Synthesis of N-(phosphonomethyl)glycine derivatives and studies of their immunotropic activity

Article abstract of DOI:10.1007/s11172-011-0111-1

The Petasis reaction between glyoxylic acid, α-amino phosphonates, and organylboronic acid afforded N-phosphonomethyl-α-amino acids. This method has an advantage of preparative simplicity and high diastereoselectivity of the reactions. Immunotropic activity of the synthesized compounds was studied using the models in vivo.

Full text of DOI:10.1007/s11172-011-0111-1

Russian Chemical Bulletin, International Edition, Vol. 60, No. 4, pp. 712—718, April, 2011
712
Synthesis of Nꢀ(phosphonomethyl)glycine derivatives
and studies of their immunotropic activity
M. V. Shevchuk,a,b L. A. Metelitsa,a L. L. Charochkina,a S. E. Mogilevich,a E. B. Rusanov,c
A. E. Sorochinsky,a V. P. Khilya,b V. D. Romanenko,a and V. P. Kukhara
aInstitute of Bioorganic Chemistry and Petrochemistry, National Academy of Sciences of Ukraine,
1 ul. Murmanskaya, 02660 Kiev 94, Ukraine.
Eꢀmail: kukhar@bpci.kiev.ua
bTaras Shevchenko Kiev National University, Department of Chemistry,
62a ul. Vladimirskaya, 01033 Kiev, Ukraine
cInstitute of Organic Chemistry, National Academy of Sciences of Ukraine,
5 ul. Murmanskaya, 02094 Kiev, Ukraine
The Petasis reaction between glyoxylic acid, αꢀamino phosphonates, and organylboronic
acid afforded Nꢀphosphonomethylꢀαꢀamino acids. This method has an advantage of preparaꢀ
tive simplicity and high diastereoselectivity of the reactions. Immunotropic activity of the
synthesized compounds was studied using the models in vivo.
Key words: the Petasis reaction, αꢀamino phosphonates, Nꢀ(phosphonomethyl)glycines,
immunotropic activity.
NꢀPhosphorylated peptides were found in the products
of natural origin, they possess useful medicobiological
properties.1,2 For instance, phosphoramidon isolated
from Actynomycetes is an efficient inhibitor of metalloproꢀ
teases,3 whereas some nucleoside phosphoramidates exꢀ
hibit antitumor4 and antiviral5 activities. However, the
P—N bond is very sensitive to hydrolysis, which limits the
use of phosphorus amides in vivo. In this connection,
Nꢀphosphonomethylamino acids and peptides containing
hydrolytically stable skeleton, whose phosphoryl group
mimics the transition state of the reaction of hydrolytic
cleavage of the peptide bond, are of significant interest.
Among this type of compounds, a large number of practiꢀ
cally useful substances were found, such as herbicide
glyphosate,6 antibacterial drug alaphosphaline,7 and highly
active antihypertensive drug CGS 24592.8
Until now, the threeꢀcomponent Kabachnick—Fields
reaction based on the reaction of a carbonyl compound,
amino acid ester, and dialkyl phosphite catalyzed by Lewis
or Brønsted acids served as the most important method for
the synthesis of Nꢀphosphonomethylglycine derivatives.2,9
The key step of this process is the addition of a hydrophosꢀ
phoryl compound to the amino acidꢀderived azomethine
generated in the course of the reaction. However, despite
numerous advantages the Kabachnick—Fields reaction
provides an access to Nꢀphosphonomethylamino acids
containing only secondary amino group. In addition,
a classic version of this reaction involving amino acid
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 4, pp. 698—703, April, 2011.
1066ꢀ5285/11/6004ꢀ712 © 2011 Springer Science+Business Media, Inc.

Nꢀ(Phosphonomethyl)glycine derivatives
Russ.Chem.Bull., Int.Ed., Vol. 60, No. 4, April, 2011
713
smoothly proceeds only with formaldehyde.10,11 The use
of aldehydes and ketones for the synthesis of Nꢀphosꢀ
phonomethylamino acids requires the use of amino acid
tertꢀbutyl esters as the amine component12 or specific catꢀ
alysts, such as aluminum tetra(tertꢀbutyl)phthalocyanine
chloride.13—15 And finally, the use of the Kabachnick—
Fields reaction for stereoselective synthesis of Nꢀphosꢀ
phonomethylamino acids with the chiral carbon atom at
the phosphorus center, >P(O)—C*—, is a rather difficult
problem.16 Among other methods for the preparation of
Nꢀphosphonomethylꢀαꢀamino acids, the following synꢀ
theses should be mentioned: through the functionalized
azomethines,17,18 1,3ꢀoxazolidinꢀ2ꢀones,19 Lꢀpyroglutꢀ
amine and 6ꢀoxoꢀLꢀpipecolinic acids,20 as well as desulfurꢀ
ization of thiocarbamoyl phosphonates21 and reactions of
1,3,5ꢀtris[Nꢀ(alkoxycarbonylalkyl)]hexahydrotriazines
with PHꢀacids.22 These methods can be used for the synꢀ
thesis of specific types of Nꢀphosphonomethylamino acids.
A possibility of application of amino phosphonates as
an amine component in the boronic version of the Manꢀ
nich reaction (the Petasis reaction) was first reported in
the patent literature.23 In the preliminary communicaꢀ
tion,24 we have shown that a threeꢀcomponent reaction of
αꢀaminophosphonates 1 with glyoxylic acid and alkenylꢀ
or arylboronic acids is an important method for the prepaꢀ
ration of Nꢀ(diethoxyphosphorylmethyl)glycine derivaꢀ
tives 2 (Scheme 1). The present work is devoted to the
synthesis of waterꢀsoluble salts of phosphonic acid 3 and
studies of their immunotropic activity.
Scheme 1
1: R2 = 4ꢀMeOC6H4, R3 = H, R4 = H (a);
R
2 = Ph, R3 = H, R4 = PhCH2 (b);
R2, R4 = –(CH2)3—, R3 = P(O)(OEt)2 (c)
2, 3: R1 = (E)ꢀPhCH=CH, R2 = 4ꢀMeOC6H4, R3 = H, R4 = H (a)
R
1 = 4ꢀMeOC6H4, R2 = Ph, R3 = H, R4 = PhCH2 (b);
The general approach to the synthesis of compounds 3
is given in Scheme 1. The threeꢀcomponent oneꢀpot reacꢀ
tion of αꢀaminophosphonates 1a—c, glyoxylic acid, and
boronic acids leads to Nꢀphosphonomethylamino acid esꢀ
ters 2a—e, whose yields depend on the nature of aminoꢀ
phosphonate, organylboronic acid, and the solvent used.
On the whole, the regularities of the reaction are similar to
those described earlier for the Petasis reaction involvꢀ
ing organylamines.25,26: (a) for organylboronic acids,
RB(OH)2, the reactivity decreases in the following order
of R: styryl > hetaryl > aryl; (b) diastereoselectivity of the
reactions is higher in the case of aminophosphonates with
secondary amino group (dr > 9 : 1) as compared to aminoꢀ
phosphonates with primary amino group (dr > 7 : 3); (c)
aprotic solvents of moderate polarity are the optimum reꢀ
action media. When the reactants are refluxed in ethyl
acetate, the reaction usually comes to completion within
2—5 h with the degree of conversion 90—95%. The use of
alcohols (methanol, ethanol), dioxane, or acetonitrile leads
to a decrease in the yields of the target products because of
side processes. In dichloromethane, the reaction proceeds
smoothly, however, at low rate (20 °C, 3—5 days). Phenylꢀ
boronic acid under indicated conditions does not react.
However, reactions involving arylboronic acids containꢀ
ing electronꢀenriched aryl groups (3,4ꢀmethylenedioxyꢀ
phenyl, 4ꢀmethoxyphenyl) lead to compounds 2 in good
R1
R1
=
=
, R2 = Ph, R3 = H, R4 = PhCH2 (c);
, R2 = Ph, R3 = H, R4 = PhCH2 (d);
R1
=
, R2, R4 = —(CH2)3—, R3 = P(O)(OEt)2 (e)
yields. All the transformations are characterized by a high
degree of diastereoselectivity (dr > 9 : 1), which was inꢀ
ferred from the 31P NMR spectra of the reaction mixtures.
In the case of Nꢀbenzylaminoꢀαꢀamino phosphonate 1b,
the diastereoselectivity is higher than 95%. Purification of
products 2 by column chromatography allowed us to isolꢀ
ate the major diastereomer in pure form.
The structures of compounds 2 were established based
on the data from elemental analysis, mass spectrometry,
and 1H, 13C, and 31P NMR spectroscopy (see Experimenꢀ
tal). The molecular structure of compound 2b was conꢀ
firmed by Xꢀray diffraction study. The general view27 of
molecule 2b and its basic geometrical parameters are givꢀ
en in Fig. 1 and in Tables 1 and 2. The central fragment of
the molecule C(12)—N(1)—C(1)—P(1) is nonplanar: the
corresponding dihedral angle is 61.1(1)°. The configuraꢀ

714
Russ.Chem.Bull., Int.Ed., Vol. 60, No. 4, April, 2011
Shevchuk et al.
C(23)
Table 2. Principal bond angles in molecule 2b
C(27)
O(1)
O(2)
C(13)
C(22)
C(24)
C(25)
Angle
Value
C(9)
C(8)
O(3)
P(1)
O(5)
C(21)
O(6)
C(12)
O(4)
N(1)
C(1)
Bond angle
ω/deg
C(26)
C(1)—N(1)—C(12)
C(1)—N(1)—C(14)
C(12)—N(1)—C(14)
118.24(1)
116.38(6)
117.99(3)
C(15)
C(16)
C(20)
C(17)
C(3)
C(4)
C(5)
C(2)
Torsional angle
ϕ/deg
C(10)
C(19)
C(18)
C(1)—N(1)—C(12)—C(13)
C(12)—N(1)—C(1)—P
90.37(1)
–61.11(7)
C(7)
C(6)
C(11)
Fig. 1. The view of molecule 2b (ORTEPꢀ3) with representation
of atoms by thermal ellipsoids (p = 50%).
its weight and cellularity were revealed, rather they signifꢀ
icantly intensify the processes of splenocyte differentiaꢀ
tion, which are responsible for the functional state of the
humoral part of immunity.28 A significant (more than
eightfold) increase in the antibodyꢀsynthesizing activity
was registered as well. At the same time, when these comꢀ
pounds were introduced into the animal body, a signifiꢀ
cant (more than twofold) decrease in the thymus weight
and cellularity was observed, whose lymphocytes funcꢀ
tionally control the state of the cellular part of immuꢀ
nity.29 The compounds had a negative effect on the nonꢀ
specific reactions of the immune response, significantly,
virtually twofold, decreasing phagocytic reactivity of the
animal polymorphonuclear blood lymphocytes. To sum up,
the biological screening of the synthesized Nꢀphosphonoꢀ
methylglycine derivatives using the model in vivo showed
that introduction of these compounds into the body leads
to a disbalance of the animal immune system.
tion of the nitrogen atom is almost planarꢀtrigonal (the
sum of bond angles is 352.6°). The phosphorus atom P(1)
has a distorted tetrahedral surrounding with the angle range
from 100.3 to 116.15°. The bond distances at the nitrogen
and phosphorus atoms are common for amines and phosꢀ
phonic acid esters, respectively. The distance P(1)—O(3)
1.466(8) Å corresponds to the double P=O bond.
Transformation of phosphonates 2 to the correspondꢀ
ing phosphonic acids, isolated as ammonium salts 3, was
performed by transesterification of compounds 2 using
bromotrimethylsilane and subsequent hydrolysis of the inꢀ
termediate trimethylsilyl esters. The reactions gave quanꢀ
titative yields in the case of phosphonates 2a—d, however,
treatment of compound 2e, containing methylenebisphosꢀ
phonate fragment incorporated into the pyrrolidine ring,
with bromotrimethylsilane was accompanied by eliminaꢀ
tion of P(OSiMe3)3 and formation of (4,5ꢀdihydropyrrolꢀ
2ꢀyl)phosphonic acid derivatives. Preliminary studies of
transesterification reactions of other Nꢀsubstituted 2,2ꢀbisꢀ
(diethoxyphosphoryl)pyrrolidines upon the action of
Me3SiBr showed that the ease of the C—P bond cleavage
in these compounds is a specific feature of pyrrolidineꢀ
2,2ꢀdiylbisphosphonates.
Experimental
All the experiments were carried out under argon. Acetoniꢀ
trile was purified by distillation over P2O5. The course of the
reactions was monitored by TLC on silica gel (Fluka). Preparaꢀ
tive flash chromatography was performed on Merck 60 silica gel.
Amino phosphonates were obtained using methods described in
the literature.30,31 Boronic acids and glyoxylic acid hydrate were
purchased from Aldrich Co and used without additional puꢀ
rification.
1H NMR spectra were recorded on Varian VXRꢀ300 and
Varianꢀ400 spectrometers. 13C and 31P NMR spectra were reꢀ
corded on a Varianꢀ400 spectrometer (100 and 162 MHz, reꢀ
spectively). Chemical shifts are given in the δꢀscale and meaꢀ
sured with respect to internal standards: hexamethyldisiloxane
(δH = 0.05), residual signal of chloroform (δH = 7.26, δC = 77.16),
or residual signal of methanol (CHD2OD, δH = 3.31); or exterꢀ
nal standard: 85% H3PO4 (δ = 0.0). Mass spectra were obtained
on an Agilent 1100 LC MSD SL instrument (chemical ionizaꢀ
tion), signals of positive ions current are labeled as pos., negative
as neg. Melting points were measured on a Nagema Boetius
PHMK05 apparatus from Rapido.
Immunotropic activity of the synthesized compounds
3c,d was studied using the models in vivo. The data obꢀ
tained (Table 3) indicate that the studied compounds unꢀ
der conditions of the experiment do not affect proliferaꢀ
tion processes in the animal spleen: no reliable changes in
Table 1. Principal bond distances in molecule 2b
Bond
d/Å
Bond
d/Å
C(1)—N(1)
C(1)—P(1)
N(1)—C(12) 1.450(2)
N(1)—C(14) 1.462(6)
1.466(2)
1.830(5)
P(1)—O(3)
P(1)—O(4)
P(1)—O(5)
C(13)—O(1) 1.201(5)
C(13)—O(2) 1.327(9)
1.466(8)
1.562(1)
1.571(9)
Single crystals of compound 2b suitable for Xꢀray diffraction
study were obtained by recrystallization from benzene. The
Xꢀray diffraction experiment for crystals with linear sizes

Nꢀ(Phosphonomethyl)glycine derivatives
Russ.Chem.Bull., Int.Ed., Vol. 60, No. 4, April, 2011
715
Table 3. Effects of compounds 3c,d on the weight and cellularity of lymphoid organs, reactions of
antibody formations, and phagocytose using the models in vivo (M m, n = 8)
Comꢀ
pound
Spleen
Amount of AT,
Thymus
M/mg
N´ b
–log2T
M/mg
Na•108
N•108
1 Day after administration of compound
Control
3c
3d
129.2 10.3
136.3 12.4
117.2 9.7
14.3 1.3
11.9 1.0
12.6 0.9
—
—
—
44.5 3.6
24.6 2.0c
52.2 4.3
17.0 1.2
10.4 0.8c
20.5 1.8
61.5 4.5
60.7 4.0
43.5 3.2c
7 Days after administration of compound
Control
3c
3d
114.3 9.8
118.2 11.0
106.4 8.9
13.4 1.2
10.6 0.8
11.0 1.0
3.0 0.2
6.3 0.4c
6.7 0.3c
45.0 3.9
49.0 4.0c
9.1 0.7c
16.2 1.4
15.4 1.1
63.3 3.8
33.6 2.9c
4.1 0.4c 19.7 1.2c
a Amount of cells per gram of an organ.
b Amount of phagocytes per 100 PMNL.
c The value reliable (p < 0.05) differs from the control one.
0.50×0.20×0.12 mm was carried out on a Smart APЕХ autoꢀ
matic diffractometer with graphite monochromator (MoꢀKαꢀ
irradiation, λ = 0.71073 Å) at room temperature. Allowance for
the absorption in crystal was made using the multiscanning method.
The structure was solved by the direct method and refined by
the least squares method in the fullꢀmatrix anisotropic approxiꢀ
mation for all the nonhydrogen atoms using the SHELXS9732
and SHELXL9733 program packages. All the hydrogen atoms
were localized geometrically. The residual electron density after
the final cycle of refinement was 0.46 and –0.37 е Å–3. The
crystallographic data, parameters of the Xꢀray diffraction experꢀ
iment and refinement for compound 2b are given in Table 4.
In biological experiments, we used female outbred mice with
the weight 18—21 g. Equimolar concentrations of compounds
under study were administered intraabdominally to the animals
in one portion in a doze 2•10–4 mol per 1 kg of body weight in
saline.34 An aliquot of saline was introduced to the control aniꢀ
mals. Testing were performed 1 and 7 days after administration
of compounds. The murine thymus and spleen were isolated and
weighed, a suspension of lymphoid cells were washed off in the
nutrient medium 199 for culturing, their amount was calculated
in the Goryaev chamber.35 The results obtained refer to 1 g of
the organ weight. The content of antibodies (AB) to the thymusꢀ
dependent antigen, the ram erythrocytes (RE), were determined
in the blood serum of immunized mice by the direact hemagglutiꢀ
nation reaction36 in the seventh day after simultaneous adminisꢀ
tration of compounds under study and RE into mice. The results
of the reaction were given as –log2T (T is the final twofold
dilution of the blood serum that manifested agglutination).
Phagocytic reactivity of the animal blood polymorphonuclear
lymphocytes (PMNL) was determined by the known method
using the Staphylococcus aureus culture.34 All the results obꢀ
tained were proceeded statistically.
for 2—2.5 h, monitoring the progress of the reaction by TLC (5%
methanol in chloroform). After the reaction has come to comꢀ
pletion, the suspension obtained was diluted with ethyl acetate
(10—15 mL), the reaction mixture was washed with brine
(2×3 mL) and dried with Na2SO4. The solvent was evaporated to
obtain a yellow oil crystallized on standing. The solid substance
formed was recrystallized from ethyl acetate—hexane (4 : 1). The
yield of compound 2a was 325 mg (75%), colorless crystals,
Table 4. Crystallographic data and principal geometric parameꢀ
ters of the Xꢀray diffraction experiment for 2b
Parameters
Value
Molecular formula
Molecular weight
C
27H32NO6P
497.51
Crystal system
Space group
Monoclinic
Р2(1)/n
Т/К
296(2)
a/Å
b/Å
c/Å
10.6098(13)
11.5274(14)
21.894(3)
103.859(4)
2599.7(6)
β/deg
V/Å3
Z4
dcalc/g cm–3
F(000)
1.271
1056
2θmax/deg
26.63
17842
5425
0.0941
2673
Number of collected reflections
Number of independent reflections
Rint
Number of observed reflections
with I > 2σ(I)
(E)ꢀ2ꢀ[(Diethoxyphosphoryl)(4ꢀmethoxyphenyl)methylamino]ꢀ
4ꢀphenylbutꢀ3ꢀenoic acid (2a) and ammonium (E)ꢀ2ꢀ[(phosꢀ
phonato)(4ꢀmethoxyphenyl)methylamino]ꢀ4ꢀphenylbutꢀ3ꢀenoate
(3a). Amino phosphonate 1a (273 mg, 1.00 mmol) and after 5 min
(Е)ꢀstyrylboronic acid (155 mg, 1.05 mmol) were added to
a suspension of glyoxylic acid monohydrate (97 mg, 1.05 mmol)
in ethyl acetate (5 mL) with stirring. The mixture was refluxed
Number of refined parameters
Absorption coefficient/mm–1
R1 (I > 2σ(I))
wR2
GOOF
322
0.147
0.0643
0.1015
0.962

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Russ.Chem.Bull., Int.Ed., Vol. 60, No. 4, April, 2011
Shevchuk et al.
m.p. 62 °C. Found (%): C, 60.96; H, 6.51; N, 3.23. C22H28NO6P.
Calculated (%): C, 60.80; H, 6.45; N, 3.19. 1H NMR (CDCl3),
δ: 1.14 (t, 3 H, OCH2CH3, J = 7.0 Hz); 1.20 (t, 3 H, OCH2CH3,
J = 7.0 Hz); 3.71 (s, 3 H, OCH3); 3.81—4.09 (m, 5 H, OCH2CH3,
CHCOOH); 4.17 (d, 1 H, ArCHP, J = 18.4 Hz); 6.12 (dd, 1 H,
PhCH=CH, J = 16.0 Hz, J = 6.4 Hz); 6.58 (d, 1 H, PhCH=CH,
J = 16.0 Hz); 6.70 (br.s, 2 H, NH); 6.80 (d, 2 H, Ar, J = 8.0 Hz);
Compound 3b was obtained similarly to compound 3a. The
yield was 63%, colorless crystals, m.p. 149—151 °C (decomp.).
1H NMR (CD3OD), δ: 3.63 (d, 1 H, PhCH2N, J = 13.9 Hz);
3.70 (s, 3 H, OCH3); 4.53 (d, 1 H, PhCHP, J = 18.8 Hz); 4.74
(s, 1 H, CHCOOH); 5.53 (d, 1 H, PhCH2N, J = 13.9 Hz); 6.50
(d, 2 H, Ar, J = 8.3 Hz); 6.72 (d, 2 H, Ar, J = 8.3 Hz); 7.01—7.11
(m, 5 H, Ar); 7.52 (m, 3 H, Ar); 7.82 (m, 2 H, Ar). 31P NMR
(CD3OD), δ: 8.9 (d, J = 18.8 Hz).
7.14—7.34 (m,
7
H, Ar). 13C NMR (CDCl3), δ: 16.3
(OCH2CH3); 55.3 (OCH3); 58.2 (d, ArCHP, J = 154.8 Hz);
61.2 (d, CHCOOH, J = 16.8 Hz); 63.3 (d, OCH2CH3, J = 7.4 Hz);
63.7 (d, OCH2CH3, J = 7.4 Hz); 114.2, 125.6, 126.6 (d, J = 5.5 Hz);
126.8, 128.1, 128.7, 130.1, 133.2, 136.5, 159.9 (Ar), 174.5
(COOH). 31P NMR (CDCl3), δ: 23.2. MS (CI), m/z (Irel (%)):
pos. 434 [M + H]+ (45), 296 [M + H – HP(O)(OEt)2]+ (100).
2,4,6ꢀTrimethylpyridine (0.33 mL, 2.5 mmol) and freshly
distilled bromotrimethylsilane (0.33 mL, 2.5 mmol) were added
to a solution of compound 2a (108 mg, 0.25 mmol) in anhydrous
CH2Cl2 (1 mL). The suspension that formed was stirred for 16 h,
then the volatile products were evaporated in vacuo (0.05 Torr).
The residue was diluted with aqueous 1 M NaOH (3 mL), the
mixture was stirred for 30 min and lyophilized. A solid substance
obtained was washed with acetone (5 mL) and diethyl ether
(10 mL), dissolved in water (5 mL), and acidified with dilute
hydrochloric acid (~1%) to pH 1. A white precipitate formed
was filtered off, dissolved in concentrated aq. ammonia (1 mL),
and concentrated in air to obtain compound 3a (87 mg,
44%), colorless crystals, m.p. 182—184 °C (decomp.). 1H NMR
(CD3OD), δ: 3.80 (s, 3 H, OCH3); 3.98 (d, 1 H, CHCOOH,
J = 9.8 Hz); 4.20 (d, 1 H, ArCHP, J = 16.1 Hz); 6.29 (dd, 1 H,
PhCH=CH, J = 15.9 Hz, J = 9.8 Hz); 6.47 (d, 1 H, PhCH=CH,
J = 15.9 Hz); 6.94 (d, 2 H, Ar, J = 8.3 Hz); 7.22—7.35 (m, 4 H,
Ar); 7.44—7.52 (m, 3 H, Ar). 31P NMR (CD3OD), δ: 10.3
(d, J = 16.1 Hz).
2ꢀ{Nꢀ[(Diethoxyphosphoryl)(phenyl)methyl]benzylamino}ꢀ2ꢀ
(2ꢀthienyl)acetic acid (2c) and ammonium 2ꢀ{Nꢀ[(phenyl)(phosꢀ
phonato)methyl]benzylamino}ꢀ2ꢀ(thiophenꢀ2ꢀyl)acetate (3c).
Compound 2c was obtained similarly to compound 2a starting
from aminophosphonate 1b (397 mg, 1.19 mmol), 2ꢀthienylboꢀ
ronic acid (153 mg, 1.20 mmol), and glyoxylic acid (110 mg,
1.20 mmol). The chloroform—ethyl acetate—methanol (35 : 55 : 5)
solvent system was used for flashꢀchromatography. The yield
was 390 mg (69%), colorless crystals, m.p. 219—220 °C (deꢀ
comp.). Found (%): C, 61.68; H, 5.95; S, 7.09. C24H28NO5PS.
Calculated (%): C, 60.88; H, 5.96; S, 6.77. 1H NMR (CDCl3), δ:
1.04 (t, 3 H, OCH2CH3, J = 7.0 Hz); 1.42 (t, 3 H, OCH2CH3,
J = 7.0 Hz); 3.74—3.85 (m, 1 H, OCH2CH3); 3.93 (m, 1 H,
OCH2CH3); 4.00 (d, 1 H, PhCH2N, J =14.2 Hz); 4.21 (m, 1 H,
OCH2CH3); 4.31 (d, 1 H, PhCHP, J = 22.0 Hz); 4.35 (m, 1 H,
OCH2CH3); 4.47 (dd, 1 H, PhCH2N, J = 14.2 Hz, J = 3.5 Hz);
5.43 (s, 1 H, CHCOOH); 6.74 (s, 1 H, Hthiophene); 6.82 (s, 1 H,
Hthiophene); 7.13 (s, 1 H, Hthiophene); 7.28—7.50 (m, 10 H, Ar);
11.00 (br.s, 1 H, COOH). 13C NMR (CDCl3), δ: 16.4
(d, OCH2CH3, J = 6.0 Hz); 16.74 (d, OCH2CH3, J = 6.0 Hz);
53.3 (d, PhCH2N, J = 4.0 Hz); 58.3 (d, PhCHP, J = 152.0 Hz);
59.8 (CHCOOH); 62.8 (d, OCH2CH3, J = 8.0 Hz); 63.3
(d, OCH2CH3, J = 8.0 Hz); 126.1, 126.3, 127.6, 127.8, 128.6,
128.6, 128.9, 130.0, 130.6 (d, J = 9.0 Hz); 134.4 (d, JC,P
=
= 8.0 Hz); 138.9, 140.1, 172.8 (COOH). 31P NMR (CDCl3), δ:
24.1. MS (CI), m/z (Irel (%)): pos. 474 [M + H]+, 336 [M + H –
2ꢀ{Nꢀ[(Diethoxyphosphoryl)(phenyl)methyl]benzylamino}ꢀ2ꢀ
(4ꢀmethoxyphenyl)acetic acid (2b) and ammonium 2ꢀ{Nꢀ[(phenꢀ
yl)(phosphonato)methyl]benzylamino}ꢀ2ꢀ(4ꢀmethoxyphenyl)acetꢀ
ate (3b). Compound 2b was obtained similarly to compound 2a
starting from aminophosphonate 1b (333 mg, 1.00 mmol),
4ꢀmethoxyphenylboronic acid (152 mg, 1.00 mmol), and glyꢀ
oxylic acid (92 mg, 1.00 mmol). The yield was 472 mg (95%),
colorless crystals, m.p. 138—139 °C (ethyl acetate). Found (%):
C, 64.95; H, 6.43; N, 2.78. C27H32NO6P. Calculated (%):
– HP(O)(OEt)2]+ (100), 196 [M + H – HP(O)(OEt)2
– (C4H3S)CHCOOH]+ (35); neg. 472 [M – H]– (100).
–
Compound 3c was obtained similarly to compound 3a. The
yield was 32%. 31P NMR (CD3OD), δ: 9.1 (d, J = 19.1 Hz).
2ꢀ{Nꢀ[(Diethoxyphosphoryl)(phenyl)methyl]benzylamino}ꢀ2ꢀ
(benzo[d][1,3]dioxolꢀ5ꢀyl)acetic acid (2d) and ammonium 2ꢀ{Nꢀ
[(phenyl)(phosphonato)methyl]benzylamino}ꢀ2ꢀ(benzo[d][1,3]ꢀ
dioxolꢀ5ꢀyl)acetate (3d). Compound 2d was obtained similarly to
compound 2a starting from aminophosphonate 1b (333 mg,
1.00 mmol), 3,4ꢀmethylenedioxyphenylboronic acid (166 mg,
1.00 mmol), and glyoxylic acid (92 mg, 1.00 mmol). The product
was recrystallized from ethyl acetate. The yield was 450 mg
(88%), colorless crystals, m.p. 176—179 °C. Found (%): C, 63.71;
H, 5.99; N, 2.79. C27H30NO7P. Calculated (%): C, 63.40; H,
5.91; N, 2.74. 1H NMR (CDCl3), δ: 1.00 (t, 3 H, OCH2CH3,
J = 7.0 Hz); 1.43 (t, 3 H, OCH2CH3, J = 7.0 Hz); 3.66—3.75
(m, 1 H, OCH2CH3); 3.85—3.94 (m, 1 H, OCH2CH3); 3.99
(d, 1 H, PhCH2N, J = 14.2 Hz); 4.18—4.27 (m, 1 H, OCH2CH3);
4.28 (d, 1 H, PhCHP, J = 20.0 Hz); 4.35—4.45 (m, 1 H,
OCH2CH3); 4.49 (dd, 1 H, PhCH2N, J = 14.2 Hz, J = 4.9 Hz);
5.04 (s, 1 H, CHCOOH); 5.85 (d, 2 H, OCH2O, J = 7.0 Hz);
6.32 (s, 1 H, Ar); 6.46 (d, 1 H, Ar, J = 7.9 Hz); 6.62 (d, 1 H,
Ar, J = 7.9 Hz); 7.14 (d, 2 H, Ar, J = 7.2 Hz); 7.16—7.33
(m, 8 H, Ar); 11.00 (br.s, COOH). 13C NMR (CDCl3), δ: 16.3
(d, OCH2CH3, J = 5.0 Hz); 16.7 (d, OCH2CH3, J = 8.0 Hz); 52.9
(PhCH2N); 58.8 (d, PhCHP, J = 150.0 Hz); 62.5 (d, OCH2CH3,
J = 7.0 Hz); 63.1 (d, OCH2CH3, J =7.0 Hz); 64.0 (CHCOOH);
1
C, 65.18; H, 6.48; N, 2.82. H NMR (CDCl3), δ: 1.01 (t, 3 H,
OCH2CH3, J = 7.0 Hz); 1.41 (t, 3 H, OCH2CH3, J = 7.0 Hz);
3.64—3.75 (m, 1 H, OCH2CH3); 3.72 (s, 3 H, OCH3); 3.84—3.92
(m, 1 H, OCH2CH3); 3.94 (d, 1 H, PhCH2N, J =14.2 Hz);
4.17—4.23 (m, 1 H, OCH2CH3); 4.29 (d, 1 H, PhCHP,
J = 20.5 Hz); 4.31—4.39 (m, 1 H, OCH2CH3); 4.44 (dd, 1 H,
PhCH2N, J = 14.2 Hz, J = 4.9 Hz); 5.09 (s, 1 H, CHCOOH);
6.70 (d, 2 H, Ar, J = 8.5 Hz); 6.84 (d, 2 H, Ar, J = 8.5 Hz); 7.12
(d, 2 H, Ar, J = 6.4 Hz); 7.16—7.33 (m, 8 H, Ar); 10.45 (br.s,
COOH). 13C NMR (CDCl3), δ: 16.4 (d, OCH2CH3, J = 5.7 Hz);
16.8 (d, OCH2CH3, J = 6.5 Hz); 53.1 (PhCH2N); 55.4 (OCH3);
58.9 (d, PhCHP, J = 150.0 Hz); 62.5 (d, OCH2CH3, J = 7.6 Hz);
63.1 (d, OCH2CH3, J =7.6 Hz); 64.1 (CHCOOH); 113.7, 127.5,
128.3, 128.4, 128.7, 129.6, 130.7, 130.7 (d, J = 9.0 Hz); 135.1
(d, J = 9.0 Hz); 159.2, 174.73 (d, COOH, J = 3.8 Hz). 31P NMR
(CDCl3), δ: 24.1. MS (CI), m/z (Irel (%)): pos. 498 [M + H]+
(70), 360 [M + H – HP(O)(OEt)2]+ (100), 196 [M + H –
–HP(O)(OEt)2 – MeOC6H4CHCOOH]+ (45), 165 [M + H –
– HP(O)(OEt)2 – PhCH2NCHPh]+ (10).

Nꢀ(Phosphonomethyl)glycine derivatives
Russ.Chem.Bull., Int.Ed., Vol. 60, No. 4, April, 2011
717
101.1 (OCH2O); 108.0, 110.0, 122.8, 127.5, 128.4, 128.5,
128.7, 129.6, 130.7 (d, J = 9.0 Hz); 131.5, 135.2 (d, J = 9.0 Hz);
139.6, 147.2, 147.6, 174.4 (d, COOH, J = 3.0 Hz). 31P NMR
(CDCl3), δ: 23.8. MS (CI), m/z (Irel (%)): pos. 512 [M + H]+
(55), 374 [M + H – HP(O)(OEt)2]+ (100), 334 [M + H –
– (CH2O2C6H3)CHCOOH]+ (10), 196 [M + H – HP(O)(OEt)2 –
– (CH2O2C6H3)CHCOOH]+ (35); neg. 510 [M – H]– (100).
Compound 3d was obtained similarly to compound 3a. The
yield was 30%, colorless crystals, m.p. 154—158 °C (decomp.).
1H NMR (CD3OD), δ: 3.65 (d, 1 H, PhCH2N, J = 13.9 Hz);
4.53 (d, 1 H, PhCHP, J = 19.3 Hz); 4.65 (s, 1 H, CHCOOH);
5.51 (d, 1 H, PhCH2N, J = 13.9 Hz); 5.75 (s, 1 H, OCH2O);
5.84 (s, 1 H, OCH2O); 6.27 (s, 1 H, Ar); 6.36 (d, 1 H, Ar, J = 8.1 Hz);
6.50 (d, 1 H, Ar, J = 8.1 Hz); 7.13 (s, 5 H, Ar); 7.51 (s, 3 H, Ar);
7.81 (s, 1 H, Ar). 31P NMR (CD3OD), δ: 9.2 (d, J = 19.7 Hz).
MS, m/z (Irel (%)): pos. 456 [M + H – NH3]+ (65), 374 [M + H –
and Proteins in Organic Chemistry, Ed. A. B. Hughes, Wileyꢀ
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– 3NH3 – HP(O)(OH)2]+ (30), 330 [M + H – 3NH3
– HP(O)(OH)2 – CO2]+ (15), 278 [M + H – 3 NH3
–(CH2O2C6H3)CHCOOH]+ (20), 196 [M + H – 3 NH3
–
–
–
– HP(O)(OH)2 – (CH2O2C6H3)CHCOOH]+ (45), 91 [M +
+ H – 3 NH3 – HP(O)(OH)2 – (CH2O2C6H3)CHCOOH –
– PhNH2]+ (40); neg. 454 [M – H]– , 276 [M – H – 3 NH3 –
– (CH2O2C6H3)CHCOOH]– (20).
11. A. A. Prishchenko, M. V. Livantsov, O. P. Novikova, L. I.
Livantsova, Yu. N. Luzikov, Zh. Obshch. Khim., 1995, 65,
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13. E. D. Matveeva, T. A. Podrugina, E. V. Tishkovskaya, N. S.
Zefirov, Synlett, 2003, 2321.
2ꢀ[2,2ꢀBis(diethoxyphosphoryl)pyrrolidino]ꢀ2ꢀ(benzo[d]ꢀ
[1,3]dioxolꢀ5ꢀyl)acetic acid (2e). Compound 2e was obtained
similarly to compound 2a starting from tetraethyl pyrrolidineꢀ
2,2ꢀdiyldiphosphonate 1c (412 mg, 1.20 mmol), 3,4ꢀmethyleneꢀ
dioxyphenylboronic acid (210 mg, 1.25 mmol), and glyoxylic
acid (112 mg, 1.22 mmol). The dichloromethane—methanol
(96 : 4) solvent systemt was used for flashꢀchromatography. The
yield was (460 mg, 73%), colorless crystals, m.p. 134—135 °C.
Found (%): C, 48.71; H, 6.23. C21H33NO10P2. Calculated (%):
C, 48.37; H, 6.38. 1H NMR (CDCl3), δ: 1.06 (t, 3 H, OCH2CH3,
J = 7.1 Hz); 1.24 (t, 3 H, OCH2CH3, J = 7.1 Hz); 1.35 (t, 3 H,
OCH2CH3, J = 7.1 Hz); 1.36 (t, 3 H, OCH2CH3, J = 7.1 Hz);
1.79—1.92 (m, 2 H, CH2); 2.29—2.53 (m, 2 H, CH2); 2.78—2.87
(m, 1 H, CH2); 3.11—3.22 (m, 1 H, CH2); 3.87—4.02 (m, 2 H,
OCH2CH3); 4.03—4.33 (m, 6 H, OCH2CH3); 5.54 (s, 1 H,
CHCOOH); 5.90 (d, 2 H, OCH2O, J = 2.0 Hz); 6.73 (d, 1 H,
5ꢀAr, J = 8.0 Hz); 6.93 (dd, 1 H, 6ꢀAr, J = 8.0 Hz, J = 1.6 Hz);
6.95 (s, 1 H, 2ꢀAr). 13C NMR (CDCl3), δ: 16.1 (d, OCH2CH3,
J = 6.0 Hz); 16.5 (d, OCH2CH3, J = 6.0 Hz); 16. 6 (d, OCH2CH3,
J = 6.0 Hz); 16.6 (d, OCH2CH3, J = 6.0 Hz); 23.9 (t, CH2,
J = 3.5 Hz); 32.1 (t, CH2, J = 3.5 Hz); 48.7 (d, CH2, J = 5.0 Hz);
62.0 (d, OCH2CH3, J = 8.0 Hz); 63.2 (d, OCH2CH3, J = 8.0 Hz);
63.6 (d, OCH2CH3, J = 8.0 Hz); 64.1 (s, CHCOOH); 64.1
(dd, PCP, J = 148.0 Hz, J = 154.0 Hz); 64.72 (d, OCH2CH3,
J = 7.0 Hz); 101.1 (s, OCH2O); 107.7, 110.6, 123.8, 130.0, 147.1,
147.2, 174.0 (s, COOH). 31P NMR (CDCl3), δ: 21.5 (d, 1 P,
J = 92.0 Hz); 23.6 (d, 1 P, J = 92.0 Hz). MS, m/z (Irel (%)): pos.
384 [M + H – HP(O)(OEt)2]+ (100), 206 [M + H –
– HP(O)(OEt)2 – CH2O2C6H3CHCOOH]+ (10); neg. 520
[M – H]–, (100), 382 [M – H – HP(O)(OEt)2]– (20), 338
[M – H – HP(O)(OEt)2 – CO2]– (10).
14. E. D. Matveeva, N. S. Zefirov, Zh. Org. Khim., 2006, 42,
1254 [J. Org. Chem. (Engl. Transl.), 2006, 42].
15. E. D. Matveeva, T. A. Podrugina, M. V. Prisyazhnoi, N. S.
Zefirov, Vestn. Mosk. Univ., Ser. 2, Khim. [Bull. Moscow
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