Synthesis and quantitative structure-activity relationships study for phenylpropenamide derivatives as inhibitors of hepatitis B virus replication

Article abstract of DOI:10.1016/j.ejmech.2015.05.032

A series of new phenylpropenamide derivatives containing different substituents was synthesized, characterized and evaluated for their anti-hepatitis B virus (HBV) activities. The quantitative structure eactivity relationships (QSAR) of phenylpropenamide compound have been studied. The 2D-QSAR models, based on DFT and multiple linear regression analysis methods, revealed that higher values of total energy (TE) and lower entropy (Sθ) enhanced the anti-HBV activities of the phenylpropenamide molecules. Predictive 3D-QSAR models were established using SYBYL multifit molecular alignment rule. The optimum models were all statistically significant with cross-validated and conventional coefficients, indicating that they were reliable enough for activity prediction.

Full text of DOI:10.1016/j.ejmech.2015.05.032

Accepted Manuscript
Synthesis and Quantitative Structure–activity Relationships Study for
Phenylpropenamide Derivatives as Inhibitors of Hepatitis B Virus Replication
Yang Jing, Ma Min, Wang Xueding, Jiang Xingjun, Zhang Yuanyuan, Yang Weiqing,
Li Zicheng, Wang Xihong, Yang bin, Ma Menglin
PII:
S0223-5234(15)30059-3
10.1016/j.ejmech.2015.05.032
EJMECH 7911
DOI:
Reference:
To appear in: European Journal of Medicinal Chemistry
Received Date: 11 November 2014
Revised Date: 29 March 2015
Accepted Date: 21 May 2015
Please cite this article as: Y. Jing, M. Min, W. Xueding, J. Xingjun, Z. Yuanyuan, Y. Weiqing, L. Zicheng,
W. Xihong, Yang bin, M. Menglin, Synthesis and Quantitative Structure–activity Relationships Study
for Phenylpropenamide Derivatives as Inhibitors of Hepatitis B Virus Replication, European Journal of
Medicinal Chemistry (2015), doi: 10.1016/j.ejmech.2015.05.032.
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ACCEPTED MANUSCRIPT
Synthesis and Quantitative Structure–activity Relationships Study for Phenylpropenamide
Derivatives as Inhibitors of Hepatitis B Virus Replication
The new phenylpropenamide derivatives were synthesized, characterized and evaluated for their
anti-HBV activities. The 2D and 3D-QSAR models of phenylpropenamide was constructed.

ACCEPTED MANUSCRIPT
Synthesis and Quantitative Structure–activity Relationships Study for Phenylpropenamide
Derivatives as Inhibitors of Hepatitis B Virus Replication
Yang Jinga, Ma Mina, Wang Xuedinga, Jiang Xingjuna, Zhang Yuanyuana, b*, Yang Weiqinga, Li
Zichengb, Wang Xihonga, Yang binc, Ma Menglina*
a Key Lab of Advanced Scientific Computation of Sichuan province, Faculty of physical and
chemistry, Xihua University, Chengdu 610039, China
b College of chemical engineering, Sichuan University, Chengdu 610065, China
c West china school of pharmacy, Sichuan University, Chengdu 610064, China
Abstract A series of new phenylpropenamide derivatives containing different substituents was
synthesized, characterized and evaluated for their anti-hepatitis B virus (HBV) activities. The
quantitative structure–activity relationships (QSAR) of phenylpropenamide compound have been
studied. The 2D-QSAR models, based on DFT and multiple linear regression analysis methods,
revealed that higher values of total energy (TE) and lower entropy (Sө) enhanced the anti-HBV
activities of the phenylpropenamide molecules. Predictive 3D-QSAR models were established
using SYBYL multifit molecular alignment rule. The optimum models were all statistically
significant with cross-validated and conventional coefficients, indicating that they were reliable
enough for activity prediction.
Keywords Phenylpropenamide, DFT, QSAR, Hepatitis B
1. Introduction
The treatment of hepatitis B virus infection represents one of the current therapeutic
challenges in virology. There are approximately 350 million chronically infected individuals,
resulting in 0.5-1.2 million deaths annually [1-3]. Although interferon-α has been a mainstay of
HBV treatment regimens, sustained response rates tend to be low (20±30%) and resistance builds
rapidly [4-5]. The other current agents approved for the treatment of HBV infection are nucleoside
analogues lamivudine, telbivudine, entecavir, adefovir and tenofovir ( Fig. 1) [6-9] .
Phenylpropenamides (Fig. 2), such as AT-61 [10] and AT-130 [11-14], were reported as a
non-nucleoside class of inhibitors of HBV replication in cell culture. It was also reported that
phenylpropenamides were specific inhibitors of HBV replication and most likely inhibited
replication by interfering with the packaging of pregenomic RNA into immature core particles.
Structure–activity relationship analysis widely applied due to their well-established
predictive power [15]. Essentially, provides an useful tool in new drugs design by correlating the
physico-chemical properties of a series of compounds with their respective biological activities. In
order to explore the relationship between structure and antiviral activity, a series of
phenylpropenamide derivatives was synthesized and their antiviral activities were then tested. The
density functional theory (DFT) and linear regression analysis method were used to construct the
2D-QSAR models [16-17]. Furthermore, three-dimensional quantitative structure-activity
relationship (3D-QSAR) method is helpful to extract a relation between biological activity and
chemical structure. Especially, the comparative molecular field analysis (CoMFA) is an effective
method based on statistical techniques [18-20]. The biological activity of new phenylpropenamide
derivatives can be predicted by using the 2D or 3D-QSAR equation, which may offers an
important reference for the future research work on novel derivative design and synthesis.
1

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2. Experimental
2.1 Chemistry
Thin layer chromatography was performed with Qingdao Ocean silica gel GF254. Nuclear
magnetic resonance (NMR) spectra were recorded on a Bruker DPX-300 FT NMR spectrometer at
300 MHz for 1H and were referenced to tetramethylsilane (δ values are given in ppm and J-values
in Hz). HRMS were measured on Shimadzu LCMS-IT-TOF. ESI-MS were measured on a Varian
CH-5 apparatus. IR spectra were measured acquired on a Nicolet 380 and obtainedby using KBr
platespellet. The X-ray diffraction data were collected on X’Pert Pro MPD diffract meter. The
synthetic route was shown in Scheme 1.
2.1.1 General procedure for the preparation of 4-arylidene-2-phenyloxazol-5(4H)-one (2a-2r)
Hippuric acid (10 g, 56 mmol), aromatic aldehydes (43 mmol ), potassium acetate (4.22 g, 44
mmol) and acetic anhydride (14mL, 17 mmol) were added to a 100 mL three-necked flask and
heated at reflux for 0.5 h. The reaction mixture was treated with ethanol (20 mL) when the
o
temperature was cooled down to 40 C. The product was further found by precipitation as the
temperature went down to ambient. The resulting solid was collected, washed with ethanol (2×10
mL) and then dried to obtain 2a-2r as yellow crystals.
2.1.2 General procedure for the preparation of N-(3-oxo-1-aryl-3-(piperidin-1-yl)prop-1-en
-2-yl)benzamide (3a-3r)
Dichloromethane (50 mL) and piperidine (2.83g, 33mmol) were added into a solution of 2
(33mmol) in dichloromethane (166mL) at 0oC, and then stirred at this temperature for 1 h to
obtain CH2Cl2 solution of 3a-3r, which were used for the next reaction without further
purification.
2.1.3 General procedure for the preparation of (Z)-N-(1-bromo-3-oxo-1-aryl-3-(piperidin-1-yl)
prop-1-en-2-yl) benzamide (4a-4r)
Solid calcium carbonate (4.16 g, 42 mmol) was added to a dichloromethane solution of 3a-3r
at 0 oC, and bromine liquid (5.32 g, 33mmol) in CH2Cl2 (50mL) was then added to the mixture.
The resulting solution was stirred at room temperature overnight, filtered and evaporated to afford
the yellow oil, which was finally crystallized from ethanol to give 4a-4r
2.1.4.1 (Z)-N-(1-chloro-3-oxo-1-phenyl-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide (5a). White
1
crystals. Yield 16%; H NMR (300MHz, CDCl3): 8.14 (s, 1H, NH), 7.92 (d, J=7.2Hz, 2H, ArH),
7.61 (m, 5H, ArH), 7.39 (t, J=3Hz, 3H, ArH), 3.65 (m, 1H, CH2), 3.33 (m, 3H, CH2), 1.56 (m, 4H,
CH2), 1.12 (m, 1H, CH2), 0.53 (m, 1H, CH2), IR (KBr, cm-1) v: 3446 (NH), 2360 (C-H), 1650,
1544 (C=O), 1268 (C=C), 1089, 790. ESI-MS 2m/z: 736.72.
2.1.4.2 (Z)-N-(1-chloro-1-(2-cyanophenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5b). Pale yellow crystals. Yield 11%; 1H NMR (300MHz, CDCl3): 8.27 (s, 1H, NH), 7.92 (d,
J=7.8Hz, 2H, ArH), 7.79 (d, J=7.5Hz, 1H, ArH), 7.68 (m, 3H, ArH), 7.54 (t, J=7.5Hz, 3H, ArH),
3.37 (s, 4H, CH2), 1.91 (m, 1H, CH2), 1.45 (m, 3H, CH2), 1.01 (m, J=3.6Hz, 2H, CH2), IR (KBr,
cm-1) v: 3293 (NH), 2882 (C-H), 2225 (CN), 1662, 1618 (C=O), 1268 (C=C), 1025, 709.
13C NMR (75Hz, CDCl3): δ 164.34, 161.89, 135.67, 133.36, 132.87. 132.61, 132.53, 132.14,
130.83, 128.75, 128.42, 127.67, 124.60, 115.88, 106.34, 47.86, 42.63, 25.61, 24.82, 24.14. HRMS
(EI) calcd. for 393.1244; C22H20ClN3O2, found (E+) 394.1323.
2.1.4.3 (Z)-N-(1-chloro-1-(2-nitrophenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide (5c).
Pale yellow crystals. Yield 28%; 1H NMR (300MHz, CDCl3): 8.23 (s, 1H, NH), 8.09 (d, J=7.5Hz,
1H, ArH), 7.93 (d, J=7.2Hz, 2H, ArH), 7.63 (d, J=6.6Hz, 1H, ArH), 7.67 (m, 5H, ArH), 3.46 (m,
2

ACCEPTED MANUSCRIPT
4H, CH2), 1.78 (m, J=0.3Hz, 1H, CH2), 1.43 (m, 5H, CH2), IR (KBr, cm-1) v: 3273 (NH), 2932
(C-H), 1652, 1618 (C=O), 1268 (C=C), 910, 740. 13C NMR (75Hz, CDCl3): δ 164.15, 162.19,
136.22, 135.44, 133.35, 132.84. 132.51, 132.63, 132.37, 130.87, 128.91, 127.45, 124.64, 106.80,
47.93, 42.67, 25.42, 24.85, 24.17. HRMS (EI) calcd. for 413.1142; C21H20ClN3O4, found (E+)
414.1221.
2.1.4.4
(Z)-N-(1-chloro-1-(4-chlorophenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
1
(5d). White crystals. Yield 3%; H NMR (300MHz, CDCl3): 8.11 (s, 1H, NH), 7.91 (d, J=7.2Hz,
2H, ArH), 7.62 (t, J=7.2Hz, 1H, ArH), 7.53 (t, J=8.4Hz, 4H, ArH), 7.37 (d, J=8.4Hz, 2H, ArH),
3.52 (m, 3H, CH2), 3.12 (m, 1H, CH2), 1.64 (s, 4H, CH2), 1.48 (m, 2H, CH2), IR (KBr, cm-1) v:
3270 (NH), 2858 (C-H), 1672, 1623 (C=O), 1267 (C=C), 1092, 707. ESI-MS 2m/z: 805.66.
2.1.4.5 (Z)-N-(1-chloro-3-oxo-3-(piperidin-1-yl)-1-o-tolylprop-1-en-2-yl)benzamide (5e). White
crystals. Yield 11%; 1H NMR (300MHz, CDCl3): 8.12 (s, 1H, NH), 7.92 (d, J=7.2Hz, 2H, ArH),
7.61 (m, 3H, ArH), 7.37 (s, 2H, ArH), 7.29 (m, 2H, ArH), 3.65 (m, 1H, CH2), 3.34 (m, 3H, CH2),
2.38 (s, 3H, CH3), 1.41 (m, 4H, CH2), 1.13 (m, 1H, CH2), 0.53 (m, 1H, CH2), IR (KBr, cm-1) v:
3234 (NH), 2921 (C-H), 1662, 1616 (C=O), 1289 (C=C), 1034, 790. ESI-MS MS 2m/z: 764.81.
2.1.4.6 (Z)-N-(1-chloro-3-oxo-3-(piperidin-1-yl)-1-p-tolylprop-1-en-2-yl)benzamide (5f). White
1
crystals. Yield 16%; H NMR (300MHz, CDCl3): 8.11 (s, 1H, NH), 7.92 (d, J=7.5Hz, 2H, ArH),
7.60 (t, J=6.9 Hz, 1H, ArH), 7.52 (dd, J=7.5Hz, J=8.1 Hz, 4H, ArH), 7.19 (d, J=8.1Hz, 2H, ArH),
3.55 (m, 3H, CH2), 3.13 (m, 1H, CH2), 2.39 (s, 3H, CH3), 1.58 (m, 4H, CH2), 1.18 (m, 1H, CH2),
0.62 (m, 1H, CH2), IR (KBr, cm-1) v: 3282 (NH), 2960 (C-H), 1662, 1635 (C=O), 1268 (C=C),
1025, 810. ESI-MS 2m/z: 764.73.
2.1.4.7 (Z)-N-(1-chloro-1-(2-methoxyphenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5g). White crystals. Yield 11%; 1H NMR (300MHz, CDCl3): 8.21 (s, 1H, NH), 7.92 (d, J=6.9Hz,
2H, ArH), 7.60 (t, J=7.2Hz, 1H, ArH), 7.52 (t, J=7.8Hz, 2H, ArH), 7.42 (t, J=7.5Hz, 2H, ArH),
6.97 (t, J＝6.9Hz, 2H, ArH), 3.89 (s, 3H, CH3), 3.33 (m, 4H, CH2), 1.53 (m, 4H, CH2), 0.97 m, 1H,
CH2), 0.78 (m, 1H, CH2), IR (KBr, cm-1) v: 3446 (NH), 2360 (-CH3), 1652, 1558 (C=O),
1268(C=C), 1093, 790. ESI-MS 2m/z: 796.72.
2.1.4.8 (Z)-N-(1-chloro-1-(4-methoxyphenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5h). White crystals. Yield 13%; 1H NMR (300MHz, CDCl3): 8.08 (s, 1H, NH), 7.92 (d, J=7.2Hz,
2H, ArH), 7.61 (m, 5H, ArH), 6.91 (d, J=8.7Hz, 2H, ArH), 3.85 (s, 3H, CH3), 3.61 (m, 1H, CH2),
3.40 (m, 2H, CH2), 3.16 (m, 1H, CH2), 1.61 (m, 4H, CH2), 1.20 (m, 1H, CH2), 0.68 (m, 1H, CH2),
IR (KBr, cm-1) v: 3446 (NH), 2360 (-CH3), 1650, 1553 (C=O), 1268 (C=C), 1089, 790. ESI-MS
2m/z: 796.79.
2.1.4.9 (Z)-N-(1-(2-bromophenyl)-1-chloro-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5i). White crystals. Yield 9%; 1H NMR (300MHz, CDCl3): 8.25 (s, 1H, NH), 7.93 (d, J=7.2Hz,
2H, ArH), 7.67 (m, 5H, ArH), 7.36 (m, 2H, ArH), 3.55 (m, 2H, CH2), 3.36 (m, 1H, CH2), 3.21 (m,
1H, CH2), 1.80 (s, 1H, CH2), 1.59 (m, 4H, CH2), 0.97 (m, 1H, CH2), IR (KBr, cm-1) v: 3446 (NH),
2360 (-CH3), 1652, 1558 (C=O), 1268 (C=C), 1091, 799. 13C NMR (75Hz, CDCl3): δ 164.13,
162.14, 135.10, 133.35, 132.65, 132.62, 132.54, 131.09, 131.03, 128.86, 127.50, 127.47, 124.12,
115.84, 47.97, 42.59, 25.35, 24.86, 24.20. HRMS (EI) calcd. for 446.0397; C21H20BrClN2O2,
found (E+) 449.0481.
2.1.4.10 (Z)-N-(1-(3-bromophenyl)-1-chloro-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5j). White crystals. Yield 15%; 1H NMR (300MHz, CDCl3): 8.19 (s, 1H, NH), 7.91 (d, J=8.1Hz,
2H, ArH), 7.71 (s, 1H, ArH), 7.61 (t, J=7.8Hz, 1H, ArH), 7.52 (dd, J=6.9Hz, J=1.2 Hz, 4H, ArH),
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7.24 (d, J=7.8Hz, 1H, ArH), 3.63 (m, 1H, CH2), 3.33 (m, 3H, CH2), 1.61 (m, 4H, CH2), 1.18 (m,
1H, CH2), 0.60 (m, 1H, CH2), IR (KBr, cm-1) v: 3243 (NH), 2921 (C-H), 1670, 1618 (C=O), 1268
(C=C), 916, 682. 13C NMR (75Hz, CDCl3): δ 164.16, 162.21, 134.78, 133.23, 132.66, 132.59,
132.47, 131.21, 131.01, 128.83, 127.74, 127.43, 124.02, 115.87, 47.86, 42.53, 25.38, 24.82, 24.24.
HRMS (EI) calcd. for 446.0397; C21H20BrClN2O2, found (E+) 449.0476.
2.1.4.11 (Z)-N-(1-(4-bromophenyl)-1-chloro-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5k). White crystals. Yield 6%; 1H NMR (300MHz, CDCl3): 8.13 (s, 1H, NH), 7.91 (d, J=7.5Hz,
2H, ArH), 7.62 (t, J=7.2Hz, 1H, ArH), 7.53 (m, 6H, ArH), 3.51 (m, 3H, CH2), 3.12 (m, 1H, CH2),
1.61 (m, 4H, CH2), 1.20 (m, 1H, CH2), 0.71 (m, 1H, CH2), IR (KBr, cm-1) v: 3322 (NH), 2903
(C-H), 1691, 1618 (C=O), 1268 (C=C), 1016 (C-H), 593 (Ar-H). 13C NMR (75Hz, CDCl3): δ
164.21, 162.28, 133.96, 132,62, 132.61, 131.48, 130.32, 129.08, 128.87, 127.47, 123.55, 116.87,
47.79, 42.51, 24.84, 24.52, 24.15. HRMS (EI) calcd. for 446.0397; C21H20BrClN2O2, found (E+)
449.0471 .
2.1.4.12
(Z)-N-(1-chloro-1-(2-fluorophenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5l). White crystals. Yield 21%; 1H NMR (300MHz, CDCl3): 8.22 (s, 1H, NH), 7.93 (d, J=7.8Hz,
2H, ArH), 7.62 (m, 5H, ArH), 7.18 (dd, J=6.6Hz, J=8.7Hz, 2H, ArH), 3.46 (m, 4H, CH2), 1.57 (m,
4H, CH2), 1.01 (m, 2H, CH2), IR (KBr, cm-1) v: 3234 (NH), 2853 (C-H), 1652, 1618 (C=O), 1268
(C=C), 1025, 761. ESI-MS 2m/z: 772.95.
2.1.4.13
(Z)-N-(1-chloro-1-(4-fluorophenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5m). White crystals. Yield 4%; 1H NMR (300MHz, CDCl3): 8.11 (s, 1H, NH), 7.91 (d, J=7.5Hz,
2H, ArH), 7.62 (s, 5H, ArH), 7.11 (t, J=8.7Hz, 2H, ArH), 3.58 (m, 1H, CH2), 3.39 (m, 2H, CH2),
3.16 (m, 1H, CH2), 1.62 (m, 4H, CH2), 1.17 (m, 1H, CH2), 0.65 (m, 1H, CH2), IR (KBr, cm-1) v:
3273 (NH), 2862 (C-H), 1691, 1618 (C=O), 1268 (C=C), 1025, 746. ESI-MS MS 2m/z: 772.95.
2.1.4.14 (Z)-N-(1-chloro-1-(2-chlorophenyl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide
(5n). White crystals. Yield 24%; 1H NMR (300MHz, CDCl3): 8.25 (s, 1H, NH), 7.93 (d, J=7.2Hz,
2H, ArH), 7.62 (t, J=7.2Hz, 1H, ArH), 7.53 (dd, J=7.8Hz, J=9Hz, 4H, ArH), 7.39 (m, 2H, CH2),
3.50 (m, 4H, CH2), 1.58 (m, 4H, CH2), 0.97 (m, 2H, CH2), IR (KBr, cm-1) v: 3270 (NH), 2931
(C-H), 1662, 1618 (C=O), 1268 (C=C), 1025, 742. 13C NMR (75Hz, CDCl3): δ 164.22, 162.15,
135.46, 134.47, 132.67, 132.58, 132.43, 131.27, 131.12, 128.98, 127.53, 127.45, 124.10, 115.89,
47.86, 42.52, 25.44, 24.85, 24.18. HRMS (EI) calcd. for 402.0902; C21H20Cl2N2O2, found (E+)
403.0981.
2.1.4.15 (Z)-N-(1-chloro-3-oxo-3-(piperidin-1-yl)-1-(yridine-3-yl)prop-1-en-2-yl)benzamide (5o)
White crystals. Yield 6%; 1H NMR (300MHz, CDCl3): 8.80 (s, 1H, NH), 8.60 (s, 1H, ArH), 8.21
(s, 1H, ArH), 7.91 (d, J=7.2Hz, 3H, ArH), 7.63 (t, J=10.2Hz, 1H, ArH), 7.53 (t, J=7.5Hz , 2H,
ArH), 7.35 (t, J=5.7Hz, 1H, ArH), 3.54 (m, 2H, CH2), 3.19 (m, 2H, CH2), 1.71 (m, 5H, CH2), 0.66
(m, 1H, CH2), IR (KBr, cm-1) v: 3648 (NH), 2931 (C-H), 1662, 1618 (C=O), 1268 (C=C), 1025,
709. 13C NMR (75Hz, CDCl3): δ 164.23, 162.24, 135.43, 132.69, 132.55, 132.40, 131.22, 131.01,
128.97, 127.52, 127.43, 124.32, 115.82, 47.83, 42.54, 25.31, 24.76, 24.25. HRMS (EI) calcd. for
369.1244; C21H20Cl2N2O2, found (E+) 370.1323.
2.1.4.16 (Z)-N-(1-chloro-3-oxo-3-(piperidin-1-yl)-1-(quinolin-3-yl)prop-1-en-2-yl)benzamide(5p)
Pale green crystals. Yield 22%; 1H NMR (300MHz, CDCl3): 9.13 (s, 1H, NH), 8.34 (s, 1H, ArH),
8.23 (s, 1H, ArH), 8.25 (d, J=6Hz, 1H, ArH), 7.97 (d, J=6Hz, 2H, ArH),7.84 (d, J=6Hz, 1H, ArH),
7.54 (t, J=6Hz, 1H, ArH), 7.35 (m, 2H, ArH), 7.21 (t, J=6Hz, 2H, ArH), 3.79 (m, 2H, CH2), 3.40
(m, 2H, CH2), 1.42 (m, 4H, CH2), 0.11 (m, 2H, CH2). IR (KBr, cm-1) v: 3236 (NH), 2939 (C-H),
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1670, 1628 (C=O), 1270 (C=C), 1030, 747. 13C NMR (75Hz, CDCl3): δ 164.15, 161.83, 150.32,
136.23, 134.78, 132.86, 132.73, 130.72, 130.63, 129.45, 129.12, 128.93, 128.18, 127.55, 127.34,
126.86, 100.98, 47.65, 42.42, 24.83, 24.52, 23.93. HRMS (EI) calcd. for 419.1401; C24H22ClN3O2,
found (E+) 420.1480.
2.1.4.17 (Z)-N-(1-chloro-1-(isoquinolin-4-yl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl) benzamide
(5q) Pale green crystals. Yield 21%; 1H NMR (300MHz, CDCl3): 9.05 (s, 1H, NH), 8.33 (s, 1H,
ArH), 8.27 (s, 1H, ArH), 8.25 (d, J=6Hz, 1H, ArH), 7.93 (d, J=6Hz, 2H, ArH),7.89 (d, J=6Hz, 1H,
ArH), 7.80 (t, J=6Hz, 1H, ArH), 7.62 (m, 2H, ArH), 7.51 (t, J=6Hz, 2H, ArH), 3.88 (m, 2H, CH2),
3.33 (m, 2H, CH2), 1.32 (m, 4H, CH2), 0.29 (m, 2H, CH2). IR (KBr, cm-1) v: 3250 (NH), 2946
(C-H), 1671, 1628 (C=O), 1266 (C=C), 1038 (C-H), 742 (Ar-H). 13C NMR (75Hz, CDCl3): δ
164.37, 161.77, 151.56, 136.34, 132.88, 132.63, 130.31, 129.29, 129.12, 129.03, 128.93, 128.42,
127.62, 127.57, 126.89, 125.94, 99.97, 47.83, 42.35, 24.84, 24.47, 23.89. HRMS (EI) calcd. for
419.1401; C24H22ClN3O2, found (E+) 420.1480.
2.1.4.17 (Z)-N-(1-chloro-1-(furan-2-yl)-3-oxo-3-(piperidin-1-yl)prop-1-en-2-yl)benzamide (5r)
White crystals. Yield 4%; 1H NMR (300MHz, CDCl3): 9.19 (s, 1H, NH), 7.96 (d, J=10.2Hz, 2H,
ArH), 7.59 (m, 4H, ArH), 6.34 (s, 1H, ArH), 5.75 (s, 1H, ArH), 3.71 (d, J=2.4Hz, 4H, CH2), 1.71
(s, 6H, CH2), IR (KBr, cm-1) v: 3322 (NH), 2931 (C-H), 1662, 1618 (C=O), 1268 (C=C), 1025
(C-H), 724 (Ar-H). 13C NMR (75Hz, CDCl3): δ 164.25, 162.10, 135.33, 133.24, 132.89, 132.43,
132.23, 131.11, 128.83, 127.57, 124.02, 116.86, 47.89, 42.53, 25.32, 24.85, 24.19. HRMS (EI)
calcd. for 358.1084; C19H19ClN2O2, found (E+) 359.1163.
2.2 Crystal data for compound 4j
The crystal data of the rearranged product were shown as follows. Crystal Data for
C21H20Br2N2O2 (M =492.21): monoclinic, space group P21/c (no. 14), a = 9.9091(7) Å, b =
11.9717(5) Å, c = 17.1808(10) Å, β = 106.799(7)°, V = 1951.2(2) Å3, Z = 4, T = 293.15 K,
ꢀ(MoKα) = 4.174 mm-1, Dcalc = 1.676 g/mm3, 11717 reflections measured (6.01 ≤ 2Θ ≤ 52.744),
3966 unique (Rint = 0.0342, Rsigma = 0.0468) which were used in all calculations. The final R1 was
0.0439 (I > 2σ(I)) and wR2 was 0.0967. The full crystallographic details of the rearranged product
have been deposited at the Cambridge Crystallographic Data Center were allocated the deposition
number CCDC 992084 .The crystal structure was shown in Fig 3.
2.3. Biological assay
Anti-HBV activity was evaluated in HepAD38 cellular assay according to literature [21]. It
has been found that this series has been found to be generally nontoxic in the HepAD38 cell line
even at the highest concentrations tested [4]. The compound 5a was studied in an additional cell line
(HepG2) and found to be similarly nontoxic [22]. This analogue also showed antiviral activity that is
specific for HBV. Theresa May Chin Tan commented that this series was able to inhibit the expression
of the viral antigens, HBsAg and HBeAg in a concentration-dependent manner with no cytotoxic
effects and without any effects on the expression of viral transcripts [23]. So the EC50 of compounds
were tested and used in QSAR study only. The DNA replication of HBV in the cell was detected
by dot blot hybridization and the results were listed in Table 1. The EC50 is the concentration of
the compound (in ꢀM) which inhibits the synthesis of viral DNA at 50%.
2.4 Computational calculations for 2D-QSAR
All computations were carried out on the Gaussian 09 [24] computer software package. The
electronic descriptors were obtained from a single-point calculation at the B3LYP/6-311+g (d)
level.
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2.4.1 The optimized molecular geometry of compound
The optimized molecular geometry of compound was computed first and the optimized
geometrical parameters, such as bond length and total charge of aromatic ring (∑Q of Ar),
benzene ring (∑Q of Ph) and a piperidine ring (∑Q of Cy) were given in Table 2.
2.4.2 The parameters for constructing the 2D-QSAR models
Structural parameters include: energy of highest occupied molecular orbital (EHOMO), energy
of lowest unoccupied molecular orbital (ELUMO), energy difference between LUMO and HOMO
(Egap)，the most positive H atom charge (qH+), molecular averaged polarizability (α) and heat of
formation (ꢁhf) for the structure at 298.15 K and 1 atm (HF). Thermodynamic parameters include:
total energy (TE), zero point energy (ZPE), enthalpy (Hө), free energy (Gө), correction value of
thermal energy (Eth), molar heat capacity at constant volume (CVө) and entropy (Sө). Some of the
quantum chemical parameters were listed in Table 3 and Table 4.
2.5 Molecular modeling and alignment of 3D-QSAR
The 3D-QSAR studies of the phenylpropenamide compounds using the CoMFA performed on
the Sybyl 8.0 package running on the Linux operating system. Partial atomic charges of all
compounds were calculated by the Gasteigere-Marsili method, and then were optimized for their
geometry using Tripos field [25] with a distance-dependent dielectric function and energy
convergence criterion of 0.21 kcal/mol Å using the maximum iterations set to 1000 [26].
2.5.1 CoMFA analysis
The improvement of QSAR may originate from the choice of DFT-optimized template and the
usage of RMSD-based molecular alignment strategy. To derive the CoMFA descriptor fields, the
aligned training set molecules were placed in a 3D cubic lattice with grid spacing of 2Å in x, y,
and z directions such that the entire set could be included in it. The CoMFA steric and electrostatic
field energies were calculated using a sp3 carbon probe atom with a van derWaals radius of 1.52
Å and a charge of +1.0. Cut-off values for both steric and electrostatic fields were set to 30.0
kcal/mol.
2.5.2 Regression analysis by PLS method
The partial least squares (PLS) methodology analysis with the leave-one-out (LOO)
cross-validation procedure was carried out to determine the optimal number of components using
the SAMPLS [27]. The cross-validated coefficient q2, as an internal statistical index of predictive
power, was subsequently obtained. The quality of the external prediction was documented using
the standard deviation of error prediction (R2).
3. Results and Discussion
3.1 SAR investigation
SAR investigation of different substitution benzene of phenylpropenamide was done firstly.
Compound 4a showed good activities with EC50 of 1.3ꢀM. When the phenylpropenamides have
same substituent but different substitution position, the ortho-substitution on the aromatic ring
provided better anti-HBV activities than Para-substitution, such as compounds o-Me 4e and p-Me
4f with EC50 of 0.9 and 2.8 ꢀM, the o-OMe 4g and p-OMe 4h with EC50 of 2.1 and 8.0 ꢀM
respectively. The same anti-HBV activities results exhibited in the vinyl chlorides series 5a-5r .
Michael J. Sofia [4] reported that the ortho-substitution on the aromatic can improve anti-HBV
activities while Para-substitution decreased the antiviral activities. In general, vinyl bromides
were slightly more active than corresponding vinyl chlorides.
6

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The compound 4o and 5o, where phenyl was replaced by pyridine, have good biological
activities with EC50 of 1.0 and 0.9 ꢀM respectively. The compound 4r and 5r containing furyl
subtitude also showed good results. But the compound 4p, 4q and 5p, 5q, which phenyl group
was replaced by quinoline and isoquinoline, have poor biological activity.
3.2 2D-QSAR
The QSAR is fast emerging as an useful tool in modern chemistry, biology, and drug
discovery. A 2D-QSAR model is a mathematical equation that correlates the biological, chemical,
or physical activity of a molecular system to its geometric and chemical characteristics. The
quantum chemical descriptors computed by density functional theory (DFT) have found
increasing use in modern QSAR analysis, so the DFT and multiple linear regression analysis
method were used to construct the 2D-QSAR models of the phenylpropenamides based on the
experimental data of –lg(1/EC50) and the parameters of computational calculations.
To obtain the optimum parameter set for establishing the QSAR model and to remove
insignificant descriptors, a preliminary screening of the 21 candidate descriptors were conducted
before multiple linear regressions. Table 5 contained the regression equations, and the correlations
between every individual descriptor and -Log(1/EC50). All the equations containing a single
independent variable were evaluated by regression coefficient R and homogeneity of variances P.
The results in Table 5 showed that TE had the highest value of regression coefficient (R 0.871)
with -Log(1/EC50), which identified that TE was the most relevant parameter to the bioactivity
indices.
The frontier orbital theory states that the energy of the HOMO and LUMO are the important
factors that determine the reactivity of a molecule [28, 29], the EHOMO, ELUMO and
△Egap was
calculated and analyzed firstly. But the single variable equations using EHOME, ELUMO and
△Egap as
parameter appeared nonlinear relationship with the R < 0.5 and P > 0.1. The result show that there
is no any association between the anti-HBV activities of phenylpropenamides and their energy of
the HOMO and LUMO.
It was found that the EC50 of phenylpropenamide compounds had a certain relation with
theoretical data of the thermodynamic parameters, the anti-HBV activities showed a downward
trend with the increasing of energy values. The results in Table 5 showed that total energy (TE),
zero point energy (ZPE), enthalpy (Hө) and free energy (Gө) have the highest value of regression
coefficient with -log(1/EC50), which identified that the these parameters are the most relevant
parameter to the bioactivity. The single variable equations using entropy (Sө) as parameter has
0.53175 and 0.61512 R and 0.02313 and 0.00659 P for 4 and 5 series respectively, implying an
association between the anti-HBV activities of phenylpropenamide and their Sө. The current study
presents a comprehensive QSAR analysis for phenylpropenamide as a inhibitors of hepatitis B
virus replication drug used energy parameters of total energy (TE) and entropy (Sө). A multiple
regression analysis was carried out and arrived at the final QSAR equation, which can be written
as Scheme 2:
where n is the number of data points, R2 is square of the correlation coefficient and represents
the goodness of fitting, F is the overall F-statistics for the addition of each successive term, and P
is the P values using the F statistics. Because total energy (TE) was more correlated with
-log(1/EC50) than the other descriptors, it is the most important descriptor the regression equations.
Besides the total energy (TE) descriptor, entropy (Sө) is another descriptor that cannot be
neglected for constructing the QSAR models.
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In series 4, phenylpropenamides of vinyl bromides compounds, the QSAR equations
possessed relative high correlation coefficient with R2=0.80692, better 31.34498 F-statistics, and
least number of variables. Some values of EC50 of chloro substituted propenamides compounds
were greater than 100 ꢀM, which has been been used to calculate -log (1/EC50) and participated
the linear regression, leading to the R2 value of series 5 (0.68535) was lower than that of series 4
(0.80692), but it still met the statistical requirements.
The model analysis suggested that the anti-HBV activities of this kind of compounds were
mainly affected by molecular total energy (TE) and entropy (Sө). The high values of energy was
likely to indicate a tendency of the molecule to donate electrons to appropriate acceptors and
lower value of energy implies high stability for the molecule in the sense of its lower sensitivity in
the biochemical processes [18, 30]. The –log(1/EC50) was directly proportional to the entropy (Sө )
due to the degree of disorder caused by Sө expression and the larger the degree of disorder the
lower activities [31].
The correlations between experimental and calculated activities values presented in Fig. 4
indicated that the selected parameters can predict the anti-HBV activities of the set
phenylpropenamide molecules with greater predictability. Thus, the new phenylpropenamide
molecules with high total energy (TE) and low entropy (Sө) may increase the anti-HBV activities.
3.3 3D-QSAR
In the study, the data-based fitting procedure was finally adopted through careful comparison
and each analog was superimposed to the template based on the common substructure of
propenamide moiety. The aligned molecules were illustrated in Fig. 5 and the statistical
parameters were listed in Table 6.
The better predictions are obtained by the 3D-QSAR/CoMFA for anti-HBV activities for
phenylpropenamide molecules from Table 6.. The model has a high R2 (0.980 and 0.986 for series
4 and 5 respectively see in Fig 6) with a low standard deviation(SE, 0.097 and 0.117, respectively)
and a high Fischer ratio (F, 116.544 and 164.602, respectively); while a QSAR model is generally
acceptable if R2 is approximately 0.9 or higher [33]. Specially, the cross-validation related
2
coefficient q is 0.696 and 0.564 respectively ( >0.5 ) [34], suggesting a good prediction ability of
this model [34].
The plots of the predicted versus actual –log(1/EC50) for the QSAR/CoMFA models were
shown in Fig. 6. It could be noted that the data points were uniformly distributed along the
regression line. Additionally, all the prediction errors in Table 6 were smaller than 0.5, suggesting
the satisfactorily predictive capability, high reliability and accuracy of the models.
To view the field effect on the target property, CoMFA contour maps were generated and
shown in Fig. 7. For steric fields, the green was bulky group favorable and yellow was bulky
group unfavorable. Similarly, the blue was electropositive charge favorable and red was
electronegative charge unfavorable.
For the compounds 4, the steric field of vinyl bromides of phenylpropenamide molecules
with green contours (64.04%) referred to sterically favored regions for anti-HBV activities and
yellow contours (35.96%) highlighted the sterically unfavorable regions. The electrostatic field
with blue contours (65.32%) represented electropositively preferrable regions to activities and red
contours (34.68%) indicated regions where more electronegative substituents are favored. For the
compounds 5, the steric field of vinyl chlorides of phenylpropenamide molecules had green
contours (39.22%) refer to sterically favored regions for anti-HBV activities and yellow contours
8

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(60.78%) highlight the sterically unfavorable regions. The electrostatic field had blue contours
(29.83%) represent electropositively preferred regions to activities and red contours (70.17%)
indicate regions where more electronegative substituents were favored.
For the vinyl bromides series 4a-4r, the steric field is the dominating factor for the anti-HBV
activities and the electrostatic field gave the least contribution. On the contrary, the electrostatic
field is the dominating factor for the vinyl chlorides series 5a-5r for the anti-HBV activities and
the steric field effect is the secondary important.
3.4 Comparison between the 2D-QSAR and 3D-QSAR models
The comparison was conducted from two viewpoints of anti-HBV activities mechanism and
prediction ability. 2D-QSAR model suggests that both the molecular total energy (TE) and entropy
(Sө) can affect the activities. Judging from 3D-QSAR model, the anti-HBV activities are mainly
affected by the steric and electrostatic properties of substituents. Thus, complementary results can
be obtained with 2D-QSAR and CoMFA models, which can provide theoretical guide to the
application of QSAR in the environmental chemical field.
4. Conclusions
In summary, a series of phenylpropenamide derivatives containing different aromatic ring
substituents were synthesized, characterized and assessed for their anti-HBV activities. The
quantum chemical parameters of phenylpropenamide derivatives were calculated at the B3LYP/6-
311G** level, based on which the 2D-QSAR model of –log(1/EC50) was proposed. The QSAR
equations developed with the two parameters total energy (TE) and entropy (Sө) provided
regression models to predict the activity of the set of phenylpropenamide molecules against DNA
replication of HBV. The QSAR equation have better stability ability from the values of R2
(0.80692 and 0.68535), F-value (31.34498 and 16.33582) and P (P<0.00010 and P<0.00017) for
vinyl bromides and vinyl chlorides respectively. In the mean time, the 3D-QSAR model was
proposed by using CoMFA based on the molecular simulation, which also exhibited good stability
and prediction ability. The optimum models were all statistically significant with cross-validated
coefficients (q2=0.696 and 0.564) >0.5 and conventional coefficients (R2=0.980 and 0.986) >0.9,
indicating they were reliable enough for activity prediction and providing some insights into the
critical structural factors which can affect the bioactivity of phenylpropenamide derivatives. The
3D equipotential map illustrated the effect of different substituent on their anti-HBV activities.
The 2D or 3D QSAR models can be utilized to predict the anti-HBV activities of the
phenylpropenamide molecules.
Acknowledgements
This work was supported by the NSFC (NO. B020601), Key Laboratory Fund of Sichuan
Province( SZJJ2014-081), and the Ministry of Education Innovation Project (No. XZD0912-09)
The authors thank the assistant of Dr. Yang of West china school of pharmacy, Sichuan University
for the work of biological assay and thank the students of Prof. Li of College of Chemical
engineering, Sichuan University for the work of computational calculations of SYBYL.
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Table 1 The anti-HBV activities of compound 4a-4r and 5a-5r
Comp. EC50
-log(1/EC50)
0.113943352
0.041392685
0.778151250
1.000000000
-0.050609993
0.447158031
0.322219295
0.903089987
1.397940009
1.544068044
1.812913357
0.176091259
0.255272505
0.903089987
0.000000000
0.845098040
0.698970004
-0.045757491
Comp.
5a
5b
5c
EC50
1.2
-log(1/EC50)
0.079181246
0.230448921
1.531478917
1.707570176
-0.004364805
0.041392685
1.322219295
1.397940009
2.000000000
2.000000000
2.000000000
0.255272505
0.342422681
1.591064607
-0.026872146
1.477121255
1.462397998
-0.096910013
1.3
1.1
4a
4b
4c
4d
4e
4f
1.7
6.0
34.0
51.0
1.0
10.0
0.9
5d
5e
2.8
1.1
5f
2.1
21.0
25.0
>100.0 a
>100.0 a
>100.0 a
1.8
4g
4h
4i
5g
5h
5i
8.0
25.0
35.0
65.0
1.5
4j
5j
4k
4l
5k
5l
1.8
2.2
4m
4n
4o
4p
4q
4r
5m
5n
5o
5p
5q
4r
8.0
39.0
0.9
1.0
30.0
29.0
0.8
7.0
5.0
0.9
Table 2 The optimized geometrical parameters of phenylpropenamide derivatives 4a-4r、5a-5r
Bond length Ǻ
∑Q
Comp
C-X
C=C
C-NH
NH-C=O
N-C=O
Ar
Ph
Cy
(X=Br、Cl)
1.95292
1.94588
1.34535
1.34332
1.39645
1.39247
1.21977
1.21853
1.22336
1.21883
-0.896151 -0.919316
0.348353 -0.844939
-1.775496
-1.688875
4a
4b
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1.34201
1.34596
1.34205
1.34530
1.34146
1.34526
1.34221
1.34597
1.34613
1.34218
1.34546
1.34219
1.34633
1.34744
1.34315
1.34807
1.34664
1.34312
1.34186
1.34712
1.34225
1.34655
1.34181
1.34644
1.34255
1.34706
1.34720
1.34228
1.34664
1.34223
1.34757
1.34840
1.34315
1.34883
1.93826
1.95194
1.96086
1.95478
1.95776
1.95799
1.95142
1.95021
1.95156
1.95228
1.95320
1.95143
1.95198
1.95151
1.95644
1.95201
1.78686
1.78174
1.77456
1.78663
1.79327
1.78866
1.78996
1.79108
1.78535
1.78488
1.78642
1.78649
1.78749
1.78548
1.78640
1.78615
1.78960
1.78393
1.39326
1.39485
1.39710
1.39694
1.39704
1.39733
1.39555
1.21925
1.21934
1.21963
1.21988
1.22015
1.22007
1.21947
1.22050
1.22326
1.22297
1.22348
1.22103
1.22358
1.21938
1.22293
1.22331
1.21945
1.22329
-1.251704 -0.837988
-1.124014 -0.969986
-0.461240 -0.825417
-0.348184 -0.980299
-1.505460 -0.837188
-0.627813 -0.959433
-0.144929 -0.846823
-0.561815 -0.908448
-0.555132 -0.959009
-1.316687 -0.841457
-1.686215
-1.784969
-1.749826
-1.767813
-1.711406
-1.760267
-1.803706
-1.742217
-1.777852
-1.827855
-1.787560
-1.782235
-1.842558
-1.838197
-1.769277
-1.822423
-1.768942
-1.748096
-1.693947
-1.773090
-1.752179
-1.760951
-1.673087
-1.755063
-1.793008
-1.740841
-1.765260
-1.822561
-1.780463
-1.788900
-1.837309
-1.834680
-1.788628
-1.835985
4c
4d
4e
4f
4g
4h
4i
1.39460 1.21921
4j
1.39462
1.39462
1.39552
1.39515
1.39455
1.39290
1.39495
1.39120
1.39763
1.39419
1.39482
1.39588
1.39846
1.39797
1.39876
1.39843
1.39659
1.39575
1.39559
1.39602
1.39661
1.39679
1.39565
1.39402
1.39654
1.39288
1.21931
1.21930
1.21952
4k
4l
-0.562458
-0.911908
4m
4n
4o
4p
4q
4r
5a
5b
5c
5d
5e
5f
1.21928 1.21927
-1.062723 -0.827646
-0.876423 -0.925590
-1.707655 -0.896693
-1.030686 -0.833105
-0.750784 -0.878430
-1.139492 -0.986517
-0.014724 -0.932297
-1.922403 -0.935734
-1.340251 -1.016954
-1.096918 -0.916345
-0.342715 -1.008493
-1.746778 -0.908333
-0.978393 -0.992218
-1.109041 -0.913557
-0.906464 -0.994730
-0.765886 -1.013198
-1.618792 -0.928978
-0.799893 -0.988045
-1.759844 -0.938263
-1.002424 -0.931401
-1.797908 -0.849097
1.21929
1.21923
1.21912
1.21876
1.21986
1.21883
1.21960
1.21951
1.21980
1.22006
1.22057
1.22026
1.21961
1.21934
1.21944
1.21947
1.21969
1.21968
1.21936
1.21933
1.21943
1.21912
1.22237
1.22302
1.22220
1.22306
1.22395
1.21923
1.22093
1.22380
1.22337
1.22386
1.22166
1.22398
1.21978
1.22370
1.22379
1.21980
1.22381
1.21993
1.22293
1.22350
1.22288
1.22348
5g
5h
5i
5j
5k
5l
5m
5n
5o
5p
5q
5r
-1.484111
-0.926502
-0.807766 -0.902821
Table 3 The calculation energy value of phenylpropenamide derivatives 4a-4r、5a-5r
EHOMO
ELUMO
.
Egap
HF
Comp.
qH+
α
Hartree
Hartree
Hartree
Hartree
-0.062979 -0.226032 0.163053 0.345575 2.5265 -3647.2554986
-0.069673 -0.201043 0.131370 0.352138 6.3080 -3739.5159911
-0.073280 -0.169556 0.096276 0.359610 6.3704 -3851.8084411
-0.068033 -0.230700 0.162667 0.344048 3.4803 -4106.8800729
-0.062275 -0.220858 0.158583 0.345015 2.4387 -3686.5788853
-0.060994 -0.222445 0.161451 0.346774 2.6379 -3686.581249
4a
4b
4c
4d
4e
4f
12

ACCEPTED MANUSCRIPT
-0.059536 -0.217935 0.158399 0.351103 1.3644 -3761.805973
-0.063359 -0.225973 0.162614 0.347432 3.9468 -3761.8106142
-0.065055 -0.167983 0.102928 0.345404 3.6920 -6220.7919548
-0.068193 -0.231952 0.163759 0.347991 4.1895 -6220.7988359
-0.071433 -0.231880 0.160447 0.343150 3.4589 -6220.7997101
-0.063786 -0.235357 0.171571 0.350402 3.7022 -3746.5204362
-0.068227 -0.231662 0.163435 0.345640 3.3396 -3746.5259104
-0.064763 -0.199450 0.134687 0.341835 3.7668 -4106.8729964
-0.069899 -0.233532 0.163633 0.350553 4.5745 -3663.2968606
-0.152916 -0.217488 0.064572 0.346016 4.3084 -3816.9741999
-0.077541 -0.218415 0.140874 0.346093 1.2262 -3816.9693588
-0.110853 -0.225450 0.114597 0.338813 2.1350 -3645.0398728
-0.062408 -0.226124 0.163716 0.343903 2.5699 -1533.3355143
-0.068916 -0.200746 0.131830 0.345178 6.2437 -1625.5956197
-0.072831 -0.169381 0.096550 0.356569 6.2551 -1737.8883378
-0.067300 -0.230864 0.163564 0.342664 3.4917 -1992.9601462
-0.061833 -0.220747 0.158914 0.343341 2.4741 -1572.6584057
-0.060514 -0.222493 0.161979 0.343891 2.7166 -1572.6612966
-0.059020 -0.218261 0.159241 0.349448 1.3783 -1647.8855821
-0.062746 -0.225889 0.163143 0.347069 4.0219 -1647.8905643
-0.064564 -0.167614 0.103050 0.344315 3.6592 -4106.8716217
-0.067453 -0.232149 0.164696 0.345872 4.1420 -4106.8789012
-0.067613 -0.230629 0.163016 0.342690 3.4627 -4106.8797531
-0.063872 -0.226796 0.162924 0.351051 3.6575 -1632.5999366
-0.065269 -0.229764 0.164495 0.343537 3.3790 -1632.6059672
-0.064172 -0.199148 0.134976 0.349224 3.6212 -1992.9526691
-0.068952 -0.233662 0.164710 0.340669 4.5732 -1549.3768395
-0.153193 -0.217783 0.064590 0.353758 4.2806 -1703.0541392
-0.077215 -0.218625 0.141410 0.339723 1.1298 -1703.0489175
-0.109331 -0.225701 0.116370 0.341666 2.1974 -1531.119129
4g
4h
4i
4j
4k
4l
4m
4n
4o
4p
4q
4r
5a
5b
5c
5d
5e
5f
5g
5h
5i
5j
5k
5l
5m
5n
5o
5p
5q
5r
Table 4 The calculation thermodynamic parameters of phenylpropenamide derivatives 4a-4r and
5a-5r
TE
ZPE
Hө
Gө
Eth
KCal/Mol Cal/Mol-Kel. Cal/Mol-Kel.
CVө
Sө
Comp.
Hartree
Hartree
kJ/mol
kJ/mol
-3646.851578 -3646.875102 -9574806.34 -9575019.262
-3739.111883 -3739.137333 -9817035.77 -9817261.198
-3851.399743 -3851.425863 -10111847.55 -10112076.16
-4106.484573 -4106.509345 -10781572.77 -10781794.36
-3686.145737 -3686.170868 -9677973.154 -9678193.559
-3686.148181 -3686.173644 -9677979.571 -9678206.112
-3761.367147 -3761.393412 -9875466.963 -9875696.569
-3761.371621 -3761.397774 -9875478.712 -9875707.267
-6220.397092 -6220.422234 -16331650.08 -16331875.56
253.464
253.582
256.462
248.180
271.805
271.754
275.368
275.472
247.780
89.559
95.662
98.177
93.421
95.485
95.648
98.839
98.781
93.975
170.684
180.710
183.267
177.634
176.684
181.604
184.058
183.217
180.748
4a
4b
4c
4d
4e
4f
4g
4h
4i
13

ACCEPTED MANUSCRIPT
-6220.403719 -6220.428801 -16331667.49 -16331893.48
-6220.404528 -6220.429537 -16331669.61 -16331893.87
-3746.123963 -3746.148383 -9835445.984 -9835664.924
-3746.129469 -3746.153873 -9835460.442 -9835679.257
-4106.477746 -4106.502637 -10781554.84 -10781778.01
-3662.904804 -3662.928227 -9616954.084 -9617166.957
-3816.533050 -3816.558987 -10020305.04 -10020531.34
-3816.528049 -3816.554110 -10020291.91 -10020519.25
-3644.667102 -3644.689886 -9569070.998 -9569281.229
-1532.931196 -1532.954411 -4024708.377 -4024917.723
-1625.191168 -1625.216357 -4266936.933 -4267159.901
-1737.479300 -1737.505198 -4561749.421 -4561977.036
247.940
93.885
93.829
92.483
92.572
93.500
88.592
100.279
100.331
85.612
89.104
95.271
97.793
92.866
95.141
95.179
98.399
98.331
93.467
93.415
93.288
92.122
92.041
93.055
88.133
99.875
99.933
85.166
181.161
179.773
175.508
175.408
178.899
170.644
181.409
182.237
168.529
167.818
178.737
182.461
174.093
175.479
178.299
181.556
180.343
177.924
178.188
176.859
173.754
172.048
175.788
167.377
180.126
180.539
165.557
4j
4k
4l
247.981
248.791
248.771
248.023
246.019
276.826
276.926
233.917
253.713
253.797
256.675
248.522
271.986
272.015
275.632
275.717
248.085
248.181
248.319
248.971
249.093
248.293
246.279
277.065
277.159
234.156
4m
4n
4o
4p
4q
4r
5a
5b
5c
5d
5e
5f
-1992.564101 -1992.588504 -5231474.57
-1572.224968 -1572.249918 -4127874.18
-1572.227813 -1572.252953 -4127881.64
-1647.446335 -1647.472304 -4325367.87
-1647.451182 -1647.477033 -4325380.60
-5231691.74
-4128093.08
-4128104.06
-4325594.36
-4325605.57
5g
5h
5i
-4106.476273 -4106.501089 -10781550.97 -10781772.93
-4106.483400 -4106.508149 -10781569.69 -10781791.97
-4106.484031 -4106.508699 -10781571.34 -10781791.97
5j
5k
5l
-1632.203176 -1632.227378 -4285346.96
-1632.209013 -1632.233064 -4285362.28
-1992.556989 -1992.581561 -5231455.90
-1548.984370 -1549.007465 -4066855.98
-1702.612608 -1702.638318 -4470206.92
-1702.607236 -1702.633038 -4470192.82
-1530.745978 -1530.768458 -4018971.08
-4285563.71
-4285576.91
-5231675.18
-4067064.78
-4470431.62
-4470418.04
-4019177.61
5m
5n
5o
5p
5q
5r
Table 5 Evaluation and preliminary screening of all the 21 descriptors.
-log(1/EC50 4a-4r)=a+bX
-log(1/EC50 5a-5r)=a+bX
Descriptors
a
b
R
P
a
b
R
P
Ǻ C=C
Ǻ C-X
1.34426
1.95331
1.39488
1.21938
2.898378*10-4 -0.07900 0.75535
1.34546
1.78713
1.39626
1.21957
-3.80595*10-4 -0.12184 0.63007
-7.6951*10-4 -0.15821 0.53065
-5.13255*10-6 -0.00258 0.99189
-0.00155
0.18125 0.47168
0.04065 0.87276
0.04325 0.86469
Ǻ C-NH
Ǻ NH-C=O
1.20929*10-4
3.08097*10-5
4.14741*10-5
-1.82031*10-4 -0.08640 0.7332
0.08373 0.74117
Ǻ N-C=O
∑Q of Ar
∑Q of Ph
∑Q of Cy
EHOMO
ELUMO
Egap
1.22187
-0.8735
1.44182*10-4
-0.02518
0.00485
0.04832 0.84901
-0.26526 0.28742
0.00552 0.98265
0.04051 0.87319
1.22264
-0.89937
-0.94342
-1.78788
-0.04809
-0.22398
0.15209
0.34433
3.35509
-0.25701
-0.00589
0.01555
-0.07189
0.00782
-0.00916
0.00154
0.16549
-0.40752 0.09323
-0.10359 0.68251
0.28459 0.25237
-0.00134 0.8497
-0.80498
-1.77528
-0.07385
-0.22149
0.14764
0.34771
3.30838
0.00326
-3.10108*10-4 -0.00782 0.97542
0.00709
0.20088 0.42414
0.32285
0.1913
-0.0074
0.14114 0.57642
-0.25187 0.31334
0.27741 0.26507
0.09901 0.69588
qH+
-9.96542*10-4 -0.12461 0.62227
0.35121 0.14401 0.56861
α
14

ACCEPTED MANUSCRIPT
HF
TE
ZPE
Hө
-3340.19581
-3339.78487
-3339.80945
-8.7686*10-6
-1374.54821
-0.83503 <0.0001 -1343.38162
-0.83502 <0.0001 -1342.97385
-0.83502 <0.0001 -1342.99793
-764.07923
-764.07666
-764.07732
-0.66501 0.00257
-0.66501 0.00260
-0.66507 0.00260
-1374.55001
-1374.55068
-3.60888*10-6 -0.83502 <0.0001 -3.52598*10-6 -2.00608*10-6 -0.66500 0.0026
Gө
-8.76882*10-6 -3.60889*10-6 -0.83502 <0.0001 -3.52619*10-6 -2.00609*10-6 -0.66500 0.0026
Eth
257.86961
93.15347
175.76684
-1.12953
2.14099
4.33901
-0.04773 0.85083
0.30715 0.21504
0.53175 0.02313
255.87533
91.74092
172.33638
1.61323
2.38453
3.75183
0.09776 0.69957
0.48783 0.04000
0.61512 0.00659
CVө
Sө
Table 6 The statistical parameters for the best 3D-QSAR CoMFA models
Statistical results
Filed contribution
Series
Methods
R2
F
SE
q2
Steric
0.829
0.454
Electrostatic
4a~4r
5a~4r
0.980
0.986
116.544
164.602
0.097
0.117
0.696
0.564
0.171
0.546
CoMFA
q2 =The leave-one-out (LOO) cross-validation coefficient; SE=Standard error of estimate. R2 =The predictive
correlation coefficient; F=test value.
q2 and R2 are calculated according to the formula of literature [32]:
Fig. 1 Agents for the treatment of hepatitis B infection
Fig. 2 Structure of phenylpropenamide derivatives
Fig 3: Crystal structure of compound 4j
15

ACCEPTED MANUSCRIPT
2.4
2.2
2.0
1.8
1.6
1.4
1.2
1.0
0.8
0.6
0.4
0.2
0.0
-0.2
-0.4
Sreise 4
Sreise 5
Serise 4: n=18, R=0.89829, SD=0.23308, P<0.0001
Serise 5: n=18, R=0.82786, SD=0.39287, P<0.0001
-0.5
0.0
0.5
1.0
1.5
2.0
Observed -Log(1/EC50
)
Fig. 4 The Relationships between the -log(1/EC50) values from experiment and prediction based
on QSAR equations.
a
b
Fig. 5 3D-views of all aligned phenylpropenamide molecules(a for series 4 and b for series 5)
congeners by RMSD-based fitting
2
2.0
1.5
1.0
0.5
0.0
Series 4, R =0.980
2
series 5, R =0.986
-0.5
0.0
0.5
1.0
1.5
2.0
Observed activity
Fig. 6 Plots of experimental activities versus the corresponding predicted activities by
QSAR/CoMFA models.
16

ACCEPTED MANUSCRIPT
a
b
c
d
Fig. 7. Stereoview of CoMFA electrostatic filed (a) and steric filed (b) of vinyl bromides of
phenylpropenamide molecules 4 and electrostatic filed (c) and steric filed (d) of vinyl chlorides of
phenylpropenamide molecules 5
Scheme 1 synthesis of phenylpropenamide derivatives: (i): aromatic aldehydes, AcOK, Ac2O,
reflux; (ii): piperidine, 0 ; (iii): Br2, 0 ; (iv): SO2Cl2, room temp.
Series 4:n=18, R2=0.80692, F=31.34498, P<0.0001
Series 5: n=18, R2=0.68535, F=16.33582, P< 0.00017
Scheme 2: The QSAR equations based on the -log(1/EC50) values
17

ACCEPTED MANUSCRIPT
New phenylpropenamide derivatives were synthesized and characterized.
Their 2D-QSAR and model 3D-QSAR were estabilished on DFT and SYBYL respectively.
QSAR model illustrated the effect of substituent on anti-HBV activities.