Synthesis, docking study and β-Adrenoceptor activity of some new oxime ether derivatives

Article abstract of DOI:10.3390/molecules19033417

A new series of oxime ethers 4a-z was designed and synthesized to test the blocking activity against β1 and β2-Adrenergic receptors. Docking of these ether derivatives into the active site of the identified 3D structures of β1 and β2-Adrenergic receptors

Full text of DOI:10.3390/molecules19033417

Molecules 2014, 19, 3417-3435; doi:10.3390/molecules19033417
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molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article
Synthesis, Docking Study and β-Adrenoceptor Activity of Some
New Oxime Ether Derivatives
Hazem A. Ghabbour 1,2,*, Eman R. El-Bendary 1, Mahmoud B. El-Ashmawy 1 and
Mohamed M. El-Kerdawy 1
1
Department of Medicinal Chemistry, Faculty of Pharmacy, Mansoura University,
Mansoura 35516, Egypt
2
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, King Saud University,
P. O. Box 2457, Riyadh 11451, Saudi Arabia
* Author to whom correspondence should be addressed; E-Mail: hghabbour@mans.edu.eg or
ghabbourh@yahoo.com; Tel.: +9-665-693-52985; Fax: +9-661-4676-220.
Received: 17 February 2014; in revised form: 5 March 2014 / Accepted: 11 March 2014 /
Published: 20 March 2014
Abstract: A new series of oxime ethers 4a–z was designed and synthesized to test the
blocking activity against β1 and β2-adrenergic receptors. Docking of these ether derivatives into
the active site of the identified 3D structures of β1 and β2-adrenergic receptors showed
MolDock scores comparable to those of reference compounds. Biological results revealed that
the inhibition effects on the heart rate and contractility are less than those of propranolol.
Nevertheless, the two compounds 4p and 4q that displayed the highest negative MolDock
score with β2-adrenergic receptors showed β2-antagonistic activity by decreasing salbutamol
relaxation of precontracted tracheal strips, which indicates the importance of a chlorothiophene
moiety in the hydrophobic region for best complementarity with β2 receptors. On other hand,
the presence of a homoveratryl moiety increases the MolDock score of the tested compounds
with the β1 receptor.
Keywords: synthesis; screening; docking; oxime ethers; adrenergic; β-receptors
1. Introduction
β-Adrenoceptor blockers represent an important class of therapeutics due to their usefulness in a
number of diseases, such as angina pectoris, arrhythmia and hypertension. They may also be used in

Molecules 2014, 19
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migraine, glaucoma, tremors, hyperthyroidism, and anxiety-provoking situations [1]. Based on the
chemical structure of the classical aryloxypropanolamine β-adrenoceptor blockers, Imbs et al. [2]
showed that the insertion of an oxime ether in place of the typical ether linkage retained β2-antagonist
activity; e.g., IPS 339 (Figure 1). Other oxime ethers were also synthesized and reported to have
β2-blocking activity with high potency, e.g., falintolol (Figure 1) [3,4]. Although the above mentioned
cardiovascular effects are mainly attributed to β1-blockage, β2-adrenoceptor antagonists might be
potentially interesting as experimental tools [5], or for testing in pulmonary arterial hypertension [6].
Figure 1. Chemical structures of IPS 339 and falintolol.
In the present investigation, we intended to further explore the general structure of oxime ethers
through different molecular modifications, and to test the β1- and β2-adrenoceptor blocking activity of
the new compounds. The manipulation strategy involves usage of aromatic, alicyclic, bicyclic and/or
heterocyclic moieties to represent the hydrophobic part of the molecule, in order to verify the
importance of these moieties in binding with the receptor, and hence on the selectivity of the resulted
compounds. Obeying the SAR of the β-blockers, the already known β-directing isopropyl and tert-butyl
group as well as the dimethoxyphenylethyl moiety were selected as the best representatives of alkyl
and arylalkylamines. The synthesis, molecular docking and preliminary biological evaluation of the
thus designed O-(3-alkylamino-2-hydroxypropyl)oxime derivatives 4a–z are reported herein.
2. Results and Discussion
2.1. Chemistry
The general method to prepare the final compounds 4a–z is outlined in Scheme 1. The oximes 2a–j
(Table 1), required in the present work, have been obtained via reaction of the corresponding ketones
1a–j with hydroxylamine hydrochloride, as previously reported [3,7–14]. The two oxime geometric
isomers were not separated, based on the previously reported absence of appreciable difference in
β- blocking activity between the E- and Z-isomers of the final oxime ether derivatives [15].
Accordingly, compounds 2a–j were reacted with epichlorohydrin, in dry DMF, in the presence of
1
NaH, to afford the epoxy compounds 3a–j (Table 1). In the H-NMR spectra, the oxirane protons
appeared mostly as multiplets at δ 2.4–3.0 ppm, while the -OCH2- protons appeared as two doublets of
doublets at δ 3.7–4.4 ppm (Experimental part). The target compounds 4a–z (Table 2) were prepared
by reaction of the epoxy compounds 3a–j with excess of isopropylamine, tert-butylamine or
homoveratrylamine in toluene, in a sealed tube, at 120 °C.

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Scheme 1. Synthesis of the oxime ethers 4a–z.
Table 1. Oximes 2a–j and oxime ethers 3a–j.
R1
R1
N
O
N
OH
O
R2
R2
2a-j
3a-j
Compound No. R1
a
R2
Compound No. R1
R2
f
CH3
Cl
S
b
c
g
h
N
OCH3
d
e
CH3
CH3
i
j
CH3
2.2. Molecular Docking
Molecular docking is a very popular method employed to investigate molecular association. It is
particularly useful in the drug discovery arena to study the binding of small molecules (ligands) to
macromolecules (receptors or enzymes). Docking-based drug design using structural biology remains

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one of the most logical and esthetically pleasing approaches in the drug discovery process. The
structured knowledge of the binding capabilities of the active site residues to specific groups on the
agonist or antagonist leads to various proposals for the synthesis of very specific agents with a high
probability of biological action [16].
Table 2. O-(3-Alkylamino-2-hydroxypropyl)oxime derivatives 4a–z.
Compound No.
4a
R1
R2
R3
Compound No.
4n
R1
R2
R3
CH3
4b
4o
CH3
4c
4d
4e
4p
4q
4r
CH3
CH3
CH3
Cl
Cl
Cl
S
S
S
4f
4s
4t
4g
4h
4i
4u
4v
4j
CH3
CH3
4w
4x
4k
4l
CH3
CH3
4y
4z
4m
CH3
Twenty six new oxime ethers 4a–z were subjected to the docking study into the identified β
1 and
β2- adrenergic receptor active sites [17–19]. The docking technique using the Molegro Virtual Docker [20]
is used to estimate the binding affinity between external ligands and structurally-known biological
macromolecules, and hence, to predict the biological importance of these ligands [19,21]. The forces
of interaction were declared and scoring functions were determined. Propranolol (a non-selective
β-antagonist), cyanopindolol (selective β1-antagonist) (Figure 2) and IPS 339 (selective β2-antagonist)
were used in this docking study as reference molecules. Snapshots for all prepared compounds were
taken to reveal their molecular interactions (hydrogen, hydrophobic and/or ionic bonds) with the amino

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acids of β1 and β2- receptor active sites. MolDock scores and the energy H-bond interactions between
these compounds and receptor were calculated (Table 3). The results showed that all tested compounds
have affinity for β1 and β2-adrenoceptors comparable to that of the reference compounds.
Figure 2. Chemical structures of propranolol and cyanopindolol.
Table 3. MolDock scores and hydrogen bond energy for compounds 4a–z and reference
compounds with the β1
β2 receptors.
and
β1-Receptor
β2-Receptor
Comp.
MolDock Score H-Bond E. MolDock Score H-Bond E.
4a
−115.13
−122.697
−142.285
−121.41
−2.5
−100.806
−99.6442
−105.6596
−86.4381
−88.3577
−95.1594
−92.7053
−85.3741
−90.616
−4.35757
−2.42254
−3.42955
−1.16172
−0.04374
0
4b
−4.58252
−5.19707
−3.59754
−3.96668
−5
4c
4d
4e
−127.903
−153.964
−125.653
−124.044
−145.546
−114.594
−120.406
−155.836
−125.091
−120.574
−167.406
−105.452
−114.132
−143.244
−108.994
−108.574
−148.747
−105.443
−105.034
−139.094
−152.107
−125.4
4f
4g
−2.5
−3.4013
−0.69094
−2.98878
0
4h
−2.83065
−5.2648
−2.62113
−2.5
4i
4j
−79.1286
−90.5964
−103.123
−74.7409
−73.6041
−91.127
4k
−2.63215
−0.16123
−0.97756
−3.78389
−2.5557
−4.03237
−4.34578
−0.41247
−3.26981
−0.78187
−0.03188
−1.98422
−0.99963
−0.47872
−2.4702
−2.16834
−3.92249
−2.87182
−2.98876
4l
−6.1153
−3.98536
−2.18542
−4.49787
−1.63375
−2.62848
−2.15101
−1.50057
−2.79887
−4.41468
−1.00129
0
4m
4n
4o
4p
−113.5079
−117.075
−102.028
−85.4672
−84.921
4q
4r
4s
4t
4u
−100.887
−84.9409
−85.1195
−99.449
4v
4w
4x
4y
−4.07406
−5.54788
−2.28939
−2.53142
−2.91744
−10.7754
−93.6354
−84.4559
−88.1631
−95.141
4z
Propranolol
IPS 339
Cyanopindolol
−110.517
−104.212
−129.204
−85.6768

Molecules 2014, 19
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In the β1-receptor, the selective antagonist cyanopindolol has a MolDock score −129.2 with H-bond
energy −10.7 and compounds 4c, 4f, 4i, 4l, 4o, 4r, 4u, 4x and 4y have better MolDock scores than
cyanopindolol.
The best compound in this study was 4o, which has MolDock score of −167.4 and
forms five hydrogen bonds with the amino acids Ser 211 and Tyr 207 in the active site of the receptor
Moreover, hydrophobic interactions probably existed between compound 4o and Phe 201, Phe 216
and Phe 307 (Figure 3a). It is worth mentioning here that all tested compounds which have better
MolDock score than the reference drug have in its structure the homoveratryl moiety which may
increase the quality of binding between the ligands and receptor.
Figure 3. (a) Docking snapshot of compound 4o with β1 receptor. (b) Docking snapshot of
compound 4q with β2 receptor. Compounds are represented as stick (thick lines) while the
amino acids of the active site of the β2-adrenoreceptor appear as (light) lines. Hydrogen
atoms in both ligands and receptors have been omitted for clarity. Blue dashed lines
indicate the hydrogen bonds.
(a)
(b)
In the β2-receptor, only six compounds 4a, 4b, 4c, 4l, 4u and 4x have slightly lower MolDock
scores than the β2-antagonist IPS 339, while compounds 4p and 4q displayed the highest negative
MolDock scores that indicated highest complementarity with β2-adrenoceptors (Table 3).
Figure 3b represents the snapshot for compound 4q and illustrates the different amino acids of the
binding site that are involved in the compound-receptor interaction, where more hydrogen bonds and
hydrophobic interactions were involved for this compound than others. Hydrogen bonding interactions
were β-hydroxyl group in 4q with the peptide carbonyl group of Ser 215 (3.29 Ǻ) and with the free
hydroxyl group of Ser 311 (3.19 Ǻ), oxygen of the oxime moiety makes a hydrogen bond with Ser 211
and Ser 212 (3.09 and 2.62 Ǻ, respectively) and the nitrogen of the oxime moiety also interacts with
Asn 310 by hydrogen bonding (2.96 Ǻ). Hydrophobic interactions may be found between the
thiophene ring and the amino acids Phe 201 and Phe 306.

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2.3. Biological Evaluation
Blocking of the β1-adrenoceptor prevents the increase in the heart rate and contractility caused by
epinephrine, while β2-adrenoceptor antagonism results in bronchospasms, uterine contraction and blood
vessel spasms [22]. In the present investigation, certain newly synthesized compounds were subjected to
in vitro biological testing for their blocking activity against β1 and β2-adrenoceptors, using isolated
guinea pig atria and trachea, respectively, according to reported procedures [23–25]. Meanwhile, the
agonistic activities at both receptors were also recorded.
2.3.1. In Vitro Screening for β1-Adrenoceptor Activity in Isolated Guinea Pig Atria
Seventeen of the newly synthesized compounds were subjected to an in vitro biological screening
for β1-adrenoceptor activities using isolated atria of guinea pigs. The available results indicate that six
of these compounds (4f, 4i, 4l, 4r, 4y and 4z), with 3,4-dimethoxyphenethyl groups at the terminal
amine produced 22%–31% inhibition in the heart rate and 30%–55% inhibition in the contractility of
the atria. Propranolol, the reference drug, produced 70 and 60% inhibition of the heart rate and
contractility, respectively (Table 4).
Table 4. The percent inhibition in the heart rate and the contractility of the isolated guinea
pig atria caused by the tested compounds.
β -antagonist
β -antagonist
1
1
Compound
No.
Compd. No.
% inhibition % inhibition in
% inhibition
% Inhibition
in heart rate
3%
contractility
0%
in heart rate
1%
in contractility
4a
4b
4c
4e
4f
4o
0%
0%
2%
0%
4p
6%
6%
1%
4q
2%
0%
0%
0%
4r
31%
2%
55%
0%
29%
2%
30%
0%
4u
4g
4i
4x
0%
0%
25%
3%
40%
1%
4y
4z
26%
22%
70%
35%
30%
60%
4k
4l
28%
45%
Propranolol
Replacement of the 3,4-dimethoxyphenethyl group with isopropyl or tert-butyl groups showed loss
of β1-adrenergic antagonist activity, as in 4g, 4e, 4k and 4p. It is worth mentioning that the dimethoxy
substitution on the phenethylamine was reported to be the preferable aralkylamine moiety required to
achieve β-adrenergic receptor blocking activity [26]. On the other hand, compounds 4c, 4o, 4u and
4x contain 3,4-dimethoxyphenethylamine groups, but they did not show any β1-adrenergic antagonistic
activity. This might be explained by the structural difference on the other side of the oxime moiety,
where the hydrophobic part might be improperly oriented to fit in the active site in the
β1-adrenergic receptor. Meanwhile, the study also indicated that none of the tested compounds has
β1-agonistic activity; whereby no increase in the heart rate or contractility of isolated guinea pig atria
was noticed.

Molecules 2014, 19
2.3.2. In Vitro Screening for β -Adrenoceptor Activity in Isolated Guinea pig Trachea
3424
2
Eleven of the newly synthesized compounds were subjected to an in vitro biological screening for
β2-adrenoceptor activities using isolated trachea of guinea pigs. The test depends principally on the
relaxation effect of salbutamol on the pre-contracted tracheal muscle induced by acetylcholine.
Compounds of potential antagonistic activity are expected to decrease the relaxation caused by
salbutamol. The available results indicated that only two compounds, 4p and 4q, decreased the
salbutamol relaxation of pre-contracted tracheal strips, while the rest of the tested compounds did not
show any β2-antagonist activity. The results also showed the absence of agonistic activity for all tested
compounds (Table 5).
Table 5. β2-Adrenergic activities of the tested compounds.
Compound No.
β -agonist β -antagonist Compound No.
β -agonist
2
β -antagonist
2
2
2
4a
4e
-
-
-
-
-
-
-
-
-
-
-
-
4p
-
-
-
-
-
-
+
+
-
4q
4g
4i
4r
4t
4v
-
4k
4m
-
Propranolol
+
(+) decreased relaxation, (−) no effect.
Taken together, docking of twenty six compounds into the β1 and β2 adrenergic receptors has
been studied, and the energy of interaction has been calculated. The energy of interaction revealed
that compounds 4p and 4q indicated the highest complementarity with β2-adrenoceptors (Table 3).
These two compounds, in particular, decreased the salbutamol relaxation effect on guinea pig trachea,
suggesting their β2-antagonist activity. The structural characteristics that might differentiate
compounds 4p and 4q from others is the presence of a chlorothiophene moiety in the hydrophobic
region. Compounds with chlorothienyloxypropanolamine moieties were previously studied by
El-Ashmawy et al. [27] for their β-blocking and partial agonist activity, but no tests were done with
β2-receptors. Compound 4r also has the same hydrophobic chlorothienyl moiety, however, the
presence of a bulky homoveratrylamine portion might explain its inactivity at the β2-adrenoceptors. It
is worth mentioning that this compound (4r) displayed the highest percentage inhibition on
β1-adrenoceptor (55% inhibition of the contractility) (Table 4). These findings confirm the
significance of a homoveratrylamine portion for β1 receptors, especially in view of the data from the
β1-adrenergic receptor docking study (Table 4); and indicate the importance of the presence of a
chlorothiophene moiety in the hydrophobic region for best complementarity with β2 receptors.
3. Experimental
3.1. General Information
Melting points (°C) were recorded using a Fisher-Johns melting point apparatus and are uncorrected.
1H-NMR spectra were obtained on FT-NMR spectrometer (200 MHz) Gemini Varian using TMS as
internal standard (chemical shifts in ppm, δ units). MS analyses were performed on JEOL JMS-600H

Molecules 2014, 19
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spectrometer. All the ketones used were commercially available from Sigma-Aldrich Chemical
Company (Munich, Germany), except benzophenone and acetophenone that were purchased from El-
Nasr Pharmaceutical Chemicals Company, Cairo, Egypt. Oximes 2a–j were prepared following the
procedures reported in the literature [3,7–14]. Two of the oxirane compounds 3i and 3j were
previously reported in the literature [14].
3.1.1. General Procedure for Synthesis of O-Oxiran-2-ylmethyl Oximes 3a–h
A mixture of the appropriate oxime 2a–h (0.01 mol) and NaH (0.26 g, 0.011 mol) was stirred in dry
DMF (30 mL) at room temperature for 10 min. A solution of epichlorohydrin (1.1 g, 0.01 mol) in dry DMF
(5 mL) was added dropwise over a period of 10 min. The resulting mixture was further stirred for 36 h
at room temperature (TLC). The reaction mixture was poured in ice water (200 mL) and extracted with
chloroform (3 × 100 mL). The organic layers were collected, washed twice with brine solution, dried
over anhydrous MgSO4, and the chloroform was evaporated under reduced pressure to obtain the
desired compounds in pure form.
1
Dicyclohexylmethanone O-oxiran-2-yl methyl oxime (3a). Yield 55%; M.p. 120–122 °C; H-NMR
(CDCl3) δ 1.15–1.70 (m, 20H, 10CH2 in cyclohexane), 2.21–2.44 (m, 2H, 2CHC=NO), 2.54–2.90 (m, 3H,
CH2 and CH oxirane), 3.85 (dd, 1H, J = 11.2, 6.4 Hz, OCH2), 4.17 (dd, 1H, J = 11.4, 2.6 Hz, OCH2).
MS m/z (%); 266 (10.6, M++1), 176 (63.2), 147 (100), 95 (48.65).
1
Cyclohexyl(phenyl)methanone O-oxiran-2-yl methyl oxime (3b). Yield 60%; oil; H-NMR (CDCl3) δ
1.25–1.70 (m, 10H, 5CH2 in cyclohexane), 2.15–2.41 (m, 1H, CHC=NO), 2.74–3.01 (m, 3H, CH2, CH
oxirane), 4.11 (dd, 1H, J = 11.5, 6.4 Hz, OCH2), 4.40 (dd, 1H, J = 11.6, 3.0 Hz, OCH2), 7.55 (t, 3H,
Ar-H), 7.91 (d, 2H, J = 7.0, 2,6 Ar-H). MS m/z (%); 260 (4.55, M++1), 186 (100), 104 (78), 77 (61.62).
1
Phenyl(Pyridin-2-yl)methanone O-oxiran-2-yl methyl oxime (3c). Yield 62%; oil; H-NMR (CDCl3) δ
2.64–2.90 (m, 3H, CH2, CH oxirane), 3.91 (dd, 1H, J = 11.4, 6.1 Hz, OCH2), 4.31 (dd, 1H, J = 11.8,
2.9 Hz, OCH2), 7.44–7.68 (m, 5H, Ar-H and pyridinyl-H), 7.79 (d, 2H, J = 7.0, 2,6-Ar-H), 7.95 (d, 1H,
J = 7.5, pyridinyl-H) 8.51 (d, 1H, J = 8.0, pyridinyl-H).
1
1-(Naphthalen-2-yl)ethanone O-oxiran-2-yl methyl oxime (3d). Yield 70%; M.p. 40–42 °C; H-NMR
(CDCl3) δ 2.54–2.77 (m, 3H, CH2, CH oxirane), 2.95 (s, 3H, CH3), 3.86 (dd, 1H, J = 11.5, 6.5 Hz,
OCH2), 4.17 (dd, 1H, J = 11.6, 2.6 Hz, OCH2), 7.42–7.71 (m, 7H, Ar-H). MS m/z (%); 242 (8.06, M++1),
154 (63.2), 127 (100), 87 (48.65).
1
1-(Biphenyl-4-yl)ethanone O-oxiran-2-yl methyl oxime (3e). Yield 70%; M.p. 72–74 °C; H-NMR
(CDCl3) δ 2.18 (s, 3H, CH3), 2.47–2.91 (m, 3H, CH2, CH oxirane), 3.74 (dd, 1H, J = 11.2, 6.6 Hz,
3',4',5'-Ar-H), 7.65 (d, 2H,
OCH2), 3.96 (dd, 1H, J = 11.4, 2.9 Hz, OCH2), 7.45 (t, 3H, J = 8.5,
2',6'-Ar-H), 7.98 (d, 2H, Ar-H), 8.24 (d, 2H, Ar-H).
J = 7.5,
J = 8.5
J = 8.5
1
1-(5-Chlorothiophen-2-yl)ethanone O-oxiran-2-yl methyl oxime (3f). Yield 65%; oil; H-NMR
(CDCl3) δ 2.16 (s, 3H, CH3), 2.56–3.04 (m, 3H, CH2, CH oxirane), 4.16 (dd, 1H, J = 11.3, 6.5 Hz,
3-thiophene-H), 7.41 (d, 1H,
OCH2), 4.30 (dd, 1H, J = 11.2, 2.7 Hz, OCH2), 7.15 (d, 1H, J = 8.0,
4-thiophene-H).
J = 8.5,

Molecules 2014, 19
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1
1,3,3-Trimethylbicyclo[2.2.1]heptan-2-one O-oxiran-2-yl methyl oxime (3g). Yield 45%; oil; H-NMR
(CDCl3) δ 1.30 (s, 6H, 2CH3), 1.35 (s, 3H, CH3), 1.52–1.70 (m, 7H, bicyclic moiety-H), 2.41–2.70
(m, 3H, CH2, CH oxirane), 3.75 (dd, 1H, J = 11.0, 6.1 Hz, OCH2), 3.94 (dd, 1H, J = 11.5, 2.4 Hz,
OCH2). MS m/z (%); 224 (5.16, M++1), 150 (56.62), 136 (100), 77 (35.04).
5-Methoxy-3,4-dihydronaphthalen-1(2H)-one O-oxiran-2-yl methyl oxime (3h). Yield 67%; oil;
1H-NMR (CDCl3) δ 2.17–2.91 (m, 9H, 3CH2 of dihydronaphthalene, CH2 oxirane, CH oxirane),
3.78 (s, 3H, OCH3), 3.85 (dd, 1H, J = 11.2, 6.0 Hz, OCH2), 4.1 (dd, 1H, J = 11.5, 2.7 Hz, OCH2), 7.14
(d, 1H, J = 8.0, 6-Ar-H), 7.35 (t, 1H, J = 8.5, 7-Ar-H), 7.56 (d, 1H, J = 8.5, 8-Ar-H).
3.1.2. General Procedure for Synthesis of O-(3-Alkylamino-2-hydroxypropyl)oxime Derivatives 4a–z
A mixture of the appropriate oxirane 3a–j (0.01 mol) and the appropriate amine (isopropylamine,
tert-butylamine or homoveratrylamine, 0.04 mol) in anhydrous toluene (10 mL) was heated in
sealed tube at 120 °C for 24 h (TLC). After cooling, the solution was evaporated under reduced
pressure to give an oily residue. The obtained product was dissolved in diethylether, and treated with
oxalic acid (0.45 g, 0.005 mol) in acetone or ethereal HCl to give either the oxalate or hydrochloride
salt. The separated solid was collected by filtration, washed with diethyl ether, dried and
crystallized from ethanol.
Dicyclohexylmethanone O-2-hydroxy-3-(isopropylamino)propyl oxime, hydrogen oxalate (4a). Yield
1
63%; M.p. 123–125 °C; H-NMR (free base in CDCl3) δ 1.06 (d, 6H, J = 6.5, 2CH3), 1.52–1.77
20H, 10CH cyclohexane), 2.42–2.65 (m, 2H, 2CHC=NO), 2.95–3.12 (m, 3H, CH N, CH(CH ) ),
(m,
2
2
3 2
3.4–
3.7 (m, 3H, CHOH, OH and NH exchangeable with D2O), 3.95 (dd, 1H, J = 10.8, 6.4 Hz,
OCH2), 4.15 (dd, 1H, J = 11.2, 2.8 Hz, OCH2). MS m/z (%); 326 (6.19, M++2), 325 (6.82, M++1),
210 (41.72), 98 (100), 83 (34.55), 72 (81.95).
Dicyclohexylmethanone O-3-(tert-butylamino)-2-hydroxypropyl oxime, hydrogen oxalate (4b). Yield
1
20H,
55%; M.p. 117–119 °C; H-NMR (free base in CDCl3) δ 1.46 (s, 9H, 3CH3), 1.59–1.72 (m,
10CH cyclohexane), 2.41–2.55 (m, 2H, CHC=NO), 2.81–2.98 (m, 2H, CH NH), 3.35–
3.54 (m, 3H,
2
2
CHOH, OH and NH exchangeable with D2O), 4.12 (dd, 1H, J = 11.2, 6.2 Hz, OCH2), 4.26 (dd, 1H,
J = 11.8, 2.1 Hz, OCH2). MS m/z (%); 340 (9.9, M++2), 339 (5.04, M++1), 225 (38.09), 113 (100),
86 (10.24), 72 (76.36).
Dicyclohexylmethanone O-3-(3,4-dimethoxyphenethylamino)-2-hydroxypropyl oxime, hydrogen oxalate
(4c). Yield 50%; M.p. 174–176 °C; 1H-NMR (free base in CDCl3) δ 1.5–1.8 (m, 20H, 10CH2
cyclohexane), 2.4–2.9 (m, 8H, 2CHC=NO, CH2NHCH2CH2), 3.50–3.71 (m, 3H, CHOH, OH and NH
exchangeable with D2O), 3.97 (s, 6H, 2OCH3), 4.08 (dd, 1H, J = 10.8, 6.0 Hz, OCH2), 4.24 (dd, 1H,
J = 11.5, 2.5 Hz, OCH2), 6.82 (d, 1H, J = 9.0, Ar-H), 7.01 (s, 1H, Ar-H), 7.21 (d, 1H, J = 8.5, Ar-H).
Cyclohexyl(phenyl)methanone O-2-hydroxy-3-(isopropylamino)propyl oxime, hydrochloride (4d).
1
Yield 70%; M.p. 157–159 °C; H-NMR (free base in CDCl3) δ 1.15 (d, 6H, J = 6.5, 2CH3), 1.52–1.77
10H, 5CH cyclohexane), 2.32–2.41 (m, 1H, CHC=NO), 2.77–2.89 (m, 3H, CH N, CH(CH ) ),
(m,
2
2
3 2
3.45–
3.62 (m, 3H, CHOH, OH and NH exchangeable with D2O), 3.98 (dd, 1H, J = 11.2, 6.1 Hz, OCH2),

Molecules 2014, 19
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4.12 (dd, 1H, J = 11.6, 2.7 Hz, OCH2), 7.45 (m, 5H, Ar-H). MS m/z (%); 320 (6.94, M++2), 319 (3.75,
M++1), 188 (68.77), 104 (70.80), 72 (100).
Cyclohexyl(phenyl)methanone O-3-(tert-butylamino)-2-hydroxypropyl oxime, hydrochloride (4e).
1
Yield 55%; M.p. 107–109 °C; H-NMR (free base in DMSO-d6) δ 1.21 (s, 9H, 3CH3), 1.47–1.68 (m,
10H, 5CH2 cyclohaxane), 2.45–2.62 (m, 1H, CHC=NO), 3.1–3.4 (m, 2H, CH2N), 3.44–3.68 (m, 3H,
CHOH, OH and NH exchangeable with D2O), 3.88 (dd, 1H, J = 11.0, 6.5 Hz, OCH2), 4.19 (dd, 1H,
J = 11.3, 2.5 Hz, OCH2), 7.55 (m, 5H, Ar-H). MS m/z (%); 334 (7.80, M++2), 333 (5.10, M++1), 146
(50.15), 172 (73.08), 72 (100).
Cyclohexyl(phenyl)methanone
O-3-(3,4-dimethoxyphenethyl
amino)-2-hydroxypropyl
oxime,
1
hydrogen oxalate (4f). Yield 48%; M.p. 140–142 °C; H-NMR (free base in CDCl3) δ 1.40–1.70 (m,
10H, 5CH2 cyclohaxane), 2.38–2.65 (m, 7H, CHC=NO, CH2NHCH2CH2), 3.40–3.65 (m, 3H, CHOH,
OH and NH exchangeable with D2O), 3.8 (s, 6H, 2OCH3), 4.00 (dd, 1H, J = 11.5, 6.2 Hz, OCH2),
4.18 (dd, 1H, J = 11.6, 2.6 Hz, OCH2), 6.9 (d, 1H, J = 8.5, Ar-H homoveratryl), 7.2 (d, 1H, J = 8.5,
Ar-H homoveratryl), 7.2 (s, 1H, Ar-H homoveratryl), 7.6 (m, 5H, Ar-H). MS m/z (%); 442 (10.12,
M++2), 441 (9.33, M++1), 289 (27.80) 186 (41.90), 104 (100).
Phenyl(pyridin-2-yl)methanone O-2-hydroxy-3-(isopropylamino)propyl oxime, hydrochloride (4g).
1
Yield 69%; M.p. 191–193 °C; H-NMR (free base in DMSO-d6) δ 1.44 (d, 6H, J = 6.5, 2CH3),
2.95–3.12 (m, 3H, CH2N, CH(CH3)2), 3.6–3.8 (m, 3H, CHOH, OH and NH exchangeable with D2O),
4.05 (dd, 1H, J = 11.5, 6.0 Hz, OCH2), 4.24 (dd, 1H, J = 11.0, 2.5 Hz, OCH2), 7.4–7.7 (m, 5H,
3,4,5-Ar-H and 3,5- pyridinyl-H), 7.9–8.1 (m, 3H, 2,6-Ar-H, 4-pyridinyl-H), 8.5 (d, 1H, J = 8.0,
6-pyridinyl-H). MS m/z (%); 315 (0.13, M++2), 314 (0.55, M++1), 313 (0.20, M+), 198 (100), 181
(40.71), 72 (44.96).
Phenyl(pyridin-2-yl)methanone O-3-(tert-butylamino)-2-hydroxypropyl oxime, hydrochloride (4h).
1
Yield 55%; M.p. 180–182 °C; H-NMR (free base in DMSO-d6) δ 1.27 (s, 9H, 3CH3), 2.94–3.08 (m,
3H, CH2N), 3.5–3.8 (m, 3H, CHOH, OH and NH exchangeable with D2O), 4.14 (dd, 1H, J = 11.5, 6.2 Hz,
OCH2), 4.33 (dd, 1H, J = 11.6, 2.8 Hz, OCH2), 7.6–7.8 (m, 5H, 3,4,5-Ar-H and 3,5- pyridinyl-H), 8.0–8.2
(m, 3H, 2,6-Ar-H, 4-pyridinyl-H), 8.5 (d, 1H, J = 8, 6-pyridinyl-H). MS m/z (%); 329 (5.38, M++2),
328 (1.35, M++1), 198 (100), 183 (70.91), 86 (46.49), 78 (42.53).
Phenyl (pyridin-2-yl)methanone O-3-(3,4-dimethoxyphenethyl amino)-2-hydroxypropyl oxime,
1
hydrogen oxalate (4i). Yield 50%; M.p. 177–179 °C; H-NMR (free base in CDCl3) δ 2.4–2.9 (m,
6H, CH2NHCH2CH2), 3.50–3.85 (m, 3H, CHOH, OH and NH exchangeable with D2O), 3.97 (s, 6H,
2OCH3), 4.10 (dd, 1H, J = 11.5, 6.0 Hz, OCH2), 4.22 (dd, 1H, J = 11.0, 2.8 Hz, OCH2), 6.8 (d, 2H,
5,6-Ar-H homoveratryl), 7.1 (s, 1H, 2-Ar-H homoveratryl), 7.4–7.7 (m, 5H, 3,4,5-Ar-H and
3,5- pyridinyl-H), 7.9 (m, 3H, 2,6-Ar-H, 4-pyridinyl-H), 8.5 (d, 1H, J = 8.5, 6-pyridinyl-H).
1-(Naphthalen-2-yl)ethanone O-2-hydroxy-3-(isopropylamino)propyl oxime, hydrochloride (4j).
Yield 67%; M.p. 117–119 °C; 1H-NMR (free base in CDCl3) δ 1.61 (d, 6H, J = 6.5, 2CH3), 2.20 (s,
3H, CH3), 2.7–2.9 (m, 2H, CH2NH), 3.44–3.65 (m, 3H, CHOH, OH and NH exchangeable with

Molecules 2014, 19
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D2O), 4.42 (dd, 1H, J = 11.6, 6.4 Hz, OCH2), 4.57 (dd, 1H, J = 11.2, 2.4 Hz, OCH2), 7.65 (t, 2H,
J = 8.0, 6,7-Ar-H), 8.06 (d, 4H, J = 7.5, 3,4,5,8- Ar-H), 8.45 (s, 1H, 1-Ar-H). MS m/z (%); 300
(1.60, M+), 168 (6.54), 127 (21.09), 73 (91.92), (100), 60 (55.21).
1-(Naphthalen-2-yl)ethanone O-3-(tert-butylamino)-2-hydroxypropyl oxime, hydrochloride (4k). Yield
1
59%; M.p. 120–122 °C; H-NMR (free base in CDCl3) δ 1.18 (s, 9H, 3CH3), 2.38 (s, 3H, 3CH3),
3.0–3.4 (m, 2H, CH2NH), 3.4–3.75 (m, 3H, CHOH, OH and NH exchangeable with D2O), 4.24 (dd,
1H, J = 11.3, 6.2 Hz, OCH2), 4.38 (dd, 1H, J = 11.8, 2.2 Hz, OCH2), 7.56 (t, 2H, J = 8.5 6,7-Ar-H),
8 (d, 4H, J = 7.5, 3,4,5,8- Ar-H), 8.4 (s, 1H, 1-Ar-H). MS m/z (%); 316 (9.25, M++2), 315 (7.18,
M++1), 167 (84.11), 127 (60.13), 86 (100), 58 (43.86).
1-(Naphthalen-2-yl)ethanone O-3-(3,4-dimethoxyphenethyl amino)-2-hydroxypropyl oxime, hydrogen
oxalate (4l). Yield 50%; M.p. 181–183 °C; 1H-NMR (free base in CDCl3) δ 2.10 (s, 3, CH3), 3.1–3.4 (m,
6H, CH2NHCH2CH2), 3.3.75 (m, 3H, CHOH, OH and NH exchangeable with D2O), 3.8 (s, 6H,
2OCH3), 4.15 (dd, 1H, J = 11.5, 6.2 Hz, OCH2), 4.30 (dd, 1H, J = 11.6, 2.6 Hz, OCH2), 6.8 (d, 2H,
J = 8.0, 5,6-Ar-H homoveratryl), 7.17 (s, 1H, 2-Ar-H homoveratryl), 7.65 (t, 2H, J = 8.5, 6,7-Ar-H),
8.14 (d, 4H, J = 7.5, 3,4,5,8-Ar-H), 8.62 (s, 1H, 1-Ar-H). MS m/z (%); 424 (12.07, M++2), 423 (9.11,
M++1), 181 (11.91), 152 (100), 137 (28.19).
1-(Biphenyl-4-yl)ethanone O-2-hydroxy-3-(isopropylamino)propyl oxime, hydrogen oxalate (4m).
Yield 60%; M.p. 169–171 °C; 1H-NMR (free base in DMSO-d6) δ 1.21 (d, 6H, J = 6.5, 2CH3), 2.16 (s,
3H, CH3C), 2.95–3.12 (m, 3H, CH2N, CH(CH3)), 3.45–3.72 (m, 3H, CHOH, OH and NH exchangeable
with D2O), 3.89 (dd, 1H, J = 10.9, 6.1 Hz, OCH2), 4.29 (dd, 1H, J = 11.0, 2.5 Hz, OCH2), 7.2–7.4
' ' '
' '
(m, 3H, 3 ,4 ,5 -Ar-H), 7.56 (d, 2H, J = 8.0, 2 ,6 -Ar-H), 7.84 (d, 4H, J = 8.5, 2,3,5,6-Ar-H).
1-(Biphenyl-4-yl)ethanone O-3-(tert-butylamino)-2-hydroxypropyl oxime, hydrochloride (4n). Yield
52%; M.p. 165–167 °C; 1H NMR (free base in DMSO-d6) δ 1.21 (s, 9H, 3CH3), 2.16 (s, 3H, CH3C),
2.95–3.12 (m, 3H, CH2N, CH(CH3)), 3.45–3.72 (m, 3H, CHOH, OH and NH exchangeable with
D2O), 3.89 (dd, 1H, J = 10.9, 6.1 Hz, OCH2), 4.29 (dd, 1H, J = 11.0, 2.5 Hz, OCH2), 7.2–7.4 (m,
' ' '
' '
3H, 3 ,4 ,5 -Ar-H), 7.56 (d, 2H, J = 8.0, 2 ,6 -Ar-H), 7.84 (d, 4H, J = 8.5, 2,3,5,6-Ar-H). MS m/z
(%); 342 (8.39, M++2), 341 (7.71, M++1), 193 (69), 153 (39.43), 86 (100).
1-(Biphenyl-4-yl)ethanone O-3-(3,4-dimethoxyphenethylamino)-2-hydroxypropyl oxime, hydrochloride
1
2.85–3.1 (m,
(4o). Yield 48%; M.p. 157–159 °C; H-NMR (free base in CDCl3) δ 2.2 (s, 3H, CH3),
6H, CH2NHCH2CH2), 3.45–3.72 (m, 3H, CHOH, OH and NH exchangeable with D2O),
3.85 (s,
6H, 2OCH3) 4.02 (dd, 1H, J = 11.2, 6.2 Hz, OCH2), 4.22 (dd, 1H, J = 11.5, 2.2 Hz, OCH2), 6.85
–
(d, 2H, J = 8.0, 5,6-Ar-H homoveratryl), 7.25 (s, 1H, 2-Ar-H homoveratryl), 7.3 7.5 (m, 3H,
' ' '
' '
3 ,4 ,5 -Ar-H), 7.59 (d, 2H, J = 8.0, 2 ,6 -Ar-H), 7.88 (d, 4H, J = 8.5, 2,3,5,6-Ar-H). MS m/z (%);
450 (11.9, M++2), 449 (6.12, M++1), 297 (34.08), 194 (100).
1-(5-Chlorothiophen-2-yl)ethanone O-2-hydroxy-3-(isopropyl amino)propyl oxime, hydrochloride (4p).
Yield 70%; M.p. 145–147 °C; 1H-NMR (free base in DMSO-d6) δ 1.03 (d, 6H, J = 7.0, 2CH3), 2.10 (s, 3H,
CH3C=NO), 2.85–3.06 (m, 3H, CH2N, CH(CH3)2)), 3.5–3.7 (m, 3H, CHOH, OH and NH exchangeable

Molecules 2014, 19
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with D2O), 4.11 (dd, 1H, J = 11.5, 6.2 Hz, OCH2), 4.39 (dd, 1H, J = 11.5, 2.2 Hz, OCH2), 7.17 (d, 1H,
J = 4.0, 3-thiophene-H), 7.35 (d, 1H, J = 4.1, 4-thiophene -H).
1-(5-Chlorothiophen-2-yl)ethanone O-3-(tert-butylamino)-2-hydroxypropyl oxime, hydrochloride (4q).
Yield 62%; M.p. 152–154 °C; 1H-NMR (free base in DMSO-d6) δ 1.33 (s, 9H, 3CH3), 2.21 (s, 3H,
CH3C=NO), 2.77–2.96 (m, 2H, CH2N) 3.47–3.66 (m, 3H, CHOH, OH and NH exchangeable with
D2O), 4.08 (dd, 1H, J = 11.4, 6.5 Hz, OCH2), 4.38 (dd, 1H, J = 11.3, 2.8 Hz, OCH2), 6.98 (d, 1H,
J = 4.2, 3-thiophene-H), 7.37 (d, 1H, J = 4.0, 4-thiophene-H). MS m/z (%); 306 (2.06, M++2), 305
(1.45, M++1), 160 (100), 145 (82.20), 72 (84.98).
1-(5-Chlorothiophen-2-yl)ethanone O-3-(3,4-dimethoxyphenethylamino)-2-hydroxypropyl oxime,
1
hydrochloride (4r). Yield 52%; M.p. 148–150 °C; H-NMR (free base in DMSO-d6) δ 2.19 (s, 3H,
2.79 3.05 (m, 6H, CH NHCH CH ),
3.4–3.6 (m, 3H, CHOH, OH and NH exchangeable
CH3C=NO),
–
2
2
2
with D2O), 4.0 (dd, 1H, J = 11.0, 6.0 Hz, OCH2), 4.32 (dd, 1H, J = 11.5, 2.6 Hz, OCH2), 6.88 (d, 2H,
J = 8.0, 5,6-Ar-H homoveratryl), 7.17 (d, 1H, J = 4.0, 3-thiophene-H), 7.29 (s, 1H, 2-Ar-H homoveratryl),
7.39 (d, 1H, J = 4.01, 4-thiophene -H). MS m/z (%); 413 (3.22, M+), 261 (77.44), 158 (100), 73 (13.79).
1,3,3-Trimethylbicyclo[2.2.1]heptan-2-one O-2-hydroxy-3-(isopropylamino)propyl oxime, hydrochloride
(4s). Yield 55%; M.p. 127–129 °C; 1H-NMR (free base in CDCl3) δ 1.14 (d, 6H, J = 6.5, 2CH3), 1.25 (s,
6H, 2CH3), 1.32 (s, 3H, CH3), 1.44–1.81 (m, 7H, CH, 3CH2 cyclic moiety), 2.7–3.0 (m, 2H, CH2N),
3.33–3.71 (m, 3H, CHOH, OH and NH exchangeable with D2O), 4.11 (dd, 1H, J = 11.2, 6.5 Hz, OCH2),
4.31 (dd, 1H, J = 11.7, 2.5 Hz, OCH2). MS m/z (%); 284 (11.25, M++2), 167 (12.71), 81 (100), 69 (36.25).
1,3,3-Trimethylbicyclo[2.2.1]heptan-2-one O-3-(tert-butylamino)-2-hydroxypropyl oxime, hydrogen
1
oxalate (4t). Yield 52%; M.p. 115–117 °C; H-NMR (free base in CDCl3) δ 1.12 (s, 9H, 3CH3), 1.25
(s, 6H, 2CH3), 1.34 (s, 3H, CH3), 1.4–1.8 (m, 7H, CH, 3CH2 cyclic moiety), 2.9–3.4 (m, 2H, CH2N),
3.65–3.9 (m, 3H, CHOH, OH and NH exchangeable with D2O), 3.88 (dd, 1H, J = 11.5, 6.5 Hz,
OCH2), 4.21 (dd, 1H, J = 11.0, 2.8 Hz, OCH2).
1,3,3-Trimethylbicyclo[2.2.1]heptan-2-one O-3-(3,4-dimethoxyphenethylamino)-2-hydroxypropyl oxime,
1
1.22 (s, 6H,
hydrogen oxalate (4u). Yield 39%; M.p. 140–142 °C; H-NMR (free base in DMSO-d6) δ
2CH ), 1
1.5 1.78 (m, 7H, CH, 3CH in cyclic moiety), 2.4 2.9 (m, 6H,
– –
2
.31 (s, 3H, CH3),
3
CH2NHCH2CH2),
3.61–3.81 (m, 3H, CHOH, OH and NH exchangeable with D2O), 3.97 (s, 6H,
OCH3), 4.10 (dd, 1H, J = 11.6, 6.4 Hz, OCH2), 4.22 (dd, 1H, J = 11.4, 2.5 Hz, OCH2), 6.9 (d, 2H,
J = 8.5, 5,6-Ar-H), 7.15 (s, 1H, 2-Ar-H). MS m/z (%); 406 (9.17, M++2), 405 (7.45, M++1), 253
(26.99), 151 (50.87), 104 (88.50), 81 (100).
5-Methoxy-3,4-dihydronaphthalen-1(2H)-one O-2-hydroxy-3-(isopropylamino)propyl oxime, hydrogen
1
oxalate (4v). Yield 76%; M.p. 180–182 °C; H-NMR (free base in CDCl3) δ 1.21 (d, 6H, J = 6.5,
2CH3), 2.22–2.61 (m, 6H, 3CH cyclic structure), 2.8–3.1 (m, 3H, CH2NCH), 3.5–3.7 (m, 3H,
2
CHOH, OH and NH exchangeable with D2O), 3.95 (dd, 1H, J = 11.5, 6.4 Hz, OCH2), 4.14 (dd, 1H,
J = 11.6, 2.6 Hz, OCH2), 4.19 (s, 3H, OCH3), 7.11 (d, 1H, J = 9.0, 6-Ar- H), 7.36 (t, 1H, J = 8.5, 7-Ar-H),
7.55 (d, 1H, J = 8.5, 8-Ar-H). MS m/z (%); 308 (6.55, M++2), 307 (3.45, M++2), 248 (35), 174 (100).

Molecules 2014, 19
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5-Methoxy-3,4-dihydronaphthalen-1(2H)-one O-3-(tert-butylamino)-2-hydroxypropyl oxime, hydrogen
1
oxalate (4w). Yield 69%; M.p. 177–179 °C; H-NMR (free base in CDCl3) δ 1.21 (s, 9H, 3CH3),
2.03–2.81 (m, 6H, 3CH2 cyclic structure), 3.1–3.4 (m, 2H, CH2N), 3.5–3.7 (m, 3H, CHOH, OH and
NH exchangeable with D2O), 3.89 (dd, 1H, J = 11.5, 6.2 Hz, OCH2), 4.08 (dd, 1H, J = 11.6, 2.4 Hz,
OCH2), 4.15 (s, 3H, OCH3), 7.07 (d, 1H, J = 9.0, 6-Ar-H), 7.31 (t, 1H, J = 8.5, 7-Ar-H), 7.52 (d, 1H,
J = 8.5, 8-Ar-H).
5-Methoxy-3,4-dihydronaphthalen-1(2H)-one O-3-(3,4-dimethoxyphenethylamino)-2-hydroxypropyl
1
oxime, hydrogen oxalate (4x). Yield 58%; M.p. 160–162 °C; H-NMR (free base in DMSO-d6) δ
2.1–2.9 (m, 12H, CH2NHCH2CH2, 3CH2 cyclic structure), 3.51–3.70 (m, 3H, CHOH, OH and NH
exchangeable with D2O), 3.82 (s, 6H, 2OCH3), 3.88 (s, 3H, OCH3), 4.11 (dd, 1H, J = 11.1, 6.4 Hz,
OCH2), 4.27 (dd, 1H, J = 11.3, 3.0 Hz, OCH2), 6.79 (d, 2H, J = 8.5, 5,6-Ar-H homoveratryl), 7.0 (s,
1H, 2-Ar-H homoveratryl), 7.25 (d, 1H, J = 8.0, 6-Ar-H), 7.47 (t, 1H, J = 9.0, 7-Ar-H), 7.62 (d, 1H,
J = 8.0, 8-Ar-H).
Diphenylmethanone O-3-(3,4-dimethoxyphenethylamino)-2-hydroxypropyl oxime, hydrochloride (4y).
Yield 52%; M.p. 166–168 °C; 1H-NMR (free base in DMSO-d6) δ 2.58–2.96 (m, 6H, CH2NHCH2CH2),
3.6–3.9 (m, 3H, CHOH, OH and NH exchangeable with D2O), 3.95 (s, 6H, 2OCH3), 4.16 (dd, 1H,
J = 10.8, 5.9 Hz, OCH2), 4.28 (dd, 1H, J = 11.4, 3.4 Hz, OCH2), 6.67 (d, 2H, J = 8.5, 5,6-Ar-H
homoveratryl), 7.18 (s, 1H, 2-Ar-H homoveratryl), 7.60–8.02 (m, 10H, Ar-H). MS m/z (%); 436 (5.85,
M++2), 435 (6.79, M++1), 152 (100), 137 (26.50).
1-Phenylethan-1-one O-3-(3,4-dimethoxyphenethylamino)-2-hydroxypropyl oxime, hydrochloride (4z).
Yield 56%; M.p. 138–141 °C; 1H-NMR (free base in DMSO-d6) δ 1.91 (s, 3H, CH3), 2.4–2.9 (m, 6H,
CH2NHCH2CH2), 3.6–3.9 (m, 3H, CHOH, OH and NH exchangeable with D2O), 3.90 (s, 6H, 2OCH3),
4.05 (dd, 1H, J = 11.2, 6.4 Hz, OCH2), 4.25 (dd, 1H, J = 11.9, 2.7 Hz, OCH2), 6.87 (d, 2H, 5,6-Ar-H
homoveratryl), 7.15 (s, 1H, 2-Ar-H homoveratryl), 7.53 (t, 3H, J = 8, Ar-H), 7.9 (d, 2H, J = 8.5,
Ar-H). MS m/z (%); 374 (5.97, M++2), 375 (7.45, M++1), 165 (19.97), 118 (100), 77 (29.65).
3.2. Molecular Docking Methodology
Automated docking simulations were conducted with the Molegro Virtual Docker 2007 (MVD
2007.2.2.5–Aug 27, 2007 [win32]) fully functional free trial version with time limiting license [20]
3D Molecular structures and energy minimization were carried out by free version of Marvinsketch
4.1.13 from Chemaxon Ltd. (Budapest, Hungary) [28]. 3D crystal structures of β1 and β2-adrenergic
receptors were obtained from Protein Data Bank (PDB); entries 2VT4 and 2R4R, respectively [18,19,29].
All the molecular modeling studies were carried out on a PC equipped with an Intel Celeron 1.2 GHz
processor, and 320 MB memory running the Windows XP operating system.
The docking study was performed, in the present investigation, following a general procedure for
docking [30,31]. The coordinates of the crystal structures adrenergic receptors were imported into
Molegro Virtual Docker. Receptor structures were checked for missing atoms, bonds and contacts. The
ligand molecules were constructed in 2D form then converted to 3D form and energy minimized by
Marvinsketch. Energy minimized conformers were imported into Molegro Virtual Docker. The

Molecules 2014, 19
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dimensions of the docking constraint were manipulated so as to accommodate all the amino acid
residues present in the active site (radius 12 Ǻ). The ligands were manually docked into the binding
cavity of β2 adrenergic receptor, guided by all the amino acids present in the active site, and snapshots
were recorded. Figure 2a,b represent the docking snapshots of compounds 4o and 4q with the β1 and
β2-adrenergic receptors, respectively, showing the forces of interaction. In this study the MolDock
score function and hydrogen bond interactions between tested compounds and the target receptors
were used to compare between the tested compounds and reference drugs. The MolDock score is a
new technique for high-accuracy molecular docking. It is an extension of the piecewise linear potential
(PLP) [32,33] including new hydrogen bonding and electrostatic terms. To further improve docking
accuracy, a re-ranking scoring function is introduced, which identifies the most promising docking
solution from the solutions obtained by the docking algorithm [34]. The lower MolDock score means
better ligand-receptor interactions. The MolDock scores and hydrogen bond were recorded (Table 3).
3.3. Biological Evaluation
Epinephrine hydrochloride injection (0.25 mg/1 mL) was obtained from Misr Co. Cairo, Egypt.
Propranolol hydrochloride (Inderal injection) B.P. 0.1%, was obtained from AstraZeneca Limited
(Cairo, Egypt). Acetylcholine chloride and salbutamol sulfate were purchased from Sigma-Aldrich
chemical Co. All other chemicals used in this study were of finest analytical grade, and obtained from
El-Nasr Pharmaceutical Chemicals Company. Tested compounds were dissolved in distilled water or
20% ethanol, just before use, in 10−5 mole concentration. Adult male Hartley guinea pigs weighing
350–500 g were used. Animals were maintained under standard conditions of temperature with regular
12 h light/dark cycles and allowed free access to standard laboratory food and water. Biological
evaluation using animals was performed in accordance with the Ethics Committee of faculty of
Pharmacy Mansoura University.
3.3.1. In Vitro Screening for β1-Adrenoceptor Activity in Isolated Guinea Pig Atria
Guinea pigs were anaesthetized by diethyl ether. The chest was opened and the heart was quickly
excised and placed in oxygenated Ringer Locke solution of the following composition (g/L): NaCl (9),
KCl (0.42), CaCl2 (0.24), NaHCO3 (0.15) and glucose (1). Atria were dissected away from the rest of the
heart, freed from connective tissue and suspended in 50 mL organ bath containing Ringer Locke
solution. This solution is thermoregulated at 37 °C and continuously bubbled with pure oxygen. The
extremities of the strip, consisting of both atria, were tied with a thread. The first thread was used to tie
the organ to a muscle holder fixed in place in a muscle chamber of an isolate organ bath; the second
thread was used to connect atria to an isomeric force-displacement transducer (model 50–7905, Harvard
Apparatus Inc, South Natick, MA, USA) situated above the muscle chamber and connected to a two-
channel oscillograph (model 50–8622, Harvard Apparatus). Atria spontaneously beating were loaded
with 0.5 g and were allowed to adjust to the bath conditions for at least 30 min prior to the experiment,
and the Ringer Locke solution was changed at 10 min intervals. Epinephrine (43 µL of epinephrine vial;
10−6 mol) was applied and allowed to act till the response was fully developed. The atria were washed
with Ringer Locke solution. Test compound (10−5 mol, 2.5–4.5 mg/L) was applied and incubated for
5 min in the organ bath to test and evaluate any possible agonistic activity on the β1-adrenergic receptor.

Molecules 2014, 19
3432
Then, epinephrine (43
−6 mol)
µL of epinephrine vial; 10
was added in the organ bath in order to evaluate
antagonistic activity of the test compound to epinephrine on the β -adrenergic receptor.
any competitive
1
Propranolol (10−5 mol) was used as reference drug.
3.3.2. In Vitro Screening for β2-Adrenoceptor Activity in Isolated Guinea Pig Trachea
Guinea pig was anaesthetized by diethylether; the trachea was carefully isolated and immersed in
oxygenated Krebs-Hanseleit solution of the following composition (g/L): NaCl (6.9), KCl (0.35),
KH2PO4 (0.16), NaHCO3 (2.1), MgSO4 (0.29), anhydrous CaCl2 (0.28) and glucose (2). The tracheal
preparation was cautiously cleaned of unnecessary adipose and connective tissues. Subsequently, the
tracheal cartilage containing smooth muscles was cut into zig-zag strips according to Emmerson and
Mackay method [35]. Zig-zag tracheal strips were tied at each end with a thread. The first thread was
used to tie the organ to a muscle holder fixed in place in a muscle chamber of an isolate organ bath; the
second thread was used to connect trachea to an isomeric force-displacement transducer (model
50–7905, Harvard Apparatus) situated above the muscle chamber and connected to a two-channel
oscillograph (model 50–8622, Harvard Apparatus). The organ bath contains 50 mL of Krebs-Hanseleit
solution at 32 °C and gassed with pure oxygen. The zig-zag tracheal strips were loaded with 1 g and were
left to stabilize for 1 h, and the Krebs-Hanseleit solution was changed at 15 min intervals. The zig-zag
tracheal strips were precontracted with acetylcholine chloride (0.9 mg/50 mL; 10−4 mol) and allowed
to act till the response was fully developed. The test compound (10−5 mol) was then added and
incubated for 20 min in the organ bath to evaluate its agonistic activity on β2-adrenergic receptor.
Salbutamol sulfate (10−5 mol) was then added in organ bath in the presence of the test compound in order
to evaluate any blocking activity of the test compound to β2- adrenergic receptor. Propranolol (10−5 mol)
was used as reference drug.
4. Conclusions
In this study, a series of oxime ether derivatives 4a–z were designed and successfully synthesized in
convenient steps. Selected compounds were tested for their biological activities against in vitro β1- and
β2-adrenergic receptors and also examine their binding energies with 3D crystal structures of β1 and
β2 receptors by doing molecular docking analysis. These findings confirm the significance of a
homoveratrylamine portion for β1 receptors and also indicate the importance of the presence of a
chlorothiophene moiety in the hydrophobic region for best complementarity with β2 receptors.
Supplementary Materials
Supplementary materials can be accessed at: http://www.mdpi.com/1420-3049/19/3/3417/s1.
Acknowledgments
Authors would like to thank Ghada M. Suddek, Associate Professor of Pharmacology, Faculty of
Pharmacy, Mansoura University for performing the biological screening. HAG thanks the Deanship of
Scientific Research and the Research Center, College of Pharmacy, King Saud University.

Molecules 2014, 19
3433
Conflicts of Interest
Authors have no conflict of interest to declare in connection with the contents of this manuscript.
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Sample Availability: Samples of the compounds 4y–z are available from the authors.
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