ADME properties, bioactivity and molecular docking studies of 4-amino-chalcone derivatives: new analogues for the treatment of Alzheimer, glaucoma and epileptic diseases

Article abstract of DOI:10.1007/s40203-021-00094-x

In this study, in vitro inhibition effects of (E)-1-(4-aminophenyl)-3-(aryl) prop-2-en-1-one (4-amino-chalcones) derivatives (3a–o) on acetylcholinesterase (AChE) enzyme and human erythrocyte carbonic anhydrase I and II isoenzymes (hCA I- II) were investigated. And also, the biological activities of 4-amino-chalcone derivatives against enzymes which names are acetylcholinesterase (PDB ID: 1OCE), human Carbonic Anhydrase I (PDB ID: 2CAB), human carbonic anhydrase II (PDB ID: 3DC3), were compared. After the results obtained, ADME/T analysis was performed in order to use 4-amino-chalcone derivatives as a drug in the future. Effective inhibitors of carbonic anhydrase I and II isozymes (hCAI and II) and acetylcholinesterase (AChE) enzymes with Ki values in the range of 2.55 ± 0.35–11.75 ± 3.57?nM for hCA I, 4.31 ± 0.78–17.55 ± 5.86?nM for hCA II and 96.01 ± 25.34–1411.41 ± 32.88?nM for AChE, respectively, were the 4-amino-chalcone derivatives (3a–o) molecules.

Full text of DOI:10.1007/s40203-021-00094-x

In Silico Pharmacology
(2021) 9:34
https://doi.org/10.1007/s40203-021-00094-x
ORIGINAL RESEARCH
ADME properties, bioactivity and molecular docking studies
of 4‑amino‑chalcone derivatives: new analogues for the treatment
of Alzheimer, glaucoma and epileptic diseases
Meliha Burcu Gürdere · Yakup Budak · Umit M. Kocyigit · Parham Taslimi · Burak Tüzün · Mustafa Ceylan1
1
1
2
3
4
Received: 17 January 2021 / Accepted: 23 April 2021
©
The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2021
Abstract
In this study, in vitro inhibition efects oꢀ (E)-1-(4-aminophenyl)-3-(aryl) prop-2-en-1-one (4-amino-chalcones) deriva-
tives (3a–o) on acetylcholinesterase (AChE) enzyme and human erythrocyte carbonic anhydrase I and II isoenzymes (hCA
I- II) were investigated. And also, the biological activities oꢀ 4-amino-chalcone derivatives against enzymes which names
are acetylcholinesterase (PDB ID: 1OCE), human Carbonic Anhydrase I (PDB ID: 2CAB), human carbonic anhydrase II
(
PDB ID: 3DC3), were compared. Aꢀter the results obtained, ADME/T analysis was perꢀormed in order to use 4-amino-
chalcone derivatives as a drug in the ꢀuture. Efective inhibitors oꢀ carbonic anhydrase I and II isozymes (hCAI and II) and
acetylcholinesterase (AChE) enzymes with Ki values in the range oꢀ 2.55± 0.35–11.75± 3.57 nM ꢀor hCA I, 4.31± 0.78–
1
7.55± 5.86 nM ꢀor hCA II and 96.01± 25.34–1411.41± 32.88 nM ꢀor AChE, respectively, were the 4-amino-chalcone
derivatives (3a–o) molecules.
Keywords 4-Amino-chalcones · Carbonic anhydrase · Acetylcholinesterase · Molecular docking
Introduction
oꢀ the nerve end and removing them ꢀrom there. Thus, a
possible malꢀunction in nerve conduction does not occur
(Göcer et al. 2017; Ökten et al. 2020; Çakmak et al. 2020).
One oꢀ the most important causes oꢀ Alzheimer’s disease
is the decrease in the amount oꢀ acetylcholine in the brain
(Göcer et al. 2017).
Acetylcholine is a neurotransmitter substance secreted ꢀrom
the synapses oꢀ autonomic ganglia and the ends oꢀ nerve
strands laying the skeletal muscle. Acetylcholinesterase
(
AChE; EC 3.1.1.7) is a nonspeciꢁc enzyme that hydrolyzes
lipotropic acetylcholine in the tissues, ꢀree or combined with
phospholipids.In other words, acetylcholinesterase ꢀunctions
around by removing the chemicals that accumulate in ꢀront
AChE inhibitors (Donepezil, Rivastigmin) are widely
used ꢀor Alzheimer’s patients to live as high quality (Göcer
et al. 2013, 2015). However, such drugs with inhibitory
efects cause side efects such as gastrointestinal disorders
and hepatotoxicity. For this reason, AChE inhibitors, which
are both efective and saꢀe, as well as especially natural, have
become more and more important in recent times (Gülçin
et al. 2020; Timur et al. 2019).
*
*
Meliha Burcu Gürdere
burcugurdere@gmail.com
Burak Tüzün
theburaktuzun@yahoo.com
Carbonic anhydrase inhibitors are among the most
potent drugs that reduce the intraocular pressure used in
the treatment oꢀ eye diseases. These have been recently
used ꢀor reducing intraocular pressure, topical and is used
systemically. Acetazolamide, a systemic inhibitor oꢀ car-
bonic anhydrase, was ꢁrst used in 1954 by Becker, Grant,
Trotter, Breinin and Gürtz to reduce intraocular pressure in
glucose disorder (Becker 2020; Kim et al. 2019; Supuran
2019). In addition, acetazolamide is the most used inhibi-
tor oꢀ the carbonic anhydrase enzyme. Recently, there has
1
2
Department oꢀ Chemistry, Faculty oꢀ Arts and Sciences,
Tokat Gaziosmanpaşa University, 60250 Tokat, Turkey
Department oꢀ Basic Pharmaceutical Sciences, Division
oꢀ Biochemistry, Faculty oꢀ Pharmacy, Sivas Cumhuriyet
University, 58140 Sivas, Turkey
3
4
Department oꢀ Biotechnology, Faculty oꢀ Science, Bartin
University, 74100 Bartin, Turkey
Department oꢀ Chemistry, Faculty oꢀ Science, Sivas
Cumhuriyet University, 58140 Sivas, Turkey
Vol.:(0
1
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O
been a search ꢀor inhibitors ꢀor hCA isoenzymes in struc-
tures other than sulꢀonamides. Inhibitor development and
synthesis studies have been increased, especially ꢀor use
in drug design development.
O
H N
2
2
Ar
3
a-o
H N
2
O
Recently, in many studies, the importance and place
oꢀ theoretical studies has increased. Experimental studies
conducted in these studies are guiding (Tüzün et al. 2018,
i
Ar
H
ii
O
1
a-o
O
O N
2
020; Bilgiçli et al. 2020a, b; Douche et al. 2020). Theo-
2
4
Ar
retical studies are preꢀerred because they are very ꢀast,
and their cost is cheap. Among the theoretical methods,
the most common and ꢀastest molecular docking method
was used to compare the biological activities oꢀ 4-amino-
chalcone derivatives (3a–o) against enzymes (Taslimi
et al. 2020). These enzymes are acetylcholinesterase
O N
2
5c,f,i,l
Scheme 1 Reagents and conditions: (i) NaOH, C H OH, 0 °C, 3 h,
2
5
(
ii) SnCl ·2H O, C H OH, reꢂux, 2 h
2 2 2 5
(
AChE) (PDB ID: 1OCE) (Cheung et al. 2013), human
Carbonic Anhydrase I (hCA I) (PDB ID: 2CAB) (Alterio
et al. 2010), human carbonic anhydrase II (hCA II) (PDB
ID: 3DC3) (Ivanova et al. 2015). ADME/T (Absorption,
distribution, metabolism, excretion, and toxicity) analysis
was then perꢀormed to investigate the drug availability
oꢀ 4-amino-chalcone derivatives (3a–o). The parameters
obtained as a result oꢀ this ADME/T analysis theoretically
examined the biological efects and reactions oꢀ this mol-
ecule in human metabolism.
Biochemical studies
Inhibition oꢀ metabolic enzymes was investigated, and their
results were reported as ꢀollows.
In this study, the compounds 3a–o were screened against
the AChE enzyme due to signiꢁcant reports on AD oꢀ phe-
nolic natural or synthetic compounds. IC and K values
5
0
i
oꢀ the reꢀerence drug Tacrin were 1143.31 nM (IC ) and
5
0
858.85± 12.11 nM (Ki) towards AChE, as shown in Table 2.
Whether hCA I and II isoenzymes and AChE, these
three enzyme systems are oꢀ great importance ꢀor human
health. Changes in the level oꢀ hCA in human erythro-
cytes are associated with many metabolic disorders such as
diabetes, edema and hypertension (Bayindir et al. 2019).
AChE enzyme, on the other hand, they are the enzymes
responsible ꢀor the emergence oꢀ many neurological dis-
eases, including Alzheimer’s (Güzel et al. 2019).
The compounds inhibited the AChE enzyme in nanomo-
lar concentration in the range oꢀ K values oꢀ 96.01 ± 25.
i
34–1411.41± 302.88 nM and with IC values oꢀ 179.71—
5
0
1048.41 nM The compounds 3a and 3d were ꢀound potent
.
AChE inhibitors with the Ki values oꢀ 96.01 ± 25.34 nM
and 156.45± 66.48 nM, respectively, while the compound
was the least inhibiting compound with the highest K value
i
oꢀ 1411.41± 302.88 nM. On the other hand, the compound
3e was also considered as one oꢀ the potent inhibitors with
the lowest IC value oꢀ 179.71 nM against AChE (Figs. 1,
In the scope oꢀ our study, the inhibition properties oꢀ
4
-amino-chalcone derivatives (3a–o) were investigated
5
0
by using them as inhibitors. Depending on the results
obtained, these molecules or their derivatives are used in
drug designs in terms oꢀ usability in the treatment oꢀ many
diseases. We anticipate that it will shed light and make
signiꢁcant contributions in the ꢁeld oꢀ pharmacology.
2). The Table 2 showed that the K values oꢀ reꢀerence
i
drug AZA were 83.90± 19.71 nM and 104.60± 27.60 nM,
whereas the IC values oꢀ AZA were 97.30 nM and
5
0
115.50 nM towards hCA I and II, respectively. Ki values
oꢀ 3a–o were calculated as 2.55± 0.35–11.75± 3.57 nM ꢀor
hCA I, 4.31± 0.78–17.55± 5.86 nM ꢀor hCA II.
Glaucoma with high intraocular pressure is one oꢀ the
most important eye diseases. It causes blindness with a rate
oꢀ 15–20%. hCA II in the eye retina is the primary cause oꢀ
intraocular pressure ꢀormation. The most important method
to eliminate this disease is to inhibit hCA II activity. For this
purpose, acetazolamide and heteroaromatic sulꢀonamides
have been used as inhibitors ꢀor many years (Ellman et al.
1961; Lineweaver and Burk 1934; Verpoorte et al. 1967).
Acetyl CoA is a product ꢀormed as a metabolic prod-
uct oꢀ pyruvate occurring in glycolysis. Alzheimer’s dis-
ease occurs as a result oꢀ decreased neurotransmitters in the
brain. The most decreasing neurotransmitter in this disease
Results and discussion
Chemistry
4
-Amino-chalcone derivatives (3a–o) were resynthesized
ꢀ
rom the Claisen-Schmidtcondensation oꢀ the benzalde-
hyde derivatives (1a–o) with 4-aminoacetophenone (2)
according to the published procedure (Koçyiğit et al. 2018;
Gürdere et al. 2020). (Scheme 1, Table 1). All spectro-
scopic data are in good agreement with the published data.
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34
Table 1 Chemical structures oꢀ
compounds
is widespread dementia and neurodegenerative disease. Alz-
heimer’s disease ꢀunction oꢀ memory.
results oꢀ 4-amino-chalcone derivatives appear to be in
great agreement with the experimental results (Aktas et al.
2020a, b; Gedikli et al. 2021). In the docking calculations
oꢀ 4-amino-chalcone derivatives, numerical values oꢀ
calculated many parameters as a result oꢀ docking cal-
culations oꢀ molecules and enzymes support this situa-
tion. The most important ꢀactor afecting the numerical
values oꢀ the parameters calculated ꢀrom the interaction
oꢀ amino-chalcone derivatives with enzymes is the inter-
action (Tüzün et al. 2021). Many chemical interactions
occur between amino-chalcone derivatives with enzymes.
As these chemical interactions increase, the biologi-
cal activities oꢀ 4-amino-chalcone derivatives appear to
increase. These interactions have many interactions such
as hydrogen bonds, polar and hydrophobic interactions,
π-π and halogen bonds (Sayin and Karakas 2017, 2018a; b;
Sayin and Üngördü 2018, 2019; Üngördü and Sayin 2019;
Jayarajan et al. 2020). As a result oꢀ calculations, the inter-
actions oꢀ molecules with enzymes are given in Figs. 3, 4,
It was ꢀound as disorder. The decrease in the level oꢀ ace-
tylcholine in the brain is the biggest biochemical ꢀactor oꢀ
this disease. There is no cure ꢀor this disease. The treat-
ments applied today are aimed at eliminating the symptoms
oꢀ this disease. There is no treatment method that eliminates
it. AChE inhibitors such as Rivastigmine and Donepezil are
generally used ꢀor this purpose (Kocyigit et al. 2017; Güzel
et al. 2019; Ökten et al. 2019).
The presence oꢀ some unwanted negative physiological
efects oꢀ these drugs in patients has made the discovery oꢀ
new inhibitors important.
Molecular docking results
Molecular docking calculations were made to compare
the biological activities oꢀ 4-amino-chalcone derivatives
enzymes against proteins. Molecular docking calculation
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Table 2 Inhibition results oꢀ novel 4-Amino-chalcone derivatives (3a-o) on carbonic anhydrase and acetylcholinesterase
Compounds
IC50 (nM)
hCA I
K (nM)
i
r2
hCA II
r2
AChE
r2
hCA I
hCA II
AChE
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
a
b
c
d
e
ꢀ
g
h
i
j
k
l
m
n
o
8.35
8.15
5.87
7.01
4.35
3.82
8.25
7.37
5.37
6.35
4.91
6.61
3.66
5.54
7.45
97.304
–
0.9717
0.9658
0.9580
0.9960
0.9575
0.9797
0.9639
0.9917
0.9991
0.9839
0.9965
0.9660
0.9722
0.9847
0.9865
0.9889
–
9.11
9.01
6.86
9.36
5.02
4.84
10.51
8.55
6.66
6.93
7.29
8.88
5.82
7.45
8.46
115.50
–
0.9810
0.9569
0.9973
0.9724
0.9953
0.9784
0.9528
0.9768
0.9896
0.9683
0.9543
0.9703
0.9773
0.9832
0.9425
0.9719
–
261.91
408.61
700.01
222.47
382.45
179.71
1048.41
548.26
758.21
898.83
327.21
564.33
595.36
756.55
488.02
–
0.9639
0.9817
0.9872
0.9848
0.9811
0.9668
0.9689
0.9905
0.9490
0.9691
0.9753
0.9692
0.9908
0.9933
0.9518
–
5.46± 1.32
11.75± 3.57
4.26± 0.85
6.94± 1.83
4.62± 0.82
2.81± 0.18
10.26± 1.47
7.31± 2.98
6.14± 1.42
3.97± 0.91
4.87± 0.83
10.04± 1.80
2.55± 0.35
6.56± 1.96
5.04± 1.55
83.390± 19.71
–
9.68± 2.51
17.55± 5.86
5.76± 0.91
13.21± 4.07
5.38± 1.23
4.31± 0.78
11.80± 3.95
7.22± 2.26
6.04± 1.52
9.93± 2.51
7.91± 1.88
6.23± 2.20
6.94± 1.55
5.91± 1.01
10.18± 0.85
104.60± 27.60
–
156.45± 66.48
259.33± 35.95
514.47± 95.66
96.01± 25.34
474.15± 70.66
159.62± 68.17
456.80± 114.41
218.63± 59.47
1411.41± 302.88
361.63± 73.80
299.77± 47.33
337.48± 99.43
495.94± 72.81
619.81± 85.46
285.20± 86.92
–
AZA
Tacrine
1143.312
0.9948
858.85± 12.11
Fig. 1 K values oꢀ novel
i
4
-Amino-chalcone derivatives
120
00
Ki values
(
3a–o) on hCA I and hCA II
1
hCA I
hCA II
8
6
4
2
0
0
0
0
0
AZA 3a
3b
3c
3d
3e
3f
3g
3h
3i
3j
3k
3l
3m 3n
3o
Compounds
and 5. In picture 3, in the interactions between the proteins
oꢀ the AChE enzyme and molecule 3d, a hydrogen bond is
121 protein. There is a hydrogen bond interaction between
the amine group in the molecule and the TRY 70 protein.
However, in the interactions between the proteins oꢀ the
hCA II enzyme and molecule 3ꢀ, there is a hydrogen bond
between the amino group in the molecule and the HIE
protein. Supplementary data ꢀor all other interactions is
given in Figure S29–S72.
ꢀ
ormed between the oxygen in the methoxy group attached
to the phenyl group in molecule 3d and the TRP 5 protein.
Again, pi-pi interactions occur between the phenyl group
in molecule 3d and HIE 64. There is a Pi-cation interac-
tion between the aniline ring and the LYS 170 protein. On
the other hand, in the interactions between the proteins
oꢀ the hCA I enzyme and the molecule 3 m, there is a
pi–pi interaction between the aniline ring and the TRY
Molecular docking calculations were perꢀormed against
enzymes oꢀ 4-amino-chalcone derivatives (3a–o) and bio-
logical activities oꢀ molecules were compared as a result oꢀ
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34
Fig. 2 K values oꢀ novel
i
1600
4
-Amino-chalcone derivatives
(
3a–o) on AChE enzymes
1
1
1
400
200
000
AChE
8
6
4
2
00
00
00
00
0
TAC 3a 3b 3c 3d 3e 3f 3g 3h 3i 3j 3k 3l 3m 3n 3o
Compounds
Fig. 3 Representation oꢀ interactions oꢀ molecules 3m and hCA I enzymes
calculations. The molecular docking parameters obtained
as a result oꢀ this comparison are given in Table 3 and S1.
The ꢁrst parameter among parameters obtained to com-
pare the biological activities oꢀ 4-amino-chalcone deriva-
tives (3a–o) is the docking scooter. this parameter is the
most important parameter to compare the biological activi-
ties oꢀ molecules. The molecular biological activity oꢀ the
Glide hbond parameter, whose numerical value gives inꢀor-
mation about the number oꢀ hydrogen bonds ꢀormed during
interactions. Glide hbond parameter is afected by the atoms
ꢀorming the hydrogen bond and the geometry oꢀ the hydro-
gen bonds. Another important parameter, that is Glide ligand
eꢃciency parameter, is a numerical value used to sort the
efectiveness oꢀ 4-amino-chalcone derivatives (3a–o) (Genc
Bilgicli et al. 2020; Bilgiçli et al. 2020a, b). Aꢀter examining
the interactions oꢀ 4-amino-chalcone derivatives (3a–o) with
enzymes, the efects and responses oꢀ these derivatives on
4
-amino-chalcone derivatives (3a–o) with the numerical
value oꢀ the docking score parameter is the most negative.
Another parameter among the parameters obtained is the
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In Silico Pharmacology
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Fig. 4 Representation oꢀ interactions oꢀ molecules 3d and AChE enzymes
Fig. 5 Representation oꢀ interactions oꢀ molecules 3f and hCA II enzymes
human metabolism are theoretically investigated in order to
study their drug availability properties. For this, ADME/T
analysis is required ꢀor 4-amino-chalcone derivatives (3a-o).
Each parameter obtained is as important as others to
explain the drug properties oꢀ 4-amino-chalcone derivatives
(3a–o). Because they each examined the efect or response oꢀ
diferent organs or tissues. In particular, it should be known
very well that one oꢀ the most important methods used
without experiment to explain the efects and responses oꢀ
4-amino-chalcone derivatives (3a–o) in human metabolism
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Table 3 Interaction value oꢀ studied molecule with enzymes
The ꢁrst parameter among the obtained parameters is Sol-
ute Molecular Weight, which tells how much the molecular
weight oꢀ the molecule should be ꢀor human metabolism.
Another parameter is Solute Hydrophobic SASA, which is
the Hydrophobic component oꢀ the SASA (saturated carbon
and attached hydrogen). Another parameter is QPlogHERG,
Molecules
Docking score
AChE
hCA I
hCA II
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
a
b
c
d
e
ꢀ
g
h
i
j
k
l
m
n
o
− 5.22
− 6.00
− 3.94
− 6.55
− 6.26
− 3.89
− 6.30
− 5.98
− 5.80
− 6.00
− 6.46
− 5.64
− 6.07
− 6.35
− 5.64
–
− 4.37
− 4.10
− 4.28
− 4.38
− 3.79
− 4.49
− 3.88
− 4.03
− 3.83
− 3.46
− 3.80
− 4.22
− 3.46
− 4.14
− 3.80
− 3.98
–
− 4.43
− 4.47
− 4.70
− 3.30
− 5.40
− 4.55
− 4.78
− 4.80
− 4.99
− 4.71
− 4.74
− 4.70
− 5.65
− 4.55
− 4.94
− 5.19
–
which is predicted IC value ꢀor blockage oꢀ HERG K chan-
50
nels. Another parameter is QPPCaco, which is Predicted
apparent Caco-2 cell permeability in nm/s. Caco-2 cells are
a model ꢀor the gut-Blood barrier. QikProp predictions are
ꢀ
or non-active transport. the numerical value oꢀ this param-
eter should be<25 poor,>500 great. Another parameter is
QPlogBB, which is Predicted brain/Blood partition coeꢃ-
cient. Another parameter is QPPMDCK, which is Predicted
apparent MDCK cell permeability in nm/sec. MDCK cells
are considered to be a good mimic ꢀor the Blood brain bar-
rier. QikProp predictions are ꢀor non-active transport. the
numerical value oꢀ this parameter should be<25 poor,>500
great. Two parameters shown among the most important
parameters obtained as a result oꢀ ADME/T analysis are
RuleOꢀFive (Lipinski 2004; Lipinski et al. 1997) and Rule-
OꢀThree (Jorgensen and Dufy 2002). These parameters are
more important than other parameters. When the numeri-
cal conditions oꢀ this parameter are not met, the molecule
is said to not be a deꢁnite drug. RuleOꢀFive is Lipinski
Rule oꢀ 5 and RuleOꢀThree is Jorgensen Rule oꢀ 3. Rule-
OꢀFive is number oꢀ violations oꢀ Lipinski’s rule oꢀ ꢁve. The
rules are: mol_MW < 500, QPlogPo/w < 5, donorHB ≤ 5,
accptHB≤10. Compounds that satisꢀy these rules are con-
sidered drug like (The “ꢁve” reꢀers to the limits, which are
AZA
TAC
− 6.06
is molecular docking calculations. By comparing the numer-
ical values oꢀ these efects and reactions, it is possible to
design more efective and active molecules (Taslimi et al.
2
020).
As a result oꢀ the ADME/T analysis, many parameters
were obtained. Some oꢀ these parameters are given in
Table 4. All remaining parameters are given in table S1.
Table 4 ADME properties oꢀ molecules
Reꢀerence
range
Solute molecu- QPlogHERG
lar weight
QPPCaco (nm/s)
QPPMDCK (nm/s)
RuleOꢀFive
RuleOꢀThree
1
30–725
(concern below − 5)
a<25 is poor and
a>500 is great
a<25 is poor and
a>500 is great
Maximum is 4
Maximum is 3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
a
b
c
d
e
ꢀ
g
h
i
j
k
l
m
n
o
237
237
237
253
253
253
258
258
258
302
302
302
229
213
223
− 5.56
− 5.64
− 5.63
− 5.67
− 5.57
− 5.55
− 5.6
− 5.63
− 5.62
− 5.62
− 5.66
− 5.65
− 5.32
− 5.09
− 5.68
971
970
970
971
970
970
971
970
970
971
970
970
969
968
971
479
479
479
479
479
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
479
1050
1180
1181
1130
1269
1269
884
478
479
1
3

34
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In Silico Pharmacology
(2021) 9:34
multiples oꢀ 5.). RuleOꢀThree is number oꢀ violations oꢀ
Jorgensen’s rule oꢀ three. The three rules are: QPlogS>-5.7,
QP PCaco > 22 nm / s, # Primary Metabolites < 7. Com-
pounds with ꢀewer violations oꢀ these rules are more likely
to be orally available (Çetiner et al. 2021).
Enzymes studies
Determination of the eꢀects of new compounds
on acetylcholinesterase enzyme
Many ADME/T parameters were calculated in the cal-
culations. Each oꢀ the ADME/T parameters ꢀound gives
inꢀormation about a diferent property oꢀ the molecules.
Molecules are desirably in the range oꢀ 130–725 g/mol.
The molecular weight oꢀ the molecules is in the range oꢀ
The efect oꢀ 4-amino-chalcone derivatives (3a–o) used in
the study on the enzyme acetylcholinesterase was investi-
gated. In this study, Acetylcholinesterase method was used.
Based on the data obtained, the inhibition types were deter-
mined by calculating the IC and K values. The principle
5
0
i
2
37–258. Another parameter ꢀor molecules to become drugs
oꢀ the method; As mentioned in the previous sections, AChE
is QPPCaco, which is expected to be less than 500. Oꢀ all the
molecules, there are 970 units. This situation shows that the
intestinal absorption oꢀ the molecules is diꢃcult. Another
parameter is QPPMDCK, the numerical value oꢀ this param-
eter is expected to be less than 500. but the chi oꢀ 3g, 3h, 3i,
is used ꢀor hydrolysis oꢀ acetylcholine.
It is responsible ꢀor the ꢀormation oꢀ thiocoline and ace-
tate, which are catalysis decomposition products. 5-thio-
2-nitrobenzoic acid, a yellow compound, is ꢀormed as a
result oꢀ the interaction oꢀ DTNB, which is used during
inhibition studies, with thiocoline, one oꢀ the disintegra-
tion products. The color intensity oꢀ the colored compound
3
j, 3k, 3l, and 3m molecules is greater than 500. These drugs
are not suitable ꢀor the brain.
ꢀ
ormed is measured at 412 nm (Ellman et al. 1961). The
absorbance oꢀ the sample and blind cuvettes at 412 nm at
baseline and at the 5th minute is measured.
Conclusions
Determination of the eꢀects of new compounds
on carbonic anhydrase
Biological activities oꢀ 4-amino-chalcone derivatives (3a–o)
against enzymes were compared. Aꢀterwards, ADME/T
analysis oꢀ this molecule was done, and it was theoretically
investigated in the ꢀuture. As a result oꢀ molecular dock-
ing and ADME/T calculations, the biological activities oꢀ
molecules were compared against my enzyme. There is a
great agreement between the experimental results and the
theoretical results. However, the most important reason
Esterase activity method was used to measure the carbonic
anhydrase enzyme activity. The method is based on the ꢀact
that CA has esterase activity. The principle oꢀ the method;
p-nitrophenylasetate oꢀ the carbonic anhydrase enzyme used
as a substrate. It is hydrolysis to p-nitrophenol or p-nitrophe-
nol to give absorption at 348 nm.
ꢀ
or some diferences is that the theoretical calculations are
made in an isolated environment. The results showed that
-amino-chalcone derivatives (3a–o) do not have any side
In this method, both p-nitrophenol and p-nitrophenolate
at 348 nm show the same absorbance. Thereꢀore, phenol or
phenolate ꢀormation does not afect the measurement during
the reaction (Lineweaver and Burk 1934; Verpoorte et al.
1967; Kocyigit et al. 2017; Ökten et al. 2019). Since 348 nm
p nitro phenyl acetate gives little absorption, it blindly.
It is used. In the measurements, an activity determination
procedure was applied by mixing the reaction mixture using
3 mL quartz cuvettes.
4
efects ꢀor human metabolism in the ꢀuture. However, it was
a good guide ꢀor ꢀuture in vivo and in vitro studies. 4-Amino-
chalcone derivatives (3a–o) had potent efects inhibiting CA
enzymes such as AZA. AZA, a well-known CA receptor, is
a positive regulation agent, such as topiramate, zonisamide,
which methazolamide, and has been approved ꢀor epilepsy
treatment and epileptic disorder.
Molecular docking method
Molecules docking is the most common method used to
compare the biological activities oꢀ 4-amino-chalcone
derivatives against enzymes, and there are many parameters
obtained by this method (Ojha et al. 2020; Koçyiğit et al.
Experimental
Measurements
General
2
020; Gezegen et al. 2020). To compare the biological activ-
ities oꢀ molecules, the numerical value oꢀ obtained many
parameters that provide important inꢀormation about mol-
[32]
4
-Amino-chalcone derivatives (3a–o) were resynthesized
ecules , is used. These parameters are used to compare the
biological activities oꢀ molecules. The calculations made ꢀor
this comparison consist oꢀ many stages. First, each molecule
according to published procedures (Scheme 1) (Kocyigit
et al. 2018; Gürdere et al. 2020).
1
3

In Silico Pharmacology
(2021) 9:34
Page 9 of 11
34
is optimized with the Gauss soꢀtware program (Frisch et al.
the carbonic anhydrase I–topiramate complex. Org Biomol
Chem 8(15):3528–3533
2
009) to prepare 4-amino-chalcone derivatives ꢀor calcula-
Bayindir S, Caglayan C, Karaman M, Gülcin İ (2019) The green syn-
thesis and molecular docking oꢀ novel N-substituted rhodanines
as efective inhibitors ꢀor carbonic anhydrase and acetylcho-
linesterase enzymes. Bioorg Chem 90:103096
tions. Maestro Molecular modeling platꢀorm (version 12.2)
by Schrödinger, LLC (Schrodinger 2019) was used ꢀor all
subsequent docking calculations.
Becker HM (2020) Carbonic anhydrase IX and acid transport in can-
cer. Br J Cancer 122(2):157–167
In the next process, it is the preparation oꢀ the studied
enzymes that are human acetylcholinesterase (PDB ID:
Bilgiçli HG, Bilgiçli AT, Günsel A, Tüzün B, Ergön D, Yarasir
MN, Zengin M (2020a) Turn-on ꢂuorescent probe ꢀor Zn2+
ions based on thiazolidine derivative. Appl Organomet Chem
4
M0E), human Carbonic Anhydrase I (PDB ID: 3LXE),
human carbonic anhydrase II (PDB ID: 5AML) in this
study. The protein preparation module (Schrödinger Release
3
4(6):e5624
Bilgiçli HG, Ergön D, Taslimi P, Tüzün B, Kuru İA, Zengin M, Gülçin
İ (2020) Novel propanolamine derivatives attached to 2-metoxiꢀe-
nol moiety: synthesis, characterization, biological properties, and
molecular docking studies. Bioorganic Chem 103969
2019b; Friesner et al. 2006) was used to prepare the calcula-
tions ꢀor enzymes. With this module, the water molecules
in the proteins oꢀ the enzymes were removed and then the
active sites oꢀ the enzymes were determined. All proteins in
this active site are given ꢀreedom oꢀ movement, because in
this way the proteins interact more easily with the 4-amino-
chalcone derivatives. In the next step, the LigPrep module
Çakmak O, Ökten S, Alımlı D, Ersanlı CC, Taslimi P, Koçyiğit ÜM
(
2020) Novel piperazine and morpholine substituted quinolines:
selective synthesis through activation oꢀ 3, 6, 8-tribromoquino-
line, characterization and their some metabolic enzymes inhibition
potentials. J Mol Struct 1220:128666
Çetiner E, Sayin K, Tüzün B, Ataseven H (2021) Could Boron-Con-
taining compounds (BCCs) be efective against SARS-CoV-2 as
anti-viral agent? Bratisl Lek Listy. https://doi.org/10.4149/BLL_
2021_44
(
Schrödinger Release 2019a; Sastry et al. 2013) was used
to prepare 4-amino-chalcone derivatives. In the next step,
calculations were made with the Glide ligand docking mod-
ule (Du et al. 2020) to interact with enzyme proteins with
Cheung J, Gary EN, Shiomi K, Rosenberry TL (2013) Structures oꢀ
human acetylcholinesterase bound to dihydrotanshinone I and
territrem B show peripheral site ꢂexibility. ACS Med Chem Lett
4
-amino-chalcone derivatives. All calculations ꢀor molecular
docking calculations were made using the OPLS3e method.
Finally, a detailed analysis oꢀ ADME / T analysis (absorp-
tion, distribution, metabolism, excretion and toxicity) was
perꢀormed ꢀor 4-amino-chalcone derivatives to be drug
molecules. Many parameters were calculated using the Qik-
prop module (Schrödinger Release, 2020-1) oꢀ the Maestro
Molecular modeling platꢀorm.
4
(11):1091–1096
Douche D, Elmsellem H, Guo L, Haꢀez B, Tüzün B, El Louzi A, et al.
(2020) Anti-corrosion perꢀormance oꢀ 8-hydroxyquinoline deriva-
tives ꢀor mild steel in acidic medium: gravimetric, electrochemi-
cal, DFT and molecular dynamics simulation investigations. J Mol
Liquids 113042
Du Q, Qian Y, Yao X, Xue W (2020) Elucidating the tight-binding
mechanism oꢀ two oral anticoagulants to ꢀactor Xa by using
induced-ꢁt docking and molecular dynamics simulation. J Biomol
Struct Dynamics 38(2):625–633
Supplementary Information The online version contains supplemen-
Ellman GL, Courtney KD, Andres V Jr, Featherstone RM (1961) A
new and rapid colorimetric determination oꢀ acetylcholinesterase
activity. Biochem Pharmacol 7(2):88–95
tary material available at https://doi.org/10.1007/s40203-021-00094-x.
Acknowledgements This work is supported by the Scientiꢁc Research
Project Fund oꢀ Sivas Cumhuriyet University under the project number
RGD-020 and ECZ-079. This research was made possible by TUBI-
TAK ULAKBIM, High Perꢀormance and Grid Computing Center (TR-
Grid e-Inꢀrastructure).
Friesner RA, Murphy RB, Repasky MP, Frye LL, Greenwood JR, Hal-
gren TA et al (2006) Extra precision glide: Docking and scoring
incorporating a model oꢀ hydrophobic enclosure ꢀor protein−
ligand complexes. J Med Chem 49(21):6177–6196
Frisch MJ, Trucks GW, Schlegel HB, Scuseria GE, Robb MA, Cheese-
man JR, Scalmani G, Barone V, Mennucci B, Petersson GA,
Nakatsuji H, Caricato M, Li X, Hratchian HP, Izmaylov AF,
BKoino J, Zheng G, Sonnenberg JL, Hada M, Ehara M, Toyota
K, Fukuda R, Hasegawa J, Ishida M, Nakajima T, Honda Y, Kitao
O, Nakai H, Vreven T, Montgomery JA, Peralta JE, Ogliaro F,
Bearpark M, Heyd JJ, Brothers E, Kudin KN, Staroverov VN,
Kobayashi R, Normand J, Raghavachari K, Rendell A, Burant JC,
Iyengar SS, Tomasi J, Cossi M, Rega N, Millam JM, Klene M,
Knox JE, Cross JB, Bakken V, Adamo C, Jaramillo J, Gomperts
R, Stratmann RE, Yazyev O, Austin AJ, Cammi R, Pomelli C,
Ochterski JW, Martin RL, Morokuma K, Zakrzewski VG, Voth
GA, Salvador P, Dannenberg JJ, Dapprich S, Daniels AD, Farkas
O, Foresman JB, Ortiz JV, Cioslowski J, Fox DJ (2009) Gaussian
Declarations
Conflict of interest The authors declare that there are no conꢂicts oꢀ
interest.
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