Bioorganic & Medicinal Chemistry Letters 29 (2019) 1909–1912

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Bioorganic & Medicinal Chemistry Letters

journal homepage: www.elsevier.com/locate/bmcl

Chalcone derivatives bearing chromen or benzo[f]chromen moieties: T

Design, synthesis, and evaluations of anti-inflammatory, analgesic, selective
COX-2 inhibitory activities
Zhi-Yang Fua, Qing-Hao Jinb, , Yu-Le Qua, Li-Ping Guana,
⁎ ⁎

a
Food and Pharmacy College, Zhejiang Ocean University, Zhejiang, Zhoushan 316022, PR China
b
Donghai Science and Technology College, Zhejiang Ocean University, Zhejiang, Zhoushan 316000, PR China

ARTICLE INFO ABSTRACT

Keywords: Thirty-eight chalcone derivatives bearing a chromen or benzo[f]chromen moiety were synthesized and eval-
Chalcone derivatives uated for their anti-inflammatory and analgesic activities. Using an ear edema model, anti-inflammatory ac-
Chromen or benzo[f]chromen tivities were observed for compounds 3a-3s (ear inflammation: 1.75–3.71 mg) and 4a-4s (ear inflammation:
Anti-inflammatory 1.71–4.94 mg). All compounds also displayed analgesic effects with inhibition values of 66.7–100% (3a-3s) and
Analgesic
96.2–100% (4a-4s). The 12 compounds that displayed excellent anti-inflammatory and analgesic effects were
COX-1/COX-2
tested for their inhibitory activity against ovine COX-1 and COX-2. Six compounds bearing a chromen moiety
were weak inhibitors of the COX-1 isozyme but showed moderate COX-2 isozyme inhibitory effects (IC50s from
0.37 μM to 0.83 μM) and COX-2 selectivity indexes (SI: 22.49–9.34). Those bearing a benzo[f]chromen moiety
were more selective toward COX-2 than those bearing a chromen moiety with IC50s from 0.25 μM to 0.43 μM and
COX-2 selectivity indexes from SI: 31.08 to 20.67.

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used
therapeutics to treat inflammation and pain from various pathological
disorders. NSAIDs work by inhibiting cyclooxygenase (COX) enzymes,
which catalyze prostaglandin biosynthesis through arachidonic acid.
COX-1 and COX-2 are two known COX isoforms.1–3 However, the use of
NSAIDs may result in severe side effects including pepticulcers, renal
dysfunction and bleeding.4–6 Interestingly selective inhibition of COX-2
over COX-1 is believed to reduce gastric ulceration side effects and
identification of clinically useful NSAIDs via selective inhibition of
COX-2 is an ongoing goal in medicinal and organic chemistry.7 Hence,
the development of efficient, new selective COX-2 inhibitory com-
pounds is essential.
Chalcones are flavonoids that possess a phenolic α,β-unsaturated
ketone structure containing a 1,3-diphenyl-2-en-1-one core and show
anti-inflammatory and analgesic effects.8–11 A previous study de-
termined that the presence of an α,β-unsaturated ketone structure in a
chalcone derivative is critical for its effects.12 Additionally, structure
activity relationship studies showed that reported COX-2 inhibitors
generally contain two aryl ring substituents on a core scaffold.13 One of
the core systems contains an α,β-unsaturated ketone and this hydro-
phobic structural feature (eC]CeC]O) may be responsible for the
observed pharmacological effects because of its ability to help facilitate
compound transport across various biological membranes.14 Previous
studies have shown that the chalcone moiety may contribute to the
anti-inflammatory effect by modulation of pro-inflammatory gene ex-
pression as exemplified by the comparable activities observed for se-
lective COX-2 inhibitory compounds and common drugs as shown Fig
1.15–17
As part of our continuous search for potential anti-inflammatory,
analgesic and selective COX-2 inhibitory compounds with the chemical
and biological properties of chalcone derivatives, we synthesized 19 3-
(2H-chromen-3-yl)-1-phenylprop-2-en-1-one derivatives (3a-3s) and 19
3-(3H-benzo [f]chromen-2-yl)-1-phenylprop-2-en-1-one derivatives
(4a-4s) (Fig. 2), and evaluated their anti-inflammatory and analgesic
activities in vivo. The COX-1 and COX-2 inhibition effects of selected
compounds were also determined in vitro.
The synthetic routes of the target compounds are illustrated in
Scheme 1. The treatment of 2-hydroxybenzaldehyde or 2-hydroxy-1-
naphthaldehyde with acrolein, a Michael acceptor, in refluxing dioxane
in the presence of potassium carbonate afforded 2H-chromene-3-car-
baldehyde 1 (yield: 96%) or 3H-benzo[f]chromene-2-carbaldehyde 2
(yield: 92%), respectively. The Claisen–Schmidt condensation of 2H-
chromene-3-carbaldehyde 1 or 3H-benzo[f]chromene-2-carbaldehyde 2
with the appropriate acetophenone in an ethanolic solution of NaOH

⁎
Corresponding authors.
E-mail addresses: qhjin2012@163.com (Q.-H. Jin), glp730@163.com (L.-P. Guan).

https://doi.org/10.1016/j.bmcl.2019.05.051
Received 23 January 2019; Received in revised form 16 May 2019; Accepted 27 May 2019
Available online 28 May 2019
0960-894X/ © 2019 Elsevier Ltd. All rights reserved.
Z.-Y. Fu, et al. Bioorganic & Medicinal Chemistry Letters 29 (2019) 1909–1912

Table 1
Anti-inflammatory effect of compounds 3a-3s and 4a-4s in vivo.
Compd Anti-inflammatory activity

Edema mean ± SD Edema mean ± SD (mg) Inhibition rate %

(mg) Inhibition rate %a

3a 2.54 ± 1.17** 64.4 4a 2.88 ± 1.12** 73.3

3b 3.40 ± 1.55* 52.4 4b 3.22 ± 0.87* 70.1
3c 2.46 ± 0.22** 65.6 4c 4.60 ± 1.96* 57.3
3d 3.41 ± 1.34* 52.3 4d 3.66 ± 1.33* 66.1
3e 3.50 ± 0.70* 51.0 4e 2.70 ± 1.50** 75.0
3f 3.14 ± 0.72* 56.1 4f 3.55 ± 0.61* 67.1
3g 3.34 ± 1.24* 53.3 4g 1.71 ± 0.89*** 84.1
3h 3.10 ± 1.30* 56.6 4h 2.18 ± 1.35** 79.8
3i 3.45 ± 1.22* 51.7 4i 3.46 ± 0.56* 67.9
3j 2.43 ± 0.75** 66.0 4j 4.58 ± 1.54* 57.5
3k 1.75 ± 0.07*** 75.5 4k 1.77 ± 0.67*** 83.6
Fig. 1. Chemical structures of reported anti-inflammatory chalcone derivatives 3l 3.52 ± 1.74** 50.8 4l 2.56 ± 0.78** 76.3
as selective COX-2 inhibitors. 3m 3.35 ± 0.35* 53.1 4m 2.74 ± 1.37** 74.6
3n 2.42 ± 0.97** 66.2 4n 2.23 ± 1.74** 79.3
3o 3.05 ± 1.02* 57.3 4o 3.54 ± 1.24* 67.2
3p 3.39 ± 0.52* 52.6 4p 3.55 ± 1.17* 67.1
3q 2.28 ± 1.20** 68.1 4q 1.82 ± 0.85*** 83.1
3r 3.71 ± 0.89* 48.1 4r 4.94 ± 0.87* 54.2
3s 2.32 ± 0.92** 67.6 4s 4.84 ± 1.17* 55.1
Idm 2.55 ± 1.3** 64.3 Idm 1.92 ± 0.9*** 82.2
control 7.15 ± 1.42 — control 10.78 ± 1.70 —

Idm: Indomethacin.
Fig. 2. The structures of designed chalcone derivatives 3a-3s and 4a-4s. *
p < 0.05, **p < 0.01, ***p < 0.001 compared with the PEG-400 (control)
group.
a
% at 1 h of anti-inflammatory activity.

inflammatory activity at a dose of 30 mg/kg administered in-

Scheme 1. The synthesis routes of target compounds 3a–3s and 4a-4s.
yielded nineteen 3-(2H-chromen-3-yl)-1-phenylprop-2-en-1-ones (3a-
3s) and nineteen 3-(3H-benzo[f]chromen-2-yl)-1-phenylprop- 2-en-1-
ones (4a-4s) in yields of 76–93% and 67–85% at room temperature,
respectively.
The structures of products 3a–3s and 4a–4s were confirmed by IR,
NMR and mass spectrometry. The IR spectra of the synthesized target
derivatives showed absorption bands at 1695–1701 cm−1 for the eC]
O stretching vibration and 3340 cm−1 for the eNH2 stretching vibra-
tion. Chalcones were always obtained as the (E)-isomer as determined
from the 1H NMR spectra. The 1H NMR coupling constants of the α and
β-vinylic protons in the chalcone derivatives had large Ja,β values of
15.0 Hz suggesting a trans-configuration. The 1H NMR data confirmed
the presence of an eNH2 group as a singlet signal at 10.3 ppm and the
eOCH2 group as singlet signals from 5.03 to 5.15 ppm. The 13C NMR
spectra showed the appearance of a new peak from 186.08 to
197.59 ppm for the O]C group.18–20
Thirty-eight of the chalcone derivatives and the reference drug in-
domethacin were examined for their anti-inflammatory activities using
an in vivo xylene-induced ear edema model.21,22 The percentage in-
hibition of the inflammation for the chalcone compounds was tested
after 1 h of treatment with xylene. The anti-inflammatory effects of
chalcone derivatives 3a-3s and 4a-4s (30 mg/kg) were comparable to
indomethacin (30 mg/kg) (Table 1). Compounds 3a-3s showed anti-
traperitoneally before the inflammatory agent xylene with ear in-
flammation values of 1.75 ± 0.07–3.71 ± 0.89 mg and inhibition of
inflammation of 49.4–75.5%. Eight compounds, 3a, 3c, 3g, 3j, 3k, 3n,
3q and 3s, displayed good anti-inflammatory activity with inhibition
values greater than 60%, which was nearly equivalent to that of in-
domethacin (64.3%). Chalcone derivatives 4a-4s also exhibited anti-
inflammatory activity at a dose of 30 mg/kg administered in-
traperitoneally before the inflammatory agent xylene with ear in-
flammation values of 1.71 ± 0.89–4.94 ± 0.87 mg and inhibition of
inflammation of 54.2–84.1%. Ten compounds, 4a, 4b, 4e, 4g, 4h, 4k-
4n, and 4q, showed anti-inflammatory activities with inhibition of in-
flammation of more than 70%, which was nearly equivalent to that of
indomethacin (82.2%).
The structure activity relationship studies for the anti-inflammatory
effects of chalcone derivatives 3a-3s and 4a-4s shown in Table 1 illu-
strated that the 38 compounds that reduced the ear inflammation and
showed anti-inflammatory activities not only contained electron-with-
drawing substituents but also electron-donating substituents on the
chromen or benzo[f] chromen moiety. Of nine compounds with elec-
tron-withdrawing with halogen atom substituent groups, the introduc-
tion of F (3a, 3b and 3c), Cl (3d, 3e and 3f), or Br (3h, 3i and 3j),
resulted in the best anti-inflammatory activity when substituted at the
p-position of the phenyl ring as demonstrated for compounds 3c, 3f and
3j, with the best compound being 3j with ear inflammation values 66%.
Within 4a-4c and 4h-4j, excellent anti-inflammatory effects were ob-
served when an electron-donating group was substituted at the o-po-
sition of the phenyl ring as shown for compounds 4a and 4h. Com-
pound 4e showed the best activity of 4d-4f with substitution at the m-
position of the phenyl ring. Thus, the substitution position of the ha-
logen atom greatly influenced the anti-inflammatory activities. The
order of activity for 3a-3f and 3h-3j compounds containing substitu-
tions on the phenyl ring was p-F > o-F > m-F, p-Cl > o-Cl > m-Cl,
and p-Br > o-Br > m-Br. The order of activity for 4a-4c and 4h-4j
compounds containing the benzo[f]chromen moiety with substitutions
on the phenyl ring was o-F > m-F > p-F and the order of activity for

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Z.-Y. Fu, et al. Bioorganic & Medicinal Chemistry Letters 29 (2019) 1909–1912

Br-substituted positions was o-Br > m-Br > p-Br. However, for 4d-4f Table 2
compounds containing the benzo[f]chromen moiety where the phenyl COX-1/COX-2 enzyme inhibition assay in vitro.
was substituted with a Cl, the order of activity was m-Cl > p-Cl > o- Compounds COX-1, IC50 (μM)a COX-2, IC50 (μM) COX-2, SIb
Cl. Compound 4g with a 2,4-dichlolo substituent showed the best anti-
inflammatory activity (84.1%). Compounds 3k, 3l, 4k and 4l with a 3c 8.32 ± 0.19 0.37 ± 0.22 22.49
3j 7.21 ± 0.25 0.42 ± 0.30 17.17
trifluoromethyl (eCF3) or nitro (eNO2) group on the phenyl or naph-
3k 7.75 ± 0.26 0.83 ± 0.10 9.34
thalene ring decreased the ear edema in mice by 75.5%, 50.8%, 83.6%, 3n 9.20 ± 0.30 0.46 ± 0.18 20.0
and 76.3%, respectively, with the best compound being 4k with an ear 3q 6.22 ± 0.28 0.39 ± 0.16 15.95
edema value of 1.75 mg. For compounds containing the electron-do- 3s 7.08 ± 0.24 0.46 ± 0.13 15.39
nating groups of eCH3, eOCH3, eN(CH3)2, eNH2, eCH2CH3, e(CH3)2, 4a 6.41 ± 0.27 0.31 ± 0.06 20.68
4g 7.92 ± 0.13 0.29 ± 0.11 27.31
and e(OCH3)2 on the phenyl ring, the anti-inflammatory activities were
4k 8.63 ± 0.17 0.33 ± 0.07 26.15
in the order of 3q > 3s > 3n > 3o > 3m > 3p > 3r. Compounds 4l 7.77 ± 0.31 0.25 ± 0.21 31.08
containing the benzo[f]chromen moiety had activities in the order of 4n 9.04 ± 0.19 0.38 ± 0.05 23.79
4q > 4n > 4m > 4o > 4p > 4s > 4r. 4q 8.89 ± 0.24 0.43 ± 0.12 20.67
Celecoxib 6.13 ± 0.08 0.22 ± 0.09 27.86
The analgesic activities of the target derivatives 3a-3s and 4a-4s
were evaluated using a hot plate technique at 60 min after in- a
IC50 value is the compound concentration required to produce 50% in-
traperitoneal injection in mice at 58 °C and compared with in- hibition of COX-1 or COX-2 for means of three determinations and deviation
domethacin as the reference.23–25 All compounds (3a-3s and 4a-4s) from the mean is < 10% of the mean value.
displayed potent analgesic effects not only with electron-donating b
SI: selectivity index (COX-1 IC50/COX-2 IC50).
substituents but also with electron-withdrawing substituents on the
phenyl ring at a dose of 30 mg/kg administered intraperitoneally with
inhibition values of 66.7–100% (3a-3s) and 96.2–100% (4a-4s) ring, the order of activity was o-F > p-F > m-F, o-Cl > p-Cl > m-Cl,
(Fig. 3). For compounds containing the electron-donor groups of eCH3, and o-Br > p-Br > m-Br. Compound 4g with a 2,4-dichlolo sub-
eOCH3, eN(CH3)2, eNH2, eCH2CH3, e(CH3)2, and e(OCH3)2 on the stituent showed the best anti-inflammatory activity with inhibition
phenyl ring, the order of the analgesic effects was 3p > 3r > 3n, values of 100%. Compounds 3k, 3l, 4k and 4l with a trifluoromethyl
3q > 3m, 3o > 3s. For compounds containing the benzo[f]chromen (eCF3) or nitro (eNO2) group substituted on the phenyl or naphthalene
moiety, the activity order was 4n, 4o, 4p, 4r > 4s > 4q > 4m. For ring decreased the ear edema in mice by 100%, 85.1%, 100%, and
nine compounds with electron-withdrawing with halogen atom sub- 100%, respectively.
stituent groups, the introduction of F (3a, 3b and 3c), Cl (3d, 3e and Traditional NSAIDs such as aspirin, indomethacin and ibuprofen
3f), or Br (3h, 3i and 3j), excellent analgesic activities were observed provide their anti-inflammatory activity through non-selective inhibi-
for those substituted at the p-position of the phenyl ring as shown for tion of both COX-1 and COX-2.26,27 The bleeding, gastrointestinal irri-
compounds 3c, 3f and 3j. Thus, the position of the halogen atom sub- tation and ulceration side effects of the classical NSAIDs are attributed
stitution greatly influenced the analgesic effects. In this assay, the order to their selectivity for COX-1 versus COX-2. Therefore, the development
of activity for compounds containing substitutions on the phenyl ring of novel selective COX-2 inhibitors through medicinal chemistry ap-
was p-F > m-F > o-F and p-Br > m-Br > o-Br. The order of activity proaches is essential.28,29 12 compounds that displayed excellent anti-
observed for compounds containing a Cl-substituent was p-Cl > o- inflammatory and analgesic effects were tested for their inhibitory ac-
Cl > m-Cl. For compounds 4a-4f and 4h-4j, those that displayed the tivity against ovine COX-1 and COX-2 (Table 2). The results indicated
best analgesic activities contained substituents at the o-position of the that six compounds containing a chromen moiety were weak inhibitors
phenyl ring as shown by compounds 4a, 4d and 4h. For derivatives of the COX-1 isozyme but showed moderate COX-2 isozyme inhibitory
bearing a benzo[f]chromen moiety, with substitutions on the phenyl effects (IC50 from 0.37 μM to 0.83 μM) and COX-2 selectivity indexes
(SI: 22.49–9.34). Furthermore, six compounds containing the benzo[f]
chromen moiety were weak inhibitors of the COX-1 isozyme but
showed moderate COX-2 isozyme inhibitory effects (IC50: from 0.25 μM
to 0.43 μM) and COX-2 selectivity indexes (SI: 31.08–20.67). The active
compounds containing the benzo[f]chromen moiety were more sensi-
tive to structural modifications in terms of their selectivity for COX-2
than those containing the chromen moiety. Additionally, the activity of
compound 4l (IC50 = 0.25 μM, SI = 31.08) was more similar to in-
hibitors of the COX-2 isozyme than the reference drug celecoxib
(IC50 = 0.22 μM, SI = 27.86).
The results indicated that the molecular hybridization of the chal-
cone pharmacophore of the COX-2 inhibitors was a useful building
block to produce effective hybrid scaffolds with improved analgesic and
anti-inflammatory activity potential.
In conclusion, a series of chalcone derivatives bearing a chromen or
benzo[f]chromen moiety were synthesized and evaluated for their anti-
inflammatory and analgesic effects in vivo. The results indicated that all
compounds exhibited good anti-inflammatory and analgesic effects.
Among them, 12 compounds that exhibited the best anti-inflammatory
and analgesic activities were tested for their inhibitory activity against
ovine COX-1 and COX-2. All target compounds were more selective
toward COX-2 compared with COX-1. The activity of compound 4l was
more similar in magnitude to inhibitors of the COX-2 isozyme than the
Fig. 3. Analgesic effect of the designed compounds 3a-3s and 4a-4s. Notes:
reference drug celecoxib. Hence, the compounds reported herein are
Idm: Indomethacin. promising starting points for the development of an inhibitor of COX-2.

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Z.-Y. Fu, et al.
Acknowledgements
This work was supported by Zhejiang Province Public Technology
Application Project (No. 2017C33131). We thank Renee Mosi, PhD,
from Liwen Bianji, Edanz Group China (www.liwenbianji.cn/ac), for
editing the English test of a draft of this manuscript.
References
1. Quach ND, Arnold RD, Cummings BS. Biochem Pharmacol. 2014;90:338.
2. Lokeshwari DM, Achutha DK, Srinivasan B, Shivalingegowda N, Krishnappagowda
LN, Kariyappa AK. Bioorg Med Chem Lett. 2017;27:3806.
3. Hassan GS, Abou-Seri SM, Kamel G, Ali MM. Eur J Med Chem. 2014;78:482.
4. Ugwu DI, Okoro UC, Ukoha PO, Gupta A, Okafor SNJ. Enzyme Inhib Med Chem.
2018;33:405.
5. Abdel-Sayed MA, Bayomi SM, El-Sherbeny MA, et al. Bioorg Med Chem.
2016;24:2032.
6. Makhdoumi P, Zarghi A, Daraei B, Karimi GJ. Pharmacopuncture. 2017;20:207.
7. Gouvea DP, Vasconcellos FA, Berwaldt, et al. Eur J Med Chem. 2016;118:259.
8. Raj R, Saini A, Gut J, Rosenthal PJ, Kumar V. Eur J Med Chem. 2015;95:230.
9. Sashidhara KV, Kumar M, Modukuri RK, et al. Bioorg Med Chem Lett. 2011;21:4480.
10. Chen WB, Ge XT, Xu FL, et al. Bioorg Med Chem Lett. 2015;25:2998.
11. Wei ZY, Chi KQ, Yu ZK, et al. Bioorg Med Chem Lett. 2016;26:5920.
12. Guan LP, Liu BY. Eur J Med Chem. 2016;121:47.
13. Ahmed HEA, Abdel-Salam HA, Shaker MA. Bioorg Chem. 2016;66:1.
14. Badrey MG, Abdel-Aziz HM, Gomha SM, Abdalla MM, Mayhoub AS. Molecules.
2015;20:15287.
15. Labib MB, Sharkawi SMZ, El-Daly M. Bioorg Chem. 2018;80:70.
16. Zarghi A, Zebardast T, Hakiminon F, Shirazi FH, Rao PNP, Knaus EE. Bioorg Med
Chem. 2006;14:7044.
17. Bano S, Javed K, Ahmad S, et al. Eur J Med Chem. 2013;65:51.
1912
Bioorganic & Medicinal Chemistry Letters 29 (2019) 1909–1912
18. Foroumadi A, Emami S, Sorkhi M, et al. Chem Biol Drug Des. 2010;75:590.
19. Nazarian Z, Emami S, Heydari S, et al. Eur J Med Chem. 2010;45:1424.
20. The synthesis of 3-(2H-chromen-3-yl)-1-(2,4-dichloro)phenyl-prop-2-en-1-one 3g
and nineteen 3-(3H-benzo[f]chromen-2-yl)-1-(2,4-dichloro)phenylprop-2-en-1-one
4g: To a solution of 2H-chromene-3-carbaldehyde 1 or 3H-benzo[f]chromene-2-
carbaldehyde 2 (1 mmol) and 2,4-dichloroacetophenone (1 mmol) in absolute
ethanol (20 mL), NaOH solution (3.5 M, 8 mL) was added and stirred overnight at
room temperature. The reaction mixture was diluted with water and the precipitate
was filtered and crystallized from ethanol to give the target compounds. 3g: Yield:
76.5%, m.p. 92.1–92.9 °C. 1H NMR (CDCl3, 300 MHz): δ 5.14 (s, 2H, –CH2), 6.81–7.
03 (m, 3H, –C6H3), 7.12 (d, 1H, J = 15 Hz, =CH), 6.89 (s, 1H, =CH), 7.17–8.34 (m,
4H, –C6H4), 7.53 (d, 1H, J = 15 Hz, =CH). 13C NMR (CDCl3, 75 MHz): 65.13, 115.
89, 120.78, 121.94, 123.88, 128.51, 129.89, 131.33, 133.22, 142.77, 150.06, 154.
83, 188.09. IR (KBr) cm−1: 1698 (C=O), 1622 (C–O). MS m/z 331.22 (M+1). 4g:
Yield: 68.4%, mp: 98.3–100.8 °C. 1H NMR (CDCl3, 300 MHz): δ 5.11 (s, 2H, –CH2),
6.49 (d, 1H, J = 15 Hz, =CH), 7.07–7.10 (d, 1H, J = 9 Hz, –C6H2), 7.38 (s, 1H,
=CH), 7.33–7.51 (m, 3H, –C6H3), 7.36–7.77 (m, 4H, –C6H4), 7.53 (d, 1H, J = 15
Hz, =CH). 7.96–7.99 (d, 1H, J = 9 Hz, –C6H2). 13C NMR (CDCl3, 75 MHz): 64.86,
114.98, 117.36, 121.22, 124.23, 124.38, 126.99, 127.39, 127.57, 128.77, 129.35,
129.64, 130.08, 130.13, 130.37, 132.05, 132.11, 136.64, 137.42, 143.82, 153.94,
192.04. IR (KBr) cm−1: 1685 (C=O), 1618 (C–O). MS m/z 381.29 (M+1).
21. Ohlsson L. Acta Pharmacol Toxicol. 1953;9:322.
22. Cordeiro KW, Felipe JL, Malange KF, et al. J Ethnopharmacol. 183 2016,;183:128.
23. Leighton G, Johnson M, Meecham K, Hill R, Hughes J. Br J Pharmacol. 1987;92:915.
24. Shaaban OG, Rizk OH, Bayad AE, El-Ashmawy IM. Open Med Chem J. 2013;7:49.
25. Guan LP, Xia YN, Jin QH, Liu BY, Wang SH. Bioorg Med Chem Lett. 2017;27:3378.
26. Shi L, Aixi H, Jiangpina X, Yiping J. Chin J Chem. 2012;30:1339.
27. Dou J, Shi L, Hu A, et al. Arch Pharm Chem Life Sci. 2014;347:89.
28. Eleftheriou P, Geronikaki A, Hadjipavlou-Litina D, et al. Eur J Med Chem.
2012;47:111.
29. El-Sayed M, Abdel-Aziz NI, Abdel-Aziz AA-M, El-Azab AS, ElTahir K. Bioorg Med
Chem. 2012;20:3306.