Design, synthesis and docking studies of some spiro-oxindole dihydroquinazolinones as antibacterial agents

Article abstract of DOI:10.1016/j.tet.2015.12.055

Two series of spiro-oxindole dihydroquinazolinone derivatives have been designed and synthesized using a catalytic amount of nano cerium oxide with excellent product yields by a convenient one-pot process. The synthesized compounds were evaluated for thei

Full text of DOI:10.1016/j.tet.2015.12.055

Accepted Manuscript
Design, synthesis and docking studies of some spiro-oxindole dihydroquinazolinones
as antibacterial agents
Jin Zhang, Jiawen Zhao, Liangpeng Wang, Jia Liu, Decheng Ren, Yangmin Ma
PII:
S0040-4020(15)30303-3
10.1016/j.tet.2015.12.055
TET 27381
DOI:
Reference:
To appear in: Tetrahedron
Received Date: 8 August 2015
Revised Date: 13 December 2015
Accepted Date: 23 December 2015
Please cite this article as: Zhang J, Zhao J, Wang L, Liu J, Ren D, Ma Y, Design, synthesis and docking
studies of some spiro-oxindole dihydroquinazolinones as antibacterial agents, Tetrahedron (2016), doi:
10.1016/j.tet.2015.12.055.
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to
our customers we are providing this early version of the manuscript. The manuscript will undergo
copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please
note that during the production process errors may be discovered which could affect the content, and all
legal disclaimers that apply to the journal pertain.

ACCEPTED MANUSCRIPT
Graphical Abstract
To create your abstract, type over the instructions in the template box below.
Fonts or abstract dimensions should not be changed or altered.
Leave this area blank for abstract info.
Design, synthesis and docking studies of
some spiro-oxindole dihydroquinazolinones as antibacterial agents
Jin Zhanga, b, Jiawen Zhaoa, Liangpeng Wanga, Jia Liua, Decheng Rena, Yangmin Maa, b, *

1
ACCEPTED MANUSCRIPT
Tetrahedron
journal homepage: www.elsevier.com
Design, synthesis and docking studies of some spiro-oxindole
dihydroquinazolinones as antibacterial agents
Jin Zhang a, b, Jiawen Zhaoa, Liangpeng Wanga, Jia Liua, Decheng Rena, Yangmin Maa, b,
aCollege of Chemistry & Chemical Engineering, Shaanxi University of Science & Technology, Xi’an, Shaanxi 710021, PR China
bKey Laboratory of Auxiliary Chemistry & Technology for Chemical Industry, Ministry of Education, Xi’an, Shaanxi 710021, PR China
ARTICLE INFO
ABSTRACT
Article history:
Received
Received in revised form
Accepted
Two series of spiro-oxindole dihydroquinazolinone derivatives have been designed and
synthesized using a catalytic amount of nano cerium oxide with excellent product yields by a
convenient one-pot process. The synthesized compounds were evaluated for their in vitro
antibacterial activity, and compounds 1i-1p showed considerable antibacterial avtivities to E.
coli. Molecular docking of Escherichia coli Biotin Carboxylase (EcBC) enzyme was also
performed in order to predict the interactions of the synthesized compounds.
Available online
Keywords:
Spiro-oxindole dihydroquinazolinone
Nano CeO2
2009 Elsevier Ltd. All rights reserved
.
Antibacterial activity
Molecular docking
1. Introduction
of cerium oxide nanoparticles has not yet evolved. Hence, it is a
long-termed theme to develop novel, efficient and practical
cerium oxide nanoparticles catalytic methods for the synthesis of
this class of spiro-oxindole dihydroquinazolinones.
Quinazolines are considered as important bioactive scaffold
due to their anti-virus,1 antimalarial,2 anti-tuberculosis,3
antioxidant,4 anti-hypertensive5,
and antiinflammatory
6
activities.7 They also possess anticancer activities8 like inhibition
of tubulin formation,9,
inhibition against VEGFR2 tyrosine
10
kinase and cell proliferation.11 And spiro-oxindoles are of great
interest due to their role as key precursors in natural product
synthesis12,13 and significant role in medicinal chemistry for their
biological activity as antibacterial, antiinflammatory, anticancer
15
and laxatives.14,
Furthermore, both of the two ring systems
showed considerable antibacterial activities.16, 17. It is noteble that
spiro-oxindole dihydroquinazolinone derivatives combine two
pharmacophores into one molecule, so our attention was attracted
to find more potent pharmaceutical active molecules (Fig 1).
Recently, cerium oxide nanoparticles have been extensively
used as heterogeneous catalysts for green organic reactions such
as N-formylation of amines,18 catalytic CO2 conversion to
organic carbonates with alcohols,19 construction of amide
bonds,20 synthesis of α-aminophosphonates21 and synthesis of
substituted benzimidazole, benzothiazole, and benzoxazole,22. A
few of different strategies have been devised for condensation to
spiro-oxindole dihydroquinazolinones with isatoic anhydride,
isatin and amine or phenylhydrazine,23, 24 however, they suffered
from some inconveniences such as corrosive solvent, special
reactor and narrow substrate scope. The preparation of
Fig 1. Some natural and synthesized bioactive molecules contained
quinazolinone or oxindole scaffolds.
2. Results and discussion
The reaction of isatoic anhydride, isatin and aniline was used
as the model in a one-pot manner to investigate experimental
conditions including catalysts and solvents. Isatoic anhydride and
aniline were refluxed in ethanol for 1 h in the presence of
different catalysts, such as nano CuO, nano Al2O3, nano Fe3O4,
corresponding quinazolinone analogues using a catalytic amount
———
Corresponding author. Tel.: +86-029-86168312; fax: +86-029-86168312; e-mail: mym63@sina.com

2
Tetrahedron
ACCEPTED MANUSCRIPT
nano CeO2 and CeO2 powder (Table 1, entry 1-5). Then isatin
was added to the reaction mixture, refluxed for another 5 hours.
We were delighted to find that nano CeO2 was superior for the
reaction, and the yield of product 1a was 76% (Table 1, entry 4).
Next, we screened protic and aprotic solvents on the reaction.
Obviously, protic solvents were better for the reaction, especially
when water and ethanol were applied (Table 1, entry 6-11).
Encouraged by this result, we carried on with examining the
effect of solvents using different ratios of EtOH/H2O. It was clear
that the highest yield of product 1a was achieved when the
reaction was performed in EtOH/H2O=1:1 (Table 1, entry 14).
Furthermore, the influence of the amount of nano CeO2 catalyst
was examined to increase the efficiency of the reaction. When the
amount was decreased to 1 mol % of nano CeO2 catalyst was
used, the 1a was obtained in 73% yield (Table 1, entry 15). As
one can see from Table 1, the highest yield of 85% when 5 mol%
of nano CeO2 was used (Table 1, entry 16). On the other hand,
the yield of compound 1a was not significantly improved when
the amount of nano CeO2 catalyst was increased to 10 mol %
(Table 1, entry 17).
phenylhydrazine exhibited good reactivity and achieved good
yields in the corresponding products (Table 2, entry 7). However,
only trace amount of corresponding product were observed when
the strong electron-withdrawing group NO2 was situated in the
para position of aniline (Table 2, entry 8).
To further assess the general applicability of the procedure,
we then chose 5-chloroisatin to perform the reactions. Methyl-,
methoxy-substituted amines was also tolerable under the present
reaction conditions and obtained in good yields(Table 2, entry 9-
13). Aniline and phenylhydrazine were produced under the same
conditions with yields of 92% respectively (Table 2, entry14, 15).
Interestingly, when p-aminoaniline possessing electron-donating
group was employed, it was found that the promising product
was synthesized in this way smoothly (Table 2, entry 16). In
addition, when p-nitroaniline were used as a substrate, the
reaction still gave a trace yield (Table 2, entry 17). Then we
chosed 5-bromoisatin to perform the reaction, and compound 1r
was abtained (Table 2, entry 19).
We went on to further expand the scope of this methodology
by using N-methylisatin and N-benzylisatin. The results were
subsequently displayed in Table 2. When the strong electron-
donating group methyl or methoxyl was situated in the ortho,
meta or para position the reactions went into the desired products
(Table 2, entry 18, 19, 20, 23, 26). Aniline were also found to be
suitable for the reaction and gave the corresponding products in
satisfied yields (Table 2, entry 21, 24). Surprisingly, we were
delighted to find that the ammonium oxalate furnished the
desired product with N-benzylisatin (Table 2, entry 25).
However, the p-nitroaniline went into the desired product just in a
trace yield (Table 2, entry 22, 27) .
Table 1 Investigation of conditions for nano CeO2-catalyzed
synthesis of spiro-oxindole dihydroquinazolinone derivatives
(1a) using isatoic anhydride, isatin and aniline as the
substratesa.
N-methylisatin and N-benzylisatin produced less products than
5-chloroisatin and isatin using the same nitrogen source. In
general, anilines bearing electron-donating groups were more
reactive than those with electron-withdrawing groups. On the
contrary, 5-chloroisatin with a electron-withdrawing group was
more active than N-methylisatin. N-benzylisatin was less active
might because of the steric hindrance.
Entry
Catalyst(mol%)
Nano CuO(3)
Nano Fe3O4(3)
Solvent
EtOH
Yield(%)b
1
2
50
67
EtOH
3
Nano Al2O3(3)
Nano CeO2(3)
EtOH
43
76
70
45
62
58
50
70
73
60
70
81
73
85
84
4
EtOH
CeO2 powder
Nano CeO2(3)
Nano CeO2(3)
Nano CeO2(3)
Nano CeO2(3)
Nano CeO2(3)
Nano CeO2(3)
Nano CeO2(3)
Nano CeO2(3)
Nano CeO2(3)
Nano CeO2(1)
Nano CeO2(5)
Nano CeO2(10)
5
EtOH
6
DMSO
All the compounds synthesized were well characterized by
spectral (NMR, IR) data, the molecular structure of 1c and1k was
7
CHCl3
8
CH2Cl2
toluene
CH3OH
H2O
characterized by X-ray crystallography
(CCDC 1057927-
9
1057931) , and the atom-labelling scheme of compound 1c and
1k are as shown in Figure 2.
10
11
12
13
14
15
16
17
Table 2. Synthesis of spiro-oxindole dihydroquinazolinone
EtOH/H2O=4:1
EtOH/H2O=2:1
derivativesa.
EtOH/H2O=1:1
EtOH/H2O=1:1
EtOH/H2O=1:1
EtOH/H2O=1:1
aConditions: isatoic anhydride 1 (1.0 mmol), aniline (1.1 mmol), isatin (1.1
mmol) in the presence of catalyst in solvent (10 mL) for 6 h under reflux
condition.
bIsolated yields.
Entry
1
R1
R2
H
R3
H
Product
Yieldb
85%
O
With the best reaction conditions in hand, we tested the
generality of this methodology with different amines,
phenylhydrazine or ammonium oxalate. Results listed in Table 2
demonstrate that most of the amines tested underwent smooth
transformation to afford the corresponding pruducts in satisfied
yields. Amines bearing electron-donating groups on the benzene
ring, such as methyl-, methoxy-substituted amines were
successfully converted and gave the corresponding products in
good yields (Table 2, entry 2, 3, 4, 5, 6). It is notable that
O
N
NH
N
H
1a

3
ACCEPTED MANUSCRIPT
OCH3
O
O
O
N
O
N
NH
14
H
H
Cl
Cl
92%
N
H
2
H
H
89%
NH
N
H
Cl
1n
1b
1c
1d
15
92%
89%
3
4
5
H
H
H
H
H
H
87%
85%
93%
1o
NH2
O
O
O
OCH3
N
O
N
NH
N
H
16
H
Cl
NH
N
H
CH3
Cl
1p
1q
O
NO2
O
N
NH
N
H
O
O
N
17
18
H
H
Cl
Br
trace
88%
1e
NH
N
H
CH3
O
Cl
O
N
CH3
6
7
8
H
H
H
H
H
H
80%
88%
trace
NH
N
H
O
O
N
1f
NH
N
H
Br
1r
CH3
1g
1h
O
NO2
O
N
19
20
CH3
CH3
H
H
85%
93%
O
N
N
H
O
N
2a
NH
N
H
OCH3
O
CH3
O
N
O
N
N
H
O
N
9
H
H
Cl
Cl
83%
88%
2b
2c
NH
N
H
Cl
1i
OCH3
21
22
CH3
CH3
H
H
87%
88%
CH3
O
O
N
10
NH
O
N
H
O
N
N
N
H
Cl
1j
1k
1l
O
2d
NO2
O
CH3
N
NH
11
12
13
H
H
H
Cl
Cl
Cl
91%
87%
81%
N
H
O
O
N
23
CH3
H
trace
Cl
N
N
H
O
2e
O
OCH3
OCH3
N
NH
N
H
O
O
N
24
25
Bn
Bn
H
H
93%
93%
Cl
N
N
H
CH3
O
2f
O
N
NH
N
H
Cl
1m
2g

4
Tetrahedron
ACCEPTED MANUSCRIPT
26
27
(NH4)2C2O4
Bn
Bn
H
H
75%
2h
CH3
O
O
N
88%
N
N
H
2i
NO2
O
O
N
28
Bn
H
trace
Scheme.1 The plausible mechanism of the reaction.
N
N
H
3. Antibacterial activity
2j
aConditions: isatoic anhydride (1.0 mmol), aniline (1.1 mmol) were first
refluxed in the presence of catalyst in solvent (10 mL) for 1 h followed by the
addition of isatin (1.1 mmol) and another 5 h.
All the synthesized compounds were screened for antibacterial
activity against two Gram-negative bacterial strains: Escherichia
coli ((ATCC 25922) and Pseudomonas aeruginosa (ATCC
85327) and two Gram-positive bacterial strains: Bacillus subtilis
(ATCC 465) and Staphylococcus aureus (ATCC 25923) by using
the minimum inhibitory concentration (MIC) method. The MICs
of the compounds against these bacteria were presented in Table
3. Standard antibacterial agent penicillin was also screened under
identical conditions for comparison.
bIsolated yields.
Table 3. Antibacterial activity of synthetic compounds.
Minimum inhibitory concentrations (µg/mL)
Compounds
Gram-negative
E.coli P.aeruginosa
Gram-positive
B. subtilis
S.aureus
62.5
1a
>100
31.2
15.6
31.2
>100
62.5
15.6
15.6
15.6
7.8
62.5
>100
>100
31.2
62.5
31.2
31.2
31.2
>100
15.6
15.6
31.2
62.5
62.5
31.2
15.6
>100
>100
62.5
31.2
>100
15.6
15.6
62.5
-
31.2
>100
62.5
31.2
31.2
31.2
>100
15.6
31.2
31.2
31.2
15.6
62.5
15.6
>100
15.6
62.5
>100
>100
62.5
62.5
31.2
31.2
>100
3.9
1b
62.5
31.2
15.6
>100
31.2
62.5
31.2
62.5
62.5
31.2
31.2
31.2
>100
62.5
31.2
62.5
62.5
31.2
62.5
31.2
62.5
62.5
62.5
3.9
1c
1d
1e
1f
1g
1i
1j
1k
1l
15.6
15.6
15.6
15.6
15.6
31.2
15.6
15.6
62.5
31.2
15.6
15.6
15.6
31.2
-
1m
1n
1o
1p
1r
2a
2b
2c
2d
Fig 2. A view of the atom-labelling scheme of compound 1c and1k.
2f
2g
According to the relevant literature,25 a plausible pathway for
nano CeO2 particles catalyzed this one-pot reaction was
suggested in Scheme 1. First, the isatoic anhydride was activated
by CeO2 nano particles in the protic solvent, then the N-
nucleophilic attack of amines, phenylhydrazine or ammonium
oxalate to the isatoic anhydride formed intermediate I with the
decarbonylation. Next, the imine intermediate II was given by
nucleophilic attack of isatin that promoted by CeO2 nano
particles. Finally, the product formed from III by 1,5-H shift
depicted.
2h
2i
penicillin
streptomycin
sulfate
7.8
7.8
-
-

5
ACCEPTED MANUSCRIPT
It was observed that many compounds exhibited interesting
antibacterial activity displaying MIC values between 62.5 and
15.6 µg/mL. Based on the data obtained, we found that
compounds 1i-1p with a chloro substitution in 5th position of
isatin ring showed MIC values in 15.6 µg/mL or 7.8 µg/mL,
while other series gave higher MIC values than this series.
Compounds 1a and 1e formed hydrophobic interactions with the
amino acid Tyr203, His438, Leu278, Lys159 and Met169,
however, there was no hydrogen bond between these compounds
and enzyme, which probably decreased the antibacterial activity.
The results explained that the quinazoline ring played an
important role in the binding interaction and the beneficial role of
the amino acid residues His236, Glu441 and Gly166 in the
binding. The docking was consistent with the results we reported
before.27 It can be conjectured that the main pharmacophore was
quinazolinone ring and amino acid residues His236, Glu441 and
Gly166 in the enzyme might be the hotspot residue to form the
hydrogen bond and hydrophobic interactions with the amino acid
residues Ile437, His236 were favorable for increasing the
antibacterial activity.
4. Molecular docking
In order to get a deep understanding of the mechanism of
antibacterial activity, docking studies were conducted between
the Escherichia coli Biotin Carboxylase (EcBC) enzyme26 and
synthesized compounds including inactive compounds 1a and 1e
and compounds 1i-1p with significant activity against
Escherichia coli. EcBC enzyme, which catalyzed the ATP-
dependent carboxylation of the vitamin biotin, was a potential
target of the fatty acid biosynthetic pathway for the development
of novel antibacterial agents. There have been reported that
quinazoline scaffold was EcBC inhibitors with mechanism-based
antibacterial activity. The binding mode of the ligand natural
origin to EcBC enzyme was previously determined by X-ray
crystallography (PDB: 2W6O), and the original ligand was
similar to the synthesized compounds. As depicted in Table 4,
most of the compounds represented a relatively good binding
energy. Hence, this might give us a reasonable description for
their good antibacterial activities. Interestingly, compound 1k
performed significant vitro antibacterial activity but with slightly
higher binding energy. We conjectured that 1k fitted well in the
hydrophobic enclosure formed by Ile437, His236 which were
pivotal to increase the antibacterial activity. And the hydrogen
bonds formed by the amino acid residues His236 and Glu441
were very important for increasing the antibacterial activity. The
other molecular basis of interactions between target enzyme and
1i-1j, 1l-1p can be understood with the help of docking analysis
and interactions as represented in Fig. 3. The carbonyl group of
the quinazolinone ring formed hydrogen bonds with the residue
GLY 166 (Fig. 3, 1n, 1j, 1m, 1o, 1p). As depicted in Fig. 3,
Table 4. Docking results of synthetic compounds.
Binding
Energy
(kcal/mol)
-7.67
Hydrogen
bond
interaction
Inhibitory
constant
Compounds
a
1a
1e
1i
2.39 µM
1.09 µM
_
a
-8.14
-8.20
-8.66
-6.76
_
a
969.08 nM
452.29 nM
11.16 µM
_
1j
GLY-166
HIS-236
GLU-441
1k
a
1l
1m
1n
1o
-7.53
-8.37
-8.51
-8.05
-8.52
3.05 µM
731.47 nM
573.66 nM
1.25 µM
_
GLY-166
GLY-166
GLY-166
GLY-166
MET-169
1p
567.23 nM
aNot formed
Fig 3. Three dimensional diagrams displaying the binding poses and interactions for the compounds 1a, 1e, 1k, 1n, 1j, 1m, 1o and 1p to the
binding site of EcBC enzyme.
were synthesized. The method offered several advantages
including high yield of products, easy experimental workup
procedure and mild reaction conditions. Most of the compounds
exhibited promising antibacterial activity especially to E. coli. In
particular 1i-1p emerged as the most active compounds to E. coli
in the series. The molecular docking results were reasonably fair
5. Conclusion
In conclusion, a convenient and efficient nano CeO2-catalyzed
one-pot method for the synthesis of quinazolinones from the
isatoic anhydride, isatin and amine, phenylhydrazine has been
achieved. This procedure showed good functional group
tolerance. A total of 24 spiro-oxindole dihydroquinazolinones

6
Tetrahedron
ACCEPTED MANUSCRIPT
1
in understanding the binding interactions of these compounds to
1478.90, 752.64 cm-1; H NMR (400 MHz, DMSO) δ 10.39 (s,
(EcBC) enzyme.
1H), 7.67 (dd, J = 7.7, 1.4 Hz, 1H), 7.59 (s, 1H), 7.56 (d, J = 7.3
Hz, 1H), 7.34 – 7.26 (m, 1H), 7.17 (d, J = 1.2 Hz, 1H), 6.95 (d, J
= 0.8 Hz, 1H), 6.90 (d, J = 9.0 Hz, 2H), 6.78 – 6.73 (m, 3H), 6.71
(d, J = 8.0 Hz, 1H), 6.66 (d, J = 7.7 Hz, 1H), 3.66 (s, 3H). 13C
NMR (101 MHz, DMSO) δ 178.80, 175.47, 163.75, 158.12,
146.09, 141.59, 133.57, 130.74, 130.53, 127.60, 127.42, 126.35,
122.13, 117.58, 114.52, 114.01, 113.74, 110.10, 76.51, 55.07.
6. Experimental section
6.1. General information
Commercial reagents were used as received without additional
purification. All reactions were monitored by thin layer
chromatography (TLC) using plates precoated with silica gel 60
GF-254. Column chromatography was performed by using
6.4.3 3'-(o-tolyl)-1'H-spiro[indoline-3,2'-quinazoline]-2,4'(3'H)-
o
dione (1c). White solid, mp 268-269 C; IR (KBr) νmax 3344.47,
neutral Al2O3 (200-300 mesh). H NMR (400 MHz) and 13C
1
1744.21, 1614.62, 1491.20, 1360.78, 1186.97, 752.18 cm-1; H
1
NMR (101 MHz) were recorded with Bruker spectrometer. IR
spectra were recorded on a FT-IR spectrometer. The mass
spectrometric identification of the products was performed on a
LC-MS with electrospray ionization.
NMR (400 MHz, DMSO) δ 10.54 (s, 1H), 7.73 – 7.63 (m, 1H),
7.60 (s, 1H), 7.35 (d, J = 6.9 Hz, 1H), 7.34 – 7.28 (m, 1H), 7.14
(ddd, J = 7.6, 4.5, 1.2 Hz, 2H), 7.04 (td, J = 7.4, 1.3 Hz, 1H),
7.02 – 6.96 (m, 1H), 6.88 – 6.81 (m, 2H), 6.80 – 6.73 (m, 2H),
6.73 – 6.68 (m, 1H), 2.22 (s, 3H). 13C NMR (101 MHz, DMSO) δ
176.18, 162.67, 146.16, 141.93, 137.81, 136.36, 133.49, 130.95,
130.29, 127.62, 127.40, 126.62, 126.31, 125.27, 125.15, 122.01,
117.73, 115.23, 114.21, 110.08, 75.95, 18.09. ESI-MS (m/z) =
356.1376 ([M+H]+); found: 356.1399.
6.2. General procedure for the synthesis of spiro-oxindole
dihydroquinazolinone derivatives.
Typically, isatoic anhydride (1.0 mmol), aniline (1.1mmol),
isatin (1.1 mmol), and CeO2 nano particles (5 mol %, OD 40 nm)
were mixed in EtOH/H2O=1:1 (10 mL). Isatoic anhydride and
aniline were first refluxed in the presence of catalyst for 1 h
followed by the addition of isatin and another 5 h. After
completion of the reaction (monitored by TLC), the CeO2 nano
particles catalyst was separated by filtration. The crude product
was purified by short column layer chromatography on neutral
Al2O3 to provide the analytically pure product 1 and 2.
6.4.4 3'-(2-methoxyphenyl)-1'H-spiro[indoline-3,2'-quinazoline]-
2,4'(3'H)-dione (1d). White solid, mp 184-186 C; IR (KBr) νmax
o
3271.64, 1724.50, 1613.65, 1477.35, 1364.36, 1266.31, 1191.92,
752.91 cm-1; 1H NMR (400 MHz, DMSO) δ 10.42 (s, 1H), 7.65
(td, J = 8.2, 1.4 Hz, 1H), 7.55 (s, 1H), 7.46 (d, J = 7.3 Hz, 1H),
7.29 (ddd, J = 9.5, 3.6, 1.8 Hz, 1H), 7.16 – 7.08 (m, 3H), 6.87
(ddd, J = 7.9, 5.5, 0.9 Hz, 3H), 6.79 – 6.68 (m, 5H), 6.62 (d, J =
7.7 Hz, 1H), 3.71 (s, 3H), 3.46 (s, 1H). 13C NMR (101 MHz,
DMSO) δ 176.08, 162.55, 155.42, 152.47, 146.22, 141.76,
133.43, 130.90, 129.33, 127.98, 127.31, 126.98, 126.76, 125.56,
121.56, 120.12, 117.47, 114.77, 113.99, 111.71, 109.75, 76.22,
55.08. ESI-MS (m/z) = 394.1146 ([M+Na]+); found: 394.1164.
6.3. Molecular docking analysis
The crystallographic 3D structure of Escherichia coli BC
(EcBC) enzyme was accessed from the Protein Data Bank (PDB
ID: 2W6O) and the initial structures of antibiotics were generated
by Chem-Bio-Office 2010 with energy optimized using Merk
Molecular Force Field 94. AutoDock 4.2 suit of packages
(http://autodock.scripps.edu/), a popular and proven robust
approach with good docking accuracy and realibility, was
utilized to study the interaction modes of BSA with
antibiotics.Gasteiger partial charges,nonpolar hydrogen atoms,
and rotatable bonds were dealt with the aid of AutoDock Tools
6.4.5 3'-(m-tolyl)-1'H-spiro[indoline-3,2'-quinazoline]-2,4'(3'H)-
dione (1e). White solid, 210-212 oC; IR (KBr) νmax 3378.25,
1722.00, 1688.02, 1476.54, 1361.39, 1192.39, 749.26 cm-1; H
1
NMR (400 MHz, DMSO) δ 10.42 (s, 1H), 7.67 (dd, J = 7.7, 1.4
Hz, 1H), 7.61 (s, 1H), 7.55 (dd, J = 7.4, 0.6 Hz, 1H), 7.31 (ddd, J
= 8.2, 7.4, 1.6 Hz, 1H), 7.16 (td, J = 7.7, 1.2 Hz, 1H), 7.08 (t, J =
7.7 Hz, 1H), 6.98 – 6.90 (m, 2H), 6.83 (s, 1H), 6.80 – 6.74 (m,
2H), 6.71 (d, J = 8.0 Hz, 1H), 6.65 (d, J = 7.7 Hz, 1H), 2.16 (s,
3H). 13C NMR (101 MHz, DMSO) δ 175.48, 163.54, 146.08,
141.66, 137.95, 137.85, 137.74, 137.66, 133.60, 130.78, 128.34,
128.23, 127.34, 126.46, 122.03, 117.64, 114.62, 114.06, 110.06,
76.33, 20.69.
1.5.6
(http://mgltools.scripps.edu).
And
spacing
(Angstrom):0.3750Å. The population size and the maximum
number of evaluations were 150 and 2 500 000, respectively.
Lamarckian Genetic Algorithm was chosen as the searching and
scoring procedure for van der Waals force, electrostatic
interaction, entropy loss upon binding process, hydrogen bonding
and salvation; and the docking results were clustered with a root-
mean-square deviation tolerance of 2.0 Å. The output results
were graphically analyzed using Pymol 1.6.0.0 and Ligplus 1.3.2.
6.4.6 3'-(p-tolyl)-1'H-spiro[indoline-3,2'-quinazoline]-2,4'(3'H)-
dione (1f). White solid,242-244 oC; IR (KBr) νmax 3840.46,
3652.26, 3158.52, 1721.24, 1646.56, 1515.87, 1478.70, 1361.72,
1193.61, 751.97 cm-1 ; 1H NMR (400 MHz, DMSO) δ 10.40 (s,
1H), 7.69 (d, J = 7.7 Hz, 1H), 7.60 (s, 1H), 7.56 (d, J = 7.4 Hz,
1H), 7.31 (t, J = 7.7 Hz, 1H), 7.17 (t, J = 7.7 Hz, 1H), 7.02 (d, J =
8.1 Hz, 2H), 6.95 (t, J = 7.5 Hz, 1H), 6.89 (d, J = 8.2 Hz, 2H),
6.80 – 6.70 (m, 2H), 6.67 (d, J = 7.8 Hz, 1H), 2.19 (s, 3H). 13C
NMR (101 MHz, DMSO) δ 175.47, 163.64, 146.09, 141.63,
136.82, 135.44, 133.59, 133.51, 130.76, 129.15, 127.51, 127.41,
126.39, 122.12, 117.62, 114.59, 114.05, 110.11, 76.41, 20.52.
6.4. Characterization data of products
6.4.1 3'-phenyl-1'H-spiro[indoline-3,2'-quinazoline]-2,4'(3'H)-
dione (1a). White solid, mp 230-232 C; IR (KBr) νmax 3766.14,
o
3200.37, 2819.18, 1720.87, 115.37, 1615.37, 1490.79, 1360.35,
753.25 cm-1; 1H NMR (400 MHz, DMSO) δ 10.42 (s, 1H), 7.68
(dd, J = 7.7, 1.5 Hz, 1H), 7.63 (s, 1H), 7.55 (d, J = 7.2 Hz, 1H),
7.31 (ddd, J = 8.1, 7.4, 1.6 Hz, 1H), 7.22 (t, J = 7.4 Hz, 2H), 7.16
(ddd, J = 9.0, 4.9, 1.2 Hz, 2H), 7.03 – 6.97 (m, 2H), 6.93 (td, J =
7.6, 0.9 Hz, 1H), 6.80 – 6.74 (m, 1H), 6.72 (d, J = 8.1 Hz, 1H),
6.64 (d, J = 7.7 Hz, 1H). 13C NMR (101 MHz, DMSO) δ 175.67,
163.56, 146.10, 141.65, 138.08, 133.64, 130.79, 128.60, 127.61,
127.59, 127.40, 127.29, 126.47, 122.08, 117.65, 114.57, 114.08,
110.07, 76.35.
6.4.7
3'-(phenylamino)-1'H-spiro[indoline-3,2'-quinazoline]-
2,4'(3'H)-dione (1g). Light yellow solid, 188-190 oC; IR (KBr)
νmax 3300.19, 1726.52, 1650.50, 1614.30, 1487.95, 1328.11,
1237.04, 1197.89, 750.22 cm-1; 1H NMR (400 MHz, DMSO) δ
10.46 (s, 1H), 7.95 (s, 1H), 7.64 (s, 2H), 7.45 (d, J = 8.7 Hz, 1H),
7.39 (s, 1H), 7.26 (s, 2H), 7.06 (s, 3H), 6.94 (d, J = 7.8 Hz, 1H),
6.85 (d, J = 7.8 Hz, 1H), 6.80 (d, J = 7.6 Hz, 1H), 6.71 (s, 3H).
13C NMR (101 MHz, DMSO) δ 163.14, 148.46, 146.26, 142.48,
6.4.2 3'-(4-methoxyphenyl)-1'H-spiro[indoline-3,2'-quinazoline]-
2,4'(3'H)-dione (1b). White solid, mp 244-246 oC; IR (KBr) νmax
3840.79, 3305.43, 1722.73, 1612.34, 1364.08, 1247.48, 1511.21,

7
139.78, 133.86, 130.62, 129.46, 128.49, 128.19, 127.64, 125.17,
122.88, 121.85, 121.13, 119.19, 118.60, 117.39, 114.05, 113.05,
110.48, 109.95, 78.21.
3329.87, 1749.42, 1641.84, 1482.06, 1362.24, 1275.65,
ACCEPTED MANUSCRIPT
1189.40, 749.61, 693.49, 537.49 cm-1; H NMR (400 MHz,
DMSO) δ 7.69 (dd, J = 10.4, 3.7 Hz, 3H), 7.33 (s, 1H), 7.26 (t, J
= 7.5 Hz, 2H), 7.21 (dt, J = 10.7, 4.8 Hz, 2H), 7.05 (d, J = 7.5 Hz,
2H), 6.78 (s, 1H), 6.71 (d, J = 8.0 Hz, 1H), 6.67 (d, J = 8.3 Hz,
1H). 13C NMR (101 MHz, DMSO) δ 175.33, 163.33, 145.81,
140.58, 137.88, 133.78, 130.73, 129.08, 128.80, 127.85, 127.44,
126.62, 126.07, 117.89, 114.51, 114.08, 111.63, 76.36. ESI-MS
(m/z) = 398.0652 ([M+Na]+); found: 398.0672.
1
6.4.8 5-chloro-3'-(p-tolyl)-1'H-spiro[indoline-3,2'-quinazoline]-
o
2,4'(3'H)-dione (1i). White solid, mp 172-174 C; IR (KBr) νmax
3340.12, 2951.55, 1747.91, 1659.24, 1516.35, 1358.28, 1196.27,
814.58, 756.62 cm-1; 1H NMR (400 MHz, DMSO) δ 7.68 (d, J =
6.9 Hz, 3H), 7.36 – 7.29 (m, 1H), 7.23 (dd, J = 8.3, 2.2 Hz, 1H),
7.06 (d, J = 8.1 Hz, 2H), 6.93 (t, J = 7.2 Hz, 2H), 6.77 (dd, J =
11.0, 4.1 Hz, 1H), 6.69 (t, J = 8.3 Hz, 2H), 2.20 (s, 3H). 13C NMR
(101 MHz, DMSO) δ 175.34, 163.44, 145.78, 140.59, 137.12,
135.20, 133.73, 130.70, 129.32, 127.44, 126.49, 126.08, 117.86,
114.50, 114.05, 111.69, 76.43, 20.53. ESI-MS (m/z) = 412.0811
([M+Na]+); found: 412.0829.
6.4.14
5-chloro-3'-(phenylamino)-1'H-spiro[indoline-3,2'-
quinazoline]-2,4'(3'H)-dione (1o). White solid, mp 192-193 oC;
IR (KBr) νmax 3618.95, 3302.46, 1757.08, 1672.42, 1485.59,
1345.43, 1185.63, 752.46, 551.86 cm-1; H NMR (400 MHz,
1
DMSO) δ 10.61 (s, 1H), 8.05 (s, 1H), 7.70 (s, 2H), 7.62 (d, J =
7.5 Hz, 1H), 7.50 (d, J = 7.9 Hz, 1H), 7.39 (s, 1H), 7.33 (d, J =
7.6 Hz, 1H), 7.28 (dd, J = 8.3, 2.2 Hz, 1H), 7.08 (s, 3H), 6.87 (d,
J = 7.9 Hz, 3H), 6.81 (d, J = 8.2 Hz, 1H), 6.77 – 6.67 (m, 3H).
13C NMR (101 MHz, DMSO) δ 175.33, 163.33, 145.81, 140.58,
137.88, 133.78, 130.73, 129.08, 128.80, 127.85, 127.44, 126.62,
126.07, 117.89, 114.51, 114.08, 111.63, 76.36. ESI-MS (m/z) =
413.0781 ([M+Na]+); found: 413.0781.
6.4.9
5-chloro-3'-(4-methoxyphenyl)-1'H-spiro[indoline-3,2'-
quinazoline]-2,4'(3'H)-dione (1j). White solid, mp 228-229 oC;
IR (KBr) νmax 3157.91, 1679.32, 1554.44, 1478.05, 1242.35,
1166.48, 812.26, 662.23 cm-1; H NMR (400 MHz, DMSO) δ
1
10.57 (s, 1H), 7.68 (d, J = 6.1 Hz, 3H), 7.32 (s, 1H), 7.24 (dd, J =
8.3, 2.2 Hz, 1H), 7.06 (d, J = 12.3 Hz, 1H), 6.95 (d, J = 8.9 Hz,
2H), 6.78 (dd, J = 14.3, 8.4 Hz, 3H), 6.72 – 6.66 (m, 2H), 3.68 (s,
3H). 13C NMR (101 MHz, DMSO) δ 175.34, 163.53, 158.27,
145.79, 140.52, 133.71, 130.68, 130.31, 129.42, 127.47, 126.49,
126.11, 119.62, 117.82, 114.76, 114.46, 113.90, 111.66, 76.53,
55.11.
6.4.15
3'-(4-aminophenyl)-5-chloro-1'H-spiro[indoline-3,2'-
quinazoline]-2,4'(3'H)-dione (1p). White solid, mp 188-190 oC;
IR (KBr) νmax 3362.90, 1731.15, 1642.32, 1512.55, 1479.34,
1362.81, 1300.08, 1191.75, 870.88, 756.33, 555.01 cm-1; H
1
NMR (400 MHz, DMSO) δ 10.49 (s, 1H), 7.65 (d, J = 7.7 Hz,
1H), 7.62 – 7.57 (m, 2H), 7.30 (s, 1H), 7.22 (dd, J = 8.3, 2.2 Hz,
1H), 6.75 (s, 1H), 6.69 – 6.62 (m, 4H), 6.34 (d, J = 8.8 Hz, 2H),
5.09 (s, 2H). 13C NMR (101 MHz, DMSO) δ 175.54, 163.56,
147.92, 145.78, 140.56, 133.46, 130.43, 129.78, 127.43, 126.45,
125.92, 125.69, 117.67, 114.73, 113.88, 113.48, 111.51, 76.57.
ESI-MS (m/z) = 413.0781 ([M+Na]+); found: 413.0781.
6.4.10 5-chloro-3'-(o-tolyl)-1'H-spiro[indoline-3,2'-quinazoline]-
2,4'(3'H)-dione (1k). White solid, mp 175-177 C; IR (KBr) νmax
o
3248.57, 1742.82, 1640.64, 1492.63, 1355.19, 1272.60, 1191.18,
754.23, 627.87 cm-1; 1H NMR (400 MHz, DMSO) δ 10.72 (s,
1H), 7.38 (d, J = 2.2 Hz, 1H), 7.36 – 7.30 (m, 2H), 7.21 (dt, J =
14.5, 4.1 Hz, 2H), 7.12 – 7.07 (m, 1H), 7.04 (dd, J = 7.7, 1.2 Hz,
1H), 6.86 (dd, J = 7.8, 1.1 Hz, 1H), 6.82 – 6.76 (m, 2H), 6.75 –
6.70 (m, 2H), 2.22 (s, 3H). 13C NMR (101 MHz, DMSO) δ
175.93, 162.50, 156.84, 145.88, 140.86, 137.42, 136.35, 133.67,
130.92, 130.43, 128.44, 127.95, 127.46, 126.48, 125.84, 125.34,
125.12, 118.02, 115.09, 114.26, 111.69, 76.05, 18.00. ESI-MS
(m/z) = 412.0829 ([M+Na]+); found: 412.0829.
6.4.16 5-bromo-3'-(p-tolyl)-1'H-spiro[indoline-3,2'-quinazoline]-
2,4'(3'H)-dione (1r). yellow solid, mp 150-151 oC; IR (KBr)
νmax 3664.67, 3556.95, 3340.12, 2951.55, 1747.91, 1659.24,
1358.28, 934.41 cm-1; 1H NMR (400 MHz, DMSO) δ 7.67 (dd,
J = 4.3, 1.9 Hz, 3H), 7.33 (dd, J = 10.3, 4.7 Hz, 2H), 7.23 (dd,
J = 8.3, 2.2 Hz, 1H), 7.06 (d, J = 8.1 Hz, 2H), 6.93 (dd, J =
6.0, 2.2 Hz, 2H), 6.77 (t, J = 7.5 Hz, 1H), 6.73 – 6.63 (m, 2H),
2.21 (s, 3H). 13C NMR (101 MHz, DMSO) δ 175.29, 163.38,
145.78, 140.56, 137.07, 135.23, 134.69, 133.68, 130.68,
130.01, 129.29, 127.43, 117.83, 117.45, 114.54, 114.05,
113.08, 111.65, 76.42, 20.52. ESI-MS (m/z) = 434.0509
([M+H]+); found: 434.0514.
6.4.11
5-chloro-3'-(2-methoxyphenyl)-1'H-spiro[indoline-3,2'-
quinazoline]-2,4'(3'H)-dione (1l). White solid, mp 274-275 oC;
IR (KBr) νmax 3211.22, 1734.05, 1642.84, 1499.39, 1365.50,
1275.68, 1191.74, 753.28, 628.63 cm-1; H NMR (400 MHz,
1
DMSO) δ 10.60 (s, 1H), 7.65 (d, J = 4.6 Hz, 2H), 7.49 (d, J = 2.2
Hz, 1H), 7.32 (s, 1H), 7.20 (s, 3H), 6.92 (s, 2H), 6.83 – 6.74 (m,
3H), 6.70 (d, J = 8.2 Hz, 1H), 6.64 (d, J = 8.3 Hz, 1H), 3.75 (s,
3H). 13C NMR (101 MHz, DMSO) δ 175.79, 162.44, 160.15,
155.26, 145.95, 140.63, 133.66, 130.74, 129.77, 128.50, 128.39,
127.38, 126.49, 125.67, 125.49, 120.35, 117.80, 114.55, 114.04,
111.76, 111.28, 76.37, 55.05. ESI-MS (m/z) = 428.0777
([M+Na]+); found: 428.0778.
6.4.17 1-methyl-3'-(p-tolyl)-1'H-spiro[indoline-3,2'-quinazoline]-
o
2,4'(3'H)-dione (2a). White solid, mp 194-196 C; IR (KBr) νmax
1
3270.24, 1727.64, 1521.24, 1366.25, 1243.24, 749.92 cm-1; H
NMR (400 MHz, DMSO) δ 7.68 (dd, J = 7.7, 1.3 Hz, 1H), 7.62
(d, J = 7.4 Hz, 1H), 7.59 (s, 1H), 7.29 (dtd, J = 8.9, 8.0, 1.4 Hz,
2H), 7.06 – 6.96 (m, 3H), 6.88 (d, J = 7.8 Hz, 1H), 6.84 – 6.79
(m, 2H), 6.76 (d, J = 7.7 Hz, 1H), 6.70 (d, J = 7.9 Hz, 1H), 3.03
(s, 3H), 2.17 (s, 3H). 13C NMR (101 MHz, DMSO) δ 173.85,
163.59, 146.02, 143.10, 136.83, 135.28, 133.62, 130.92, 129.16,
127.44, 126.73, 126.13, 122.79, 117.76, 114.56, 114.08, 108.96,
76.11, 25.92, 20.51.
6.4.12
5-chloro-3'-(m-tolyl)-1'H-spiro[indoline-3,2'-
o
quinazoline]-2,4'(3'H)-dione (1m). White solid, mp 188-189 C;
IR (KBr) νmax 3286.87, 1750.81, 1646.74, 1486.78, 1355.25,
1261.38, 1191.14, 750.94, 691.50, 627.40, 552.36 cm-1; 1H NMR
(400 MHz, DMSO) δ 7.74 – 7.63 (m, 3H), 7.36 – 7.30 (m, 1H),
7.22 (dd, J = 8.3, 2.2 Hz, 1H), 7.13 (t, J = 7.7 Hz, 1H), 7.01 (d, J
= 7.6 Hz, 1H), 6.89 (s, 1H), 6.84 (d, J = 7.7 Hz, 1H), 6.78 (t, J =
7.5 Hz, 1H), 6.69 (dd, J = 15.0, 8.2 Hz, 2H), 2.19 (s, 3H). 13
C
6.4.18
3'-(4-methoxyphenyl)-1-methyl-1'H-spiro[indoline-3,2'-
NMR (101 MHz, DMSO) δ 175.36, 163.31, 145.80, 140.63,
138.03, 137.78, 133.72, 130.68, 129.12, 128.52, 127.42, 126.65,
126.04, 117.88, 114.61, 114.09, 111.61, 76.35, 20.68. ESI-MS
(m/z) = 412.0823 ([M+Na]+); found: 412.0829.
quinazoline]-2,4'(3'H)-dione (2b). White solid, mp 224-225 oC;
IR (KBr) νmax 3265.93, 1720.72, 1640.30, 1514.87, 1361.31,
1243.31, 749.71 cm-1; 1H NMR (400 MHz, DMSO) δ 7.68 (d, J =
7.7 Hz, 1H), 7.63 (d, J = 7.4 Hz, 1H), 7.58 (s, 1H), 7.34 – 7.26
(m, 2H), 7.04 (dd, J = 11.0, 4.1 Hz, 1H), 6.89 (d, J = 7.8 Hz, 1H),
6.84 (d, J = 9.1 Hz, 2H), 6.77 (s, 1H), 6.74 (s, 1H), 6.71 (d, J =
6.4.13 5-chloro-3'-phenyl-1'H-spiro[indoline-3,2'-quinazoline]-
o
2,4'(3'H)-dione (1n). White solid, mp 179-180 C; IR (KBr) νmax

8
Tetrahedron
2.8 Hz, 1H), 3.65 (s, 3H), 3.02 (s, 3H). 13C NMR (101 MHz,
128.71, 128.53, 127.38, 127.25, 126.87, 125.22, 123.04, 117.32,
ACCEPTED MANUSCRIPT
DMSO) δ 173.84, 163.71, 158.11, 146.03, 143.07, 133.62,
130.90, 130.35, 127.46, 126.87, 126.06, 122.81, 117.72, 114.49,
114.05, 113.72, 108.97, 76.24, 55.06, 25.91.
114.28, 113.93, 109.56, 70.78.
6.4.24 1-benzyl-3'-(p-tolyl)-1'H-spiro[indoline-3,2'-quinazoline]-
2,4'(3'H)-dione (2i). White solid, mp 180-182 C; IR (KBr) νmax
o
6.4.19
1-methyl-3'-(m-tolyl)-1'H-spiro[indoline-3,2'-
3289.94, 1731.82, 1642.40, 1507.22, 1348.45, 1173.30, 753.83
cm-1; 1H NMR (400 MHz, DMSO) δ 7.73 (s, 1H), 7.70 (d, J = 6.9
Hz, 1H), 7.65 (d, J = 7.4 Hz, 1H), 7.36 – 7.31 (m, 2H), 7.30 (d, J
= 1.7 Hz, 1H), 7.28 (d, J = 1.8 Hz, 2H), 7.23 – 7.18 (m, 1H), 7.16
– 7.12 (m, 2H), 7.02 (t, J = 7.5 Hz, 1H), 6.94 (d, J = 8.1 Hz, 2H),
6.82 – 6.77 (m, 4H), 6.74 (d, J = 8.7 Hz, 2H), 4.79 (dd, J = 60.6,
15.8 Hz, 3H), 2.19 (s, 3H). 13C NMR (101 MHz, DMSO) δ
173.65, 163.58, 145.95, 142.12, 136.98, 135.40, 135.05, 133.73,
130.85, 129.17, 128.43, 127.48, 127.34, 127.02, 126.13, 122.93,
117.83, 114.41, 114.15, 109.66, 76.30, 42.69, 20.57. ESI-MS
(m/z) = 468.1666 ([M+Na]+); found: 468.1688.
o
quinazoline]-2,4'(3'H)-dione (2c). White solid, mp 185-187 C 1;
IR (KBr) νmax 3286.05, 1724.03, 1643.31, 1440.20, 1338.59,
751.00, 693.80 cm-1; 1H NMR (400 MHz, DMSO) δ 7.69 (dd, J =
7.7, 1.3 Hz, 1H), 7.61 (t, J = 3.3 Hz, 2H), 7.32 (td, J = 8.2, 1.6 Hz,
1H), 7.27 (td, J = 7.8, 1.2 Hz, 1H), 7.03 (dt, J = 7.6, 4.2 Hz, 2H),
6.95 (d, J = 7.6 Hz, 1H), 6.87 (d, J = 7.8 Hz, 1H), 6.78 (dd, J =
11.6, 4.3 Hz, 2H), 6.70 (d, J = 7.9 Hz, 2H), 3.03 (s, 3H), 2.14 (s,
3H). 13C NMR (101 MHz, DMSO) δ 173.84, 163.46, 146.05,
143.12, 137.87, 137.80, 133.65, 130.95, 128.34, 128.24, 127.42,
126.61, 126.16, 122.71, 117.78, 114.56, 114.10, 108.91, 76.06,
25.87, 20.63.
Acknowledgments
6.4.20
1-methyl-3'-(m-tolyl)-1'H-spiro[indoline-3,2'-
quinazoline]-2,4'(3'H)-dione (2d). White solid, mp 192-194 oC;
IR (KBr) νmax 3330.93, 3259.12, 1714.64, 1659.46, 1607.85,
The work was supported by Scientific Research Project Item
for Key Laboratory of Shaanxi Province Education Department,
China (No. 2010JS056), Natural Science Basic Research Plan in
Shaanxi Province of China (No.2014JQ2064) and the Foundation
for Young Scholars of Shaanxi University of Science &
Technology (No. BJ12-26). Training Programs of Innovation and
Entrepreneurship for Undergraduates of Shaanxi Province
(NO.1219).
1487.78, 1342.32, 757.71, 689.54 cm-1; H NMR (400 MHz,
1
DMSO) δ 7.69 (s, 1H), 7.62 (d, J = 6.2 Hz, 2H), 7.33 (s, 1H),
7.26 (d, J = 1.0 Hz, 1H), 7.20 (dd, J = 11.3, 4.7 Hz, 2H), 7.15 (d,
J = 7.1 Hz, 1H), 7.02 (t, J = 7.3 Hz, 1H), 6.94 (d, J = 7.5 Hz, 2H),
6.87 (d, J = 7.8 Hz, 1H), 6.79 (s, 1H), 6.72 (d, J = 8.0 Hz, 1H),
3.03 (s, 3H). 13C NMR (101 MHz, DMSO) δ 173.80, 163.52,
146.06, 143.11, 137.90, 133.70, 130.96, 128.62, 127.64, 127.46,
126.58, 126.16, 122.78, 117.81, 114.53, 114.14, 108.93, 76.10,
25.91.
References and notes
1. Li, H.; Huang, R.; Qiu, D.; Yang, Z.; Liu, X.; Ma, J.; Ma, Z. Prog. Nat.
Sci. 1998, 8, 359.
2. Lakhan, R.; Singh, O. P.; Singh, R. L. J. Indian Chem. Soc. 1987, 64,
316.
3. Nandy, P.; Vishalakshi, M. T.; Bhat, A. R. Indian J. Heterocycl. Chem.
2006, 15, 293.
4. Saravanan, G.; Alagarsamy, V.; Prakash, C. R. Int. J. Pharm. Pharm.
Sci. 2010, 2, 83.
5. Honkanen, E.; Pippuri, A.; Kairisalo, P.; Nore, P.; Karppanen, H.;
Paakkari, I. J. Med. Chem. 1983, 26, 1433.
6. Xue, S.; McKenna, J.; Shieh, W.-C.; Repic, O. J. Org. Chem. 2004, 69,
6474.
7. Erlanson, D. A.; McDowell, R. S.; O'Brien, T. J. Med. Chem. 2004, 47,
3463.
8. Xia, Y.; Yang, Z. Y.; Hour, M. J.; Kuo, S. C.; Xia, P.; Bastow, K. F.;
Nakanishi, Y.; Nampoothiri, P.; Hackl, T.; Hamel, E.; Lee, K. H.
Bioorg. Med. Chem. Lett. 2001, 11, 1193.
9. Hour, M.-J.; Huang, L.-J.; Kuo, S.-C.; Xia, Y.; Bastow, K.; Nakanishi,
Y.; Hamel, E.; Lee, K.-H. J. Med. Chem. 2000, 43, 4479.
10. Kamal, A.; Bharathi, E. V.; Reddy, J. S.; Ramaiah, M. J.; Dastagiri, D.;
Reddy, M. K.; Viswanath, A.; Reddy, T. L.; Shaik, T. B.; Pushpavalli,
S. N. C. V. L.; Bhadra, M. P. Eur. J. Med. Chem. 2011, 46, 691.
11. Sun, J.; Li, D.-D.; Li, J.-R.; Fang, F.; Du, Q.-R.; Qian, Y.; Zhu, H.-L.
Org. Biomol. Chem. 2013, 11, 7676.
6.4.21
1-benzyl-3'-(4-methoxyphenyl)-1'H-spiro[indoline-3,2'-
quinazoline]-2,4'(3'H)-dione (2f). White solid, mp 196-198 oC;
IR (KBr) νmax 3280.68, 1730.64, 1610.05, 1509.74, 1361.35,
1241.23, 1178.22, 756.47 cm-1; 1H NMR (400 MHz, DMSO) δ
7.74 – 7.64 (m, 3H), 7.36 – 7.31 (m, 1H), 7.28 (dd, J = 5.0, 1.7
Hz, 3H), 7.22 (td, J = 7.8, 1.2 Hz, 1H), 7.12 (dd, J = 6.7, 2.8 Hz,
2H), 7.04 (td, J = 7.6, 0.7 Hz, 1H), 6.82 (d, J = 8.9 Hz, 2H), 6.78
(d, J = 7.6 Hz, 2H), 6.74 (d, J = 8.0 Hz, 1H), 6.66 (d, J = 9.1 Hz,
2H), 4.88 (d, J = 15.7 Hz, 1H), 4.69 (d, J = 15.7 Hz, 1H), 3.66 (s,
3H). 13C NMR (101 MHz, DMSO) δ 173.57, 163.66, 158.25,
145.97, 142.03, 135.37, 133.72, 130.80, 130.12, 128.41, 127.51,
127.44, 127.37, 127.25, 125.94, 122.95, 117.81, 114.34, 114.13,
113.72, 109.67, 76.47, 55.08. ESI-MS (m/z) = 484.1626
([M+Na]+); found: 484.1637.
6.4.22 1-benzyl-3'-phenyl-1'H-spiro[indoline-3,2'-quinazoline]-
2,4'(3'H)-dione (2g). White solid, mp 228-230 C; IR (KBr) νmax
o
3278.47, 1735.28, 1640.68, 1506.23, 1365.93, 1173.66, 755.92,
693.79 cm-1; 1H NMR (400 MHz, DMSO) δ 7.77 (s, 1H), 7.72
(dd, J = 7.7, 1.2 Hz, 1H), 7.66 (d, J = 6.8 Hz, 1H), 7.38 – 7.24 (m,
5H), 7.22 – 7.09 (m, 7H), 7.01 (t, J = 7.5 Hz, 1H), 6.93 (d, J =
6.5 Hz, 2H), 6.84 – 6.73 (m, 3H), 4.86 (d, J = 15.7 Hz, 1H), 4.72
(d, J = 15.7 Hz, 1H). 13C NMR (101 MHz, DMSO) δ 173.66,
163.53, 145.98, 142.15, 137.70, 135.41, 133.79, 130.87, 128.63,
128.50, 127.74, 127.46, 127.40, 126.80, 126.23, 122.91, 117.88,
114.43, 114.20, 109.64, 76.24. ESI-MS (m/z) = 454.1228
([M+Na]+); found: 454.1231.
12. Lin, H.; Danishefsky, S. J. Angew. Chem. 2003, 115, 38.
13. Ding, K.; Lu, Y.; Nikolovska-Coleska, Z.; Qiu, S.; Ding, Y.; Gao, W.;
Stuckey, J.; Krajewski, K.; Roller, P. P.; Tomita, Y.; Parrish, D. A.;
Deschamps, J. R.; Wang, S. J. Am. Chem. Soc. 2005, 127, 10130.
14. Arun, Y.; Bhaskar, G.; Balachandran, C.; Ignacimuthu, S.; Perumal, P.
T. Bioorg. Med. Chem. Lett. 2013, 23, 1839.
15. Rojas-Duran, R.; Gonzalez-Aspajo, G.; Ruiz-Martel, C.; Bourdy, G.;
Doroteo-Ortega, V. H.; Alban-Castillo, J.; Robert, G.; Auberger, P.;
Deharo, E. J. Ethnopharmacol. 2012, 143, 801.
16. Bhaskar, G.; Arun, Y.; Balachandran, C.; Saikumar, C.; Perumal, P. T.
Eur. J. Med. Chem. 2012, 51, 79.
6.4.23 1-benzyl-1'H-spiro[indoline-3,2'-quinazoline]-2,4'(3'H)-
o
dione (2h). White solid, mp 162-163 C; IR (KBr) νmax 3274.34,
1
17. Parhi, A. K.; Zhang, Y.; Saionz, K. W.; Pradhan, P.; Kaul, M.; Trivedi,
K.; Pilch, D. S.; LaVoie, E. J. Bioorg. Med. Chem. Lett. 2013, 23, 4968.
18. Sajadi, S. M.; Maham, M.; Rezaei, A. Lett. Org. Chem. 2014, 11, 49.
19. Honda, M.; Tamura, M.; Nakagawa, Y.; Tomishige, K. Catal. Sci.
Technol. 2014, 4, 2830.
20. Tamura, M.; Tonomura, T.; Shimizu, K.-i.; Satsuma, A. Appl. Catal., A
2012, 417-418, 6.
21. Agawane, S. M.; Nagarkar, J. M. Tetrahedron Lett. 2011, 52, 3499.
22. Shelkar, R.; Sarode, S.; Nagarkar, J. Tetrahedron Lett. 2013, 54, 6986.
3458.80, 1266.26, 1189.25, 1100.20, 1053.83, 749.54 cm-1; H
NMR (400 MHz, DMSO) δ 8.52 (s, 1H), 7.64 (dd, J = 7.7, 1.2
Hz, 1H), 7.61 – 7.56 (m, 1H), 7.44 – 7.32 (m, 6H), 7.31 – 7.22
(m, 2H), 7.13 (t, J = 7.5 Hz, 1H), 6.91 (d, J = 7.8 Hz, 1H), 6.72 (t,
J = 7.5 Hz, 1H), 6.66 (d, J = 7.9 Hz, 1H). 13C NMR (101 MHz,
DMSO) δ 174.45, 163.94, 146.67, 135.74, 133.35, 130.82,

9
23. Engen, K.; Sävmarker, J.; Rosenström, U.; Wannberg, J.; Lundbäck, T.;
ACCEPTED MANUSCRIPT
Jenmalm-Jensen, A.; Larhed, M. Org. Process Res. Dev 2014, 18,
1582.
24. Mohammadi, A. A.; Askari, S.; Rohi, H.; Soorki, A. A. Synth. Commun
2013, 44, 457.
25. Zhang, J.; Ren, D.; Ma, Y.; Wang, W.; Wu, H. Tetrahedron 2014, 70,
5274.
26. Mochalkin, I.; Miller, J. R.; Narasimhan, L.; Thanabal, V.; Erdman, P.;
Cox, P. B.; Prasad, J. V. N. V.; Lightle, S.; Huband, M. D.; Stover, C.
K. ACS Chem. Biol. 2009, 4, 473.
27. Ma, Y.; Ren, D.; Zhang, J.; Liu, J.; Zhao, J.; Wang, L.; Zhang, F.
Tetrahedron Lett 2015, 56, 4076.
Supplementary Material
Supplementary material that may be helpful in the review
process should be prepared and provided as a separate electronic
file. That file can then be transformed into PDF format and
submitted along with the manuscript and graphic files to the
appropriate editorial office.