1.25 (s, 28H, (CH2)14), 0.90–0.86 (t, 3H, CH3(CH2)14).
31P NMR (CDCl3, 121.5 MHz): −5.61. Anal Calcd for
C29H51N2O7P: C 61.03, H 9.01, N 4.91; found: C 61.19,
H 8.99, N 5.01.
O-Ethyl O-(3-methylphenyl) O-2-(stearamido)-
ethyl phosphate (1l). White solid, mp 35–36◦C, yield
30%. 1H NMR (CDCl3, 300 MHz) δ: 7.25–7.20 (t, 1H,
ArH), 7.02–6.99 (d, 3H, ArH), 6.14 (bs, 1H, NH),
4.26–4.18 (m, 4H, NHCH2CH2O, CH3CH2OP), 3.57–
3.53 (dd, 2H, NHCH2CH2O), 2.35 (s, 3H, ArCH3),
2.15–2.10 (t, 2H, CH2CONH), 1.60–1.56 (t, 2H,
CH2CH2CONH), 1.39–1.34 (t, 3H, CH3CH2OP), 1.25
(s, 28H, (CH2)14), 0.90–0.86 (t, 3H, CH3(CH2)14). 31P
NMR (CDCl3, 121.5 MHz): −4.83. Anal Calcd for
C29H52NO5P: C 66.26, H 9.97, N 2.66; found: C 66.28,
H 9.92, N 2.70.
O-Propyl O-phenyl O-2-(stearamido)ethyl phos-
phate (1q). White solid, mp 37–38◦C, yield 43%.
1H NMR (CDCl3, 300 MHz) δ: 7.38–7.33 (t,
2H, ArH), 7.22–7.20 (d, 3H, ArH), 6.18 (bs,
1H, NH), 4.25–4.19 (m, 2H, NHCH2CH2O), 4.16–
4.09 (dd, 2H, CH3CH2CH2OP), 3.57–3.52 (dd, 2H,
NHCH2-CH2O), 2.15–2.09 (t, 2H, CH2CONH), 1.78–
1.67 (m, 2H, CH3CH2CH2OP), 1.61–1.56 (t, 2H,
CH2CH2CONH), 1.25 (s, 28H, (CH2)14), 0.98–0.93 (t,
3H, CH3CH2CH2OP), 0.90–0.86 (t, 3H, CH3(CH2)14).
31P NMR (CDCl3, 121.5 MHz): −4.75. Anal Calcd for
C29H52NO5P: C 66.26, H 9.97, N 2.66; found: C 66.28,
H 10.01, N 2.71.
O-Ethyl O-(3,4-dimethylphenyl) O-2-(stearamido)-
ethyl phosphate (1m). Yellowy solid, mp 34–35◦C,
1
yield 51%. H NMR (CDCl3, 300 MHz) δ: 7.09–7.06
(d, 1H, ArH), 6.98 (s, 1H, ArH), 6.94–6.92
(d, 1H, ArH), 6.20 (bs, 1H, NH), 4.27–4.17 (m,
4H, NHCH2CH2O, CH3CH2OP), 3.56–3.51 (dd, 2H,
NH-CH2CH2O), 2.24 (s, 3H, ArCH3), 2.22 (s, 3H,
ArCH3), 2.14–2.09 (t, 2H, CH2CONH), 1.60–1.55
(t, 2H, CH2CH2CONH), 1.38–1.33 (t, 3H,
CH3CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–0.86
(t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5 MHz):
−4.61. Anal Calcd for C30H54NO5P: C 66.76, H 10.08,
N 2.60; found: C 66.67, H 9.98, N 2.68.
O-(2-Chloroethyl) O-phenyl O-2-(stearamido)-
ethyl phosphate (1r). Yellowy solid, mp 53–55◦C,
1
yield 42%. H NMR (CDCl3, 300 MHz) δ: 7.39–7.34
(t, 2H, ArH), 7.24–7.21 (d, 3H, ArH), 6.08 (bs, 1H,
NH), 4.41–4.33 (m, 2H, NHCH2CH2O), 4.29–4.23 (m,
2H, ClCH2CH2OP), 3.72–3.68 (t, 2H, ClCH2CH2OP),
3.59–3.54 (dd, 2H, NHCH2CH2O), 2.16–2.11 (t, 2H,
CH2CONH), 1.61–1.56 (t, 2H, CH2CH2CONH), 1.25
O-Ethyl O-(2,6-dimethylphenyl) O-2-(stearamido)-
ethyl phosphate (1n). Ivory solid, mp 50–52◦C, yield
38%. 1H NMR (CDCl3, 300 MHz) δ: 7.06–6.97
(m, 3H, ArH), 6.07 (bs, 1H, NH), 4.25–4.18 (m,
4H, NHCH2-CH2O, CH3CH2OP), 3.54–3.50 (dd, 2H,
NHCH2CH2O), 2.36 (s, 6H, ArCH3), 2.14–2.09 (t, 2H,
CH2CONH), 1.60–1.55 (t, 2H, CH2CH2CONH), 1.37–
1.32 (t, 3H, CH3-CH2OP), 1.25 (s, 28H, (CH2)14), 0.90–
0.86 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5
MHz): −4.19. Anal Calcd for C30H54NO5P: C 66.76,
H 10.08, N 2.60; found: C 66.61, H 10.08, N 2.64.
(s, 28H, (CH2)14), 0.90–0.86 (t, 3H, CH3-(CH2)14). 31
P
NMR (CDCl3, 121.5 MHz): −5.29. Anal Calcd for
C28H49ClNO5P: C 61.58, H 9.04, N 2.56; found: C
61.59, H 9.09, N 2.58.
O-(2-Benzyloxyethyl) O-phenyl O-2-(stearamido)-
ethyl phosphate (1s). Off-white solid, mp <30◦C,
1
yield 51%. H NMR (CDCl3, 300 MHz) δ: 7.34–7.30
O-Ethyl O-(4-tertbutylphenyl) O-2-(stearamido)-
(m, 7H, ArH), 7.22–7.16 (m, 3H, ArH), 6.12 (bs,
1H, NH), 4.55 (s, 2H, PhCH2), 4.41–4.25 (m, 2H,
NHCH2CH2O), 4.23–4.13 (m, 2H, BnOCH2-CH2OP),
3.72–3.69 (t, 2H, BnOCH2CH2OP), 3.52–3.47 (dd, 2H,
NHCH2CH2O), 2.09–2.04 (t, 2H, CH2CONH), 1.58–
1.53 (t, 2H, CH2CH2CONH), 1.25 (s, 28H, (CH2)14),
0.90–0.86 (t, 3H, CH3(CH2)14). 31P NMR (CDCl3,
121.5 MHz): −4.88. Anal Calcd for C35H56NO6P: C
68.04, H 9.14, N 2.27; found: C 67.96, H 9.11, N 2.27.
ethyl phosphate (1o). Yellowy solid, mp 54–55◦C,
1
yield 48%. H NMR (CDCl3, 300 MHz) δ: 7.37–7.34
(d, 2H, ArH), 7.13–7.10 (d, 2H, ArH), 6.13 (bs, 1H,
NH), 4.28–4.19 (m, 4H, NHCH2CH2O, CH3CH2OP),
3.58–3.53 (dd, 2H, NHCH2CH2O), 2.16–2.11 (t, 2H,
CH2CONH), 1.62–1.57 (t, 2H, CH2CH2CONH), 1.39–
t
1.34 (t, 3H, CH3- CH2OP), 1.30 (s, 9H, BuH), 1.25
(s, 28H, (CH2)14), 0.90–0.85 (t, 3H, CH3(CH2)14). 31P
NMR (CDCl3, 121.5 MHz): −4.53. Anal Calcd for
C32H58NO5P: C 67.69, H 10.30, N 2.47; found: C
67.77, H 10.38, N 2.34.
O-((Tetrahydrofuran-2-yl)methyl) O-phenyl O-2-
(stearamido)ethyl phosphate (1t). Yellowy solid, mp
Heteroatom Chemistry DOI 10.1002/hc

608 Han, Gao, and Li
70–72◦C, yield 60%. 1H NMR (CDCl3, 300 MHz)
δ: 7.38–7.33 (t, 2H, ArH), 7.23–7.21 (d, 3H, ArH),
5.98 (bs, 1H, NH), 4.34–4.04 (m, 3H, NHCH2CH2O,
THFOCHaHb), 3.93–3.79 (m, 1H,THFOCHaHb),
(g) Molina-Holgado, F.; Lledo´, A.; Guaza, C. Neurore-
port 1997, 8, 1929.
[2] (a) Chapman, K. D.; Venables, B.; Blair, R. Jr.;
Berringer, C. Plant Physiol 1999, 120, 1157; (b)
Schmid, H. H.; Berdyshev, E. V. Prostaglandins
Leukot Essent Fatty Acids 2002, 66, 363; (c) Di Marzo,
V.; De Petrocellis, L.; Fezza, F.; Ligresti, A; Bisogno,
T. Prostaglandins Leukot Essent Fatty Acids 2002, 66,
377; (d) Chapman, K. D.; Tripathy, S.; Venables, B.;
Desouza, A. Plant Physiol 1998, 116, 1163; (e) Stella,
N.; Schweitzer, P.; Piomelli, D. Nature(London) 1997,
388, 773; (f) Tripathy, S.; Venables, B. J.; Chapman,
K. D. Plant Physiol 1999, 121, 1299; (g) Chapman,
K. D.; Venables, B. J.; Dian, E. E.; Gross, G. W.
3.75–3.72 (t, 2H,
CH2CH2CH2OCH ), 3.57–
3.53 (t, 1H, CH2CH2CH2OCH ), 3.46–3.41 (dd,
2H, NHCH2CH2O), 2.24–2.18 (t, 2H, CH2CONH),
2.16–2.11 (t, 2H, CH2CH2CH2OCH ), 2.05 1.81
(t, 2H,
CH2CH2CH2OCH ), 1.66–1.61 (t, 2H,
CH2CH2CONH), 1.25 (s, 28H, (CH2)14), 0.90–0.86
(t, 3H, CH3(CH2)14). 31P NMR (CDCl3, 121.5 MHz):
−4.92. Anal Calcd for C31H54NO5P: C 67.48, H 9.86,
N 2.54; found: C 67.45, H 9.56, N 2.89.
J
Am Oil Chem Soc (JAOCS) 2003, 80, 223;
(h) Giuffrida, A.; Rodriguez de Fonseca, F.; Piomelli,
D. Anal Biochem 2000, 280, 87; (i) Schmid, P. C.;
Schwartz, K. D.; Smith, C. N.; Krebsbach, R. J.;
Berdyshev, E. V.; Schmid, H. H. Chem Phys Lipids
2000, 104, 185.
ACKNOWLEDGMENTS
We thank Prof. Wei-Ming Guo for her helpful advice
and kind assistance.
[3] (a) Chapman, K. D. Chem Phys Lipids 2000, 108, 221;
(b) Tripathy, S.; Kleppinger-Sparace, K.; Dixon, R. A.;
Chapman, K. D. Plant Physiol 2003, 131, 1781.
[4] Blancaflor, E. B.; Hou, G.; Chapman, K. D. Planta
2003, 217, 206.
[5] Chapman, K. D. Prog Lipid Res 2004, 43, 302.
[6] Motes, C. M.; Pechter, P.; Yoo, C. M.; Wang, Y. S.;
Chanpman, K. D.; Blancaflor, E. B. Protoplasma
2005, 226(3–4), 109.
[7] Chapman, K. D. WO 01/30143 A2, 2001.
[8] Zhang, Y.; Guo, W. M.; Chen, S. M.; Han, L.; Li, Z.
M. J Plant Physiol 2007, 164, 993.
[9] Han, L.; Li, Z. M.; Zhang, Y.; Guo, W. M. Chin J Org
Chem 2006, 26, 242.
REFERENCES
[1] (a) Kuehl, F. F.; Jacob, T. A.; Ganley, O. H.; Ormond,
R. E.; Meisinger, M. A. P. J Am Chem Soc 1957, 79,
5577; (b) Parinandi, N. L.; Schmid, H. H. O. FEBS
Lett 1988, 237, 49; (c) Gulaya, N. M.; Kuzmenko,
A. I.; Margitich, V. M.; Govseeva, N. M.; Melnichuk,
S. D; Goridko, T. M.; Zhukov, A. D. Chem Phys Lipids
1998, 97, 49; (d) Skaper, S. D.; Buriani, A.; Dal Toso,
R.; Petrelli, L.; Romanello, S.; Facci, L.; Leon, A.
Proc Natl Acad Sci USA 1996, 93, 3984; (e) Facci, L.;
Dal Toso, R. Proc Natl Acad Sci USA 1995, 92, 3376;
(f) Mazzari, S.; Canella, R.; Petrelli, L.; Marcolongo,
G.; Leon, A. Eur J Pharmacol 1996, 300, 227;
[10] Han, L.; Gao, J. R.; Li, Z. M.; Zhang, Y.; Guo, W. M.
Bioorg Med Chem Lett 2007, 17, 3231.
[11] Roe, E. T.; Miles, T. D.; Swern, D. J Am Chem Soc
1952, 74, 3442.
Heteroatom Chemistry DOI 10.1002/hc