Emami et al. BMC Chemistry(2024) 18:125
https://doi.org/10.1186/s13065-024-01230-2

BMC Chemistry

RESEARCH

Open Access

6-Bromo quinazoline derivatives as cytotoxic
agents: design, synthesis, molecular docking
and MD simulation
Leila Emami1, Maryam Hassani2, Pegah Mardaneh2,3, Fateme Zare1, Maryam saeedi2, Mina Emami1,
Soghra Khabnadideh1,2* and Sara Sadeghian2*
Abstract
Based on unselectively, several side effects and drug resistance of available anticancer agents, the development
and research for novel anticancer agents is necessary. In this study, a new series of quinazoline-4(3H)-one
derivatives having a thiol group at position 2 of the quinazoline ring (8a-8 h) were designed and synthesized as
potential anticancer agents. The Chemical structures of all compounds were characterized by 1H-NMR, 13C-NMR,
and Mass spectroscopy. The antiproliferative activity of all derivatives were determined against two cancer cell lines(MCF-7 and SW480) and one normal cell lines (MRC-5) by the MTT method. Cisplatin, Erlotinib and Doxorubicin
were used as positive controls. The results of in vitro screening showed that 8a with an aliphatic linker to SH group
was the most potent compound with IC50 values of 15.85 ± 3.32 and 17.85 ± 0.92 µM against MCF-7 and SW480
cell lines, respectively. 8a indicated significantly better potency compared to Erlotinib in the MCF-7 cell line. The
cytotoxic results obtained from testing compound 8a on the normal cell line, revealing an IC50 value of 84.20 ± 1.72
µM, provide compelling evidence of its selectivity in distinguishing between tumorigenic and non-tumorigenic celllines. Structure–activity relationship indicated that the variation in the anticancer activities of quinazoline-4(3H)-one
derivatives was affected by different substitutions on the SH position. Molecular docking and MD simulation werecarried out for consideration of the binding affinity of compounds against EGFR and EGFR-mutated. The binding
energy of compounds 8a and 8c were calculated at -6.7 and − 5.3 kcal.mol− 1, respectively. Compounds 8a and 8c
were found to establish hydrogen bonds and some other important interactions with key residue. The DFT analysis
was also performed at the B3LYP/6–31 + G(d, p) level for compounds 8a, 8c and Erlotinib. Compound 8a wasthermodynamically more stable than 8c. Also, the calculated theoretical and experimental data for the IR spectrum
were in agreement. The obtained results delineated that the 8a can be considered an appropriate pharmacophore
to develop as an anti-proliferative agent.
Keywords Cancer, Quinazoline-4(3H)-one, MTT, Docking, MD, DFT*Correspondence:
Soghra Khabnadideh
khabns@sums.ac.ir
Sara Sadeghians_sadeghian@sums.ac.ir

1

Pharmaceutical Sciences Research Center, Shiraz University of Medical
Sciences, Shiraz, Iran2
Department of Medicinal Chemistry, Faculty of Pharmacy, Shiraz
University of Medical Sciences, Shiraz, Iran3
Medicinal and Natural Products Chemistry Research Center, Shiraz
University of Medical Sciences, Shiraz, Iran
© The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use,sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) andthe source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in thisarticle are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not includedin the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you willneed to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The
Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available
in this article, unless otherwise stated in a credit line to the data.

Emami et al. BMC Chemistry(2024) 18:125

Introduction
Cancer remains one of the major health problems aroundthe world which is associated with a high mortality rateand is the second cause of death after cardiovascular diseases [1, 2]. Cancer is a group of diseases characterized
by uncontrolled growth and proliferation of abnormal
cells that have the potential to spread to other tissues
throughout the body [3, 4]. Unfortunately, there has beena steady increase in the annual incidence rate of cancer
globally. In 2020, 19.3 million new cases of cancer andapproximately 10.0 million deaths were reported. Furthermore, it is estimated that the number of new cancer
cases might reach 28.4 million in 2040 [5, 6]. This has
raised an urgent need for the development of preventiveor therapeutic strategies. Different approaches including
immunotherapy, hormone therapy, surgery, radiotherapy,
and chemotherapy have been used to treat cancer, among
which chemotherapy is the most common and effective
approach [7, 8]. Chemotherapy agents kill cancer cellsby disrupting the cell cycle by one or multiple pathways,
either by directly inhibiting DNA or RNA synthesis or byaffecting key proteins involved in the cell cycle [7, 9].
A wide range of chemotherapeutic drugs alone or in
combination with other drugs are mainly used for thetreatment of various cancers. Despite the wide range ofanticancer drugs that are available in clinics, several significant challenges including lack of selectivity, inductionof multi-drug resistance and toxic side effects hindered
the efficiency of chemotherapy [3, 10]. Therefore, thediscovery and development of novel anticancer agentsto address the challenges above of chemotherapy areurgently needed. On this basis, substantial efforts have

Fig. 1 FDA-approved quinazoline-based drugs

Page 2 of 18

been made by various researchers to develop novel anticancer agents by incorporating various active functionalities into chemical scaffolds [11–13].
Quinazoline is one of the excellent scaffolds used in
the design and synthesis of new biologically active compounds with diverse biological activities [14–16]. Indeed,quinazoline derivatives have shown various biologicalactivities such as anticancer, antimicrobial, antihypertensive, antihyperlipidemic, anti-inflammatory, and anticonvulsant activities [16–19].
In recent years, several quinazoline based drugs, like
Gefitinib, Erlotinib, and Lapatinib have been approved by
the FDA as anticancer drugs (Fig. 1) [10]. Among various
quinazoline derivatives 2-thioxo-3-substituted quinazolinones and their S-methyl thioether counterparts, as wellas 6-substituted quinazolinone derivatives showed promising anticancer activity [16, 20–22]. According to theliterature, 3-position substituted quinazoline derivatives
can exert their cytotoxic activities through inhibition of
EGFR as an important target in cancer [23–25].
Quinazoline and quinazolinone scaffolds as active
frameworks in the design of anticancer agents have been
the subject of numerous studies [26, 27]. Currently, anumber of quinazoline-based drugs such as Gefitinib,
Erlotinib, Afatinib, etc. have been approved by FDA for
the treatment of cancer. Studies have also shown that thepresence of halogen atom at the 6-position of quinazoline
ring can improve anticancer effects [28, 29]. Therefore, inthe present study, a rational template with quinazoline4-one scaffold as pharmacophoric group was designed
which includes a thio-benzyl moiety at 2-position ofquinazoline ring and a phenyl moiety at 3-position of theEmami et al. BMC Chemistry(2024) 18:125

quinazoline ring to improve the electronic and hydrophobic interactions with the active site of enzyme (Fig. 2).
We also synthesized and evaluated some quinazoline
derivatives as anticancer agents in our previous works[29–32]. In continuation of our work on the synthesis ofquinazoline derivatives, herein, several new quinazoline4(3H)-one derivative (8a-8 h) were synthesized and evaluated for their in vitro cytotoxicity against MCF-7 andSW480 cell lines. Cisplatin, Erlotinib and Doxorubicin
were used as positive controls. Additionally, a molecular docking study and MD simulation were performed to
predict the possible binding mode of these compounds
in the active site of EGFR as the possible receptor. DFT
analysis was also carried out to provide useful information about the reactivity of the molecule in the various
reactions.

Results and discussion
Chemistry

The reaction of anthranilic acid (1) and N-bromosuccinimide (NBS) (2) in acetonitrile afforded compound 3
which was then reacted with phenyl isothiocyanate (4) inethanol to obtain the key intermediate 6-bromo-2-mercapto-3-phenylquinazolin-4(3H)-one (5). The reaction ofintermediate 5 with various alkyl halides or substituted
benzyl bromides in DMF and the presence of K2CO3
resulted in the corresponding quinazoline-4-one derivatives (8a-8 h) in high yields (Fig. 3). The structures of thetarget compounds were identified using 1H-NMR, 13CNMR, and Mass spectroscopy. In the 1H-NMR spectraof the synthesized compounds, the two benzylic hydrogens of compounds 8c-8 h were observed as a singlet
signal at δ = 4.4 ppm with the integration of two protons.
Three protons of methyl group in compounds 8d and 8e
appeared at δ = 2.3 ppm. In the 13C-NMR spectra, carbonyl peak appeared at δ = 160 ppm. Furthermore, thebenzylic CH2 group of compounds 8c-8 h appeared at

Fig. 2 The design strategy of the target compounds

Page 3 of 18

δ = 37 ppm and the methyl group of compounds 8d and8e appeared at δ = 21 ppm. In addition, in the Mass spectra of the synthesized compounds, the molecular ion is inagreement with the molecular weight of the synthesizedcompounds. The details of spectroscopic data are listed
in the supplementary data.
Biological activity

In vitro cytotoxic activity of the novel quinazoline4(3H)-one derivatives were assayed by the MTT standard method. The results are presented in Table 1; Fig. 4.
All compounds represented the less cytotoxic effect on
normal cell (MRC-5) compared to MCF-7 and SW480
cell lines, that indicated appropriate selectivity between
tumorigenic and non-tumorigenic cell lines. These resultsdelineate that compound 8a IC50 (15.85 ± 3.32) against
MCF-7 Cell line is more potent than Erlotinib with IC50
(9.9 ± 0.14). One possible reason for this observation
could be the better accommodation of compound 8a withalkyl ring into EGFR enzyme that is agreement with thebinding energies obtained from docking experiments.
To better explain the structure-activity relationship, thesynthesized compounds were divided into two categories
based on substitutions on SH fragments (aliphatic or aromatic substitutions). Analogs with aliphatic chain (8a-8b)
especially with 4 carbons spacer had strong potency in
MCF-7 and SW480 cell lines and also, increased linker
length leading to a decrease in the activity. The second
category (8c-8 h) containing aryl linker represented thatcompound 8e with methyl substitution on the para position of the phenyl ring had higher potency. As dedicatedin Table 1, the unsubstituted derivative (8c) had the least
activity which revealed that the existence of substitution on the phenyl ring is necessary for antiproliferative
activity.
The presence of electron-donating groups on thephenyl ring (8e) resulted in an improvement in theEmami et al. BMC Chemistry(2024) 18:125

Page 4 of 18

Fig. 3 Synthesis of compounds 8a-8 h. Reagents and conditions: (a) CH3CN, r.t., 2 h; (b) EtOH, Et3N, reflux, 20 h; (c) DMF, K2CO3, reflux, 24 h; (d) DMF,
K2CO3, reflux, 24 h

Emami et al. BMC Chemistry(2024) 18:125

Page 5 of 18

Table 1 Anticancer activity (IC50 ± SD (µM)) of all the synthesized compounds

antiproliferative activity compared to electron-withdrawing groups (8f-8 h). However, (8e) bearing methyl at para
position of phenyl demonstrated better results with an
IC50 value of 35.14 ± 6.87 µM and 63.15 ± 1.63 µM compared to the meta derivative (8d) with an IC50 value of
59.15 ± 5.73 µM and 72.45 ± 2.90 µM, against MCF-7 andSW480 cell lines, respectively. This finding indicatedthat placement of a methyl group at para position of thephenyl moiety increased the potency of the compound
more than meta position. Evaluation of the electron

withdrawing substitutions on the phenyl ring (Br, Cl, andNO2) represented that no significant differences wereobserved among them. In series of 8a-8 h, the aliphatic
chain-containing derivatives (8a and 8b) showed cytotoxicity activity 2–8 folds more than non-substituted
and compounds with aromatic chain. On the other hand,
8a exhibited better potency vs. Cisplatin and Erlotinib
as positive control. It was understood that the aromatic
chain at thiol position is not tolerated. Overall, compounds, containing alkyl linker were more potent thanEmami et al. BMC Chemistry(2024) 18:125

Page 6 of 18

Fig. 4 Cytotoxic activity of all the synthesized compounds against MCF-7, SW480 and MRC-5 cell lines∗, ∗∗, and ∗∗∗ indicate p < 0:05, p < 0:01, andp < 0:001 respectively compared to cisplatin

Fig. 5 Superimposition of Erlotinib in the active site of EGFR (PDB: 1M17): yellow color indicated the redocked model and the purple color illustrated the
crystal orientation

compounds bearing aryl linker and the compounds are
more effective for MCF-7 in comparison with the SW480
cell line, too.
Molecular docking study

Molecular docking study was performed to understandthe binding sites and interactions of the studied ligands
in the active site of the EGFR target as a plausible mechanism. Redocking of the co-crystal ligand (Erlotinib)
was done to evaluate the docking process and RMSD of

docking was found to be 1.78. The result is presented inFig. 5.
The types of interactions and binding poses of Erlotinib
as an internal ligand in the active site of 1M17 protein are
shown in Fig. 6. Hydrogen bond interactions with the residues of Cys773, and Met769 and also π-sigma interactions with Leu820 and Leu694 were observed. Some alkyl
and π-alkyl interactions with Val702, Ala719, Leu764,
Lys721, and Met742 were also, seen in Fig. 6.

Emami et al. BMC Chemistry(2024) 18:125

Page 7 of 18

Fig. 6 Interactions and orientation of Erlotinib in the active site of 1M17 (green: hydrogen bond, light green: van der Waals, purple: pi-pi, pink: pi-alkyl)

As it is shown in, Figs. 7 and 8, based on docking
results, common interactions such as π - π or π -alkyl andhydrophobic interactions with Thr 766, Thr 830, Leu 764,
Phe 699, Val 702, Gly 772, Ala 719, Leu 694 and Leu 820
were seen in all compounds which had benzyl moiety atthio group (8c-8 h), showed different interaction depend
on substitution on benzyl ring. Compounds 8e and 8f,
which had methyl and chlorine substitution at para position on benzyl moiety, they interacted through similarhydrogen bond interaction between carbonyl group andLys 721 and π-anion interaction was also, seen. On theother hand, compounds 8a and 8b with alkyl chain werestabilized by hydrogen interactions between the thiogroup of the alkyl ring and Lys 721 residue and anotherπ-sulfur and halogen bond were observed. These common and extra strong and desirable interactions with
EGFR target made the 8a and 8b as potential inhibitorsof EGFR target among all the tested compounds. Aminoacid residues Lys 721, Leu 694, Leu 820, Met 742, Cys 773
are very important for the active conformation of Erlotinib. Compounds 8a and 8b also showed desirable interactions with the same residue similar to Erlotinib. This
results indicate that these two compounds are well targeted the EGFR active sites and showed more inhibitory
effects than the other compounds.
The Reports showed that mutation in EGFR increased
the activity of EGFR and as a result caused a variety ofcancers and resistance to EGFR inhibitor drugs, such as
Erlotinib and Gefitinib. Therefore, it is necessary to consider the binding affinity of synthesized EGFR inhibitor
compounds towards mutated EGFR. The interactionsof the potent EGFR inhibitor, 8a, and co-crystal ligand(Neratinib) of EGFR-mutated with the pdb code of 3w2q
are displayed in Fig. 9. The total binding energy values

for co-crystal ligand and 8a were calculated at -7.1 and− 6.8 kcal.mol-1, respectively.
The Naritinib surrendered by residues of Leu 718, Met
793, Leu 844, Ala 743, Met 790, Val 726, Lys 745, Glu 762,
Leu 788, Mt 766, Ala 763 and Asp 855. The key interactions of Naritinib were hydrogen bonding with residuesof Met 793 and Lys 745 and also π contacts with Glu
762 and mutant gatekeeper residue, Met 790. Moleculardocking results of the mutated-EGFR showed a well-fit
binding pattern of compound 8a with several key residues in the active site (Fig. 9). Quinazolinone ring interacted through the π interactions with Lys 745, Met 790(the critical residues of the active site) whereas, the phenyl moiety formed π interaction with Asp 855 [33]. As
can be seen, compound 8a interacted with the Met 790,
mutant gatekeeper residue as π-sulfur interaction similar
to Neratinib [33].
The results indicated that the selected compound was
located in the binding pocket EGFR-mutated. Consequently, the compound 8a can inhibit both EGFR and
EGFR-mutated.
No similar residues were showed in both protein so,
perhaps mutant protein can change the binding pocket.
The placement position of compound 8a in the active siteof EGFR-mutated (10a) and EGFR protein kinase wasshown in Fig. 10.
Molecular dynamics simulation

Another effective approach to studying the structuralmodification of biomedicine molecules and intermolecular interaction patterns is molecular dynamic (MD)simulation [34]. In this study, MD simulation was conducted over a 100 ns period, exploring interactions
between the atoms of compounds (8a, 8c) and the native

Emami et al. BMC Chemistry(2024) 18:125

Page 8 of 18

Fig. 7 Interactions and orientation of the compounds 8a-8d in the active site of 1M17 (green: hydrogen bond, light green: van der Waals, blue: halogen
bond, orange: π-sulfur, purple: π-π, pink: π -alkyl)

ligand Erlotinib with the EGFR receptor (1M17), as wellas compound 8a with EGFR-mutated (3W2Q). The main
purpose of the simulation was to assess the conformational stability of the enzyme. Conformational stability was evaluated by calculating the Root-Mean-Square
Deviation (RMSD) values, which measure the deviationof atomic positions in different frames of the simulationcompared to a reference frame. The alignment was performed by comparing the backbone atoms of each frameto those of the first frame in the simulation. Based on theregular RMSD profile, compound 8a, 8c, and Erlotinib
achieved the equilibrium phase at approximately 30, 20,
and 30 ns of the simulation, respectively (Fig. 11). Additionally, compound 8a attained the equilibrium phaseat 25 ns (Fig. 15). This indicates that the ligands havesettled into stable positions within the active site of the
EGFR and EGFR-mutated kinase protein. The regularityof the RMSD profile suggests that the ligands remained

relatively stable throughout the simulation. Furthermore, the RMSD values for both compounds 8a and 8c,
as well as Erlotinib with 1M17, and compound 8a withthe 3W2Q protein (presumably a reference structure),
demonstrate significant stability. Additionally, the RMSD
values of both compound 8a and 8c and Erlotinib with
1M17 protein (presumably a reference structure) aresignificantly stable. This implies that the ligands maintain a consistent conformation and interaction with the
EGFR kinase receptor throughout the simulation. The
root mean mean square deviation (RMSD) of 8a and 8ccompared to the original ligand (Erlotinib) when interacting with 1M17 was 0.7 and 1.03 Å, while for the native
ligand, it was 0.9 Å throughout the simulation periods.
The results indicate that ligand 8a demonstrates betterstability within the protein’s active site compared to thenative ligand. In contrast, ligand 8c exhibits less stability
in comparison to Erlotinib, the native ligand [35].

Emami et al. BMC Chemistry(2024) 18:125

Page 9 of 18

Fig. 8 Interactions and orientation of the compounds 8e-8 h in the active site of 1M17 (green: hydrogen bond, light green: van der Waals, purple: π- π,pink: π-alkyl)

Fig. 9 2D interactions and orientations of Naritinib (a) and compound 8a (b) in the active site of EGFR-mutated (pdb code: 3w2q) (green: hydrogenbond, light green: van der Waals, purple: π - π, pink: pi-alkyl, orange: π -cation)

Based on the analysis of the root mean square fluctuation (RMSF) of the backbone, the flexibility of EGFR and
EGFR-mutated protein was investigated to ligand binding
throughout the simulation (Figs. 12 and 15). The findings

indicate that complexes with 8a, 8c, and with the native
ligand (Erlotinib) exhibited a comparable distributionof RMSF values. This suggests that the binding of either
ligand does not significantly alter the overall flexibility ofEmami et al. BMC Chemistry(2024) 18:125

Page 10 of 18

Fig. 10 The orientations of compound 8a in the in the active site of EGFR-mutated (pdb code: 3w2q) (a) and EGFR (1M17) (b)Fig. 11 RMSD evaluation of a complex involving compound 8a (shown in violet), 8c (shown in green), and Erlotinib (shown in red) with EGFR kinase
enzymeboth the proteins. Furthermore, it was observed that noresidues within the active site of the protein kinase displayed an RMSF value greater than 0.3 nm. This impliedthat the binding of the ligands did not induce any substantial fluctuations or changes in the flexibility of theresidues within the active site. The active site remained
relatively stable throughout the simulation. The RMSF
values suggest that the interaction of 8a and 8c with theprotein induces flexibility across all regions, similar to theeffect observed with the native co-crystal ligand during
a 100 ns simulation. The RMSF plot supports the lackof structural changes in 1M17 upon binding 8a and 8ccompounds.
The parameter known as the radius of gyration (Rg) isused to assess the compactness changes in a ligand-protein complex [36]. The analysis revealed that the protein

accumulated during the simulation time, indicating somecompression of the protein. The Rg plot shows that theErlotinib, compound 8a and 8c complexes have the sameRg platform, which was calculated as 2.06, 2.11, and 2.05
respectively, with the EGFR protein. The Rg values for
all complexes exhibit minimal variation and maintaina low average Rg value throughout a 100 ns simulation.
This consistency serves as a reliable indicator of theirremarkable stability and compactness within the system.
Additionally, the 8a compound demonstrated a rangeof hydrogen bonds between 0 and 3 with the EGFRmutated protein. Studying hydrogen bonding during
MD simulations offers valuable insights into the interactions between molecules in ligand-enzyme systems. This
analysis can greatly contribute to our comprehension ofFig. 12 RMSF values of the complexes between 8a (in violet), 8c (in green) and Erlotinib (in red) in the simulation time (100 ns)

Emami et al. BMC Chemistry(2024) 18:125

Page 11 of 18

Overall, compound 8a showed greater MD simulation
items such RMSD, Rg, RMSF and hydrogen bond interaction compared to 8c which confirmed higher inhibitory activity for 8a.
DFT analysis

Fig. 13 The radius of gyration (Rg) for EGFR receptors in complex with
Erlotinib, 8a, and 8c during the simulation timebinding strength and the stability of molecular structures(Fig. 13).
Hydrogen bonding is crucial in determining thestrength of the interaction between the ligand andenzyme. To evaluate the stability of the ligand’s conformation, the number of hydrogen bonds was determinedthroughout the MD simulation (Fig. 14). The investigation revealed that compound 8a, 8c, and the native ligand
exhibited a range of hydrogen bonds between 0 and 2,
0 and 1, and 0 and 3, respectively. Studying hydrogenbonding during MD simulations offers valuable insights
into the interactions between molecules in ligandenzyme systems. This analysis can greatly contribute to
our comprehension of binding strength and the stabilityof molecular structures. In general, the examination ofhydrogen bond interactions between erlotinib, 8a, and 8cligands and 1M17 was computed and depicted in Fig. 14.
Throughout the 100 ns simulation, 8c displayed minimal
interactions with residues in the active site compared to
the reference molecule (Erlotinib). On the other hand,the 8a compound exhibited greater stability with an
RMSD of 0.2 Å.
To evaluate the reproducibility of MD simulation, we
performed two replica simulations on 8a compound [37].
The results are justifiable and almost adaptable, suggesting that the findings have a reasonable foundation while
also hinting at the potential for flexibility in their application (Fig. 16).

Fig. 14 The total number of hydrogen bonds of Erlotinib, compound 8aand 8cThe measurement of the electronic parameters of a molecule can provide useful information about the behaviorof the molecule in various types of reactions and also justify the results obtained from its biological activity [38].
Density functional theory (DFT) for 8a, and 8c as themost active and less active compounds according to the
biological results and Erlotinib as reference compound
was carried out with Gaussian 09 at the B3LYP/6–31 + G(d, p) level of theory. B3LYP is a hybrid density functional
method that uses the Becke three-parameter exchange
functional and the Lee-Yang-Parr correlation functional.
It is a very popular and accurate method for quantum
chemical calculations, especially for molecules with N, O,
C, and H atoms. B3LYP is a GGA method, which means
it includes both exact exchange and GGA corrections inaddition to LDA electron-electron and electron-nuclei
energy. The exact exchange term is manually adjusted
to enforce the Pauli Exclusion Principle, while the GGA
corrections are used to account for various effects suchas dispersion, polarization, and relativistic effects. The
6–31 + G (d, p) basis set is a split-valence basis set that
includes polarization functions, which are important for
accurately describing the electronic structure of molecules [39–41]. The molecular orbitals, HOMO and
LUMO, and their energies are shown in Fig. 16. The
HOMO for both compounds 8a and 8c were located on
two rings such as bromobenzene and quinazolinone, andsulfur atom while The LUMO distributed the charges
throughout the molecule for both. As can be observed inFig. 17, the HOMO and LUMO orbitals for Erlotinib arelocated in the over this molecule. The energy gap between
HOMO and LUMO is a suitable measure to determine
the reactivity or kinetic stability of compounds, therefore a compound with a higher energy gap is more stable
and less reactive. The energy gaps between HOMO andLUMO were calculated at 4.71 and 4.57 and 4.18 ev for
8a, 8c, and Erlotinib respectively which indicates thatcompound 8a is more stable than 8c and Erlotinib.
The electrostatic potential (ESP) map is a tool to identify the active sites of the compound for electrophilic
and nucleophilic attacks. The ESP maps for compounds
8a and 8c are given in Fig. 18. The areas with negativeelectrostatic potential are indicated by red and yellow
spheres, which are preferred sites for electrophilic attack,while blue and green areas have positive electrostatic
potential and are suitable sites for nucleophilic attack.
As can be seen, in the compounds, the oxygen atomand the carbonyl group are in the red region, so they areEmami et al. BMC Chemistry(2024) 18:125

Page 12 of 18

Fig. 15 The Molecular dynamics analysis and interactions of compound 8a with EGFR-mutated for 100 ns. (A) Assessment of the Root-Mean-Square Deviation (RMSD) for the complex consisting of 8a in conjunction with the EGFR-mutated kinase (PDB: 3W2Q). (B) The RMSF values for the complex formedby the 8a molecule were analyzed. (C) Rg plot for 8a ligand in complex with EGFR-mutated during the simulation. (D) The total number of hydrogenbonds of 8a complex

interested in electrophilic attack. Also, the placement of
nitrogen in blue areas indicates a more positive charge

on nitrogen, which makes it susceptible to nucleophilic
attack.

Fig. 16 The RMSD (A), RMSF (B), Rg (C), and H-bonds (D) values are calculated for both ligand 8a and its repetitions

Emami et al. BMC Chemistry(2024) 18:125

Page 13 of 18

Fig. 17 DFT calculated LUMO, HOMO, and their energies for (a) 8a (left) and 8c (right) and (b) Erlotinib, at the B3LYP/6–31 + G (d, p) level of theory

Fig. 18 Electrostatic potential maps for 8a, 8c and Erlotinib at B3LYP/6–31 + G (d, p) level of theory

Emami et al. BMC Chemistry(2024) 18:125

Page 14 of 18

Table 2 The chemical reactivity indices of 8a and 8c at B3LYP/6–31 + G (d, p) level of theory
Entry
8a
8c
Erlotiniba

Etota-3946.24-3833.11-1309.54

Ha-3946.24-3833.11-1309.54Ga-3946.32-3833.16-1309.63

Sb
162.181122.36184.46’

ɳc
2.35
2.28
2.092

σd
0.21
0.22
0.24

Ac
1.41
1.57
1.48

in Hartree/particle. bin cal/mol.K. cin ev. din ev− 1

Fig. 19 Calculated IR spectra for 8a (red), 8c (yellow) and Erlotinib (blue), at B3LYP/6–31 + G(d, p) level of theory

The chemical reactivity indices for 8a and 8c are shownin Table 2. According to this Table, the calculated valuesof total energy (E), enthalpy (H), and Gibbs free energy(G) showed that 8a was more thermodynamically stable
than 8c. Hardness and softness parameters are obtained
from HOMO and LUMO energies, as a result, compounds with a higher energy gap between HOMO andLUMO have high hardness and kinetic stability. The
results shown in Table 2 stated that compound 8a had agreater hardness value compared to compounds 8c and
Erlotinib, therefore it is more stable and less reactive.
The theoretical analysis of the IR spectrum for 8a, 8c,
and Erlotinib was computed by B3LYP/6–31 + G (d, p)level of theory. is indicated in Fig. 19. The IR peaks of
C = C, C = N, C = O, C-N, and C-H aliphatic and aromatic
excellently agree with the experimental results.

Material and method
Chemistry

All commercially available materials and solvents from
Merck and Sigma-Aldrich Chemical Co. were used without further purification. Thin-layer chromatography(TLC) was performed on Merck silica gel (60 F254, Merck)
plates to monitor the progress of the reactions. Melting points were determined on an electrothermal 9200
instrument and were uncorrected. The 1H-NMR and 13CNMR spectra were recorded in CDCl3 on a Bruker 500
and 125 MHz spectrometer, respectively. Chemical shifts(δ) are reported in ppm scale toward tetramethylsilane(TMS) as an internal standard. Mass analyses were carried out with an Agilent Technologies (HP) mass spectrometer operating at an ionization potential of 70 eV.

Synthesis of 5-bromoanthranilic acid (3)Anthranilic acid (1) (2.74 g, 20 mmol) was dissolved in 20
mL acetonitrile. Then, a solution of N-bromosuccinimide(2) (3.73 g, 21 mmol) in the acetonitrile (30 mL) was
added dropwise to the reaction mixture. The resulting
mixture was stirred at room temperature for 2 h. Finally,
the obtained precipitate was filtered, washed with acetonitrile, and dried at room temperature to afford 5-bromoanthranilic acid (3) with a 61.85% yield.
Synthesis of 6-bromo-2-mercapto-3-phenylquinazolin4(3H)-one (5)A mixture of 5-bromoanthranilic acid (3) (2.16 g, 10mmol), phenyl isothiocyanate (4) (1.8 mL, 15 mmol), andtriethylamine (2 mL) in absolute ethanol (30 mL) was
refluxed at 65 ˚C for 20 h. After completion of the reaction (checked by TLC), the reaction mixture was filteredand the obtained residue was recrystallized with ethanol to give the key intermediate 6-bromo-2-mercapto3-phenylquinazolin-4(3H)-one (5) in 83.2% yield.
Synthesis of compounds 8a-8 h

For the preparation of compounds 8a-8 h, intermediate 5(1 mmol) was dissolved in 15 mL DMF, and then, K2CO3(1.2 mmol) was added over 5 min to it. In the following,
various alkyl halides (1.5 mmol) or substituted benzyl
bromides (1.5 mmol) were added to the reaction mixture.
The resulting mixture was refluxed for 24 h. After completion of the reaction (checked by TLC), the reaction
mixture was poured onto crushed ice and the obtained
precipitate was filtered, dried, and recrystallized with

Emami et al. BMC Chemistry(2024) 18:125

ethanol to obtain the quinazoline-4-one derivatives
8a-8 h in 68.5–92.3% yields [21, 22].
1) 1) 6-bromo-2-(butylthio)-3-phenylquinazolin4(3H)-one (8a)White solid; yield: 74.6%, mp: 128–130 ˚C, 1H-NMR(500 MHz, CDCl3) δ (ppm): 8.37 (d, J = 2.0 Hz, 1H, quinazoline), 7.82 (dd, J = 8.5 Hz, J = 2.5 Hz, 1H, quinazoline),
7.58–7.56 (m, 3 H, phenyl), 7.51 (d, J = 8.5 Hz, 1H, quinazoline), 7.33–7.31 (m, 2 H, phenyl), 3.17 (t, J = 7.5 Hz, 2 H,
CH2), 1.71–1.65 (m, 2 H, CH2), 1.48–1.41 (m, 2 H, CH2),
0.96 (t, J = 7.5 Hz, 3 H, CH3).13C-NMR (125 MHz, CDCl3)
δ (ppm): 160.71, 158.48, 146.72, 137.66, 135.79, 130.01,
129.72, 129.65, 129.04, 128.07, 121.25, 118.78, 32.37,
30.64, 22.04, 13.65. MS: m/z (%): 390.1 [M+], 333.0 (100),301.0 (21.95), 252.1 (22.99), 224.0 (16.23), 192.1 (26.88),
170.0 (33.48), 133.1 (21.13), 91.1 (30.40), 77.1 (59.09),
51.1 (15.61).
2) 6-bromo-2-(pentylthio)-3-phenylquinazolin4(3H)-one (8b)White solid; yield: 68.5%, mp: 99–101 ˚C, 1H-NMR(500 MHz, CDCl3) δ (ppm): 8.37 (d, J = 2.5 Hz, 1H, quinazoline), 7.82 (dd, J = 8.5 Hz, J = 2.0 Hz, 1H, quinazoline),
7.58–7.57 (m, 3 H, phenyl), 7.51 (d, J = 8.5 Hz, 1H, quinazoline), 7.33–7.31 (m, 2 H, phenyl), 3.16 (t, J = 7.5 Hz, 2 H,
CH2), 1.73–1.68 (m, 2 H, CH2), 1.43–1.33 (m, 4 H, CH2CH2), 0.93 (t, J = 7.0 Hz, 3 H, CH3). 13C-NMR (125 MHz,CDCl3) δ (ppm): 160.70, 158.49, 146.74, 137.65, 135.80,
130.01, 129.72, 129.65, 129.05, 128.08, 121.26, 118.77,
32.64, 31.07, 28.26, 22.18, 13.93. MS: m/z (%): 404.2[M+] (13.40), 357.2 (19.25), 333.1(100), 301.1 (23.97),
272.1 (3.30), 252.2 (18.46), 224.2 (11.04), 192.2 (16.29),
170.1 (17.95), 133.1 (7.94), 104.2 (3.28), 91.2 (10.66), 77.2
(17.35), 51.2 (2.34).
3) 2-(benzylthio)-6-bromo-3-phenylquinazolin4(3H)-one (8c)White solid; yield: 81.9%, mp: 140–142 ˚C, 1H-NMR(500 MHz, CDCl3) δ (ppm): 8.39 (d, J = 2.0 Hz, 1H, quinazoline), 7.85 (dd, J = 9.0 Hz, J = 2.0 Hz, 1H, quinazoline),
7.59–7.54 (m, 5 H, phenyl), 7.40 (d, J = 7.0 Hz, 2 H, benzyl), 7.33–7.32 (m, 3 H, benzyl), 7.28 (d, J = 7.0 Hz, 1H,
quinazoline), 4.43 (s, 2 H, CH2).13C-NMR (125 MHz,CDCl3) δ (ppm): 160.63, 157.92, 146.63, 137.78, 136.12,
135.51, 130.14, 129.79, 129.74, 129.38, 129.05, 128.59,
128.07, 127.59, 121.36, 119.03, 37.23. MS: m/z (%): 424.1[M+] (14.18), 391.1 (31.87), 333.1 (14.17), 313.1 (26.29),
274.1 (3.25), 252.1 (26.32), 224.1 (12.10), 197.1 (12.48),
167.2 (100), 133.1 (10.38), 91.2 (72.98), 65.2 (14.09).

Page 15 of 184) 6-bromo-2-((3-methylbenzyl)thio)-3phenylquinazolin-4(3H)-one (8d)White solid; yield: 78.5%, mp: 101–103 ˚C, 1H-NMR(500 MHz, CDCl3) δ (ppm): 8.36 (s, 1H, quinazoline),
7.83 (d, J = 9.0 Hz, 1H, quinazoline), 7.56 (d, J = 8.5 Hz,
1H, quinazoline), 7.53–7.52 (m, 3 H, phenyl), 7.31–7.29(m, 2 H, phenyl), 7.17 (br, 3 H, benzyl), 7.06 (d, J = 5.0 Hz,
1H, benzyl), 4.38 (s, 2 H, CH2), 2.31 (s, 3 H, CH3).13CNMR (125 MHz, CDCl3) δ (ppm): 160.60, 158.04, 146.60,
138.26, 137.75, 135.77, 135.49, 130.13, 130.09, 129.75,
129.71, 129.02, 128.48, 128.37, 128.01, 126.42, 121.32,
118.99, 37.28, 21.34. MS: m/z (%): 438.1 [M+] (13.09),
403.1 (37.90), 346.9 (5.56), 329.0 (24.16), 313.0 (8.22),
272.0 (3.78), 252.1 (28.42), 224.1 (11.63), 197.0 (11.46),
181.1 (96.56), 165.1 (14.63), 133.0 (15.89), 105.1 (100),77.1 (47.74), 51.1 (11.00).
5) 6-bromo-2-((4-methylbenzyl)thio)-3phenylquinazolin-4(3H)-one (8e)White solid; yield: 79.4%, mp: 164–166 ˚C, 1H-NMR(500 MHz, CDCl3) δ (ppm): 8.39 (d, J = 2.0 Hz, 1H, quinazoline), 7.85 (dd, J = 8.5 Hz, J = 2.0 Hz, 1H, quinazoline),
7.58 (d, J = 8.5 Hz, 1H, quinazoline), 7.55–7.54 (m, 3 H,
phenyl), 7.32–7.31 (m, 2 H, phenyl), 7.29 (d, J = 8.0 Hz,
2 H, benzyl), 7.12 (d, J = 8.0 Hz, 2 H, benzyl), 4.40 (s, 2 H,
CH2), 2.34 (s, 3 H, CH3).13C-NMR (125 MHz, CDCl3) δ(ppm): 160.64, 158.07, 146.66, 137.76, 137.38, 135.53,
132.93, 130.10, 129.76, 129.72, 129.29, 129.04, 128.07,
121.35, 118.98, 37.04, 21.15. MS: m/z (%): 438.1 [M+](22.25), 403.2 (40.20), 327.1 (29.40), 301.1 (6.68), 274.1(2.65), 252.1 (27.11), 224.1 (12.05), 181.2 (100), 133.1(7.89), 105.2 (73.17), 77.1 (18.72), 51.1 (3.13).
6) 6-bromo-2-((4-chlorobenzyl) thio)-3phenylquinazolin-4(3H)-one (8f)White solid; yield: 83.1%, mp: 145–147 ˚C, 1H-NMR(500 MHz, CDCl3) δ (ppm): 8.38 (d, J = 2.0 Hz, 1H, quinazoline), 7.85 (dd, J = 8.5 Hz, J = 2.0 Hz, 1H, quinazoline),
7.56–7.55 (m, 4 H, quinazoline (1H) + phenyl (3 H)),
7.34–7.30 (m, 4 H, benzyl), 7.27 (d, J = 8.5 Hz, 2 H, phenyl), 4.37 (s, 2 H, CH2).13C-NMR (125 MHz, CDCl3) δ(ppm): 160.55, 157.51, 146.51, 137.83, 135.41, 134.93,
133.42, 130.68, 130.21, 129.81, 129.77, 129.03, 128.71,
127.99, 121.35, 119.17, 36.34. MS: m/z (%): 458.2 [M+](33.12), 425.2 (50.05), 388.2 (2.50), 367.1 (4.35), 333.1(21.56), 313.2 (14.87), 272.1 (5.20), 252.2 (38.18), 224.2(17.28), 201.2 (100), 165.2 (15.82), 125.1 (92.49), 89.1
(24.52), 63.1 (10.47).
7) 6-bromo-2-((4-bromobenzyl) thio)-3phenylquinazolin-4(3H)-one (8 g)Emami et al. BMC Chemistry(2024) 18:125

White solid; yield: 92.3%, mp: 175–177 ˚C, 1H-NMR(500 MHz, CDCl3) δ (ppm): 8.35 (s, 1H, quinazoline),
7.82 (d, J = 8.5 Hz, 1H, quinazoline), 7.53 (br, 4 H, quinazoline (1H) + phenyl (3 H)), 7.40 (d, J = 8.0 Hz, 2 H, phenyl), 7.28–7.27 (m, 2 H, benzyl), 7.25 (d, J = 8.5 Hz, 2 H,
benzyl), 4.32 (s, 2 H, CH2).13C-NMR (125 MHz, CDCl3)
δ (ppm): 160.51, 157.44, 146.47, 137.80, 135.45, 135.37,
131.63, 130.99, 130.19, 129.78, 129.74, 129.00, 127.95,
121.49, 121.32, 119.14, 36.34. MS: m/z (%): 502.1 [M+](58.74), 469.1 (87.76), 411.0 (9.19), 388.2 (6.36), 314.2(44.69), 272.1 (8.21), 245.2 (100), 223.2 (11.03), 199.1(25.71), 197.1 (25.26), 169.1 (71.72), 133.1 (18.06), 90.2
(37.94), 63.2 (9.51).
8) 6-bromo-2-((4-nitrobenzyl) thio)-3phenylquinazolin-4(3H)-one (8 h)Yellow solid; yield: 87.9%, mp: 231–233 ˚C, 1H-NMR(500 MHz, CDCl3) δ (ppm): 8.37 (d, J = 2.0 Hz, 1H,
quinazoline), 8.17 (d, J = 9.0 Hz, 2 H, benzyl), 7.86 (dd,J = 8.5 Hz, J = 2.5 Hz, 1H, quinazoline), 7.60–7.56 (m, 5 H,
phenyl), 7.54 (d, J = 9.0 Hz, 1H, quinazoline), 7.33–7.30(m, 2 H, benzyl), 4.46 (s, 2 H, CH2).13C-NMR (125 MHz,CDCl3) δ (ppm): 160.45, 156.76, 146.33, 144.56, 137.94,
135.25, 130.36, 130.11, 129.88, 129.84, 129.01, 127.90,
127.49, 124.27, 123.72, 121.36, 119.42, 35.97. MS: m/z(%): 469.1 [M+] (19.56), 436.1 (17.33), 378.0 (3.34), 333.0(30.15), 301.1 (19.81), 274.1 (8.20), 252.1 (39.01), 212.1(100), 199.0 (32.00), 197.0 (34.45), 170.0 (25.17), 133.1(19.18), 106.1 (9.06), 77.1 (44.57), 51.2 (11.70).
Cytotoxic assay

The antiproliferative activity of all the synthesizedcompounds (8a-8 h) was done by MTT (standard
3-(4,5-dimethylthiazol-yl)-2,5-diphenyl-tetrazolium bromide) assay according to our previous protocols [31, 42].
MCF-7 (breast carcinoma) and SW480 (Colorectal carcinoma) cell lines were choose [29, 43–45] and purchasedfrom the National Cell Bank of Iran (NCBI, Pasteur Institute, Tehran, Iran). RPMI 1640 culture media with 10%
fetal bovine serum (FBS) and 1% penicillin-streptomycin(Gibco, USA) were applied as culture. Trypsin/EDTA
0.5% solution (Gibco/USA) was used to harvest cells andthen, the cells were seeded at a density of 1 × 104 cells perwell in 96-well microplates [46]. Five different concentrations of the derivatives and Cisplatin as the positive control (1 to 200 µM) were used for treatment in triplicate
times. Three untreated wells were applied as the negative control. After 72 h, the media was changed by 100µL fresh MTT solution and incubated for 4 h at 37 °C in
the incubator to achieve formazan purple crystals [47].
Finally, the media was removed and 150 µL of DMSO
was added and incubated at 37 °C in the dark for 10 min
to dissolve the crystals. A Microplate ELISA reader wasPage 16 of 18

applied to read the absorbance of individual wells at
490 nm. Analysis of the data was obtained by Excel 2016
and Curve Expert 1.4. The data was expressed as themean ± SD for each analysis. GraphPad Prism software
was used to perform one-way ANOVA statistical analyses, followed by Tukey’s multiple comparison test.
Molecular docking study

The crystal structure of the EGFR target was downloaded
from the RCSB protein data bank site (PDB ID: 1M17)[48]. AutoDock Vina was used to run the molecular docking procedure. The structure of compounds was minimized in terms of energy and converted to pdbqt format.
A grid box of 70 × 70 × 70 Å and an exhaustiveness of 100
were set for docking analysis. To visualize the interaction
and orientation of the compounds, the Discovery Studio
2016 client was used.
Molecular dynamic simulation

The Gromacs molecular dynamics package on a Centos
Linux server equipped with GPUs was used to performthe molecular dynamics simulation of compounds 8a, 8c
and native ligand (Erlotinib) in complex with EGFR protein kinase (Pdb: 1M17) ), as well as the 8a compoundwith EGFR-mutated protein (3W2Q). The Amber99sb
force field was employed to define the atom types andsimulate the dynamics of the system. ACPYPE webserver
was used to generate the compounds 8a, 8c and native
ligand topology parameters. An octahedral box was
defined around the solute (compounds 8a, 8c and Erlotinib) in a complex with EGFR and EGFR-mutated kinase.
TIP3P water molecules were added to solvate the protein
complex model, ensuring it was surrounded by water.
The appropriate number of Na+ ions replaced water molecules to ensure the system neutralization. The system
was subjected to NVT heating. Restraints were applied tothe solute (compounds 8a, 8c and Erlotinib) to prevent
large conformational changes during the initial equilibration phase. The particle-mesh Ewald (PME) algorithm
was used to efficiently handle long-range electrostaticinteractions. The system’s pressure was stabilized at an
average pressure of 1 atm. This step ensures that the density of water molecules reaches an equilibrium state during the equilibration phase. The system was equilibrated
under the NPT ensemble (constant Number of particles,
Pressure, and Temperature) for 500 ps. In general, molecular dynamics simulations require to be adequately long
to be able to draw reliable conclusions. The production
MD run was performed during 100 ns to obtain the best
equilibration point of the system with favorable temperature and pressure. After the MD run was completed,
the trajectory (the sequence of atomic coordinates overtime) was corrected for periodic boundary conditions.
To determine the equilibrium time range within the MD

Emami et al. BMC Chemistry(2024) 18:125

simulation, the root-mean-square deviation (RMSD) was
considered. RMSD measures the deviation of atomic
positions in each snapshot of the trajectory from a reference frame, typically the starting structure [49].
DFT analysis

Density functional theory was used to investigate thereactivity descriptors of the compounds with the highest(8a) and lowest (8c) biological activity at the B3LYP/6–
31 + G (d, p) level of theory. The molecular orbitals(HOMO and LUMO), electrostatic surface potential
energy, and thermochemical parameters were also studied in detail.

Conclusion
In this study, a series of quinazoline-4(3H)-one derivatives were synthesized and evaluated as cytotoxic agents.
The SARs were presented by analyzing the impact ofvarying substitutions on SH and phenyl fragments.
6-bromo-2-(butylthio)-3-phenylquinazolin-4(3H)-one(8a) displayed the best cytotoxic activity and is about 2times more potent than Erlotinib in MCF-7 cell line.
Cytotoxic results on normal cell line showed that all of
the compounds had appropriate selectivity between
tumorigenic and non-tumorigenic cell lines. Molecular docking studies against EGFR and EGFR-mutatedwere applied to determine the binding conformationof all derivatives. Based on these values, it can be mentioned that compound 8a was located in the binding
pocket of EGFR and EGFR-mutated and bonded strongly
to both receptors. The DFT analysis was performed formore active and less active compounds. The reactivity descriptors, the energy gap between HOMO andLUMO revealed that 8a is more stable than 8c. Also, agood agreement between theoretical and experimental
IR spectra was observed. Finally, the outcomes obtained
from the MD simulation, which encompassed parameters such as RMSD, RMSF, Rg, and the count of hydrogen bonds demonstrated the stability of compound 8a, 8c
and Erlotinib complexes within the active site of EGFR
and EGFR-mutated enzymes throughout the entire simulation duration.
Abbreviations
EGFR	Epidermal Growth Factor Receptor
MDR	Multidrug Resistance
ADME	Adsorption, Distribution, Metabolism and Discretion
Rg	Radius of gyration
RMSD	Root Mean Square Deviation
RMSF	Root Mean Square Fluctuation
TPSA	Total polar surface area
TEA	Triethylamine
TK	Tyrosine kinase
MTT	3-(4, 5 dimethylthiazol-yl)-2,5-diphenyl-tetrazolium bromide

Page 17 of 18

Supplementary Information

The online version contains supplementary material available at https://doi.
org/10.1186/s13065-024-01230-2.
Supplementary Material 1
Acknowledgements
Not applicable.
Author contributions
L.E. prepare the manuscript, and supervise the study. M. H. contributed to thesynthesis of compounds. P. M. performed and written the simulation section.
F. Z. performed and written DFT section. M. S. performed the Docking section.
M. E. performed the biological assay. S. Kh contributed to the preparation of
the manuscript. S. S. edit the manuscript and supervise the study. All authors
read and approved the final manuscript.
Funding
Financial assistance from the Shiraz University of Medical Sciences by way ofgrant number 28186 is gratefully acknowledged.
Data availability
The data sets used and analyzed during the current study are available fromthe corresponding author on reasonable request. We have presented all datain the form of Tables and Figure. The PDB code (1M17) was retrieved fromprotein data bank (www.rcsb.org). https://www.rcsb.org/structure/1m17.

Declarations
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare no competing interests.
Received: 6 April 2024 / Accepted: 24 June 2024

References
1. Dhuguru J, Ghoneim OA. Quinazoline based HDAC dual inhibitors as potential anti-cancer agents. Molecules. 2022;27(7):2294.
2. Wang Q, et al. Novel approaches for the solid-phase synthesis of Dihydroquinazoline-2 (1H)-One derivatives and biological evaluation as potential
anticancer agents. Molecules. 2022;27(23):8577.
3. Bansal R, Malhotra A. Therapeutic progression of quinazolines as targetedchemotherapeutic agents. Eur J Med Chem. 2021;211:113016.
4. Khalifa MM, et al. Topo II inhibition and DNA intercalation by newphthalazine-based derivatives as potent anticancer agents: design, synthesis,anti-proliferative, docking, and in vivo studies. J Enzyme Inhib Med Chem.
2022;37(1):299–314.
5. Niu Z, et al. Discovery of Novel quinazoline derivatives as potent antitumor
agents. Molecules. 2022;27(12):3906.
6. Ferlay J, et al. Cancer statistics for the year 2020: an overview. Int J Cancer.
2021;149(4):778–89.
7. Behranvand N, et al. Chemotherapy: a double-edged sword in cancer treatment. Cancer Immunol Immunother. 2022;71(3):507–26.
8. Alanazi AM, et al. Synthesis, antitumor and antimicrobial activity of somenew 6-methyl-3-phenyl-4 (3 H)-quinazolinone analogues: in silico studies. J
Enzyme Inhib Med Chem. 2016;31(5):721–35.
9. Dickens E, Ahmed S. Principles of cancer treatment by chemotherapy. Surg
(Oxford). 2021;39(4):215–20.
10. Lin S, et al. Design, synthesis and biological evaluation of quinazoline–phosphoramidate mustard conjugates as anticancer drugs. Eur J Med Chem.
2017;127:442–58.

Emami et al. BMC Chemistry(2024) 18:125

11. Hawash M, et al. Synthesis, chemo-informatics, and anticancer evaluation offluorophenyl-isoxazole derivatives. Open Chem. 2021;19(1):855–63.
12. Hawash M, et al. Design and synthesis of novel substituted indole-acrylamide
derivatives and evaluation of their anti-cancer activity as potential tubulintargeting agents. J Mol Struct. 2022;1254:132345.
13. Hawash M. Recent advances of tubulin inhibitors targeting the colchicine
binding site for cancer therapy. Biomolecules. 2022;12(12):1843.
14. Haghighijoo Z, et al. Therapeutic potential of quinazoline derivatives
for Alzheimer’s disease: a comprehensive review. Eur J Med Chem.
2022;227:113949.
15. Zhang Y, et al. Enrichment of novel quinazoline derivatives with highantitumor activity in mitochondria tracked by its self-fluorescence. Eur J Med
Chem. 2019;178:417–32.
16. Khabnadideh S, Sadeghian S. A review on current synthetic methods of
4-aminoquinazoline derivatives. J Chem. 2022;2022.
17. Alqahtani AS, et al. Cytotoxicity of newly synthesized quinazoline–sulfonamide derivatives in human leukemia cell lines and their effect on hematopoiesis in zebrafish embryos. Int J Mol Sci. 2022;23(9):4720.
18. Wang C-J, et al. Discovery of penipanoid C-inspired 2-(3, 4, 5-trimethoxybenzoyl) quinazolin-4 (3H)-one derivatives as potential anticancer agentsby inhibiting cell proliferation and inducing apoptosis in hepatocellular
carcinoma cells. Eur J Med Chem. 2021;224:113671.
19. Alagarsamy V, et al. An overview of quinazolines: pharmacological significance and recent developments. Eur J Med Chem. 2018;151:628–85.
20. Chang J, et al. Development of a series of novel 4-anlinoquinazoline derivatives possessing quinazoline skeleton: design, synthesis, EGFR kinase inhibitory efficacy, and evaluation of anticancer activities in vitro. Eur J Med Chem.
2017;138:669–88.
21. Khodair AI, Alsafi MA, Nafie MS. Synthesis, molecular modeling and anticancer evaluation of a series of quinazoline derivatives. Carbohydr Res.
2019;486:107832.
22. Abuelizz HA, et al. Synthesis and anticancer activity of new quinazoline
derivatives. Saudi Pharm J. 2017;25(7):1047–54.
23. Le Y, et al. Design, synthesis and in vitro biological evaluation of quinazolinone derivatives as EGFR inhibitors for antitumor treatment. J Enzyme Inhib
Med Chem. 2020;35(1):555–64.
24. Allam HA, et al. Design and synthesis of some new 2, 4, 6-trisubstituted
quinazoline EGFR inhibitors as targeted anticancer agents. Bioorg Chem.
2020;98:103726.
25. Tu Y, et al. Design, synthesis, and docking studies of quinazoline analogues
bearing aryl semicarbazone scaffolds as potent EGFR inhibitors. Bioorg Med
Chem. 2017;25(12):3148–57.
26. Awad MK, et al. Design, synthesis, molecular modeling, and biologicalevaluation of novel α-aminophosphonates based quinazolinone moiety as
potential anticancer agents: DFT, NBO and vibrational studies. J Mol Struct.
2018;1173:128–41.
27. Ghorab MM, et al. Design, synthesis and anticancer evaluation of novel
quinazoline-sulfonamide hybrids. Molecules. 2016;21(2):189.
28. Zayed MF, et al. Design, synthesis, cytotoxic evaluation and moleculardocking of new fluoroquinazolinones as potent anticancer agents withdual EGFR kinase and tubulin polymerization inhibitory effects. Int J Mol Sci.
2018;19(6):1731.
29. Zare S, et al. Design, synthesis, computational study and cytotoxic evaluationof some new quinazoline derivatives containing pyrimidine moiety. Sci Rep.
2023;13(1):14461.
30. Ataollahi E et al. Novel quinazolinone derivatives as Anticancer agents:
design, synthesis, Biological evaluation and computational studies. J Mol
Struct. 2023:136622.
31. Emami L, et al. Design, synthesis, molecular simulation, and biological activities of novel quinazolinone-pyrimidine hybrid derivatives asPage 18 of 18

32.
33.
34.
35.
36.
37.
38.
39.

40.

41.
42.
43.
44.

45.
46.
47.
48.
49.

dipeptidyl peptidase-4 inhibitors and anticancer agents. New J Chem.
2020;44(45):19515–31.
Emami L, et al. Synthesis, biological evaluation, and computational studies
of some novel quinazoline derivatives as anticancer agents. BMC Chem.
2022;16(1):1–14.
Ye L, et al. Discovery of aminopyridine-containing spiro derivatives as EGFR
mutations inhibitors. J Enzyme Inhib Med Chem. 2019;34(1):1233–46.
Arshia AH, et al. De novo design of novel protease inhibitor candidates inthe treatment of SARS-CoV-2 using deep learning, docking, and molecular
dynamic simulations. Comput Biol Med. 2021;139:104967.
Hawash M, et al. Anticancer activity of thiophene carboxamide derivatives
as CA-4 biomimetics: synthesis, biological potency, 3D spheroid model, andmolecular dynamics simulation. Biomimetics. 2022;7(4):247.
Emami L, et al. Novel n-substituted isatin‐ampyrone schiff bases as a newclass of antiproliferative agents: design, synthesis, molecular modeling and invitro cytotoxic activity. J Heterocycl Chem. 2022;59(7):1144–59.
Knapp B, Ospina L, Deane CM. Avoiding false positive conclusions inmolecular simulation: the importance of replicas. J Chem Theory Comput.
2018;14(12):6127–38.
Hawash M, et al. New Thiazole Carboxamide derivatives as COX inhibitors:
design, synthesis, Anticancer Screening, in Silico Molecular Docking, andADME Profile studies. ACS Omega. 2023;8(32):29512–26.
Hirao H. Which DFT functional performs well in the calculation of methylcobalamin? Comparison of the B3LYP and BP86 functionals and evaluation of the impact of empirical dispersion correction. J Phys Chem A.
2011;115(33):9308–13.
Kavitha T, Velraj G. Structural, spectroscopic (FT-IR, FT-Raman, NMR) andcomputational analysis (DOS, NBO, Fukui) of 3, 5-dimethylisoxazole and4-(chloromethyl)-3, 5-dimethylisoxazole: a DFT study. J Theoretical Comput
Chem. 2016;15(05):1650039.
Bruna-Larenas T, Gomez-Jeria JS. A DFT and semiempirical model-based
study of opioid receptor affinity and selectivity in a group of molecules witha morphine structural core. Int J Med Chem. 2012;2012.
Su S, et al. Novel penta-1, 4-diene-3-one derivatives containing quinazoline
and oxime ether fragments: design, synthesis and bioactivity. Bioorganic Med
Chem. 2021:115999.
Faraj FL et al. Synthesis, characterization, and anticancer activity of newquinazoline derivatives against MCF-7 cells. Sci World J. 2014:2014.
Bathula R, et al. Evaluation of antitumor potential of synthesized novel
2-substituted 4-anilinoquinazolines as quinazoline-pyrrole hybrids in MCF-7
human breast cancer cell line and A-549 human lung adenocarcinoma celllines. Future J Pharm Sci. 2020;6:1–11.
Zahedifard M, et al. Synthesis, characterization and apoptotic activity of quinazolinone Schiff base derivatives toward MCF-7 cells via intrinsic and extrinsic
apoptosis pathways. Sci Rep. 2015;5(1):11544.
Faghih Z, et al. Synthesis of some novel dibromo-2-arylquinazolinone derivatives as cytotoxic agents. Res Pharm Sci. 2019;14(2):115.
Hashemi S, et al. Two new cytotoxic ursane triterpenoids from the aerial partsof Salvia Urmiensis Bunge. Fitoterapia. 2021;154:105030.
Amelia T, et al. Computational prediction of resistance induced alaninemutation in ATP site of epidermal growth factor receptor. Int J Mol Sci.
2022;23(24):15828.
Zare F, et al. A combination of virtual screening, molecular dynamics simulation, MM/PBSA, ADMET, and DFT calculations to identify a potential DPP4
inhibitor. Sci Rep. 2024;14(1):7749.

Publisher’s NoteSpringer Nature remains neutral with regard to jurisdictional claims inpublished maps and institutional affiliations.