Synthesis, Src kinase inhibitory and anticancer activities of 1-substituted 3-(N-alkyl-N-phenylamino)propane-2-ols

Article abstract of DOI:10.1016/j.biochi.2010.04.022

A series of 3-(N-alkyl-N-phenylamino)propan-2-ol derivatives were synthesized from epichlorohydrine in a multi-step strategy and were evaluated as Src kinase inhibitors. First, epoxy ring opening of epichlorohydrine was carried out in the presence of N-alkylanilines to yield 3-(N-alkyl-N-phenylamino)-1-chloro-propan-2-ol derivatives using Ca(OTf)₂ as catalyst based on our previous studies [1]. Second, ring closure was performed under basic conditions to afford N-epoxymethyl N-alkylaniline derivatives. Finally, the epoxide ring opening with four different secondary amines and three nucleobases afforded the final products, i.e., a series of β-amino alcohols. All compounds were screened for their inhibitory activity against Src kinase and anticancer activity on human breast carcinoma cells, BT-20 cell line. Among all compounds, 3-N-methyl-N-phenylamino-1-(pyrrolidin-1-yl)propan-2-ol (13b) exhibited the highest inhibitory potency (IC₅₀ = 66.1 μM) against Src kinase. Structure-activity relationship studies suggested that the incorporation of bulky groups at position 1 and N-substitution with groups larger than methyl moiety, reduced the inhibitory potency of the compound significantly. Compounds 3-(N-ethyl-N-phenylamino-)-1-(4-methylpiperazin-1-yl)propan-2-ol (14c) and 3-(N-ethyl-N-phenylamino)-1-(thymine-1-yl)propan-2-ol (17) were found to inhibit the growth of breast carcinoma cells by approximately 45-49% at concentration of 50 μM.

Full text of DOI:10.1016/j.biochi.2010.04.022

Biochimie 92 (2010) 1164e1172
Contents lists available at ScienceDirect
Biochimie
journal homepage: www.elsevier.com/locate/biochi
Research paper
Synthesis, Src kinase inhibitory and anticancer activities of 1-substituted
3-(N-alkyl-N-phenylamino)propane-2-ols
Deepti Sharma a, Raman K. Sharma a, Sumati Bhatia a, Rakesh Tiwari b, Deendayal Mandal b,
**
,
c
a
a,
*
Jessica Lehmann b, Keykavous Parang b , Carl E. Olsen , Virinder S. Parmar , Ashok K. Prasad
a Bioorganic Laboratory, Department of Chemistry, University of Delhi, Delhi 110 007, India
b Department of Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, RI 02881, USA
c University of Copenhagen, Faculty of Life Sciences, Department of Natural Sciences, DK-1871 Frederiksberg C, Denmark
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of 3-(N-alkyl-N-phenylamino)propan-2-ol derivatives were synthesized from epichlorohydrine
in a multi-step strategy and were evaluated as Src kinase inhibitors. First, epoxy ring opening of epi-
chlorohydrine was carried out in the presence of N-alkylanilines to yield 3-(N-alkyl-N-phenylamino)-1-
chloro-propan-2-ol derivatives using Ca(OTf)2 as catalyst based on our previous studies [1]. Second, ring
closure was performed under basic conditions to afford N-epoxymethyl N-alkylaniline derivatives.
Finally, the epoxide ring opening with four different secondary amines and three nucleobases afforded
the final products, i.e., a series of b-amino alcohols. All compounds were screened for their inhibitory
activity against Src kinase and anticancer activity on human breast carcinoma cells, BT-20 cell line.
Among all compounds, 3-N-methyl-N-phenylamino-1-(pyrrolidin-1-yl)propan-2-ol (13b) exhibited the
Received 21 January 2010
Accepted 27 April 2010
Available online 4 May 2010
Keywords:
b-Amino alcohol
Src kinase
Cancer
Cell proliferation
Epoxide
highest inhibitory potency (IC50 ¼ 66.1
mM) against Src kinase. Structure-activity relationship studies
suggested that the incorporation of bulky groups at position 1 and N-substitution with groups larger
than methyl moiety, reduced the inhibitory potency of the compound significantly. Compounds 3-(N-
ethyl-N-phenylamino-)-1-(4-methylpiperazin-1-yl)propan-2-ol (14c) and 3-(N-ethyl-N-phenylamino)-
1-(thymine-1-yl)propan-2-ol (17) were found to inhibit the growth of breast carcinoma cells by
approximately 45e49% at concentration of 50 mM.
Ó 2010 Elsevier Masson SAS. All rights reserved.
1. Introduction
regulating various cellular functions such as cell proliferation and
cell differentiation [4]. Src kinase is a protooncogenic tyrosine
kinase [5] and has been implicated in the genesis and progression
of multiple types of human cancer including colon, breast, lung, and
other cancers [6].
Protein tyrosine kinases (PTKs) are enzymes responsible for the
phosphorylation of other proteins and can catalyze the transfer of
g-phosphate group of ATP to the protein phenolic groups (on Tyr).
PTKs play a central role in signal transduction pathways and are
involved in immune, endocrine, and nervous system physiology
and pathology. On the other hand, the abnormal phosphorylation
could be a trigger of different diseases such as cancer [2]. PTKs are
believed to be important in the development of many different
types of cancers [3].
The Src family of PTKs is divided into 9 subfamilies, Src, Yes, Lck,
Fyn, Lyn, Hck, Frg, Blk, and Yrk, based on catalytic domain sequence.
Src kinases play crucial roles in signal transduction pathways and
Thus, designing Src kinase inhibitors is a subject of major
interest by pharmaceutical industry. Most of the attention has been
on designing Src kinase inhibitors through blocking Src-dependent
phosphorylation of substrate proteins. The kinase domain inhibi-
tors are designed to inhibit the binding of ATP (ATP-binding
site inhibitors) [7] or the protein substrate (substrate binding site
inhibitors) [8]. High affinity ligands targeting the ATP-binding site
have been developed [9]. Unfortunately, most of the ATP-binding
sites within protein kinases are highly conserved. This makes it very
difficult to design a compound that inhibits a specific protein kinase
by targeting the ATP-binding site.
Designing small-molecule inhibitors specific for the Src kinase
domain is challenging due to the highly homologous nature of this
domain within the Src family. Several experimental compounds
[10], such as PP1, PP2, CGP76030, SKI606, PD173955, PD180970,
* Corresponding author. Tel.: þ91 11 27662486.
** Corresponding author. Tel.: þ1 401 874 4471; fax: þ1 401 874 5787.
E-mail addresses: kparang@uri.edu (K. Parang), ashokenzyme@yahoo.com
(A.K. Prasad).
0300-9084/$ e see front matter Ó 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.biochi.2010.04.022

D. Sharma et al. / Biochimie 92 (2010) 1164e1172
1165
and SU6656 have been used for targeting the Src family of kinases.
Only a few inhibitors have been tested without provoking toxicity
in animal models [11]. Thus, there is opportunity to identify other
small-molecule inhibitors or pharmacophore fragments that can
inhibit Src kinase domain.
hematological malignancies such as acute myeloid leukemia, have
also amino alcohol moiety.
Mukaiyama et.al. [22] reported the synthesis of potent pyrazolo
[1,5-a]pyrimidine derivatives containing b-amino alcohols as Src
kinase inhibitors. Lombardo et. al. [23a] and Das et. al. [23b] in
Bristol-Myers Squibb Pharmaceutical research institute reported
BMS-354825 (Dasatinib, trade name Sprycel), a 2-(aminopyrimidinyl)
thiazol-5-carboxamides containing N-2-hydroxyethylpiperidine
group, as a nanomolar Src/Abl kinase inhibitor.
Herein, we report the synthesis and evaluation of a series of 1-
substituted 3-(N-alkyl-N-phenylamino)propane-2-ols as Src kinase
inhibitors to establish the structural requirements surrounding the
b-Amino alcohol fragment attracts interest as an important
moiety present in vast range of biologically active natural and
synthetic compounds [12], unnatural amino acids [13] and as chiral
auxiliaries for asymmetric synthesis [14]. Either the amine or the
alcohol can be acylated, alkylated or contained within rings in
b
b
-amino alcohols. Compounds like bestatin [15], a syn-
-amino acid belonging to this group is an aminopeptidase inhib-
a-hydroxyl-
itor that exhibits immunomodulatory activity [16] and is used as an
adjuvant in cancer chemotherapy [17].
b
-amino alcohols in small molecules for producing inhibitory
potency. These compounds contain a nitrogen atom at the 3-posi-
tion of propanol moiety. Furthermore, in some compounds
There are various anticancer drugs containing
b-amino alcohol
b-
moiety as an important pharmacophore. Daunorubicin (1) and
doxorubicin (2) are the two such examples that are known as
potent and clinically useful agents in cancer chemotherapy.
Daunorubicin and elsamicin A (3) contain vicinal amino alcohol
moiety as sugar which is essential for their biological activity.
Elsamicin A is an antitumor antibiotic in which the presence of
amino sugar enhances the biological activity and the solubility of
the antibiotic. Another example is bestatin (4) which contains
amino alcohol moiety was combined with purine and pyrimidine
nucleobases in one molecule. All synthesized compounds were also
evaluated for their anticancer activity against human breast carci-
noma cells.
2. Materials and methods
2.1. Materials
a syn-a-hydroxy-b-amino acid (Fig. 1). Bestatin is an aminopepti-
dase inhibitor that exhibits immunomodulatory activity and is used
clinically as an adjuvant in cancer chemotherapy [18].
Reactions were conducted under an atmosphere of nitrogen
when anhydrous solvents were used. Column chromatography was
carried out using silica gel (100e200 mesh). Analytical TLCs were
performed on precoated Merck silica-gel 60F254 plates; the spots
were detected under UV light. Silica gel (100e200 mesh) was used
for column chromatography. Dry DMSO was kept over molecular
sieves (4 Å) prior to use.
There are certain form of cancers, cardiovascular and cerebro-
vascular diseases and ischemia/reperfusion injuries which are
known to be caused by free radicals [19]. Antioxidant prevent
oxidative damage and protects body from the harmful effects of
free radicals, a way to prevent cancer. In recent years diphenylalkyl
piperazine derivatives containing the thio- or aminopropanol
moiety have been evaluated for their calcium antagonistic and
antioxidative activities [20]. Phenylamino and benzylamino deriv-
atives of diphenylalkyl piperizine were found to possess inhibitory
activity against auto-oxidative lipid peroxidations in canine brain
homogenates [20]. 9-Substituted adenine and 1-substituted
pyrimidine derivatives which incorporated the phenoxy-2-prop-
anol moiety have also shown hypolipidimeic activity [21]. A
number of nucleobase- and nucleoside-based drugs, like cytar-
2.2. Physical measurements
Melting points were determined using H2SO4 bath and are
uncorrected. The 1H NMR spectra were recorded on a Bruker
Avance spectrometer at 300 and 400 MHz, while 13C NMR spectra
were recorded on a Bruker Avance spectrometer at 75.5 MHz. The
chemical shift values are reported as d ppm relative to TMS used as
internal standard and the coupling constants (J) are measured in
Hz. The ESI-MS spectra of all the compounds were recorded on
a JEOL JMS-AX505W high-resolution mass spectrometer in positive
mode using the matrix HEDS (bishydroxyethyldisulphide) doped
with sodium acetate. The IR spectra were recorded on a Per-
kineElmer model 2000 FT-IR spectrometer by making KBr disc for
solid samples and thin film for oils. The peak of OH group in the 1H
NMR spectra is ascertained on the basis of comparison of spectrum
of the compound in the neat deuterated solvent and in the
deuterated solvent plus D2O.
abine,
a chemotherapeutic agent used for the treatment of
O
OH
O
O
OH
O
OH
OH
OH
O
OMe O
OH O
O
O
OH
O
CH3
OH
H3C
O
2.3. General procedure for the synthesis of 3-(N-alkyl-N-phenyl)
amino-1-chloropropane-2-ols (8 and 9)
H2N
NH2
OH
Daunorubicin (1)
Doxorubicin (2)
To a mixture of N-alkylaniline (5 and 6, 93 mmol) and Ca(OTf)2
(46 mmol) in acetonitrile was added epichlorohydrin (7, 93 mmol)
dropwise. The reaction was stirred at room temperature for 6 h. The
reaction mixture was added to water and extracted with ethyl
acetate. The organic phase was dried over Na2SO4 and evaporated
under reduced pressure. It was further purified by column chro-
matography eluting with 6e8% ethyl acetateepetroleum ether
containing 0.5% of triethylamine to afford compound 8 and 9.
Compounds 8 and 9 were obtained as racemic mixture in 80e87%
yield, as racemic epichlorohydrin has been used for their synthesis
and chiral conditions have not been used in any of the steps for the
synthesis of these compounds.
HO
O
HO
O
O
O
NH2
O
Ph
HOOC
N
H
H
O
OH
NH2
O
OH
O
Elsamicin A (3)
Bestatin (4)
Fig. 1. Some of the anticancer drugs containing b- aminoalcohol moiety.

1166
D. Sharma et al. / Biochimie 92 (2010) 1164e1172
2.3.1. 1-Chloro-3-(N-methyl-N-phenylamino)propan-2-ol (8)
It was obtained as brown colored oil (14.8 g) in 80% yield.
Rf ¼ 0.4 in 15% ethyl acetate in petroleum ether. IR (cmꢀ1, nujol):
3413 (OH), 2906 (NeCH3), 1600, 1505, 750 and 694; 1H NMR
(NeCH2CH3), 48.08 and 48.14 (NeCH2CH3 and C-3), 53.30 (C-1),
54.45 (C-2), 114.72 (C-40), 118.92 (C-30 and C-50), 131.87 (C-20 and C-
60) and 150.59 (C-10); HRMS (ESI positive mode): m/z 178.1228
[M þ H]þ, calculated for C11H15NO þ H 178.1226.
(400 MHz, CDCl3):
d 2.57 (1H, br s, OH), 2.90 (3H, s, NeCH3), 3.35
(2H, d, J ¼ 6.4 Hz, Ce3H), 3.51 (1H, dd, J ¼ 5.4 and 11.2 Hz, Ce1Ha),
3.60 (1H, dd, J ¼ 4.0 and 11.2 Hz, Ce1Hb), 4.05 (1H, br s, Ce2H),
6.66e6.71 (3H, m, Ce30H, Ce40H and Ce50H) and 7.15e7.17 (2H, m,
2.5. General procedure for the synthesis of 1-substituted-3-(N-
alkyl-N-phenyl)amino propane-2-ols (13ae13d and 14ae14d)
Ce20H and Ce60H); 13C NMR (75.5 MHz, CDCl3):
d
39.96 (NeCH3),
To a mixture of secondary cyclic amines (12ae12d, 5.6 mmol)
and Ca(OTf)2 (2.8 mmol) in acetonitrile was added N-Epoxymethyl-
N-alkyl anilines (10 and 11, 5.6 mmol) dropwise. The reaction was
stirred at room temperature for 6 h. The reaction mixture was
added to water and extracted with ethyl acetate. The organic phase
was dried over Na2SO4 and evaporated under reduced pressure. It
was further purified by column chromatography eluting with 2e4%
methanol-chloroform containing 0.5% of triethylamine to afford
compound 13ae13d and 14ae14d in 75e90% yield. Compounds
13ae13d and 14ae14d were obtained as racemic mixture as
racemic epichlorohydrin has been used for their synthesis and
chiral conditions have not been used in any of the steps for the
synthesis of these compounds.
48.16 (C-3), 56.94 (C-1), 69.41 (C-2), 113.35 (C-40), 117.93 (C-30 and
C-50), 129.70 (C-20 and C-60) and 148.22 (C-10); HRMS (ESI positive
mode): m/z 200.0839 [M þ H]þ, calculated for C10H14ClNO þ H
200.0837.
2.3.2. 1-Chloro-3-(N-ethyl-N-phenylamino)propan-2-ol (9)
It was obtained as brown color oil (17.3 g) in 87% yield. Rf ¼ 0.3 in
15% ethyl acetate in petroleum ether. IR (cmꢀ1, nujol): 3430 (OH),
2972 (NeCH2CH3), 1599, 1505, 748 and 694; 1H NMR (300 MHz,
CDCl3):
d
1.12 (3H, t, J ¼ 7.0 Hz, NeCH2CH3), 2.60 (1H, br s, OH),
3.32e3.45 (4H, m, Ce3H and NeCH2CH3), 3.58 (1H, dd, J ¼ 5.4 and
11.2 Hz, Ce1Ha), 3.66 (1H, dd, J ¼ 4.3 and 11.2 Hz, Ce1Hb), 4.07e4.11
(1H, m, Ce2H), 6.71e6.78 (3H, m, Ce30H, Ce40H and Ce50H) and
7.20e7.25 (2H, t, J ¼ 8.5 Hz, Ce20H and Ce60H); 13C NMR (75.5 MHz,
2.5.1. 3-(N-Methyl-N-phenylamino)-1-(piperidin-
1-yl)propan-2-ol (13a)
It was obtained as brown colored oil (1.1 g) in 80% yield. Rf ¼ 0.5
in 5% methanol in chloroform. IR (cmꢀ1, nujol): 3311 (OH), 2937
(NeCH3), 1599, 1507, 749 and 694; 1H NMR (300 MHz, CDCl3):
CDCl3):
d
11.81 (NeCH2CH3), 46.61 (NeCH2CH3), 47.96 (C-3), 54.16
(C-1), 69.09 (C-2), 113.75 (C-40), 117.64 (C-30 and C-50), 129.59 (C-20
and C-60) and 148.22 (C-10); HRMS (ESI positive mode): m/z
214.0994 [M þ H]þ, calculated for C11H16ClNO þ H 214.0993.
d
1.41e1.43 (2H, m, Ce400H), 1.55 (4H, br s, Ce300H and Ce500H),
2.4. General procedure for the synthesis of N-epoxymethyl-
N-alkyl anilines (10 and 11)
2.27e2.36 (4H, m, Ce200H and C-600), 2.54e2.56 (2H, m, Ce1H), 2.90
(3H, s, NeCH3), 3.32 (2H, d, J ¼ 5.5 Hz, C-3 H), 3.97e3.99 (1H, m,
Ce2H), 4.20 (1H, br s, OH), 6.65e6.67 (3H, m, Ce30H, Ce40H and
Ce50H) and 7.17e7.22 (2H, Ce20H and Ce60H); 13C NMR (75.5 MHz,
To a solution of 3-(N-Alkyl-N-phenyl)amino-1-chloropropane-
2-ols (8 and 9, 50 mmol) in toluene (75 mL) was added 5% aq. NaOH
(75 mL) solution. The reaction mixture was stirred at room
temperature for 5 h and then extracted with ethyl acetate. The
organic phase was washed with water, was dried over Na2SO4, and
evaporated under reduced pressure. It was further purified by
column chromatography eluting with 2e3% ethyl acetateepetro-
leum ether containing 0.5% of triethylamine to afford compound 10
and 11 in 75e80% yield.
CDCl3):
d
24.57 (C-400), 26.34 (C-500 and C-300), 39.98 (NCH3), 55.18
(C-200 and C-600), 57.84 (C-1), 63.27 (C-3), 65.78 (C-2), 112.76 (C-40),
116.84 (C-30 and C-50), 129.55 (C-20 and C-60) and 150.15 (C-10);
HRMS (ESI positive mode): m/z 249.1962 [M þ H]þ, calculated for
C15H24N2O þ H 249.1961.
2.5.2. 3-(N-Methyl-N-phenylamino)-1-(pyrrolidin-
1-yl)propan-2-ol (13b)
It was obtained as brown colored oil (1.1 g) in 83% yield. Rf ¼ 0.4
in 5% methanol in chloroform. IR (cmꢀ1, nujol): 3394 (OH), 2966
(NeCH3), 2928, 1599, 1506, 746 and 693; 1H NMR (400 MHz,
2.4.1. N-Epoxypropyl-N-methylaniline (10)
It was obtained as light brown colored oil (6.1 g) in 75% yield.
Rf ¼ 0.4 in 5% ethyl acetate in petroleum ether. IR (cmꢀ1, nujol):
2993 (NeCH3), 2920, 1600, 1505, 1255, 1125 (CeO), 750 and 693; 1H
CDCl3):
d
1.82e1.85 (4H, m, Ce300H and Ce400H), 2.63 (2H, dd, J ¼ 2.4
and J ¼ 12.4 Hz, Ce1H), 2.80e2.93 (7H, m, NeCH3,Ce200H and
Ce500H), 3.31 (2H, dd, J ¼ 4.8 and 13.2 Hz, Ce3H), 4.09e4.12 (1H, br
s, Ce2H), 6.62e6.67 (3H, m, Ce30H, Ce40H and Ce50H) and
7.13e7.19 (2H, m, Ce20H and Ce60H); 13C NMR (75.5 MHz, CDCl3):
NMR (300 MHz, CDCl3):
d
2.60 (1H, dd, J ¼ 2.6 Hz and 4.9 Hz,
Ce1Ha), 2.80e2.83 (1H, m, Ce1Hb), 3.03 (3H, s, NeCH3), 3.17e3.20
(1H, m, Ce2H), 3.43 (1H, dd, J ¼ 4.8 and 15.6 Hz, Ce3Ha), 3.68 (1H,
dd, J ¼ 3.1 and 15.6 Hz, Ce3Hb), 6.75e6.81 (3H, m, Ce30H, Ce40H
and Ce50H) and 7.25e7.31 (2H, m, Ce20H and Ce60H); 13C NMR
d
23.58 (C-300 and C-400), 45.97 (NeCH3), 54.25 (C-200 and C-500),
55.01 (C-1), 60.14 (C-3), 67.13 (C-2), 112.69 (C-40), 116.31 (C-30 and
C-50), 129.24 (C-20 and C-60) and 148.29 (C-10); HRMS (ESI positive
mode): m/z 235.1805 [M þ H]þ, calculated for C14H22N2O þ H
235.1805.
(75.5 MHz, CDCl3): d 41.45 (NCH3), 47.51 (C-3), 52.93 (C-1), 56.68 (C-
2), 114.88 (C-40), 119.33 (C-30 and C-50), 131.61 (C-20 and C-60) and
151 (C-10); HRMS (ESI positive mode): m/z 164.1066 [M þ H]þ,
calculated for C10H13NO þ H 164.1070.
2.5.3. 3-(N-Methyl-N-phenylamino)-1-(4-methylpiperazin-
1-yl)propan-2-ol (13c)
2.4.2. N-Epoxypropyl-N-ethylaniline (11)
It was obtained as light brown colored oil (7 g) in 80% yield.
Rf ¼ 0.45 in 5% ethyl acetate in petroleum ether. IR (cmꢀ1, nujol):
2973 (NeCH2CH3), 1599, 1505, 1352, 1193 (CeO), 748 and 693; 1H
It was obtained as brown colored oil (1.4 g) in 78% yield. Rf ¼ 0.5
in 5% methanol in chloroform. IR (cmꢀ1, nujol): 3374 (OH), 2938
(NeCH3), 2805, 1600, 1506, 1455, 749 and 693; 1H NMR (400 MHz,
NMR (300 MHz, CDCl3):
d
1.15 (3H, t, J ¼ 6.9 Hz, NeCH2CH3), 2.56
CDCl3): d
2.22 (3H, s, 400NeCH3), 2.29e2.39 (8H, m, Ce200H, C-300,
(1H, dd, J ¼ 2.6 and 4.9 Hz, Ce1Ha), 2.76e2.79 (1H, m, Ce1Hb),
3.11e3.13 (1H, m, Ce2H), 3.34e3.45 (3H, m, Ce3Ha and
NeCH2CH3), 3.60 (1H, dd, J ¼ 3.1 and 15.8 Hz, Ce3Hb), 6.66e6.74
(3H, m, Ce30H, Ce40H and Ce50H) and 7.21 (2H, dd, J ¼ 7.2 Hz and
Ce500H and Ce600H), 2.61 (2H, br s, Ce1H), 2.92 (3H, s, NeCH3),
3.27e3.29 (2H, m, Ce3H), 3.39 (1H, br s, OH), 3.90e3.94 (1H, m,
Ce2H), 6.62e6.69 (3H, m, Ce30H, Ce40H, and Ce50H) and 7.13e7.17
(2H, m, Ce20H and Ce60H); 13C NMR (75.5 MHz, CDCl3):
d
39.57
8.7 Hz, Ce20H and Ce60H); 13C NMR (75.5 MHz, CDCl3):
d14.47
(400NeCH3), 45.81 (NeCH3), 53.08 (C-300 and C-500), 55.02 (C-200 and

D. Sharma et al. / Biochimie 92 (2010) 1164e1172
1167
Ce600H), 57.38 (C-1), 61.90 (C-3), 65.48 (C-2), 112.44 (C-40), 116.62
(NeCH2CH3), 45.54 (400NeCH3), 46.00 (NeCH2CH3), 52.73 (C-300 and
C-500), 54.76 (C-200 and C-600), 54.88 (C-1), 61.93 (C-3), 65.49 (C-2),
112.69 (C-40), 116.38 (C-30 and C-50), 129.23 (C-20 and C-60) and
148.19 (C-10); HRMS (ESI positive mode): m/z 278.2233 [M þ H]þ,
calculated for C16H27N3O þ H 278.2227.
(C-30 and C-50), 129.13 (C-20 and C-60) and 149.74 (C-10); HRMS (ESI
positive mode): m/z 264.2077 [M
C15H25N3O þ H 264.2070.
þ
H]þ, calculated for
2.5.4. 3-(N-Methyl-N-phenylamino)-1-morpholinopropan-
2-ol (13d)
2.5.8. 3-(N-Ethyl-N-phenylamino)-1-morpholinopropan-2-ol (14d)
It was obtained as brown colored oil (1.2 g) in 86% yield. Rf ¼ 0.5
in 5% methanol in chloroform. IR (cmꢀ1, nujol): 3445 (OH), 2963
(NeCH3), 2855, 1599, 1505, 1117, 866, 748 and 695; 1H NMR
It was obtained as brown colored oil (1.2 g) in 90% yield. Rf ¼ 0.6
in 5% methanol in chloroform. IR (cmꢀ1, nujol): 3345 (OH), 2966
(NeCH3), 2865, 1599, 1505, 1117, 866, 748 and 695; 1H NMR
(400 MHz, CDCl3):
d
2.37e2.44 (4H, m, Ce200H and Ce600H),
(400 MHz, CDCl3):
d
1.07 (3H, t, J ¼ 6.8 Hz, NeCH2CH3), 2.28e2.42
2.57e2.64 (2H, m, Ce1H), 2.98 (3H, s, NeCH3), 3.34 (2H, d,
J ¼ 5.6 Hz, Ce3H), 3.43 (1H, s, OH), 3.69 (4H, br s, Ce200H and
Ce500H), 4.09e4.12 (1H, br s, Ce2H), 6.70e6.76 (3H, m, Ce30H,
Ce40H and Ce50H) and 7.19e7.24 (2H, m, Ce20H and Ce60H); 13C
(4H, m, Ce200H and Ce600H), 2.53e2.58 (2H, m, Ce1H), 3.10 (1H, br s,
OH), 3.25 (2H, d, J ¼ 6.0 Hz, Ce3H), 3.30e3.43 (2H, m, NeCH2CH3),
3.59e3.68 (4H, br s, Ce300H and Ce500H), 3.88e3.94 (1H, br s,
Ce2H), 6.59e6.67 (3H, m, Ce30H, Ce40H and Ce50H) and 7.12e7.15
NMR (75.5 MHz, CDCl3):
d
39.98 (NeCH3), 54.18 (C-1), 57.75 (C-200
(2H, m, Ce20H andCe60H); 13C NMR (75.5 MHz, CDCl3):
d 11.56
and C-600), 63.01 (C-3), 65.79 (C-300 and C-500), 67.35 (C-2), 112.86 (C-
40), 117.08 (C-30 and C-50), 129.55 (C-20 and C-60) and 150.0 (C-10);
HRMS (ESI positive mode): m/z 251.1755 [M þ H]þ, calculated for
C14H22N2O2þH 251.1754.
(NeCH2CH3), 46.02 (NeCH2CH3), 53.82 (C-1), 54.92 (C-200 and C-600),
62.69 (C-3), 65.39 (C-300 and C-500), 67.01 (C-2), 112.67 (C-40), 116.37
(C-20 and C-60), 129.25 (C-30 and C-50) and 148.25 (C-10); HRMS (ESI
positive mode): m/z 265.1912 [M
C15H24N2O2þH 265.1911.
þ
H]þ, calculated for
2.5.5. 3-(N-Ethyl-N-phenylamino)-1-(piperidin-
1-yl)propan-2-ol (14a)
2.6. General procedure for the synthesis of 3-(N-alkyl-N-phenyl)
It was obtained as brown colored oil (1.2 g) in 85% yield. Rf ¼ 0.5
in 5% methanol in chloroform. IR (cmꢀ1, nujol): 3399 (OH), 2936
(NeCH2CH3), 1598, 1505, 1246, 1031, 748 and 638; 1H NMR
amino-(thymine-1-yl)propane-2-ols (16 and 17)
To
a mixture of thymine (15) (7.36 mmol) and K2CO3
(400 MHz, CDCl3):
d
1.06 (3H, t, J ¼ 6.8 Hz, NeCH2CH3), 1.37e1.39
(61.3 mmol) in DMSO (20 mL), compound N-epoxymethyl-N-alkyl
anilines (10 and 11, 6.13 mmol) was added dropwise under N2
atmosphere. The reaction mixture was heated to 60 ꢁC for 48 h. It
was then added to ice cold water and extracted with ethyl acetate.
The organic phase was dried over Na2SO4 and removed under
pressure. The crude product was further purified by silica gel
column chromatography eluting with 2e4% methanol-chloroform
containing 0.5% of triethylamine to afford compound 16 and 17 in
45e50% yield.
(2H, m, Ce400H), 1.53e1.56 (4H, m, C-300 and Ce500H), 2.30e2.42 (4H,
m, Ce200H and Ce600H), 2.55 (2H, br s, Ce1H), 3.21e3.26 (2H, m,
NeCH2CH3), 3.33 (1H, br s, OH), 3.53 (2H, br s, Ce3H), 3.94e3.97
(1H, br s, Ce2H), 6.57e6.66 (3H, m, Ce30H, Ce40H and Ce50H) and
7.11e7.18 (2H, m, Ce20H and Ce60H); 13C NMR (75.5 MHz, CDCl3):
d
11.52 (NeCH2CH3), 23.15 (C-400), 24.60 (C-500 and C-300), 46.19
(NeCH2CH3), 49 (C-200 and C-600), 54.83 (C-1), 62.38 (C-3), 64.81 (C-
2), 113.12 (C-40), 116.89 (C-30 and C-50), 129.34 (C-20 and C-60) and
148.08 (C-10); HRMS (ESI positive mode): m/z 263.2119 [M þ H]þ,
calculated for C16H26N2O þ H 263.2118.
2.6.1. 3-(N-Methyl-N-phenylamino)-1-(thymine-
1-yl)propane-2-ol (16)
2.5.6. 3-(N-Ethyl-N-phenylamino)-1-(pyrrolidin-1-yl)propan-
2-ol (14b)
It was obtained as brown colored oil (1.1 g) in 80% yield. Rf ¼ 0.4
in 5% methanol in chloroform. IR (cmꢀ1, nujol): 3366 (OH), 2967
(NeCH2CH3), 2802, 1598, 1505, 1191, 746 and 694; 1H NMR
It was obtained as white solid (0.88 g) in 50% yield. Rf ¼ 0.5 in 5%
methanol in chloroform; M. Pt. ¼ 180e184 ꢁC. IR (cmꢀ1, KBr) 3349
(OH), 3191 (NH), 3059, 2978 (NeCH3), 2947, 1697 (CO), 1660 (CO),
1506 and 745; 1H NMR (300 MHz, CDCl3):
d
1.83 (3H, s, C-500CH3),
2.99 (3H, s, NeCH3), 3.40e3.45 (2H, m, Ce3H), 3.99e4.09 (3H, m,
Ce1H and Ce2H), 5.12 (1H, d, J ¼ 5.4 Hz, OH), 6.61e6.73 (3H, m,
Ce30H, Ce40H and Ce50H), 7.13e7.18 (2H, m, Ce20H and Ce60H),
7.26 (1H, s, Ce600H) and 11.11 (1H, s, NH); 13C NMR (75.5 MHz,
(400 MHz, CDCl3):
d
1.06 (3H, t, J ¼ 6.8 Hz, NeCH2CH3), 1.70e1.74
(4H, m, Ce300H and Ce400H), 2.40e2.65 (6H, m, Ce200H, Ce500H, and
Ce1H), 3.23e3.43 (5H, m, NCH2CH3, Ce3H and OH), 3.90e3.93 (1H,
br s, Ce2H), 6.58e6.67 (3H, m, Ce30H, Ce40H and Ce50H) and
7.12e7.15 (2H, m, Ce20H and Ce60H); 13C NMR (75.5 MHz, CDCl3):
CDCl3): d
12.24 (500CeCH3), 40.47 (NeCH3), 52.05 (C-3), 56.70 (C-1),
67.41 (C-2), 108.47 (C-500), 112.12 (C-40), 116.09 (C-30 and C-50),
128.99 (C-20 and C-60), 142.36 (C-600), 149.41 (C-10), 151.38 (C-200)
and 164.93(C-400); HRMS (ESI positive mode): m/z 290.1491
[M þ H]þ, calculated for C15H19N3O3þH 290.1499.
d
11.51 (NeCH2CH3), 23.55 (C-300 and C-400), 45.99 (NCH2CH3), 54.28
(C-200 and C-500), 54.97 (C-1), 60.19 (C-3), 67.04 (C-2), 112.72 (C-40),
116.36 (C-30 and C-50), 129.25 (C-20 and C-60) and 148.26 (C-10);
HRMS (ESI positive mode): m/z 249.1961 [M þ H]þ, calculated for
C15H24N2O þ H 249.1961.
2.6.2. 3-(N-Ethyl-N-phenylamino)-1-(thymine-
1-yl)propane-2-ol (17)
2.5.7. 3-(N-Ethyl-N-phenylamino)-1-(4-methylpiperazin-
1-yl)propan-2-ol (14c)
It was obtained as brown colored oil (1.1 g) in 75% yield. Rf ¼ 0.5
in 5% methanol in chloroform. IR (cmꢀ1, nujol): 3365 (OH), 2937
(NeCH2CH3), 1598, 1506, 747 and 694; 1H NMR (300 MHz, CDCl3):
It was obtained as white sticky mass (0.83 g) in 45% yield.
Rf ¼ 0.5 in 5% methanol in chloroform. IR (cmꢀ1, KBr): 3386 (OH),
2928 (NeCH2CH3), 1686 (CO), 1598, 1505 and 749; 1H NMR
(300 MHz, CDCl3):
d
1.06 (3H, t, J ¼ 6.9 Hz, NeCH2CH3), 1.75 (3H, s,
C-500CH3), 3.21e3.40 (4H, m, Ce3H, NeCH2CH3), 3.92 (3H, br s,
Ce1H and Ce2H), 5.23 (1H, d, J ¼ 5.4 Hz, OH), 6.56e6.68 (3H, m,
Ce30H, Ce40H and Ce50H), 7.10e7.15 (2H, m, Ce20H and Ce60H),
7.41 (1H, d, J ¼ 1 Hz, Ce600H) and 11.19 (1H, s, NH); 13C NMR
d
1.14 (3H, t, J ¼ 6.9 Hz, NeCH2CH3), 2.36 (3H, s, 400NeCH3),
2.39e2.57 (8H, m, Ce200H, Ce300H, Ce500H and Ce600H), 2.76 (2H, br
s, Ce1H), 3.31 (2H, d, J ¼ 5.7 Hz, Ce3H), 3.35e3.51 (2H, m,
NeCH2CH3), 3.69 (1H, br s, OH), 3.95e4.01 (1H, m, Ce2H),
6.66e6.75 (3H, m, Ce30H, Ce40H and Ce50H) and 7.18e7.26 (2H, m,
(75.5 MHz, CDCl3):
d
11.27 (NeCH2CH3), 11.89 (500CeCH3), 44.89
(NeCH2CH3), 54.07 (C-3), 59.73 (C-1), 66.71 (C-2), 107.46 (C-500),
Ce20H and Ce60H); 13C NMR (75.5 MHz, CDCl3):
d
11.51
111.79 (C-40), 115.23 (C-30 and C-50), 128.98 (C-20 and C-60), 142.75

1168
D. Sharma et al. / Biochimie 92 (2010) 1164e1172
(C-600), 147.78 (C-10), 150.97 (C-200) and 164.41 (C-400); HRMS (ESI
d
40.73 (NeCH3), 47.67 (C-3), 56.77 (C-1), 67.57 (C-2), 112.11 (C-40),
positive mode): m/z 304.1638 [M
C16H21N3O3þH 304.1656.
þ
H]þ, calculated for
115.96 (C-30 and C-50), 118.97 (C-500), 129.19 (C-20 and C-60), 141.81
(C-800), 149.49 (C-10), 149.98 (C-400), 152.59 (C-200) and 156.25 (C-600);
HRMS (ESI positive mode): m/z 299.1611 [M þ H]þ, calculated for
C15H18N6O þ H 299.1615.
2.7. General procedure for the synthesis of 3-(N-alkyl-N-phenyl)
amino-(uracil-1-yl) propane-2-ols and 3-(N-alkyl-N-phenyl)amino-
(adenine-9-yl) propane-2-ols (19ae19b and 20ae20b)
2.7.4. 3-(N-Ethyl-N-phenylamino)-1-(adenine-
9-yl)propan-2-ol (20b)
To a mixture of nucleobase (18a and 18b, 7.36 mmol) and K2CO3
(61.3 mmol) in DMF (20 mL) was added dropwise N-epoxymethyl-
N-alkyl anilines (10 and 11, 6.13 mmol) under N2 atmosphere.The
reaction temperature was raised to 80 ꢁC and stirred for 55 h in case
of 18a and 18 h in case of 18b. DMF was removed under reduced
pressure and the residue was added to water and extracted with
ethyl acetate. The organic phase was dried over Na2SO4 and
removed under pressure. It was further purified by silica gel column
chromatography eluting with 3e5% methanol-chloroform con-
taining 0.5% of triethylamine to afford compound 19ae19b and
20ae20b.
It was obtained as white solid (1.3 g) in 72% yield. Rf ¼ 0.5 in 5%
methanol in chloroform; M. Pt. ¼ 158e162 ꢁC. IR (cmꢀ1, nujol):
3298 (OH), 3144 (NH2), 2969 (NeCH2CH3), 2927, 1685, 1598, 1504,
748 and 649; 1H NMR (300 MHz, CDCl3):
d
1.05 (3H, t, J ¼ 6.9 Hz,
NeCH2CH3), 2.49e2.51 (2H, m, NeCH2CH3), 3.19e3.27 (2H, m,
Ce3H), 4.07e4.10 (2H, m, Ce1H), 4.26 (1H, m, Ce2H), 5.35 (1H, d,
J ¼ 4.8 Hz, OH), 6.55e6.65 (3H, m, Ce30H, Cee40H and Ce50H),
7.08e7.16 (4H, m, C-500NH2, Ce20H and Ce60H), 8.07 (1H, s, Ce800H)
and 8.12 (1H, s, Ce200H); 13C NMR (75.5 MHz, CDCl3):
d 11.23
(NeCH2CH3), 44.89 (NeCH2CH3), 47.34 (C-3), 54.13 (C-1), 67.25 (C-
2), 111.79 (C-40), 115.27 (C-30 and C-50), 118.51 (C-500), 128.99 (C-20
and C-60), 141.59 (C-800), 147.75 (C-10), 149.59 (C-400), 152.26 (C-200)
and 155.85 (C-600); HRMS (ESI positive mode): m/z 313.1759
[M þ H]þ, calculated for C16H20N6O þ H 313.1771.
2.7.1. 3-(N-Methyl-N-phenylamino)-1-(uracil-
1-yl)propan-2-ol (19a)
It was obtained as white solid (1.18 g) in 70% yield. Rf ¼ 0.5 in 5%
methanol in chloroform; M. Pt. ¼ 196e200 ꢁC. IR (cmꢀ1, KBr): 3346
(OH), 3163, 3037 (NH), 2902 (NeCH3), 2826, 1668 (CO), 1600
2.8. General procedure for the acetylation of 3-(-(N-ethyl-N-
phenyl)amino-1-)morpholinopropan-2-ol (14d) and 3-(N-ethyl-
N-phenylamino)(thymine-1-yl)propan-2-ol (17) to prepare
acetates 21 and 22, respectively
(CO),1505 and 749; 1H NMR (300 MHz, CDCl3):
d 2.99 (3H, s,
NeCH3), 3.19e3.29 (2H, m, Ce3H), 3.93 (3H, br s, Ce1H and Ce2H),
5.24 (1H, d, J ¼ 5.7 Hz, OH), 5.50 (1H, d, J ¼ 7.8 Hz, Ce500H),
6.60e6.70 (3H, m, Ce30H, Ce40H and Ce50H), 7.12e7.17 (2H, m,
Ce20H and Ce60H), 7.52 (1H, d, J ¼ 7.8 Hz, Ce600H) and 11.19 (1H, s,
To a mixture of 14d (0.500 g, 1.89 mmol) and DMAP (23.05 mg,
0.189 mmol) in acetonitrile (10 mL) acetic anhydride was added.
The reaction was stirred at room temperature for 3 h. Acetonitrile
was removed under reduced pressure and the residue added to
water and extracted with ethyl acetate. The organic phase was
dried over Na2SO4 and removed under pressure. It was further
purified by silica gel column chromatography eluting with 40%
ethyl acetateepetroleum ether containing 0.5% of triethylamine to
afford compound 21. Similarly compound 22 was obtained by
acetylation of compound 17 in the similar fashion. It was further
purified by silica gel column chromatography eluting with 2%
methanol-chloroform containing 0.5% of triethylamine to afford
compound 22.
NH); 13C NMR (75.5 MHz, CDCl3):
d 40.26 (NeCH3), 51.89 (C-3),
56.15 (C-1), 66.51 (C-2), 100.02 (C-500), 111.80 (C-40), 115.60 (C-30 and
C-50), 128.88 (C-20 and C-60), 146.89 (C-600), 149.13 (C-10), 151.0 (C-
200) and 163.88 (C-400); HRMS (ESI positive mode): m/z 276.1347
[M þ H]þ, calculated for C14H17N3O3þH 276.1343.
2.7.2. 3-(N-Ethyl-N-phenylamino)-1-(uracil-
1-yl)propan-2-ol (20a)
It was obtained as white solid 70% yield. Rf ¼ 0.5 in 5% methanol
in chloroform; M. Pt. ¼142e146 ꢁC. IR (cmꢀ1, KBr): 3408 (OH), 3186
(NH), 3045, 2937 (NeCH2CH3), 1670 (CO), 1598, 1459 and 748; 1H
NMR (300 MHz, CDCl3):
d
1.06 (3H, t, J ¼ 6.9 Hz, NeCH2CH3),
2.8.1. 2-Acetoxy-3-(N-ethyl-N-phenyl)amino-1-
morpholinopropane-2-ol (21)
It was obtained as yellow color oil in 90% yield. Rf ¼ 0.6 (40%
ethyl acetateepetroleum ether). IR (cmꢀ1, Film): 2927 (NeCH2CH3),
1737 (CO), 1599, 1506, 748 and 695; 1H NMR (300 MHz, CDCl3):
3.16e3.31 (5H, m, Ce2H, Ce3H, NeCH2CH3), 3.92e3.96 (2H, m,
Ce1H), 5.26 (1H, d, J ¼ 5.1 Hz, OH), 5.51 (1H, d, J ¼ 7.8 Hz, Ce500H),
6.560 (1H, t, J ¼ 7.2 Hz, Ce40H), 6.67 (2H, d, J ¼ 6.6 Hz, Ce30H and
Ce5 H), 7.10e7.16 (2H, m, Ce20H and Ce60H), 7.52 (1H, d, J ¼ 7.8 Hz,
Ce600H) and 11.19 (1H, s, NH); 13C NMR (75.5 MHz, CDCl3):
d
11.29
d
1.04 (3H, t, J ¼ 6.9 Hz, NeCH2CH3), 1.923 (3H, s, CH3CO), 2.41e2.49
(NeCH2CH3), 44.90 (NeCH2CH3), 51.95 (C-3), 54.05 (C-2), 66.61 (C-
1), 100.02 (C-500), 111.79 (C-40), 115.25 (C-30 and C-50), 128.99 (C-20
and C-60), 146.93 (C-600), 147.76 (C-10), 151.0 (C-400) and 163.89 (C-
200); HRMS (ESI positive mode): m/z 290.1489 [M þ H]þ, calculated
for C15H19N3O3þH 290.1499.
(4H, m, Ce200H and Ce600H), 3.32e3.58 (8H, m, Ce1H, Ce3H,
NeCH2CH3), 5.09e5.13 (1H, m, Ce2H), 6.57 (1H, t, J ¼ 5.2 Hz,
Ce40H), 6.73 (2H, d, J ¼ 8.1 Hz, Ce20H and Ce60H) and 7.14 (2H, dd,
J ¼ 7.2 and 8.1 Hz, Ce20H and Ce60H); 13C NMR (75.5 MHz, CDCl3):
d
11.60 (NeCH2CH3), 20.83 (CH3CO), 44.67 (NeCH2CH3), 51.59 (C-1),
53.76 (C-200 and C-600), 59.07 (C-3), 66.26 (C-300 and C-500), 69.19 (C-
2), 111.78 (C-40), 115.45 (C-20 and C-60), 128.98 (C-30 and C-50), 147.51
(C-10) and 169.83 (CO). HRMS (ESI positive mode): m/z 348.0915
[M þ H]þ, calculated for C17H26N2O3þH 348.0902.
2.7.3. 3-(N-Methyl-N-phenylamino)-1-(adenine-
9-yl)propan-2-ol (19b)
It was obtained as white solid (1.2 g) in 68% yield. Rf ¼ 0.6 in 5%
methanol in chloroform; M. Pt. ¼172e176 ꢁC. IR (cmꢀ1, nujol): 3309
(OH), 3143 (NH2), 2928 (NeCH3), 1680, 1603, 1507, 748 and 692; 1H
2.8.2. 2-Acetoxy-3-(N-ethyl-N-phenylamino)-1-(thymine-
1-yl)propane-2-ol (22)
It was obtained as white solid in 88% yield. Rf ¼ 0.5 (60% ethyl
acetate- petroleum ether). M.pt. ¼ 194e196 ꢁC; IR (cmꢀ1, Film):
2928 (NeCH2CH3), 1730 (CO), 1686 (CO), 1590, 1500 and 749; 1H
NMR (400 MHz, CDCl3):
d
2.93 (3H, s, NeCH3), 3.23 (1H, dd, J ¼ 6.4
and 15.0 Hz, Ce3Ha), 3.43 (1H, dd, J ¼ 4.6 and 15.0 Hz, Ce3Hb),
4.04e4.09 (2H, m, Ce1H), 4.24 (1H, d, J ¼ 10.8 Hz, Ce2H), 5.34(1H,
d, J ¼ 5.2 Hz, OH), 6.56e6.66 (3H, m, Ce30H, Ce40H and Ce50H), 7.12
(2H, t, J ¼ 7.8 Hz, Ce20H and Ce60H), 7.24 (2H, s, C-600NH2), 8.07 (1H,
s, Ce800H) and 8.11 (1H, s, Ce200H); 13C NMR (75.5 MHz, CDCl3):
NMR (300 MHz, CDCl3):
d
1.05 (3H, t, J ¼ 6.6 Hz, NeCH2CH3), 1.74
(3H, s, C-500CH3), 1.85 (3H, s, CH3CO), 3.34e3.54 (4H, m, Ce3H,

D. Sharma et al. / Biochimie 92 (2010) 1164e1172
1169
NeCH2CH3), 3.73 (1H, dd, J ¼ 9.3 and 14.4 Hz, Ce1Ha), 4.01e4.06
(1H, m, Ce1Hb), 5.30 (1H, brs, Ce2H), 6.60 (1H, t, J ¼ 7.2 Hz, Ce40H),
6.71 (2H, d, J ¼ 8.1 Hz, Ce30H and Ce50H), 7.15 (2H, t, J ¼ 7.8 Hz, C-20
and C-60), 7.45(1H, s, Ce600H) and 11.25 (1H, s, NH); 13C NMR
grown on 75 cm2 cell culture flasks with EMEM (Eagle’s minimum
essential medium), supplemented with 10% fetal bovine serum, and
1% penicillin/streptomycin solution (10,000 units of penicillin and
10 mg of streptomycin in 0.9% NaCl) in a humidified atmosphere of
5% CO2, 95% air at 37 ꢁC.
(75.5 MHz, CDCl3):
d
11.54 (NeCH2CH3), 11.78 (500CeCH3), 20.50
(CH3CO), 44.61 (NeCH2CH3), 48.75 (C-3), 50.69 (C-1), 69.63 (C-2),
108.33 (C-500), 112.09 (C-40), 115.81 (C-30 and C-50), 129.0 (C-20 and
C-60), 141.54 (C-600), 147.48 (C-10), 150.89 (C-200), 164.20 (C-400) and
169.80 (CO). HRMS (ESI positive mode): m/z 346.1761 [M þ H]þ,
calculated for C18H23N3O4þH 346.1751.
2.11. Cell proliferation assay
Cell proliferation assay was carried out using CellTiter 96
aqueous one solution cell proliferation assay kit (Promega, USA).
Briefly, upon reaching about 75e80% confluency, 5000 cells/well
2.9. c-Src kinase activity assay
were seeded in 96-well microplate in 100
for 24 h, the cells were treated with 50 M compound in triplicate.
At the end of the sample exposure period (24 h), 20 L CellTiter 96
aqueous solution was added. The plate was returned to the incu-
bator for 1 h in a humidified atmosphere at 37 ꢁC. The absorbance of
the formazan product was measured at 490 nm using microplate
reader. The blank control was recorded by measuring the absor-
bance at 490 nm with wells containing medium mixed with Cell-
Titer 96 aqueous solution but no cells. Results were expressed as
the percentage of the control (without compound set at 100%).
mL media. After growth
m
The effect of synthesized compounds on the activity of c-Src
kinase was determined by HTScan Src Kinase Assay Kit, catalogue
number 7776 from Cell Signaling Technology; according to manu-
facturer’s protocol. Streptavidin coated plates was purchased from
Pierce. In summary, the kinase reaction was started with the
m
incubation of the 12.5
GSTeSrc kinase in 1.25 mM DTT) with 12.5
compounds (dissolved in 1% DMSO) for 5 min at room temperature.
ATP/substrate (25 L, 20 M/1.5 M) cocktail was added to the
mL of the reaction cocktail (0.5 ng/
mL of
L of prediluted
m
m
m
m
mixture. The biotinylated substrate (catalogue number 1366)
contains the residues surrounding tyrosine 160 (Tyr160) of signal
transduction protein with a sequence of EGIYDVP. The reaction
mixture was incubated for 30 min at room temperature. The kinase
3. Results and discussion
3.1. Chemistry
reaction was stopped with the addition of 50
8.0). The reaction solution (25 L) was transferred into 96-well
streptavidin plates (Pierce, part number 15125), diluted with 75
double distilled water, and incubated at room temperature for
60 min. At the end of the incubation, the wells were washed three
mL of 50 mM EDTA (pH
The synthesis of 3-N-alkyl-N-phenylaminopropan-2-ol deriva-
tives were achieved in three steps. In the first stage epoxy ring in
epichlorohydrine (7) was opened with N-methylaniline (5) and
N-ethylaniline (6) in the presence of freshly prepared calcium
trifluoromethanesulfonate [Ca(OTf)2] to afford selectively 1-chloro-
3-(N-methyl-N-phenylamino)propan-2-ol (8) and 1-chloro-3-
(N-ethyl-N-phenylamino)propan-2-ol (5) in 80 and 87% yields,
respectively, according to the previously reported procedure
[24]. Ca(OTf)2 was synthesized from the reaction of calcium
carbonate suspended in toluene with two equivalents of tri-
fluoromethanesulfonic acid [24].
In the second step ring closure in compounds 8 and 9 were
achieved under basic condition to yield 10 and 11 in 75 and 80%
yields, respectively (Scheme 1). Finally, epoxide ring in N-methyl-
aniline (10) was opened by different secondary amines, such as
piperidine (12a), pyrrolidine (12b), N-methylpiperazine (12c), and
morpholine (12d) in the presence of Ca(OTf)2 in acetonitrile
resulting in the formation of 3-(N-methyl,N-phenylamino)-1-
(piperidin-1-yl)propan-2-ol (13a), 3-(N-methyl,N-phenylamino)
-1-(pyrrolidin-1-yl)propan-2-ol (13b), 3-(N-methyl,N-phenyl-
amino)-1-(4-methylpiperazin-1-yl)propan-2-ol (13c), and 3-(N-
methyl,N-phenylamino)-1-morpholinopropan-2-ol (13d) in 78e90%
yields, respectively [24]. Under similar conditions, reaction of N-
ethylaniline 11 with 12ae12d afforded 3-(N-ethyl,N-phenylamino)
-1-(piperidin-1-yl)propan-2-ol (14a), 3-(N-ethyl,N-phenylamino)-
1-(pyrrolidin-1-yl)propan-2-ol (14b), 3-(N-ethyl,N-phenylamino-)
m
ml
times with 200
mL of 0.05% Tween-20 in PBS buffer (PBS/T). After
that to the each well was added 100
mL of phosphotyrosine anti-
body (P-Tyr-100) (1:1000 dilution in PBS/T with 1% BSA) and the
wells were incubated for another 60 min. After washing three times
with 0.05% Tween-20 in PBS/T, the wells were incubated with
100
mL secondary anti-mouse IgG antibody, which was HRP-
conjugated (1:500 dilution in PBS/T with 1% BSA) for next 30 min at
room temperature. The wells were washed five times with 0.05%
Tween-20 in PBS and then were incubated with 100
tetramethylbenzidine dihydrochloride (TMB) substrate for 5 min.
The reaction was stopped by adding 100 L/well of stop solution to
m
L of 3,30,5,50-
m
each well and mixed well and read the absorbance at 450 nm using
a microplate reader (Molecular devices, spectra Max M2). IC50
values of the compounds were calculated using ORIGIN 6.0 (origin
lab) software. IC50 is the concentration of the compound that
inhibited enzyme activity by 50%. All the experiments were carried
out triplicate.
2.10. Cell culture
Human breast carcinoma cell line BT-20 was obtained from
American Type Culture Collection (ATCC no. HTB-19). Cells were
1-(4-methylpiperazin-1-yl)propan-2-ol
(14c),
and
3-(N-
R
R
N
R
OH
2
O
2'
N
N
Cl
O
H
ii
i
1'
3
Cl
1
+
4'
10, R = CH3
5, R = CH3
8, R = CH3
9, R = CH2CH3
7
11, R = CH2CH3
6, R = CH2CH3
Reagents and conditions: (i) Ca(OTf)2, CH3CN, r.t.(ii) 5% aq. NaOH solution in toluene (1:1)
Scheme 1. Synthesis of b-amino epoxides 10 and 11.

1170
D. Sharma et al. / Biochimie 92 (2010) 1164e1172
H3C
CH3 OH
Ca(OTf)2,
CH3CN, 28 oC
OH
R
N
N
R
and
R-H (12a-12d)
13a-13d
14a-14d
OH
R
N
O
O
H3C
O
DMSO
CH3 OH
N
K2CO3, 60 oC
N
N
NH
N
NH
and
thymine (15)
O
O
10, R = CH3
17
16
11, R = CH2CH3
H3C
CH3 OH
N
DMF
OH
K2CO3, 80 oC
R
N
R
and
nucleobases
(18a-18b)
19a-19b
20a-20b
18b, 19b & 20b
NH2
12d, 13d, 14d 18a, 19a & 20a
12a, 13a & 14a 12b, 13b & 14b 12c, 13c & 14c
Compds.
O
CH3
N
O
N
4"
4"
NH
R_
7"
N
5"
1"
N
1"
N
6"
2"
N
N
N
O
N
N
9"
3"
Scheme 2. Synthesis of
b
-amino alcohols 13e20.
ethyl,N-phenylamino)-1-morpholinopropan-2-ol (14d) in 75e86%
yields, respectively (Scheme 2).
epoxide ring opening of 10 and 11 with nucleobases was facilitated
when DMSO/DMF was used as solvent (Scheme 2).
Furthermore, the epoxide ring in compounds 10 and 11 were
also opened by different nucleobases, such as thymine (15) in
DMSO containing potassium carbonate leading to the formation
of compounds 3-(N-methyl-N-phenylamino)-1-(thymine-1-yl)
propan-2-ol (16) and 3-(N-ethyl-N-phenylamino)-1-(thymine-1-
yl)propan-2-ol (17) in 50 and 45% yields, respectively. Similarly
reactions of 10 and 11 with uracil (18a) and adenine (18b) in DMF
containing potassium carbonate resulted in the formation of 3-(N-
methyl-N-phenylamino)-1-(uracil-1-yl)propan-2-ol (19a) and 3-
(N-methyl-N-phenylamino)-1-(adenin-9-yl)propan-2-ol (19b) in
70 and 68% yields, respectively, and 3-(N-ethyl-N-phenylamino)-1-
(uracil-1-yl)propan-2-ol (20a) and 3-(N-ethyl-N-phenylamino)-1-
(adenine-9-yl)propan-2-ol (20b) in 70 and 72% yields, respectively
(Scheme 2). Purine and pyrimidine bases, i.e. uracil, adenine and
thymine did not react with epoxides 10 and 11 in presence of Ca
(OTf)2 as catalyst in acetonitrile, possibly because of the very poor
solubility of these bases under this condition. The nucleophilic
The selective formation of secondary alcohol on epoxide ring
opening in the presence of Ca(OTf)2 [24] was further confirmed by
acetylation of the alcohol and comparison of the chemical shift
values of the proton attached to C-2 in alcohol and the corre-
sponding acetate (Scheme 3). After acetylation of 14d, a chemical
shift from 3.88 to 3.94 deshielding to 5.09e5.13 ppm was observed
for the proton attached to C-2. Similarly, a chemical shift transition
from 3.92 to 5.30 ppm for the proton attached to C-2 was detected
in alcohol 17.
The chemical structures of all synthesized compounds, i.e., 8e11,
13ae13d, 14ae14d, 16, 17, 19ae19b, 20ae20b, 21, and 22 were
confirmed unambiguously by their spectral data (1H-,13C-, 1He1H
COSY NMR, IR, HRMS spectroscopy) analysis and by derivatization
whenever required. Thirteen new compounds, i.e, 13ae13d,
14ae14c,16,17,19ae19b and 20ae20b, were synthesized using this
strategy. The use of unsubstituted aniline as reagent for epoxide
ring opening was challenging. When
2
equivalents of
H3C
H3C
OH
O
AcO
O
O
O
DMAP
+
N
N
O
CH3CN
21
14d
O
O
H3C
H3C
OH
DMAP
O
O
AcO
NH
O
NH
O
+
N
N
N
N
CH3CN
O
22
17
Scheme 3. Acetylation of secondary alcoholic group in b-amino alcohols 14d and 17.

D. Sharma et al. / Biochimie 92 (2010) 1164e1172
1171
epichlorohydrin were used in reaction with unsubstituted aniline, it
led to the formation of diepoxide.
120
100
80
60
40
20
0
3.2. Src kinase inhibitory activity
Table 1 shows the inhibitory potency of the synthesized
compounds compared to control general protein kinase inhibitor,
staurosporine, and src kinase inhibitor, PP2. Among all the
compounds, 3-(N-methyl-N-phenylamino)propan-2-ol derivatives
substituted at C-1 position with piperidine 13a, pyrrolidine 13b, N-
methylpiperazine 13c, morpholine 13d, and uracil 19a exhibited
Compounds
Fig. 2. Inhibition of BT-20 cell proliferation by the compounds 13ae13d, 14ae14d, 16,
17, 19ae19b, and 20ae20b (50 M). The results are shown as the percentage of the
control that does not contain test compound (set at 100%). All the experiments were
performed in triplicate.
modest inhibitory activity (IC50 ¼ 66.1e88.1
tives showed no inhibition or were weak Src kinase inhibitors
(IC50 > 100 M).
mM). All other deriva-
m
m
The data suggest that N-methyl derivatives (13aed) are more
potent than the corresponding N-ethyl substituted compounds
(14aed). Furthermore, the incorporation of bulky nucleobases,
such as thymine and adenine at C-1 position as shown in
compounds 16, 17, 19b, and 20b significantly reduced the inhibitory
potency. N-Methyl substituted compound 13b with small pyrroli-
dine ring at C-1 position showed the highest inhibitory potency
when compared with 14c and 17. The data suggest that other
mechanisms may be involved in the relatively modest anticancer
activities of these compounds.
In conclusion, thirteen novel b-aminoalcohol derivatives were
synthesized and evaluated as Src kinase and BT-20 cell proliferation
inhibitors. The data indicate that designing small molecules with
modest inhibitory activity against Src kinase is possible by using
(IC50 ¼ 66.1
mM) among all the compounds, suggesting that the Src
kinase inhibition was reduced significantly by the incorporation of
bulky groups at C-1 position and N-substitution with groups larger
than methyl moiety. Some of the previously reported hetero-
b-amino alcohols incorporated into 3-(N-methyl-N-phenylamino)
propan-2-ol derivatives. The compounds may have potential to be
used in fragment-based discovery of Src kinase inhibitors. Further
optimization is required to generate lead compounds with optimal
inhibitory potency. The results indicate that different mechanisms
may be involved for Src kinase inhibition and reducing BT-20 cell
proliferation.
aromatic
Src kinase also had unsubstituted amino groups and were primary
alcohols [22]. Thus, further application of -amino alcohol frag-
b-amino alcohols with potent inhibitory potency against
b
ments in other complex structures for Src kinase activity may
require the presence of less bulky functional groups surrounding
alcohol and amino moieties.
The effect of the inhibitors on the cell proliferation of cancer
cells that overexpress c-Src was also evaluated in BT-20 cell line
[25]. The human breast carcinoma cells BT-20 overexpresses c-Src
Acknowledgements
We acknowledge the financial support from the National
Medicinal Plant Board, New Delhi and American Cancer Society
Grant # RSG-07-290-01-CDD. DS, RKS and SB thank University of
Delhi, Department of Biotechnology and University Grant Commi-
sion (New Delhi, India) respectively, for the fellowships.
kinase.
3-(N-Ethyl-N-phenylamino)-1-(4-methylpiperazin-1-yl)
propan-2-ol (14c) and 3-(N-Ethyl-N-phenylamino)-1-(thymine-1-
yl)propane-2-ol (17) were able to inhibit the growth of cancer cells
by approximately 45e49% at a concentration of 50
mM (Fig. 2).
There were no correlation between Src kinase inhibitory potency of
the compounds and the growth inhibition of cancer cells since
compounds 14c and 17 were not potent Src kinase inhibitors.
Furthermore, other compounds with modest Src kinase inhibitory
activities, such as 13b, demonstrate weaker anticancer activities
References
[1] A.K. Prasad, P. Kumar, A. Dhawan, A.K. Chhillar, D. Sharma, V. Yadav,
M. Kumar, H.N. Jha, C.E. Olsen, G.L. Sharma, V.S. Parmar, Synthesis and anti-
microbial activity of 3-arylamino-1-chloropropan-2-ols. Bioorg. Med. Chem.
Lett. 18 (2008) 2156e2161.
[2] T. Hunter, Signalinge2000 and beyond. Cell 100 (2000) 113e127.
[3] J. Schlessinger, New roles for Src kinases in control of cell survival and
angiogenesis. Cell 100 (2000) 293e296.
Table 1
Src Kinase inhibitory activity of compounds 13ae13d, 14ae14d, 16, 17, 19ae19b, and
20ae20b.
[4] F.A. al-Obeidi, J.J. Wu, K.S. Lam, Protein tyrosine kinases: structure, substrate
specificity, and drug discovery. Biopolymers 47 (1998) 197e223.
[5] K. Parang, G. Sun, Recent advances in the discovery of Src kinase inhibitors.
Expert Opin. Ther. Patents 15 (2005) 1183e1207.
[6] R.B. Irby, T.J. Yeatman, Role of Src expression and activation in human cancer.
Oncogene 19 (2000) 5636e5642.
[7] (a) L.M. Toledo, N.B. Lydon, D. Elbaum, The structure-based design of ATP-Site
Directed protein kinase inhibitors. Curr. Med. Chem. 6 (1999) 775e805;
(b) S. Schenone, O. Bruno, A. Ranise, F. Bondavalli, C. Brullo, P. Fossa, L. Mosti,
G. Menozzi, F. Carraro, A. Naldini, C. Bernini, F. Manetti, M. Botta, New pyr-
azolo[3,4-d]pyrimidines endowed with A431 antiproliferative activity and
inhibitory properties of Src phosphorylation. Bioorg. Med. Chem. Lett. 14
(2004) 2511e2517.
[8] (a) R.J. Budde, N.U. Obeyesekere, S. Ke, J.S. McMurray, Use of synthetic
peptides and copolymers to study the substrate specificity and inhibition of
protein tyrosine kinase pp60c-src. Biochim. Biophys. Acta 1248 (1995) 50e56;
(b) K.S. Lam, J. Wu, Q. Lou, Identification and characterization of a novel
synthetic peptide substrate specific for Src-family protein tyrosine kinases.
Int. J. Pept. Protein Res. 45 (1995) 587e592;
Compounds
IC50 (m
M)a
Staurosporin
PP2
13a
13b
13c
13d
14a
14b
14c
14d
16
17
19a
0.3 ꢂ 0.0
2.8 ꢂ 0.0
83.5 ꢂ 0.2
66.1 ꢂ 0.9
85.9 ꢂ 0.6
81.4 ꢂ 0.9
NAb
>100
>140
>190
>190
>100
88.1 ꢂ 1.2
20a
19b
20b
>100
NA
>150
(c) J.R. Kamath, R. Liu, A.M. Enstrom, Q. Lou, K.S. Lam, Development and
characterization of potent and specific peptide inhibitors of p60c-src protein
tyrosine kinase using pseudosubstrate-based inhibitor design approach.
J. Pept. Res. 62 (2003) 260e268.
a
The concentration of the compound that inhibited enzyme activity by 50%.
Mean ꢂ SEM.
b
No enzyme activity.

1172
D. Sharma et al. / Biochimie 92 (2010) 1164e1172
[9] H.D.H. Showalter, A.J. Kraker, Small molecule inhibitors of the platelet-
choriocarcinoma in nude mice through its direct cytostatic activity. Anti-
cancer Res. 15 (1995) 2081e2087.
[18] (a) J.A. Beisler, Chartreusin, a glycosidic antitumour antibiotic from strepto-
myces. Prog. Med. Chem. 19 (1982) 242e268;
derived growth factor receptor, the fibroblast growth factor receptor, and Src
family tyrosine kinases. Pharmacol. Ther. 76 (1997) 55e71.
[10] (a) L. Tatoon, G.M. Molrey, R. Chopra, A. Khwaja, The Src-selective kinase
inhibitor PP1 also inhibits Kit and Bcr-Abl tyrosine kinases. J. Biol. Chem. 278
(2003) 4847e4853;
(b) B.E. Leach, K.M. Calhoun, L.E. Johnson, C.M. Teeters, W.G. Jackson, Char-
treusin, a new antibiotic produced by Streptomyces chartreusis, a new species.
J. Am. Chem. Soc. 75 (1953) 4011e4012;
(c) H. Sugawara, M. Tsunakawa, M. Konishi, H. Kawaguchi, B. Krishnan,
H. Cun-heng, J. Clardy, Elsamicins A and B, new antitumor antibiotics related
to chartreusin. 2. Structures of Elsamicins A and B. J. Org. Chem. 52 (1987)
996e1001;
(b) J.F. Dorsey, R. Jove, A.J. Kraker, J. Wu, Thepyrido[2,3-d]pyrimidine deriva-
tive PD180970 inhibits p210Bcr-Abl tyrosinekinase and induces apoptosis of
K562 leukemic cells. Cancer Res. 60 (2000) 3127e3131;
(c) N. Rucci, M. Susa, A. Teti, Inhibition of protein kinase c-Src as a therapeutic
approach for cancer and bone metastases. Anticancer Agents Med. Chem. 8
(2008) 342e349.
(d) W. Lown, Anthracycline and Anthracendione-Based Anticancer Agents.
Elsevier, Amsterdam, 1988.
[11] M. Warmuth, R. Damoiseaux, Y. Liu, D. Fabbro, N. Gray, Src family kinases:
potential targets for the treatment of human cancer and leukemia. Curr.
Pharm. Des. 9 (2003) 2043e2059.
[19] (a) G. van Poppel, H. van den Berg, Vitamins and cancer. Cancer Lett. 114
(1997) 195e202;
[12] (a) E.J. Corey, F. Zhang, re- and si-Face-selective nitroaldol reactions catalyzed
by a rigid chiral quaternary ammonium salt: a highly stereoselective synthesis
of the HIV protease inhibitor amprenavir (vertex 478). Angew. Chem. Int. Ed.
38 (1999) 1931e1934;
(b) G. Shklar, Mechanism of cancer inhibition by anti-oxidant nutrients. Oral
Oncol. 34 (1998) 24e29;
(c) E. Nagel, A. Meyer zu Vilsendorf, M. Bartels, R. Pichlmayr, Antioxidative
vitamins in prevention of ischemia/reperfusion injury. Int. J. Vitam. Nutr. Res.
67 (1997) 298e306;
(b) C.W. Johannes, M.S. Visser, G.S. Weatherhead, A.H. Hoveyda, Zr-catalyzed
kinetic resolution of allylic ethers and Mo-catalyzed chromene formation in
synthesis. Enantioselective total synthesis of the antihypertensive agent (S, R,
R, R)-Nebivolol. J. Am. Chem. Soc. 120 (1998) 8340e8347;
(c) G.A. Rogers, S.M. Parsons, D.C. Anderson, L.M. Nilsson, B.A. Bahr,
W.D. Kornreich, R. Kaufman, R.S. Jacobs, B. Kirtman, Synthesis, in vitro
acetylcholine-storage-blocking activities, and biological properties of deriva-
tives and analogs of trans-2-(4-phenylpiperidino)cyclohexanol (vesamicol).
J. Med. Chem. 32 (1989) 1217e1230;
(d) A.T. Diplock, Antioxidant Nutrients and diseases. Nutr. Health 9 (1993)
37e42.
[20] M. Kimura, T. Masuda, K. Yamada, N. Kubota, N. Kawakatsu, M. Mitani,
K. Kishii, M. Inazuy, T. Namiki, Novel diphenylalkyl piperizine derivatives with
dual calcium antagonistic and antioxidative activities. Bioorg. Med. Chem.
Lett. 12 (2002) 1947e1950.
[21] W.S. DiMenna, C. Piantadosi, R.G. Lamb, Synthesis of potential hypolipidemic
agents. Reaction of substituted phenyl 2,3-epoxypropyl ethers with adenine,
uracil, and thymine. J. Med. Chem. 21 (1978) 1073e1076.
[22] H. Mukaiyama, T. Nishimura, S. Kobayashi, Y. Komatsu, S. Kikuchi, T. Ozawa,
N. Kamada, H. Ohnota, Novel pyrazolo[1,5-a]pyrimidine as c-Src kinase inhib-
itors that reduce IKr channel blockade. Bioorg. Med. Chem. 16 (2008) 909e921.
[23] (a) L.J. Lombardo, F.Y. Lee, P. Chen, D. Norris, J.C. Barrish, K. Behnia,
S. Castaneda, L.A.M. Cornelius, J. Das, A.M. Doweyko, C. Fairchild, J.T. Hunt,
I. Inigo, K. Johnston, A. Kamath, D. Kan, H. Klei, P. Marathe, S. Pang, R. Peterson,
S. Pitt, G.L. Schieven, R.J. Schmidt, J. Tokarski, M.-L. Wen, J. Wityak,
R.M. Borzilleri, Discovery of N-(2-chloro-6-methylphenyl)-2-(6-(4-(2-
hydroxyethyl)-piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-car-
boxamide (BMS-354825), a dual Src/Abl kinase inhibitor with potent anti-
tumor activity in preclinical assays. J. Med. Chem. 47 (2004) 6658e6661;
(b) J. Das, P. Chen, D. Norris, R. Padmanabha, J. Lin, R.V. Moquin, Z. Shen,
L.S. Cook, A.M. Doweyko, S. Pitt, S. Pang, D.R. Shen, Q. Fang, H.F. de Fex,
K.W. McIntyre, D.J. Shuster, K.M. Gillooly, K. Behnia, G.L. Schieven, J. Wityak,
J.C. Barrish, 2-Aminothiazole as a novel kinase inhibitor Template. Structure-
activity relationship studies toward the discovery of N-(2-Chloro-6-methyl-
phenyl)-2-[[6-[4-(2-hydroxyethyl)-1-piperazinyl)]-2-methyl-4-pyrimidinyl]
(d) L. Radesca, W.D. Bowen, L.D. Paolo, B.R. de Costa, Synthesis and receptor
binding of enantiomeric N-substituted cis -N-[2 -(3,4-dichloropheny1)eth yll-
2-(1-pyrrolidinyl)cyclohexylamines as high-affinity
Chem. 34 (1991) 3058e3065.
s receptor ligands. J. Med.
[13] (a) P. O’Brien, Sharpless asymmetric aminohydroxylation: scope, limitations,
and use in synthesis. Angew. Chem. Int. Ed. 38 (1998) 326e329;
(b) G. Li, H.-T. Chang, K.B. Sharpless, Catalytic asymmetric amino-
hydroxylation (AA) of Olefins. Angew. Chem. Int. Ed. 35 (1996) 451e454.
[14] (a) D.J. Ager, I. Prakash, D.R. Schaad, 1,2-Amino alcohols and their heterocyclic
derivatives as chiral auxiliaries in asymmetric synthesis. Chem. Rev. 96 (1996)
835e875;
(b) J.D. Bloom, M.D. Dutia, B.D. Johnson, A. Wissner, M.G. Burns, E.E. Largis,
J.A. Dolan, T.H. Claus, Disodium (R, R)-5-[-2-{[2-(3-Chlorophenyl)-2-hydrox-
yethyl]-amino}propyl]-1,3-benzodioxole-2,2-dicarboxylate (CL 316,243).
potent
ꢀadrenergic agonist virtually specific for b3 receptors. A promising
antidiabetic and antiobesity agent. J. Med. Chem. 35 (1992) 3081e3084.
A
b
[15] J.F. Evans, S. Kargman, Bestatin inhibits covalent coupling of (3H)LTA4 to
human leukocyte LTA4 hydrolase. FEBS Lett. 297 (1992) 139e142.
[16] (a) H. Umezawa, T. Aoyagi, H. Suda, M. Hamada, T. Takeuchi, Bestatin, an
inhibitor of minopeptidase B, produced by actinomycetes. J. Antibiot. 29
(1976) 97e99;
(b) H. Nakamura, H. Suda, T. Takita, T. Auyagi, H. Umezawa, Y. Iitaka, X-ray
structure determination of (2s, 3R)-3-amino-2-hydroxy-4-phenylbutanoic
acid, a new amino acid component of bestatin. J. Antibiot. 29 (1976) 102e103.
[17] K. Ino, S. Goto, S. Nomura, K.-I. Isobe, A. Nawa, T. Okamoto, Y. Tomoda,
Aminopeptidase inhibitor ubenimex (bestatin) inhibits the growth of human
amino)]-1,3-thiazole-5-carboxamide (Dasatinib, BMS-354825) as
a potent
pan-src kinase inhibitor. J. Med. Chem. 49 (2006) 6819e6832.
ꢀ
ꢁ
ꢁ ꢀ
ꢀ
[24] I. Cepanec, M. Litvic, H. . Mikuldas, A. Bartolincic, V. Vinkovic, Calcium tri-
fluorosulfonate-catalysed aminolysis of epoxides. Tetrahedron 59 (2003)
2435e2439.
[25] A.P. Belsches-Jablonski, J.S. Biscardi, D.R. Peavy, D.A. Tice, D.A. Romney,
S.J. Parsons, Src family kinases and HER2 interactions in human breast cancer
cell growth and survival. Oncogene 20 (2001) 1465e1475.