Novel deoxyxylulosephosphate-reductoisomerase inhibitors: Fosmidomycin derivatives with spacious acyl residues

Article abstract of DOI:10.1002/ardp.200700149

1-Deoxy-D-xylulose-5-phosphate reductoisomerase (Dxr) represents an essential enzyme of the mevalonate-independent pathway of the isoprenoid biosynthesis. Using fosmidomycin as a specific inhibitor of Dxr, this enzyme was previously validated as target for the treatment of malaria and bacterial infections. The replacement of the formyl residue of fosmidomycin by spacious acyl residues yielded inhibitors active in the micromolar range. As predicted by flexible docking, evidence was obtained for the formation of a hydrogen bond between an appropriately placed carbonyl group in the acyl residue and the main-chain NH of Met214 located in the flexible catalytic loop of the enzyme.

Full text of DOI:10.1002/ardp.200700149

Arch. Pharm. Chem. Life Sci. 2007, 340, 483–490
R. Ortmann et al.
483
Full Paper
Novel Deoxyxylulosephosphate-Reductoisomerase Inhibitors:
Fosmidomycin Derivatives with Spacious Acyl Residues
Regina Ortmann1, Jochen Wiesner2, Katrin Silber1, Gerhard Klebe1, Hassan Jomaa2, and
Martin Schlitzer1
1 Institut fꢀr Pharmazeutische Chemie, Philipps-Universitꢁt Marburg, Marburg, Germany
2 Institut fꢀr Klinische Chemie und Pathobiochemie, Universitꢁtsklinikum Giessen und Marburg GmbH,
Gießen, Germany
1-Deoxy-D-xylulose-5-phosphate reductoisomerase (Dxr) represents an essential enzyme of the
mevalonate-independent pathway of the isoprenoid biosynthesis. Using fosmidomycin as a spe-
cific inhibitor of Dxr, this enzyme was previously validated as target for the treatment of
malaria and bacterial infections. The replacement of the formyl residue of fosmidomycin by spa-
cious acyl residues yielded inhibitors active in the micromolar range. As predicted by flexible
docking, evidence was obtained for the formation of a hydrogen bond between an appropriately
placed carbonyl group in the acyl residue and the main-chain NH of Met214 located in the flexi-
ble catalytic loop of the enzyme.
Keywords: 1-Deoxy-D-xylulose-5-phosphate-reductoisomerase inhibitors / Non-mevalonate biosynthesis / Structure-
activity relationships /
Received: March 6, 2007; accepted: July 12, 2007
DOI 10.1002/ardp.200700149
well-known inhibitors of Dxr [2]. The efficacy of fosmido-
mycin for the treatment of acute uncomplicated malaria
was recently proven in several clinical trials [3], in addi-
tion to some earlier studies on the treatment of urinary
tract infections. However, relatively high doses of fosmi-
domycin were required to achieve the desired results.
This seems in part to be due to the highly polar nature of
this molecule.
Therefore, we addressed the question whether it might
be possible to obtain more lipophilic Dxr inhibitors by
replacing the formyl or acetyl residue of fosmidomycin
or FR900098, respectively, by more spacious residues [4].
Introduction
Isopentenyldiphosphate is the common precursor of a
wide variety of isoprenoid compounds. The biosynthesis
of isopentenyldiphosphate can be achieved by two differ-
ent pathways. Higher animals and some other organisms
exclusively use the well-known mevalonate pathway. In
contrast, the alternative pathway, the 1-deoxy-D-xylulose-
5-phosphate (DOXP) pathway is unique to a variety of
microorganism including such important pathogens as
the malaria parasite Plasmodium falciparum and Mycobac-
terium tuberculosis, the causative agent of tuberculosis.
Therefore, enzymes of the DOXP pathway provide attrac-
tive targets for the development of novel antimicrobial
agents. One important step of the DOXP pathway is the
reductive isomerisation of DOXP to 2-C-methyl-D-eryth-
riol 4-phosphate, catalyzed by DOXP-reductoisomerase
(Dxr) [1]. Fosmidomycin and its derivative FR900098 are
Chemistry
Synthesis of the target compounds 7a–j started with the
substitution of the bromine of the acetal 1 by dibenzyl-
phosphite 2 to yield the intermediate 3 (Scheme 1). Acidic
hydrolysis of the acetalic substructure of intermediate 3
yielded the free aldehyde 4 which underwent a reductive
amination with O-benzylhydroxylamine to obtain the
protected hydroxylamine 5. This was acylated by appro-
Correspondence: Martin Schlitzer, Institut fꢀr Pharmazeutische Chemie,
Philipps-Universitꢁt, Marbacher Weg 6, D-35037 Marburg, Germany
E-mail: Schlitzer@staff.uni-marburg.de
Fax: +49 6421 28-25978
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R. Ortmann et al.
Arch. Pharm. Chem. Life Sci. 2007, 340, 483–490
308C for 5 min. The reaction was started by the addition
of DOXP. The decrease of absorption was monitored at
340 nm using a SpectraMax 340PC microplate reader
(Molecular Devices, Ismaning, Germany).
It is well known that fosmidomycin and its analogues
are slow binding inhibitors of DXR. With respect to the
slow, tight-binding properties of fosmidomycin, the
inhibitors were pre-incubated with the enzyme and
NADPH before the reaction was started by addition of
DOXP.
Flexible docking
The protein structure was taken from pdb-entry 1ONP [6].
Ligands and solvent molecules were removed, but the
metal ion was included as part of the protein. To suggest a
reasonable cofactor binding mode NADPH coordinates
were transferred from pdb ID: 1JVS [7] after both protein
structures were superimposed, based on Ca coordinates. A
subsequent minimization of the transferred cofactor
with the MAB force field, as implemented in MOLOC [8],
revealed no significant movements. For the use within
AutoDock 3.0 [9], polar hydrogens were added with the
PROTONATE utility from AMBER [10], AMBER united
atom force field charges were assigned [11], and solvation
parameters were added using the ADDSOL utility from
AutoDock 3.0. Ligand structures were built in mol2-for-
mat, partial atomic charges were assigned (AM1 Hamilto-
nian [12]) within the semiempirical package MOPAC 6.0
[13] and all bonds except the one in the C(=O)–NO-frag-
ment, were kept rotatable. Docking runs were performed
using the Lamarckian genetic algorithm as implemented
in AutoDock 3.0, using an initial population of 50 ran-
domly placed individuals, a maximum number of
1.56106 energy evaluations, a mutation rate of 0.02, a
crossover rate of 0.80 and an elitism value of 1. Generated
ligand docking solutions, mutually differing by rmsd
f1 ꢀ were clustered together and the lowest docking
energy found for one entry of a cluster was used as repre-
sentative. The results of the highest ranked clusters were
chosen to compare structure-activity relationships. This
protocol was initially validated by docking fosmidomycin
back into its original protein crystal structure (1ONP) [14].
(I) NaH, DMF; (II) HCl/H2O; (III) O-benzylhydroxylamine; (IV) Na[B(CN)H3], HCl,
MeOH; (V) R-CO-Cl (glutaric anhydride/pyridine in case of 6d); (VI) H2, Pd/C, MeOH.
Scheme 1. Synthesis route of compounds1–7.
priate acyl chlorides to yield the fully benzyl protected
intermediate 6. The yields of the acylation were different
and in most cases not satisfying. The preparation was not
optimized because it was our main intention to get the
compounds for biological testing. The final step in the
synthesis was the hydrogenolytic removal of the benzyl
protective groups providing the target compounds 7.
Under these conditions, the keto groups of 7b and 7c are
stable.
DOXP reductoisomerase inhibition assay
Recombinant Dxr of E. coli was prepared as described pre-
viously [5]. In brief, the dxr gene was PCR-amplified from
genomic DNA and inserted into the pQE9 expression vec-
tor (Qiagen, Hilden, Germany). The resulting protein
with an amino-terminal His6-tag was purified by immobi-
lized metal affinity chromatography on Talon superflow
resin (Clontech, Heidelberg, Germany) followed by anion
exchange chromatography on source 15Q (GE Health-
care, Munich, Germany). Final purification was achieved
by gel permeation chromatography using an XK 16/60
Superdex 75 column (GE Healthcare). The protein was
concentrated to 15 mg/mL by ultrafiltration and stored
in aliquots at –208C until use. The assay was performed
in a reaction mixture containing 100 mM Tris HCl
(pH 7.5), 0.2% bovine serum albumin, 1 mM MnCl2, 1 mM
DOXP, 0.3 mM NADPH, and 1 lg/mL recombinant DOXP
reductoisomerase of E. coli in a final volume of 0.25 mL.
The mixture without substrate was pre-incubated with a
serial dilution of the test compounds on a 96-well plate at
Results and discussion
Aiming at the development of more lipophilic inhibitors,
a series of fosmidomycin derivatives with variable acyl
residue was synthesized. As a general finding, the intro-
duction of all alternative acyl residues tested led to a sig-
nificant loss of Dxr inhibitory activity compared to the
lead compounds fosmidomycin and FR900098 (Table 1).
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Arch. Pharm. Chem. Life Sci. 2007, 340, 483–490
Fosmidomycin Derivatives with Spacious Acyl Residues
485
Table 1. Structure and activity of compounds 7a–e.a)
Compd.
R-CO-
IC50(lM)
fosmidomycin
0.035
FR900098
0.035
10
7a
7b
7c
20
5.4
7d
7e
A30
9.0
Figure 1. Docking solution of compound 7c (light grey) showing
a possible hydrogen bond between the carbonyl oxygen and the
main-chain NH of Met214 (heavy atom distance 2.8 ꢂ) superim-
posed with the fosmidomycin crystal structure as observed in
1ONP (dark grey). The picture shows the cofactor in sticks at the
right side. The sphere represents the metal. The protein is cov-
ered by a surface but the flexible loop is shown in thin sticks.
Residues interacting to the phosphonate group are labeled.
7f
5.6
7g
A 30
7h
7i
15
5.1
tion to Met214 as well as on the chemical nature of the
modification. The derivative 7d showed decreased activ-
ity although the interaction to Met214 is comparable to
7c. Considering the desolvation needed to get the ligand
inside the protein cavity, it is obvious that binding of the
5-oxohexanoyl derivative 7c is more favorable than the
diacid derivate 7d.
7j
1.3
a)
IC50 values were determined for the inhibition of recombi-
nant Dxr from E. coli.
A comparable increase in activity as with the introduc-
tion of the 5-carbonyl group into the acyl chain could
also be obtained by the elongation of the acyl chain by 10
As observed via flexible docking of all compounds, the
acyl substituents prevent the formation of the desired
hydroxamate-metal interaction geometry. This results in
a loss of binding affinity. However, it seems possible that
some activity could be regained again by an appropri-
ately placed hydrogen-bond acceptor forming a hydro-
gen bond with the backbone NH of Met214 (Figure 1). It
was predicted by crystallographic studies (pdb entry
1ONP [6]) that Met214 is part of a flexible loop which cov-
ers the active-site cleft after substrate binding. Indeed,
the 5-oxohexanoic acid derivative 7c displayed a twofold
enhanced activity in comparison to the hexanoic acid
derivative 7a (IC50 = 5.4 lM vs. IC50 = 10 lM). This effect
proved to be dependent on the length of the substitution
as the docking mode of 7b shows a less favorable interac-
methylene units (hexadecanoic acid derivative 7f: IC50
=
5.6 lM), while the dodecanoyl derivative 7e displayed
similar activity as the hexanoyl derivative 7a. Further
elongation of the acyl chain, however, led to a virtually
complete loss of activity (7g: IC50 A 30 lM). Inhibitor 7i
(IC50 = 5.1 lM) demonstrated that a part of the alkyl chain
can be replaced by a phenyl residue, provided, the whole
molecule has a critical length since the shorter naphthy-
lacetyl derivative 7h was threefold less active. In contrast,
the pentafluoro benzoyl derivative was the best com-
pound in this series with an IC50 of 1.3 lM.
In the light of the relatively low activity of the com-
pounds against E.coli Dxr no further assays against P. falci-
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Arch. Pharm. Chem. Life Sci. 2007, 340, 483–490
P-CH2); 26.78 (d, 2JC,P = 3.9 Hz, P-CH2–CH2); 65.05 (26O-CH2); 67.11
parum Dxr or cultured parasites will be performed. Our
results are in accordance to other work published in
which no improvement on inhibitory activity could be
achieved by modifying the acyl residue [14, 15].
2
3
(d, JC,P = 6.6 Hz, 26P-O-CH2); 103.20 (d, JC,P = 19.1 Hz, O-CH);
127.84, 128.32, 128.55 (10 Ar-C); 136.41 (d, 3JC,P = 5.6 Hz, 2 q Ar-C).
31P-NMR (CDCl3, 202 MHz), d (ppm): 33.81. CI-MS, m/z: 363
([M+1]+).
In summary, several Dxr inhibitors active in the micro-
molar range were obtained by replacing the formyl
group of fosmidomycin by more lipophilic and spacious
acyl residues. Introduction of a carbonyl group into the
acyl residue resulted in a slight increase in activity in
comparison to the compound lacking this moiety, pre-
sumably by forming a hydrogen bond to a main-chain
NH of a flexible loop of the target enzyme located over
the active site. However, with respect to the reduced
activity compared to the parent compound alternative
modifications of the fosmidomycin lead structure need
to be investigated in order to develop new inhibitors
with improved therapeutic potential.
(3-Oxopropyl)phosphonic acid dibenzyl ester 4
An amount of 5.5 g (15 mmol) of (2-[1,3]-dioxolan-2-yl-ethyl)phos-
phonic acid dibenzyl ester 3 was dissolved in a few mL acetone
and 30 mL 2 N HCl were added. After stirring at room tempera-
ture for three days, the solution was extracted three times with
dichloromethane. The organic extracts were combined, dried
(Na2SO4), and then the solvent was removed by rotary evapora-
tion. Yield 71%. IR (film), cm– 1: 3090, 3065, 3033, 2953, 2894,
2831, 2732, 1724 (C=O).1H-NMR (CDCl3, 500 MHz), d (ppm): 2.01–
2.07 (m, 2 H, CH2); 2.66–2.72 (m, 2 H, CH2); 4.94–5.08 (m, 4 H,
26P-O-CH2); 7.32–7.38 (m, 10 H, Ar-H); 9.69 (s, 1 H, H–C=O). 13C-
1
NMR (CDCl3, 125 MHz, COM), d (ppm): 18.47 (d, JC,P = 144 Hz, P-
2
2
CH2); 36.80 (d, JC,P = 3.9 Hz, P-CH2-CH2); 67.51 (d, JC,P = 5.6 Hz,
3
26P-O-CH2); 128.10, 128.65, 128.75 (10 Ar-C); 136.24 (d, JC,P
=
5.8 Hz, 2 q Ar-C), 199.03 (d, 3JC,P = 16.5 Hz (C=O). 31P-NMR (CDCl3,
202 MHz), d (ppm): 32.58. CI-MS, m/z: 319 ([M+1]+).
This work was supported under the thematic programme
,Quality of life and management of living resources' (QoL) of
the 5th EU Framework Program, contract number QLK2-CT-
2002-00887. Technical assistance was provided by Dajana
Henschker.
3-[(Benzyloxyamino)propyl]phosphonic acid
dibenzyl ester 5
To a stirred solution of 3.2 g (10 mmol) 4 in 10 mL methanol was
added dropwise a solution of 1.6 g (10 mmol) O-benzylhydroxyl-
ammonium chloride and 1 g (10 mmol) triethylamine in 10 mL
methanol. The reaction mixture was refluxed for 1 h, cooled to
room temperature, and 20 mL methanol were added. After addi-
tion of 1.9 g (30 mmol) sodium cyanoborohydride, 20 mL conc.
HCl were added dropwise and the mixture was stirred overnight.
Half of the methanol was removed by rotary evaporation. The
remaining residue was treated with ice-water, adjusted to pH 10
with aqueous KOH solution and extracted with dichlorome-
thane. The combined organic layers were dried over Na2SO4 and
concentrated. The remaining oil was purified by column chro-
matography with ethylacetate : methanol 95 : 5 as eluent. Yield
71%. IR (film), cm– 1: 3247 (NH), 3088, 3963, 3031.1H-NMR (CDCl3,
500 MHz), d (ppm): 1.77–1.85 (m, 4 H, 26CH2); 2.91 (m, 2 H, N-
CH2); 4.64 (s, 2 H, N-O-CH2); 4.94–5.07 (m, 4 H, 26P-O-CH2); 5.52
(bs, 1 H, NH); 7.27–7.40 (m, 15 H, Ar-H). 13C-NMR (CDCl3,
100 MHz, COM), d (ppm): 20.50 (d, 2JC,P = 4.6 Hz, P-CH2-CH2); 23.56
(d, 1JC,P = 141.4 Hz, P-CH2); 52.01 (d, 3JC,P = 16.9 Hz, N-CH2); 67.07 (d,
2JC,P = 6.5 Hz, 2 P-O-CH2); 76.36 (N-O-CH2); 127.93, 127.90, 128.38,
Experimental
1H- and 13C-NMR (COM: Complete decoupling, DEPT: Distortion-
less Enhancemant by Polarisation Transfer) spectra were
recorded on a Jeol Eclipse 400 and a Jeol Eclipse 500 spectrome-
ter (JEOL, Tokyo, Japan). Mass spectra were obtained with a PE
Biosystems API 2000 (Applied Biosystems, Foster City, CA, USA)
or with a Jeol MStation JMS 700 (in case of FAB spectra). IR spec-
tra were recorded on a JASCO FT/IR – 410 Fourier Transform
Infrared Spectrometer (JASCO Germany, Gross-Umstadt, Ger-
many). Microanalyses were obtained from an elementar vario el.
Melting points were obtained with a Reichert Austria micro-
scope (C. Reichert, Vienna, Austria) and are uncorrected. Liquid
chromatography was carried out using silica gel 60 from ICM Sil-
itech.
3
128.48 128.58 (15 Ar-C); 136.42 (d, JC,P = 6.1 Hz, 2 P-O-CH2-C);
(2-[1,3]-Dioxolan-2-yl-ethyl)phosphonic acid dibenzyl
ester 3
137.80 (1 q Ar-C). 31P-NMR (CDCl3, 202 MHz), d (ppm): 34.00. CI-
MS, m/z (%): 426 (46, [M+H]+).
Sodium hydride (528 mg; 22 mmol) was suspended in DMF
under argon atmosphere and the mixture was cooled with an
ice bath. Then, 4.4 mL (20 mmol) dibenzylphosphite were added
dropwise. After 1 h, 3.0 mL (25 mmol) 2-(2-bromoethyl)-1,3-diox-
olan were added with further cooling. The mixture was stirred
overnight at room temperature. The mixture was diluted with
dichloromethane and washed with water and brine, and dried
(Na2SO4). After evaporation of the solvent, the residue was puri-
fied by column chromatography eluting with dichloromethane
methanol 98 : 2. Yield 90%. IR (film), cm– 1: 3090, 3064, 3033,
General procedure for the preparation of benzyl
protected derivatives 6
Method A (Synthesis of acid chlorides):
The appropriate carboxylic acid was dissolved or suspended in
a small amount of dichloromethane. After addition of oxalyl
chloride (0.2 mL per mmol acid) and DMF (two drops per mmol
acid) the mixture was stirred at room temperature for 2 h. Vola-
tiles were evaporated in vacuo and the resulting residue was used
without further purification.
1
2953, 2887. H-NMR (CDCl3, 500 MHz), d (ppm): 1.85–1.96 (m, 4
H, 26CH2); 3.81–3.93 (m, 4 H, 26O-CH2); 4.88–4.90 (m, 1 H, O-
CH); 4.95–5.07 (m, 4 H, 26P-O-CH2); 7.29–7.37 (m, 10 H, Ar-H).
13C-NMR (CDCl3, 125 MHz, COM), d (ppm): 19.93 (d, 1JC,P = 144 Hz,
Method B:
1 mmol 3-[(Phenylmethoxyamino)propyl]phosphonic acid di-
benzyl ester 5 and 1.1 mmol triethylamine were dissolved in
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Arch. Pharm. Chem. Life Sci. 2007, 340, 483–490
Fosmidomycin Derivatives with Spacious Acyl Residues
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dichloromethane. The appropriate acid chloride (1.1 mmol) was
added dropwise and the reaction mixture was stirred overnight.
The solution was washed with aqueous NaHCO3 and water and
was dried (Na2SO4). The solvent was removed and the residue was
purified by column chromatography on silica gel, eluting with
dichloromethane methanol mixtures.
CH2); 46.23 (N-CH2); 61.52 (d, 2JC,P = 6.7 Hz, 26P-O-CH2); 76.63 (N-O-
CH2); 127.91, 128.22, 128.48, 128.54, 128.56, 129.48 (15 Ar-C);
138.00 (d, 3JC,P = 5.4 Hz, 2 x P-O-CH2-C); 139.95 (1 q Ar-C); 173.59 (N-
C=O); 207.82 (H3C-C=O).31P-NMR (CDCl3, 161 MHz), d (ppm): 33.96.
HRFAB-MS m/z 538.2352 [M+H]+ (calcd. 538.2359 for
C30H36NO6PH).
Method C:
1 mmol 3-[(Phenylmethoxyamino)propyl]phosphonic acid di-
benzyl ester 5 and 2 mmol of the appropriate acid chloride were
dissolved in toluol and refluxed for 2 h. The solution was washed
with aqueous NaHCO3 and water and was dried (Na2SO4). The sol-
vent was removed and the residue was purified by column chro-
matography on silica gel, eluting with dichloromethane metha-
nol mixtures.
{3-[Glutaryl(benzyloxy)amino]propyl}phosphonic acid
dibenzyl ester 6d
1 mmol 3-[(Phenylmethoxyamino)propyl]phosphonic acid di-
benzyl ester 5 and 1.5 mmol glutaric anhydride were dissolved
in pyridine and heated at 808C under argon atmosphere for 2 h.
The pyridine was removed under vacuum. The residue was dis-
solved in ethylacetate/water, the pH was adjusted to 1–2 with
2 N HCl and the solution was extracted with ethyl acetate. The
combined organic layers were dried (Na2SO4), the solvent was
removed, and the remaining residue was purified by column
chromatography on silica gel, eluting with dichloromethane
methanol 95 : 5. Yield 63%. IR (film), cm– 1: 3064 (OH), 1726
{3-[Hexanoyl(benzyloxy)amino]propyl}phosphonic acid
dibenzyl ester 6a
Prepared according to general method B from commercially
available hexanoylchloride. Yield 18%. IR (film), cm– 1: 1661
3
1
(C=O). 1H-NMR (CDCl3, 500 MHz), d (ppm): 0.88 (t, J = 7 Hz, 3 H,
(C=O), 1659 (N-C=O). H-NMR (CDCl3, 500 MHz), d (ppm): 1.73–
CH3); 1.22–1.32 (m, 4 H, 26CH2); 1.54–1.61 (m, 2 H, CH2); 1.71-
1.94 (m, 4 H, 26CH2); 2.35 (t, 3J = 7.5 Hz, 2 H, CH2); 3.67-3.62 (m, 2
H, N-CH2); 4.75 (s, 2 H, N-O-CH2); 4.92–5.07 (m, 4 H, 26P-O-CH2);
1.80 (m, 2 H, CH2); 1.85–1.93 (m, 4 H, 26CH2); 2.35 (t, 3J = 7.1 Hz,
2 H, CH2); 2.44 (t, 3J = 6.9 Hz, 2 H, CH2); 3.64 (m, 2 H, N-CH2); 4.73
(s, 2 H, N-O-CH2); 4.92–5.05 (m, 4 H, 26P-O-CH2); 7.28–7.36 (m,
15 H, Ar-H). 13C-NMR (CDCl3, 125 MHz, COM), d (ppm): 19.58
7.30–7.37 (m, 15 H, Ar-H). 13C-NMR (CDCl3, 125 MHz, COM), d
1
1
(ppm): 13.93 (CH3); 20.13 (P-CH2-CH2); 22.40 (CH2); 23.36 (d, JC,P
=
(CH2); 20.13 (CH2); 23.28 (d, JC,P = 141 Hz, P-CH2); 31.15 (CH2);
143 Hz, P-CH2); 24.23 (CH2); 31.54 (CH2); 32.28 (CH2); 67.14 (d, 2JC,P
= 6.7 Hz, 2 P-O-CH2); 76.34 (N-O-CH2); 127.89, 128.36, 128.38,
128.47, 128.55, 128.57, 128.67, 128.90, 129.09 (15 Ar-C); 136.27,
136.32, 136.35 (3 q Ar-C); 172.01 (N-C=O). 31P-NMR (CDCl3, 202
MHz), d (ppm): 33.17. HRFAB-MS m/z 524.2564 [M+H]+ (calcd.
524.2566 for C30H38NO5PH).
33.18 (CH2); 45.68 (N-CH2); 67.33 (d, JC,P = 6.7 Hz, 26P-O-CH2);
2
76.37 N-O-CH2); 127.94, 128.39, 128.55, 128.68, 128.95, 129.19
(15 Ar-C); 134.20 (1 q Ar-C); 136.18 (d, 3JC,P = 5.6 Hz, 26P-O-CH2-C);
174.22 (N-C=O); 177.06 (COOH). 31P-NMR (CDCl3, 202 MHz), d
(ppm): 33.16. HRFAB-MS m/z 540.2136 [M+H]+ (calcd. 540.2151 for
C29H34NO7PH).
{3-[(4-Oxopentanoyl)-(benzyloxy)amino]propyl}-
{3-[Dodecanoyl(benzyloxy)amino]propyl}phosphonic acid
phosphonic acid dibenzyl ester 6b
dibenzyl ester 6e
Prepared acording to the general methods A and B from levu-
linic acid. Yield 12%. IR (film), cm– 1: 1716 (C=O), 1657 (N-C=O).1H-
NMR (CDCl3, 500 MHz), d (ppm): 1.71–1.78 (m, 2 H, CH2); 1.87–
1.97 (m, 2 H, CH2); 2.17 (s, 3 H, CH3); 2.66–2.71 (m, 4 H, C=O-CH2-
CH2); 3.62–3.66 (m, 2 H, N-CH2); 4.82 (s, 2 H, N-O-CH2); 4.92-5.06
(m, 4 H, 26P-O-CH2); 7.30–7.35 (m, 15 H, Ar-H). 13C-NMR (CDCl3,
100 MHz, COM, DEPT), d (ppm): 20.13 (d, 2JC,P = 4.5 Hz, P-CH2-CH2);
Prepared according to general method B from commercially
available lauroyl chloride. Yield 30%. IR (film), cm– 1: 1662 (C=O).
1H-NMR (CDCl3, 500 MHz), d (ppm): 0.88 (t, 3J = 7.0 Hz, 3 H, CH3);
3
1.25 (m, 16 H, 86CH2); 1.56 (t, J = 7.4 Hz, 2 H, CH2); 1.72–1.78
(m, 2 H, CH2); 1.87-1.96 (m, 2 H, CH2); 2.35 (t, 3J = 7.4 Hz, 2 H, CH2);
3.65 (m, 2 H, N-CH2); 4.74 (s, 2 H, N-O-CH2); 4.92–5.05 (m, 4 H,
26P-O-CH2); 7.30–7.37 (m, 15 H, Ar-H). 13C-NMR (CDCl3,
125 MHz, COM), d (ppm): 14.07 (CH3); 20.20 (d,2JC,P = 5.6 Hz, P-CH2-
CH2); 22.64 (CH2-CH3); 23.37 (d, 1JC,P = 142 Hz, P-CH2); 25.55, 29.29,
29.36, 29.39, 29.46, 29.59, 31.87, 32.35 (96CH2); 45.42 (N-CH2);
1
23.25 (d, JC,P = 141 Hz, P-CH2); 26.29 (N-C=O-CH2); 29.89 (CH3);
37.43 (H3C-C=O-CH2); 45.58 (N-CH2); 67.12 (d, 2JC,P = 6.7 Hz, 26P-O-
CH2); 76.38 (N-O-CH2); 127.86, 128.30, 128.51, 128.63, 128.84,
129.13 (15 Ar-C); 134.39 (1 q Ar-C); 136.32 (d, 3JC,P = 5.6 Hz, 2 P-O-
CH2-C); 173.57 (N-C=O); 207.26 (H3C-C=O). 31P-NMR (CDCl3,
202 MHz), d (ppm): 33.16. HRFAB-MS m/z 524.2199 [M+H]+ (calcd.
524.2202 for C29H34NO6PH).
2
67.12 (d, JC,P = 5.8 Hz, 26P-O-CH2); 76.36 (N-O-CH2); 127.87,
128.33, 128.54, 128.65, 128.87, 129.06 (15 Ar-C); 134.42 (1 q Ar-C);
3
136.31 (d, JC,P = 6.6 Hz, 26P-O-CH2-C); 175.16 (C=O). 31P-NMR
(CDCl3, 202 MHz), d (ppm): 33.18. CI-MS m/z 608 [M+H]+. Anal.
calcd. for C36H50NO5P (%): C 71.14, H 8.29, N 2.30. Found (%): C
71.06, H 8.40, N 2.31.
{3-[(5-Oxohexanoyl)-(benzyloxy)amino]propyl}-
phosphonic acid dibenzyl ester 6c
{3-[Hexadecanoyl(benzyloxy)amino]propyl}phosphonic
Prepared acording to the general methods A and B from 4-acetyl
butyric acid. Yield 5% [16]. IR (film), cm– 1: 1713 (C=O), 1658 (N-
C=O). 1H-NMR (CDCl3, 500 MHz), d (ppm): 1.70–1.93 (m, 6 H,
36CH2); 2.08 (s, 3 H, CH3); 2.37 (t, 3J = 7.0 Hz, 2 H, CH2); 2.43 (t, 3J =
7.1 Hz, 2 H, CH2); 3.64 (t, 3J = 6.4 Hz, 2 H, N-CH2); 4.73 (s, 2 H, N-O-
acid dibenzyl ester 6f
Prepared according to general method B from commercially
available palmitoyl chloride. Yield 41%. IR (film), cm– 1: 1661
1
(C=O). H-NMR (CDCl3, 500 MHz), d (ppm): 0.86 (t, 3J = 7 Hz, 3 H,
CH2); 4.92-5.06 (m, 4 H, 26P-O-CH2); 7.30–7.36 (m, 15 H, Ar-H).
CH3); 1.24–1.28 (m, 2 H) (CH2); 1.53–1.55 (m, 24 H, CH2); 1.72–
1.77 (m, 2 H, CH2); 1.89–1.90 (m, 2 H, CH2); 2.32–2.35 (m, 2 H,
CH2); 3.60–3.64 (m, 2 H, N-CH2); 4.72 (s, 2 H, N-O-CH2); 4.92–5.06
(m, 4 H, 26P-O-CH2); 7.30–7.35 (m, 15 H, Ar-H). 13C-NMR (CDCl3,
2
13C-NMR (CDCl3, 100 MHz, COM, DEPT), d (ppm): 16.42 (d, JC,P
=
=
1
4.1 Hz, P-CH2-CH2); 18.87 (H3C-C=O-CH2-CH2); 23.25 (d, JC,P
141 Hz, P-CH2); 26.04 (N-C=O-CH2); 29.90 (CH3); 42.39 (H3C-C=O-
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488
R. Ortmann et al.
Arch. Pharm. Chem. Life Sci. 2007, 340, 483–490
125 MHz, COM, DEPT), d (ppm): 14.05 (CH3); 20.15 (d, 2JC,P = 4.5 Hz,
P-CH2-CH2); 22.61 (CH2-CH3); 23.32 (d, 1JC,P = 142 Hz, P-CH2); 24.52,
29.27, 29.32, 29.35, 29.43, 29.56, 29.58, 29.61, 31.84, 32.30
{3-[(Benzyloxy)(pentafluorophenyl)amino]propyl}-
phosphonic acid dibenzyl ester 6j
Prepared according to general method B from commercially
available pentafluorobenzoic acid chloride. Yield 68%. IR (film),
cm– 1: 1667 (C=O). 1H-NMR (CDCl3, 500 MHz), d (ppm): 1.81–1.88,
(m, 2 H, CH2); 2.00–2.07 (m, 2 H, CH2); 3.84 (t, 3J = 6.7 Hz, 2 H, N-
CH2); 4.59 (s, 2 H, N-O-CH2); 4.95–5.08 (m, 4 H, 26P-O-CH2); 7.24–
7.35 (m, 15 H, Ar-H). 13C-NMR (CDCl3, 125 MHz, COM, DEPT), d
(ppm): 19.99 (d, 2JC,P = 5.0 Hz, P-CH2-CH2); 23.15 (d, 1JC,P = 143 Hz, P-
CH2); 45.71 (d,3JC,P = 17.1 Hz, N-CH2); 67.26 (d, 2JC,P = 5.8 Hz, 26P-O-
CH2); 77.20 (N-O-CH2); 110.73 (m, N-(C=O)-C); 127.93, 128.43,
128.59, 128.66, 129.03, 129.21 (15 Ar-C); 133.28 (N-O-CH2-C);
136.27 (d, 3JC,P = 5.8 Hz, 26P-O-CH2-C); 138.18 (m, 1 q CF); 142.12
(m, 2 q CF); 144.18 (m, 2 q CF) 175.06 (C=O). 31P-NMR (CDCl3,
202 MHz), d (ppm): 32.57. HRFAB-MS m/z 620.1624 [M+H]+ (calcd.
620.1626 for C31H27F5NO5PH).
2
(136CH2); 45.38 (N-CH2); 67.09 (d, JC,P = 5.9 Hz, 26P-O-CH2);
76.31 (N-O-CH2); 127.83, 128.29, 128.49, 128.61, 128.82, 129.03
(15 Ar-C); 134.36 (1 q Ar-C); 136.26 (d, 3JC,P = 5.7 Hz, 26P-O-CH2-C);
175.06 (C=O). 31P-NMR (CDCl3, 202 MHz), d (ppm): 33.20. CI-MS
m/z 664 [M+H]+. Anal. calcd. for C40H58NO5P (%): C 72.37, H 8.81, N
2.11. Found (%): C 72.63, H 8.72, N 2.06.
{3-[Icosanoyl(benzyloxy)amino]propyl}phosphonic acid
dibenzyl ester 6g
Prepared according to the general methods A and C from arachi-
dic acid. Yield 8% [16]. IR (KBr), cm– 1: 1652 (C=O). 1H-NMR (CDCl3,
500 MHz), d (ppm): 0.88 (t, 3J = 6.8 Hz, 3 H, CH3); 1.19–1.30 (m, 32
H, 166CH2); 1.54–1.65 (m, 2 H, CH2); 1.72–1.79 (m, 2 H, CH2);
1.86–1.95 (m, 2 H, CH2); 2.28–2.36 (m, 2 H, CH2); 3.65 (m, 2 H, N-
CH2); 4.74 (s, 2 H, N-O-CH2); 4.92–5.08 (m, 4 H, 26P-O-CH2); 7.28–
7.36 (m,15 H, Ar-H). 13C-NMR (CDCl3, 125 MHz, COM, DEPT), d
(ppm): 14.08 (CH3); 22.65, 24.55, 24.87, 25.17, 29.32, 29.38, 29.41,
29.43, 29.50, 29.62, 29.68, 31.86, 33.44, 35.33, 37.27 (206CH2);
General procedure for the preparation of phosphonic
acids 7
An amount of 1 mmol of the appropriate benzyl-protected com-
pound was dissolved in methanol (10 mL) and hydrogenated at
atmospheric pressure with 10% Pd/C (20 mg) as a catalyst. The
mixture was filtered, and the solvent was removed by rotary
evaporation.
2
67.16 (d, JC-P = 6.5 Hz, 26P-O-CH2); 76.37 (N-O-CH2); 127.88,
127.92, 128.35, 128.44, 128.55, 128.58, 128.60, 128.65, 128.78,
128.87, 129.07 (15 Ar-C); 134.32, 136.28, 136.36 (3 q Ar-C); 173.31
(C=O). 31P-NMR (CDCl3, 202 MHz), d (ppm): 33.18. HRFAB-MS m/z
720.4761 [M+H]+ (calcd. 720.4757 for C44H66NO5PH).
[3-(Hexanoylhydroxyamino)propyl]phosphonic acid 7a
Yield 93%. IR (film), cm– 1: 3131 (OH), 1615 (C=O). 1H-NMR (DMSO-
d6, 400 MHz), d (ppm): 0.86 (t, 3J = 7 Hz, 3 H, CH3); 1.22–1.31 (m, 4
H, 26CH2); 1.40–1.53 (m, 4 H, 26CH2); 1.67–1.82 (m, 2 H, CH2);
2.33 (t, 3J = 7.4 Hz, 2 H, C=O-CH2); 3.50 (t, 3J = 6.9 Hz, 2 H, N-CH2).
13C-NMR (DMSO-d6, 100 MHz, COM), d (ppm): 13.79 (CH3); 20.49 (P-
CH2-CH2); 21.87 (CH2); 23.82 (CH2); 25.10 (d, 1JC,P = 134 Hz, P-CH2);
30.98 (CH2); 31.55 (CH2); 47.82 (d, 3JC,P = 16.1 Hz, N-CH2); 172.65 (N-
C=O). 31P-NMR (DMSO-d6, 161 MHz), d (ppm): 26.26. HRFAB-MS m/z
254.1182 [M+H]+ (calcd. 254.1180 for C9H20NO5PH).
{3-[(2-Naphthalen-2-yl-acetyl)(benzyloxy)amino]propyl}-
phosphonic acid dibenzyl ester 6h
Prepared according to general method C from commercially
available 2-(2-naphthyl)acetyl chloride. Yield 42%. IR (film), cm–1:
1
1659 (C=O). H-NMR (CDCl3, 500 MHz), d (ppm): 1.69–1.76 (m, 2
H, CH2); 1.90–1.97 (m, 2 H, CH2); 3.74 (m, 2 H, N-CH2); 3.87 (s, 2 H,
C=O-CH2); 4.74 (s, 2 H, N-O-CH2); 4.87–4.98 (m, 4 H, 26P-O-CH2);
7.29–7.45 (m, 18 H, Ar-H); 7.66 (s, 1 H, Ar-H); 7.74–7.80 (m, 3 H,
2
Ar-H). 13C-NMR (CDCl3, 125 MHz, COM), d (ppm): 19.95 (d, JC,P
=
4.2 Hz, P-CH2-CH2); 22.93 (d, 1JC,P = 142 Hz, P-CH2); 39.63 (C=O-CH2);
{3-[Hydroxy(4-oxopentanoyl)amino]propyl}phosphonic
2
45.02 (N-O-CH2); 66.94 (d, JC,P = 7.2 Hz, 26P-O-CH2); 76.17 (N-O-
acid 7b
CH2); 125.46, 125.85, 127.29, 127.43, 127.45, 127.72, 127.95,
128.19, 128.38, 128.57, 128.84, 129.03 (23 Ar-C); 132.10, 132.18,
133.25, 134.05, 136.07, 136.13 (7 q Ar-C); 172.64 (C=O). 31P-NMR
(CDCl3, 162 MHz), d (ppm): 33.04. HRFAB-MS m/z 594.2411 [M+H]+
(calcd. 594.2409 for C36H36NO5PH).
1
Yield 99%. IR (film), cm– 1: 1716 (C=O), 1652 (N-C=O). H-NMR
(DMSO-d6, 500 MHz), d (ppm): 1.42–1.52 (m, 2 H, P-CH2); 1.68–
1.79 (m, 2 H, P-CH2-CH2); 2.10 (s, 3 H, CH3); 2.57–2.64 (m, 2 H, N-
C=O-CH2); 3.41–3.64 (m, 4 H, H3C-C=O-CH2 + N-CH2). 13C-NMR
(Methanol-d4, 100 MHz, COM, DEPT), d (ppm): 18.89 (P-CH2-CH2);
1
21.65 (N-C=O-CH2); 28.08 (d, JC,P = 134 Hz, P-CH2); 29.85 (CH3);
38.73 (H3C-C=O-CH2), 53.39 (d,3JC,P = 7.7 Hz, N-CH2). 31P-NMR (Meth-
anol-d4, 202 MHz), d (ppm): 27.15. HRFAB-MS m/z 254.0807 [M+H]+
(calcd. 254.0794 for C8H16NO6PH).
{3-[(Benzyloxy)(8-phenyl-octanoyl)amino]propyl}-
phosphonic acid dibenzyl ester 6i
Prepared according to the general methods A and B from 8-phe-
nyl octanoic acid. Yield 30%. IR (film), cm– 1: 1661 (C=O). 1H-NMR
(CDCl3, 500 MHz), d (ppm): 1.29–1.33 (m, 6 H, 36CH2); 1.54–
1.60 (m, 4 H, 26CH2); 1.72–1.79 (m, 2 H, CH2); 1.86–1.93 (m, 2 H,
CH2); 2.31-2.34 (m, 2 H, CH2); 2.56–2.61 (m, 2 H, CH2); 3.65 (m, 2
H, N-CH2); 4.74 (s, 2 H, N-O-CH2); 4.92–5.05 (m, 4 H, 26P-O-CH2);
7.15–7.36 (m, 15 H, Ar-H). 13C-NMR (CDCl3, 125 MHz, COM), d
(ppm): 20.18 (d, 2JC,P = 5.6 Hz, P-CH2-CH2); 23.35 (d, 1JC,P = 142 Hz, P-
CH2), 24.52, 28.99, 29.12, 29.22, 29.31, 31.44, 35.90 (7 x CH2);
{3-[Hydroxy(5-oxohexanoyl)amino]propyl}phosphonic
acid 7c
Yield 90%. IR (film), cm– 1: 1713 (C=O), 1633 (N-C=O). 1H-NMR
(DMSO-d6, 400 MHz), d (ppm): 1.35–1.45 (m, 4 H, 26CH2); 1.61–
1.73 (m, 2 H, CH2); 2.06 (s, 3 H, CH3); 2.32 (t, 3J = 7.3 Hz, 2 H, CH2);
3
2.44 (t, 3J = 7.2 Hz, 2 H, CH2); 3.50 (t, J = 6.5 Hz, 2 H, N-CH2). 13C-
2
NMR (DMSO-d6, 125 MHz, COM, DEPT), d (ppm): 19.16 (d, JC,P
=
2
67.09 (d, JC,P = 6.8 Hz, 26P-O-CH2); 76.36 (N-O-CH2); 125.52,
3.8 Hz, P-CH2-CH2), 21.16 (C=O-CH2-CH2), 25.69 (d, 1JC,P = 137 Hz, P-
CH2), 30.38 (CH3), 31.54 (N-C=O-CH2), 42.84 (H3C-C=O-CH2), 48.69
127.90, 128.19, 128.37, 128.56, 128.68, 128.91, 129.09 (20 Ar-C);
136.25, 136.30 (4 q Ar-C). 31P-NMR (CDCl3, 202 MHz), d (ppm):
33.33. HRFAB-MS m/z 628.3190 [M+H]+ (calcd. 628.3192 for
C38H46NO5PH).
3
(d, JC,P = 18.2 Hz, N-CH2), 173.22 (N-C=O), 209.00 (H3C-C=O). 31P-
NMR (DMSO-d6, 202 MHz),
d (ppm): 27.15. HRFAB-MS m/z
268.0945 [M+H]+ (calcd. 268.0950 for C9H18NO6PH).
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2007, 340, 483–490
Fosmidomycin Derivatives with Spacious Acyl Residues
489
{3-[Glutaryl(hydoxyamino)propyl}phosphonic acid 7d
Yield 90%. IR (Film), cm– 1: 3150 (OH), 1713 (C=O), 1616 (N-C=O).
1H-NMR (DMSO-d6, 500 MHz), d (ppm): 1.43-1.50 (m, 2 H, CH2);
1.66–1.74 (m, 4 H, 26CH2); 2.23 (t, 3J = 7.3 Hz, 2 H, CH2); 2.36–
2.39 (m, 2 H, CH2); 3.51 (t, 3J = 6.7 Hz, 2 H, N-CH2).13C-NMR (DMSO-
d6, 125 MHz, COM), d (ppm): 19.58 (CH2); 20.38 (CH2); 21.24 (d,1JC,P
= 141 Hz, P-CH2); 30.77 (CH2); 32.98 (CH2); 53.71 (N-CH2); 170.78
(N-C=O); 174.13 (COOH) 31P-NMR (DMSO-d6, 202 MHz), d (ppm):
26.72. HRFAB-MS m/z 268.0588 [M-H]+ (calcd. 268.0590 for
C8H15NO7P).
{3-[Hydroxy(8-phenyl-octanoyl)amino]propyl}-
phosphonic acid 7i
Yield 66%, Mp. 1028C. IR (KBr), cm– 1: 3118 (OH), 1710 (C=O). H-
NMR (DMSO-d6, 500 Hz), d (ppm): 1.24–1.28 (m, 6 H, 36CH2);
1.44–1.50 (m, 6 H, 36CH2); 1.68–1.76 (m, 2 H, CH2); 2.33 (t, 3J =
7.3 Hz, 2 H, CH2); 2.56 (t, 3J = 7.7 Hz, 2 H, CH2); 3.51 (t, 3J = 6.8 Hz, 2
H, N-CH2); 7.15–7.19 (m, 3 H, Ar-H); 7.25–7.28 (m, 2 H, Ar-H). 13C-
NMR (DMSO-d6, 125 MHz, COM), d (ppm): 21.06, 22.29, 24.36,
24.52, 28.47, 28.60, 28.69, 30.89, 31.57, 35.04 (9 x CH2); 125.45,
128.08, 128.13 (5 Ar-C); 142.22 (q Ar-C). 31P-NMR (DMSO-d6,
202 MHz), d (ppm): 26.86. HRFAB-MS m/z 358.1801 [M+H]+ (calcd.
358.1783 for C17H28NO5PH).
1
{3-[Dodecanoyl(hydoxyamino)propyl}phosphonic acid 7e
Yield 97%. IR (KBr), cm– 1: 3127 (OH), 1589 (C=O). 1H-NMR (DMSO-
d6, 400 MHz), d (ppm): 0.86 (t, 3J = 6.8 Hz, 3 H, CH3); 1.24 (s, 16 H,
86CH2); 1.42–1.50 (m, 4 H, 26CH2); 1.68–1.75 (m, 2 H, CH2);
2.32 (t, 3J = 7.5 Hz, 2 H, CH2); 3.51 (t, 3J = 6.9 Hz, 2 H, N-CH2). 13C-
NMR (DMSO-d6, 125 MHz, COM, DEPT), d (ppm): 13.87 (CH3);
22.00, 28.62, 28.76, 28.81, 28.86, 28.95, 31.20, 31.58 (176CH2);
173.57 (C=O). 31P-NMR (DMSO-d6, 161 MHz), d (ppm): 26.26.
HRFAB-MS m/z 338.2097 [M+H]+ (calcd. 338.2096 for
C15H32NO5PH).
{3-[Hydroxy(pentafluorophenyl)amino]propyl}-
phosphonic acid 7j
Yield 96%, Mp. 1028C. IR (KBr), cm– 1: 3118 (OH), 1710 (C=O). 1H-
NMR (CDCl3, 500 MHz), d (ppm): 1.53–1.60 (m, 2 H, CH2); 1.81–
1.89 (m, 2 H, CH2); 3.75 (t, 3J = 6.7 Hz, 2 H, N-CH2). 13C-NMR (DMSO-
d6, 100 MHz, COM), d (ppm): 20.17 (d, 2JC,P = 3.9 Hz, P-CH2-CH2);
1
3
24.69 (d, JC,P = 138 Hz, P-CH2); 48.50 (d, JC,P = 18.4 Hz, N-CH2);
111.46 (m, N-(C=O)-C); 135.62-143.42 (m, 5 q CF); 156.99 (C=O). 31P-
NMR (DMSO-d6, 202 MHz),
d (ppm): 26.86. HRFAB-MS m/z
350.0217 [M+H]+ (calcd. 350.0210 for C10H10F5NO5PH).
[3-Hexadecanoyl(hydroxyamino)propyl]phosphonic acid
7f
Yield 99%. IR (film), cm– 1: 3065 (OH), 1661 (C=O). 1H-NMR (DMSO-
d6, 500 MHz), d (ppm): 0.85 (t, 3J = 6.8 Hz, 3 H, CH3); 1.23 (s, 24 H,
126CH2); 1.44–1.50 (m, 4 H, 26CH2); 1.68–1.73 (m, 2 H, CH2);
References
2.32 (t, 3J = 7.0 Hz, 2 H, CH2); 3.51 (t, J = 6.7 Hz, 2 H, N-CH2). 13C-
3
[1] W. Eisenreich, A. Bacher, D. Arigoni, F. Rohdich, Cell. Mol.
Life Sci. 2004, 61, 1401–1426.
NMR (DMSO-d6, 125 MHz, COM, DEPT), d (ppm): 13.82 (CH3);
20.28, 20.34, 21.98, 24.12, 24.29, 25.36, 28.58, 28.75, 28.79,
28.85, 28.89, 28.94, 31.18, 31.57 (166CH2); 45.58 (d, 3J = 14.4 Hz,
N-CH2); 172.73 (C=O). 31P-NMR (CDCl3, 202 MHz), d (ppm): 27.35.
HRFAB-MS m/z 394.2697 [M
C19H40NO5PH).
[2] H. Jomaa, J. Wiesner, S. Sanderbrand, B. Altincicek, et al.,
Science 1999, 285, 1573–1576.
+
H]+ (calcd. 394.2722 for
[3] S. Borrmann, S. Issifou, G. Esser, A. A. Adegnika, et al., J.
Infect. Dis. 2004, 190, 1534–1540; S. Borrmann, A. A.
Adegnika, P.-B. Matsiegui, S. Issifou, et al., J. Infect. Dis.
2004, 189, 901–908; B. Lell, R. Ruangweerayut, J. Wies-
ner, M. A. Missinou, et al., Antimicrob Agents Chemother.
2003, 47, 735–738; M. A. Missinou, S. Borrmann, A.
Schindler, S. Issifou, et al., Lancet 2002, 360, 1941–1942; J.
Wiesner, D. Henschker, D. B. Hutchinson, E. Beck, H.
Jomaa, Antimicrob Agents Chemother. 2002, 46, 2889–2894.
{3-[Hydroxy(icosanoyl)amino]propyl}phosphonic acid 7g
Yield 99%. IR (KBr), cm– 1: 1652 (C=O). 1H-NMR (DMSO-d6,
500 MHz), d (ppm): 0.85 (t, 3J = 7.0 Hz, 3 H, CH3); 1.18–1.27 (m, 36
H, 186CH2); 1.47–1.54 (m, 2 H, CH2); 1.60–1.67 (m, 2 H, CH2);
1.73–1.85 (m, 2 H, CH2); 2.63–2.64 (m, 2 H, CH2); 3.57 (m, 2 H, N-
CH2). 13C-NMR (DMSO-d6, 125 MHz, COM), d (ppm): 12.29 (CH3);
17.66, 21.30, 25.53, 25.86, 28.86, 28.88, 28.90, 28.95, 30.77, 32.37
(166CH2); 174.80 (C=O). 31P-NMR (DMSO-d6, 202 MHz), d (ppm):
26.85.
[4] R. Ortmann, J. Wiesner, P. Heidler, W. Thimann, et al.,
Poster presentation at the DPhG Joint Meeting, Regensburg,
October 6–9, 2004.
[5] K. Reuter, S. Sanderbrand, H. Jomaa, J. Wiesner, et al., J.
Biol. Chem. 2002, 277, 5378–5384.
{3-[Hydroxy(2-naphthalen-2-yl-acetyl)amino]propyl}-
[6] S. Steinbacher, J. Kaiser, W. Eisenreich, R. Huber, et al., J.
Biol. Chem. 2003, 278, 18401–18407.
phosphonic acid 7h
Yield 62%, Mp. 1618C. IR (KBr), cm– 1: 3049 (OH), 1591 (C=O). H-
1
[7] S. Yajima, T. Nonaka, T. Kuzuyama, H. Seto, K. Ohsawa, J.
Biochem. (Tokyo) 2002, 131, 313–317.
NMR (DMSO-d6, 500 Hz), d (ppm): 1.48–1.55 (m, 2 H, CH2); 1.71–
1.79 (m, 2 H, CH2); 3.57–3.60 (m, 2 H, N-CH2); 3.89 (s, 2 H, C=O-
CH2); 7.39–7.50 (m, 3 H, Ar-H); 7.72 (s, 1 H, Ar-H); 7.83–7.88 (m, 3
H, Ar-H). 13C-NMR (DMSO-d6, 125 MHz, COM, DEPT), d (ppm):
[8] P. R. Gerber, K. Mꢁller, J. Comput. Aided Mol. Des. 1995, 9,
251–268.
2
1
22.70 (d, JC,P = 6.7 Hz, P-CH2-CH2); 24.91 (d, JC,P = 138 Hz, P-CH2);
3
[9] G. M. Morris, D. S. Goodsell, R. S. Halliday, R. Huey, et al.,
J. Comput Chem. 1998, 19, 1639–1662; G. M. Morris, D. S.
Goodsell, R. Huey, A. J. Olson, J. Comput. Aided Mol. Des.
1996, 10, 293–304; D. S. Goodsell, G. M. Morris, A. J.
Olson, J. Mol. Recognit. 1996, 9, 1–5.
38.56 (C=O-CH2); 48.11 (d, JC,P = 19.3 Hz, N-CH2); 125.34, 125.86,
127.26, 127.32, 127.51, 128.00 (7 Ar-C); 131.64, 132.89, 133.60 (3
q Ar-C); 170.60 (C=O). 31P-NMR (DMSO-d6, 202 MHz), d (ppm):
26.97. HRFAB-MS m/z 324.0966 [M+H]+ (calcd. 324.1000 for
C15H19NO5PH).
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

490
R. Ortmann et al.
Arch. Pharm. Chem. Life Sci. 2007, 340, 483–490
[10] D. A. Case, D. A. Pearlman, J. W. Caldwell, T. E. Cheatham
III, et al., 2002 AMBER 7; University of California, San
Francisco, CA, USA.
[14] K. Silber, T. Kurz, P. Heidler, G. Klebe, J. Med. Chem. 2005,
48, 3547–3563.
[15] L. Mercklꢂ, A. De Andrꢂs-Gomꢂz, B. Dick, R. J. Cox, C. R. A.
Godfrey, ChemBioChem 2005, 6, 1866–1874.
[11] S. J. Weiner, P. A. Kollman, D. A. Case, U.C. Singh, et al., J.
Am. Chem. Soc. 1984, 108, 765–784.
[16] The reason for the low yields of the acylation is
unknown. No further investigations of the reaction have
been performed.
[12] M. Dewar, E. Zoebisch, E. Healy, J. Stewart, J. Am. Chem.
Soc. 1985, 107, 3902–3909.
[13] Limited, D. J. J. P. S. a. F. MOPAC 6.0, Quantum Chemical
Program Exchange; #455 ed.; Indiana University: Tokyo; J.
Stewart, J. Comput. Aided Mol. Des. 1990, 4, 1–105.
i 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com