NHC-catalyzed green synthesis of functionalized chromones: DFT mechanistic insights and: In vitro activities in cancer cells

Article abstract of DOI:10.1039/c9nj02650a

An efficient synthesis of 3-aminochromones and 3-alkylchromones by a N-heterocyclic carbene (NHC) catalyzed intramolecular hydroacylation reaction of corresponding salicylaldehyde derived nitriles and activated alkynes respectively in ionic liquids under microwave conditions is reported. This protocol has the advantages of environmental friendliness, higher yields, shorter reaction times, and convenient operation using the commercially available thiazolium catalyst. The origin of the chemical reactivity of the NHC-catalyzed intramolecular hydroacylation reaction of nitriles is studied using Density Functional Theory (DFT). The results suggest that 3-aminochromone formation occurs via an acyl anion intermediate called a Breslow intermediate (INT2) through TS2. The Breslow intermediate (INT2) forms a carbon-carbon bond with the nitrile carbon to produce an imine intermediate INT3viaTS3, which further undergoes imine to amine tautomerism to give the end product. Some of the derivatives of 3-aminochromone are subjected to amine functionalization in one pot to obtain a library of compounds for anticancer activity. Among the investigated compounds, 2c (SVM-2), 4c (SVM-4) and 2d (SVM-9) show IC₅₀ values of 5.18, 4.89 and 27.3 μM respectively in HeLa S3 cancer cells. Compound 5c (SVM-5) shows IC₅₀ values of 13.3 and 14.2 μM in A549 and HeLa S3 cancer cells, respectively. Compounds 2c (SVM-2) and 4c (SVM-4) produce morphological changes and control the colony formation in HeLa S3 cells, which indicates that these small molecules are potential candidates for anticancer drugs.

Full text of DOI:10.1039/c9nj02650a

View Article Online
View Journal
NJC
New Journal of Chemistry
Accepted Manuscript
A journal for new directions in chemistry
This article can be cited before page numbers have been issued, to do this please use: N. Murugesh, J.
Haribabu, K. Arumugam, C. Balachandran, R. Swaathy, S. Aoki, S. Anandaram, R. Karvembu and S.
Vedachalam, New J. Chem., 2019, DOI: 10.1039/C9NJ02650A.
Volume 42
This is an Accepted Manuscript, which has been through the
Number
6
21 March 2018
Pages 3963-4776
Royal Society of Chemistry peer review process and has been
accepted for publication.
NJC
New Journal of Chemistry
rsc.li/njc
A journal for new directions in chemistry
Accepted Manuscripts are published online shortly after acceptance,
before technical editing, formatting and proof reading. Using this free
service, authors can make their results available to the community, in
citable form, before we publish the edited article. We will replace this
Accepted Manuscript with the edited and formatted Advance Article as
soon as it is available.
You can find more information about Accepted Manuscripts in the
Information for Authors.
Please note that technical editing may introduce minor changes to the
text and/or graphics, which may alter content. The journal’s standard
Terms & Conditions and the Ethical guidelines still apply. In no event
shall the Royal Society of Chemistry be held responsible for any errors
or omissions in this Accepted Manuscript or any consequences arising
from the use of any information it contains.
ISSN 1144-2546
PAPER
Chechia Hu, Tzu-Jen Lin et al.
Yellowish and blue luminescent graphene oxide quantum dots
prepared via a microwave-assisted hydrothermal route using
H2O2 and KMnO4 as oxidizing agents
rsc.li/njc

Please do not adjust margins
Page 1 of 17
New Journal of Chemistry
1
2
3
4
View Article Online
DOI: 10.1039/C9NJ02650A
Journal Name
5
6
7
8
9
ARTICLE
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
NHC-catalyzed green synthesis of functionalized chromones: DFT
mechanistic insights and in vitro activities in cancer cells
Nithya Murugesh,a Jebiti Haribabu,a Krishnamoorthy Arumugam,a Chandrasekar Balachandran,b
Rajagopal Swaathy,a Shin Aoki,b, c Anandaram Sreekanth,a Ramasamy Karvembu*a and Seenuvasan
Vedachalam*a
Received 00th January 20xx,
Accepted 00th January 20xx
DOI: 10.1039/x0xx00000x
An efficient synthesis of 3-aminochromones and 3-alkylchromones by N-heterocyclic carbene (NHC) catalyzed
intramolecular hydroacylation reaction of corresponding salicylaldehyde derived nitrile and activated alkynes respectively
in ionic liquid under microwave condition is reported. This protocol has the rewards of environmental friendliness, higher
yields, shorter reaction times, and convenient operation using the commercially available thiazolium catalyst. The origin of
chemical reactivity of NHC-catalyzed intramolecular hydroacylation reaction of nitrile is studied with Density Functional
Theory (DFT). The results suggest that 3-aminochromone formation occurs via an acyl anion intermediate called the Breslow
intermediate (INT2) through TS2. Breslow intermediate (INT2) forms a carbon-carbon bond with the nitrile carbon to
produce an imine intermediate INT3 via TS3, which further undergoes imine to amine tautomerism to give an end product.
Some of the derivatives of 3-aminochromone are subjected to amine functionalization in one pot to obtain a library of
compounds for anticancer activity. Among the investigated compounds, 2c (SVM-2), 4c (SVM-4) and 2d (SVM-9) show IC50
values of 5.18, 4.89 and 27.3 µM respectively in HeLa S3 cancer cells. Compound 5c (SVM-5) shows IC50 values of 13.3 and
14.2 µM in A549 and HeLa S3 cancer cells, respectively. Compounds 2c (SVM-2) and 4c (SVM-4) produced morphological
changes and controlled the colony formation in HeLa S3 cells, which indicate that these small molecules are potential
candidates for anticancer drugs.
www.rsc.org/
starting materials, and several reports highlight the recent advances
in this field.14,16-21 Green approaches to diversely substituted
chromones in shorter reaction time are highly desirable as
Introduction
N-heterocyclic carbene (NHC) catalysis has emerged as an efficient
and selective strategy for organic transformations in modern
chemistry.1 They are heavily used owing to their umpolung reactivity
and usefulness in forming important carbon-carbon bonds.2-4 Of the
heterocycles, chromones are among the most prevalent scaffolds
found in natural products and pharmaceuticals.5-8 In the last few
decades, a wealth of research has been focused on the construction
of these chromones.9 Nonetheless, access to chromones with
desirable substitution patterns using sustainable route remains a
challenge when traditional multi-step approaches are applied.10-11
Though classic synthetic protocols are available for chromone core
construction, substituent patterns of these chromones are largely
dictated by the activating groups that are required for reactivity.12-15
Recently, chromones derived by intramolecular hydroacylation of
alkene, nitrile, alkyne and activated alkyne have received much
attention because of broad availability of the salicylaldehyde based
chromones are well-known for their wide-range of pharmacological
and biological activities. This heterocyclic motif especially 3-
aminochromone has repeatedly been found in compounds exhibiting
anti-inflammation, anti-rheumatic, anti-leukemic, anti-mutagenic
and selectively inhibition of v-abl tyrosine protein kinases properties
(Figure 1).22-33 These motivate us to devise sustainable
methodologies for its synthesis using NHC catalysis. In this regard,
based on the earlier communications,34,35 an intramolecular
hydroacylation was employed using commercially available
thiazolium salt in ionic liquid under microwave condition. From
which, 3-aminochromones were generated, which were highly
valuable building blocks for further functionalization that leads to a
series of 3-aminochromone derivatives for anticancer activity.22-33
Figure 1. Biologically active 3-aminochromones.
a. Department of Chemistry, National Institute of Technology, Tiruchirappalli 620
015, India. Corresponding authors: kar@nitt.edu; seenuvasanv@gmail.com
b. Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki,
Noda 278-8510 Japan
Recently, the combinatorial syntheses of various new small organic
molecules using NHC based organocatalytic approach were
experimented with considerable success using various organic
solvents.34-38 To avoid the disadvantages such as volatility and
toxicity of many organic solvents, inherently, we have employed
c. Research Institute of Science and Technology, Tokyo University of Science, 2641
Yamazaki, Noda 278-8510 Japan
Electronic supplementary information (ESI) available: For detailed experimental
procedures and characterization data See DOI: 10.1039/x0xx00000x
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 1
Please do not adjust margins

Please do not adjust margins
New Journal of Chemistry
Page 2 of 17
ARTICLE
Journal Name
1
2
3
4
5
6
7
8
9
ionic liquids as
a green medium for this intramolecular
View Article Online
hydroacylation reaction. Ionic liquids have attracted our interest as Table 1. Optimization of the reaction conditiDoOn.I: 10.1039/C9NJ02650A
using it is one of the four principal methods to avoid conventional
organic solvents.39,40 As organic salts with melting points below
ambient temperature, they lack a measurable vapor pressure and
have high chemical and thermal stabilities, which mark them as an
environmentally benign reaction media.41-43 Microwave activation is
another green synthetic approach that has emerged as a powerful
promoter of a variety of reactions, including organic reactions.44-47
The irradiation of microwaves on the reaction mixture significantly
reduces reaction time, enhances conversions and boasts
environmental advantages as well as the induction of reactions
under dry conditions.44 In this article, we are presenting the achieved
sustainable synthesis of chromone derivatives in line with the earlier
communications on intramolecular hydroacylation chemistry34,35
using commercially available thiazolium catalyst in ionic liquids under
microwave condition. In earlier studies, chromone synthesis has
been achieved using dry organic solvent with longer reaction time.
Hence, we have undertaken an investigation using commercially
available thiazolium (1) catalyst, salicyladehyde derived nitrile or
activated alkyne as the reacting species in ionic liquid under
microwave condition. Our computational studies elucidated the
origin of intramolecular hydroacylation reactivity of nitrile. We also
provide insights on the Breslow intermediate (INT2) favored reaction
pathway. The resulting 3-aminochromones were further subjected
to amine functionalization using one pot protocol or reaction with
isolated 3-aminochromones to obtain a library of 3-aminochromone
derivatives in good to excellent yields. In this case, a series of amide
linked compounds was prepared by the reaction of amine with
bromoacetylbromide followed by N-alkylation with morpholine or
piperidine at 0 ℃ to rt. Successively, the isolated 3-aminochromones
reacted with 3-formylchromone in various alcohols, formed a new
class of three component addition products. Some of the amine
functionalized compounds were subjected to in vitro anticancer
activity using HeLa S3 and A549 cancer cell lines. Additionally,
morphological changes and controlled colony formation in HeLa S3
cells have been studied using the active compounds. Furthermore,
the developed sustainable reaction condition was applied to
salicylaldehyde derived activated alkynes which delivered the 3-
alkylchromones in good to excellent yields.
HO
O
O
Ph
N
S
N
NH2
1
Cl
Microwave condition
Et3N, [Ionic liquid]
O
O
1b
1a
S.
1
Et3N
Tem.
Time
[min]
Yield
[%]b
Solventa
No. [mol %]
[mol %]
20
[℃]
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
1
2
0
[bmim][BF4]
[bmim][BF4]
[bmim][BF4]
[bmim][BF4]
[bmim][BF4]
[bmim][BF4]
[bmim][BF4]
[bmim][BF4]
[bmim][Cl]
80
80
80
80
80
80
80
100
80
80
60
60
60
60
60
15
30
60
60
60
5
5
5
65
77
83
92
63
75
91
82
80
3
10
15
20
20
20
20
20
20
10
4
15
5
20
6
20
7
20
8
20
20
20
9
10
[bmim][Br]
Reaction condition: a1 mole equiv. of ionic liquid was used w.r.t. starting material.
bIsolated yield of 1b
With other ionic liquids such as [bmim][Cl] and [bmim][Br] under
entry 5 condition, the reaction provided moderate yields (entries 9
and 10). Finally, 0.2 equiv. of catalyst 1 and 0.2 equiv. of
triethylamine in 1 equiv. of [bmim][BF4] were employed at 80 °C over
60 min microwave irradiation to achieve 1b (entry 5). With the
optimized sustainable reaction condition in hand (Table 1, entry 5)
for the synthesis of 3-aminochromone, we explored the generality of
this reaction (Table 2). A broad range of differently substituted
salicylaldehyde derived nitriles bearing electron-neutral, electron-
donating, and electron-withdrawing substituent on aromatic rings
were examined. Our results showed that the reaction can be
effectively adopted for various salicylaldehyde derivatives (Table 2).
The electron-neutral alkyl substituted aldehydes exhibited
prominently good yields (1b–5b, 83–92 %). An electron-donating
methoxy substituent in the aromatic ring was suitable for the
reaction with no apparent change in reactivity and provided good to
excellent yields (6b–10b, 80–94 %). Substrate with an electron-
withdrawing nitro substituent in the aromatic ring showed a
moderate yield of 78 % (11b). Further exploration revealed that the
substrates with halo substituent(s) on the ring can give good to
excellent yields (12b–20b, 81–93 %). Thus, scope of the reaction is
Results and discussion
We were pleased to find the green intramolecular hydroacylation
reaction of 2-(2-formylphenoxy)acetonitrile (1a) using 3-benzyl-5-(2-
hydroxyethyl)-4-methylthiazolium chloride (1) as a catalyst, Et3N as a
base and 1 equivalent of commercially available ionic liquid as
solvent under microwave condition (Table 1). Without the catalyst,
in 1 equivalent of [bmim][BF4], the reaction afforded only a trace
amount of the product (1b) under microwave condition at 80 ℃
(entry 1) in one hour. When the amount of thiazolium salt 1 was
increased along with the equal ratio of triethylamine (5, 10, 15 and
20 mol %), the yield of 3-aminochromone 1b was improved (65, 77,
83 and 92 %) (entries 2-5). The use of 20 mol% of the catalyst (1) gave
good yield (92 %) (entry 5). Further, decrease of reaction time (30
and 15 min) lowered the yield of 1b from 92 % to 75 % and 63 %
respectively (entries 6 and 7). Raising the reaction temperature from
80 to 100 °C did not improve the yield of 1b (entry 8).
wide, allowing the facile generation of
a variety of 3-
aminochromones in a sustainable manner with excellent yields. Our
results showed that the intramolecular hydroacylation of nitrile can
be adopted for various salicylaldehyde derived nitrile compounds.
Furthermore, the reaction proceeded well with the compounds
having active alkyne group (1e–15e) to obtain the corresponding 3-
alkyl chromone derivatives (1f–15f) in good to excellent yields (Table
6). But the compound with 1-adamantane carbonyl group (16e) did
not produce the desired product due to steric hindrance.
Table 2. Substrate scope of 3-aminochromones.a
O
O
HO
N
Ph
S
N
R3
R2
NH2
R4
R3
R2
1
Cl
O
O
R4
Et3N (20 mol %),
R1
R1
1 equiv. [Bmim][BF4
]
Microwave condition, 80 C
1b-20b
1a-20a
2 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 3 of 17
New Journal of Chemistry
Please do not adjust margins
Journal Name
ARTICLE
1
2
3
4
5
6
7
8
9
intramolecular hydroacylation reaction within the Breslow
intermediate (INT2) catalyzed by NHC. The stable conformation
(minimum free energy) of the starting materials, products,
intermediates and transition states are shown in Figures 3-6. We
View Article Online
DOI: 10.1039/C9NJ02650A
O
O
O
O
O
O
O
NH2
NH2
NH2
NH2
NH2
have
studied
the
hydroacylation
reaction
of
2-(2-
O
O
O
1b, 92 %
4b, 86 %
2b, 91 %
3b, 87 %
5b, 83 %
formylphenoxy)acetonitrile (1a) using benzyl thiazolium catalyst (1).
The mechanism proposed for this NHC-catalyzed transformation
contained three intermediates and three transition states.
O
O
O
O
O
NH2
NH2
NH2
MeO
NH2
NH2
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
MeO
O
O
Ph
O
O
O
8b, 93 %
10b, 82 %
9b, 80 %
OMe
Cl
OMe
Cl
7b, 90 %
6b, 94 %
OMe
O
O
O
O
O
NH2
O2
N
NH2
Br
NH2
NH2
Cl
NH2
O
O
O
O
O
11b, 78 %
13b, 88 %
14b, 91 %
15b, 92 %
Br
12b, 87 %
Cl
O
O
O
O
O
NH2
F
NH2
I
NH2
NH2
F
NH2
F
O
O
O
O
O
16b, 93 %
19b, 88 %
20b, 81 %
OMe
18b, 89 %
F
F
17b,
92 %
aIsolated yields of 1b-20b
Density functional theory (DFT) calculation
Umpolung reactivity is an essential synthetic strategy in organic
chemistry because it can offer an unconventional technique for
conventional bond formation. Numerous NHC-catalyzed reactions
have been developed by using the umpolung strategy such as
Figure 2. The calculated Gibbs free energy (kcal/mol) profile of the possible reaction
pathway.
benzoin
condensation,
Stetter
reaction
and
cycloaddition/annulation reactions.1-4 In accordance with the
mechanistic studies of those reactions, we understood that the
umpolung which triggered the whole catalytic cycle, did it by
nucleophilic addition of the carbene carbon to the electrophilic
carbon of the aldehyde, ketene, etc.3b-j The initial step of the title
reaction was proposed to be the nucleophilic addition of the carbene
carbon in the NHC (generated in situ from the deprotonation of the
thiazolium cation by base) to the aldehyde, which resulted in the
formation of a NHC-aldehyde adduct INT1 (INT-intermediate) as
shown in Scheme 1.
During the C–C bond formation between the catalyst and aldehyde,
TS1 (TS = Transition state) occurred, which showed two strong
hydrogen bonding [H∙∙∙O∙∙∙H] interactions between the catalyst and
reactant. Nucleophilic addition to the aldehyde formed INT1 and
negative charge on oxygen was stabilized through a hydrogen bond.
Furthermore, an acyl anion intermediate (INT2) (8.96 kcal/mol) was
formed via TS2 (35.08 kcal/mol). The barrier energy of TS2 was found
to be higher; however, the inclusion of solvation effect (Et3N)
substantially reduced the barrier to 19.46 kcal/mol. Also, we tracked
down the base (Et3N) incorporated transition state (See the stabilized
TS in the SI). From this, we confirmed that the polarity of the carbonyl
carbon had changed and umpolung strategy was attained. A close
look on INT2 revealed that this intermediate had two strong
hydrogen bonding interactions involving methylene hydrogen atom
present in the catalyst and the nitrile counterpart, which in turn
stabilized the intermediate (Figure 5). Then hydroacylated product
was formed through TS3 (14.11 kcal/mol). Partial CC bond cleavage
was explained in INT3 structure. After the tautomerization, INT3
gave thermodynamically stable hydroacylated product 3-
aminochromone which had the lowest Gibbs free energy (18.62
kcal/mol) compared with the starting materials.
R2
R1
O
N
N
N
S
R2
S
N
R''
N
R''
R''
N
O
H
OH
Base
R1
R1
O
R1
O
O
S
1a
S
R2
R2
INT1
N
(NHC addition
to aldehyde)
INT2
R''
(Proton
Carbanion
intermediate
transfer)
Proton shift
NHC elemination
O
O
HN
O
R''
O
NH2
N
R1
NH
O
S
R2
1b
Product
O
H
n
tio
iza
er
om
ut
ta
INT3
Imine
Scheme 1. Reaction mechanism based on the DFT calculation.
The reaction was accomplished via three stages, i.e. the
formation of Breslow intermediate INT2, intramolecular
hydroacylation of nitrile to give imine INT3, and imine-amine
tautomerization. The calculated Gibbs free energy profile of the
mechanism is shown in Figure 2. The free energy of the aldehyde +
NHC was set as 0.0 kcal/mol. All the optimized structures and
geometrical parameters are displayed in SI.48 The B3LYP-DFT method
has been successfully applied to gain better insights on the
Figure 3. Minimum energy stable conformation of the nitrile, thiazolium catalyst and 3-
aminochromone calculated at the B3LYP/6-31g (d,p) level of theory.
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 3
Please do not adjust margins

Please do not adjust margins
New Journal of Chemistry
Page 4 of 17
ARTICLE
Journal Name
1
2
3
After the formation of 3-aminochromones (Table 2 condition), they
View Article Online
were reacted with 1.1 equiv. of bromoacetyDlbOroI:m10i.d10e3i9n/Cp9reNsJe0n2c6e50oAf
2 equiv. of Et3N for 30 min. The amine compounds were converted
to amide derivatives which further reacted with morpholine or
piperidine for another 2-10 h at room temperature to form
morpholine or piperidine based N-functionalized 3-aminochromones
in moderate yield. In this process, we have synthesized a series of
compounds (1c-12c) easily in one pot using various 3-
aminochromone derivatives under Table 2 condition (Table 3).
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Figure 4. Minimum energy stable conformation of the intermediates (INT1 and INT2)
that occur during the formation of Breslow intermediate, calculated at the B3LYP/6-31g
(d,p) level of theory.
Three component reactions
During the course of amine functionalization, we intended to
synthesize an another series of molecules for cancer treatment by
carrying out one-pot three component addition reactions (Table 4).
In this case, 1 equiv. of 3-aminochromone, 1 equiv. of 3-
formylchromone and an excess amount of alcohol were refluxed
(~80 ℃) for 2 h to produce (Z)-3-(((2-alkoxy-4-oxochroman-3-
ylidene)methyl)amino)-4H-chromen-4-one as yellow precipitate
upon cooling. The compound was obtained selectively in one
diasteromeric form which has been characterized by NMR and single
crystal X-ray analyses. Crystal structure of 1d is given in Figures 7 and
8. We explored the generality of the reaction by using various 3-
formylchromones, 3-aminochromones and alcohols to obtain the
desired three-component products (1d12d) in moderate yields (65-
78 %) (Table 4). The isolation of pure products was achieved by
simple filtration of the precipitate and then washing with cold
ethanol.
Figure 5. Minimum energy stable conformation of the imine intermediate (INT3) and
imine calculated at the B3LYP/6-31g (d,p) level of theory .
Table 4. Substrate scope for the amine functionalized 3-
aminochromone.a
O
1d, (SVM-8)
2d, (SVM-9)
68 % R=Me;R1=R2=R3=H
78 %; R= Et, R1=R2=R3=H
Figure 6. Minimum energy stable conformation of transition states TS1, TS2 and TS3,
O
O
O
NH2
R1
calculated at the B3LYP/6-31g (d,p) level of theory.
3d, (SVM-19)
4d, (SVM-10)
5d, (SVM-11)
6d, (SVM-12)
7d, (SVM-13)
8d, (SVM-14)
9d, (SVM-15)
10d, (SVM-16)
11d, (SVM-17)
12d, (SVM-18)
, 68 %; R= Isopropyl,R1=R2=R3=H
, 72 %; R= Et, R1=Me;R2=R3=H
, 77 %; R= Me, R1=R3=H; R2=OMe
, 73 %; R= R1=Me, R3=H; R2=OMe
, 75 %; R= Et; R1=R3=H; R2=OMe
, 65 %; R= Et, R1=Me, R3=H; R2=OMe
+
R3
O
R2
R1
ROH, 80 o
2-3 h
C
Formation of 3-aminochromone derivatives
Table 3. Substrate scope for the amine functionalized 3-
aminochromone.a
O
O
, 72 %; R= Me, R1=R2 H, R3=OMe
=
H
N
O
OR
, 71 %; R= Et, R1=R2 H, R3=OMe
=
O
O
, 76 %; R= R1=Me, R2 H, R3=OMe
=
N
H
N
R3
R3
1. Table 2 condition
R3
O
, 69 %; R= Et, R1=Me, R2 H, R3=OMe
=
N
R2
2. 1.1 equiv. BrCOCH2Br
0 C, 1.1 equiv. Et3N
3. Morpholine or Piperidine
O
X
R2
O
R4
R2
O
R4
aIsolated yields of 1d-12d
X=O or CH2
R1
O
R1
O
HCl
O
O
H
N
H
H
N
Cl
Cl
N
The possible reaction pathway is depicted in Scheme 2. The reaction
of 3-formylchromone with alcohol (methanol) produced acetal or
hemiacetal (I) which further eliminated methanol or water molecule
to obtain the intermediate (II). The intermediate (II) reacted with the
amine part of 3-aminochromone to produce an another
intermediate (III). Amine donated its lone pair of electrons to the
chromone ring, which favoured a ring opening intermediate (IV)
which on ring closure furnished the corresponding product (1d) at 80
℃. In order to identify the reaction pathway, we have prepared the
methylacetal protected 3-formylchromone and carried out the same
reaction with 3-aminochromone to obtain similar three component
product (1d). The result confered that the reaction might proceeded
through acetal or hemiacetal formation48 from 3-formylchromone
and alcohol, followed by 3-aminochromone addition (Path A). Also,
imine formation followed by Michael addition with alcohol can
produce three component adduct 1d through path way B. We have
tested this possibility using high molecular weight alcohol. The
F
N
N
N
O
O
O
O
O
O
O
O
1c, SVM-1, 91 %
Cl
2c, SVM-2, 65 %
3c, SVM-3, 61 %
O
H
N
O
H
N
O
H
N
MeO
N
N
N
O
O
O
O
O
O
O
O
O
O
4c, SVM-4, 63 %
5c, SVM-5, 63 %
6c, SVM-6, 80 %
OMe
O
H
N
O
O
O
H
N
H
N
N
Cl
N
N
O
O
O
O
O
O
8c, SVM-20, 64 %
O
N
7c, SVM-7, 60 %
9c, 66 %
Cl
O
O
O
H
N
O
H
N
H
N
F
N
N
N
O
O
O
O
O
O
MeO
O
OMe
12c, 64 %
11c, 62 %
10c, 66 %
aIsolated yields of 1c-12c
Functionalized 3-aminochromones showed various biological and
pharmaceutical activities22-33 which envisage us to make a new class
of compounds for cancer treatment by functionalizing the amine part.
4 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 5 of 17
New Journal of Chemistry
Please do not adjust margins
Journal Name
ARTICLE
1
2
3
4
5
6
7
8
9
reaction did not produce the desired three component adduct, which Table 5. IC50 values of the compounds against HeLaViSew3 Aartnicdle AOn5l4in9e
DOI: 10.1039/C9NJ02650A
supported that the path A was highly desirable
cancer cells.
Compo-
und
OMe
O
A549
IC50 (µM)
>100
>100
>100
75.2
HeLa S3
IC50 (µM)
12.1
Compou-
nd
A549
IC50 (µM)
71.2
HeLa S3
IC50 (µM)
41.6
43.2
44.7
36.6
36.2
47.7
42.3
44.3
82.3
21.3
-
MeO
O
N
Path B
1b
O
O
O
O
Path A
MeOH
OHC
O
O
O
MeOH
OH
or
80 
C
SVM-1
SVM-2
SVM-3
SVM-4
SVM-5
SVM-6
SVM-7
SVM-8
SVM-9
SVM-10
SVM-11
SVM-12
SVM-13
SVM-14
SVM-15
SVM-16
SVM-17
SVM-18
SVM-19
SVM-20
Cisplatina
-
80 
C
MeOH
MeO
5.18
>100
73.7
1d
O
(I)
MeO
MeO
>100
4.89
O
O
O
O
H
N
MeO
H
N
O
O
O
O
>100
>100
>100
86.7
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
NH2
O
O
O
O
13.3
14.2
(III)
(II)
1b
(IV)
>100
>100
>100
>100
>100
>100
70.7
Scheme 2. Plausible reaction pathway.
>100
42.9
>100
>100
15.1
27.3
91.8
42.6
-
a Literature values49,50
Figure 7. Solid thermal ellipsoid plot of the compound 1d with 50 % probability level.
Selected bond lengths (Å) and angles (°): O1C11.2311(15), O2C10 1.2489(15), O3C3
1.3689(15), O3C4 1.3520(16), O4C12 1.4349(15), O4C13 1.3710(15), O5C12
1.4073(14), O5C19 1.4312(16), N1C5 1.3945(16), N1C20 1.3453(16), O1C1C5
121.79(12), O1C1C2 124.30(11), C3O3C4 118.78(10), C5N1C20 125.76(11),
C12O4C13 116.14(9), C12O5C19 113.13(10), O2C10C11 123.01(12),
O2C10C14 121.14(11). Hydrogen bond distance (Å): N1H…O2 2.6638(14)
Figure 9. Anticancer property of the compounds (SVM-1 to SVM-20) against HeLa S3 cells
after 24 h. The experiments were conducted in triplicate and data was calculated by
mean ± SD.
Figure 8. (a) Crystal packing viewed down the b-axis showing N1H…O2 intramolecular
interaction in compound 1d. (b) Molecular packing diagram of compound 1d exhibiting
 stacking interaction.
The results obtained here are comparable with those of cisplatin
which has an IC50 value of 21.3 µM in HeLa S3, and 15.1 µM in A549
cancer cell lines.49,50 Based on the structure-activity relationship
(SAR), compound SVM-4 which has a benzene ring substituent at 5th
and 6th carbons of 3-aminochromone showed enhanced activity in
HeLa S3 cancer cells. Analogously, SVM-2 that has a chloro
substituent at 6th carbon of 3-aminochromone showed the lowest
IC50 value in HeLa S3 cells. Similarly, the compound with methoxy
substituent at 6th carbon of 3-aminochromone showed an excellent
in vitro activity against both HeLa S3 and A549 cells. Further,
morpholine attached compounds showed enhanced activity towards
cancer cell lines compared with the three component products.
However, few three component products showed moderate activity
when compared to the standard drug cisplatin.
In vitro anticancer activity of the functionalized 3-aminochromones
(SVM-1 to 20)
The anticancer activity of the selected compounds (SVM-1 to SVM-
20) was tested against human cervical (HeLa S3) and human lung
(A549) cancer cell lines by MTT assay. The plots of % of cell survival
versus log concentration are displayed in Figures 9 and 10; the
obtained IC50 values are given in Table 5. The IC50 values clearly
indicated that the compounds SVM-2 (5.18 µM) and SVM-4 (4.89 µM)
can exhibit potential anticancer activity against HeLa S3 cells. In the
same cell line, compounds SVM-1, SVM-5, and SVM-9 displayed
moderate anticancer activity with IC50 values of 12.1, 14.2 and 27.3
µM respectively, while the other compounds showed less activity
(IC50 > 36 µM). Besides, compound SVM-5 showed a significant
anticancer activity in A549 cancer cells with an IC50 value of 13.3 µM
while all the other compounds displayed moderate activity (Table 5).
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 5
Please do not adjust margins

Please do not adjust margins
New Journal of Chemistry
Page 6 of 17
ARTICLE
Journal Name
1
2
3
4
5
cells. Therefore, SVM-2 and SVM-4 not only showVeiedwaAnrtitcilceaOnncliener
activity in HeLa S3 cells but also controlled the colony formation
ability.
DOI: 10.1039/C9NJ02650A
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Figure 10. Anticancer property of the compounds (SVM-1 to SVM-20) against A549 cells
after 24 h. The experiments were conducted in triplicate and data was calculated by
mean ± SD.
Figure 12. Colony formation study in HeLa S3 cells after the exposure of SVM-2 and SVM-
4. The data was calculated by mean±SD using ImageJ software with three replications.
Significance was compared with treated and untreated groups **** p < 0.0001, *** p <
0.0008 and ** p<0.0091.
Morphological changes in HeLa S3 cancer cells
Table 6. Substrate scope of 3-alkylchromones.a
Ph
N
R
O
O
Cl
S
O
1
R3
R2
R
R3
R2
OH
O
O
1f-16f
O
R4
O
R4
Et3N(20 mol %),
[Bmim][BF4]
Microwave condition
R1
R1
1e-16e
O
CO2Et
CO2Et
O
O
CO2Et
O
CO2Et
O
CO2Et
O
O
O
O
1f, 93 %
4f, 95 %
2f, 96 %
3f, 94 %
5f, 85 %
O
O
CO2Et
O
CO2Et
CO2Et
O
CO2Et
O
O
CO2Et
MeO
Cl
O
CO2Et
O
O
MeO
Cl
OMe
O
Cl
O
6f, 94 %
8f, 88 %
7f, 90 %
10f, 86 %
9f, 89 %
O
CO2Et
O
CO2Et
O
14f, R=Me, 84 %
15f, R=Ph, 81 %
O
CO2Et
R
O
F
O
O
O
16f, R=Admandyl, 0 %
O
12f, 90 %
Figure 11. Morphological changes and nuclear condensation were observed by
fluorescence probes after the treatment of SVM-2 and SVM-4 in HeLa S3 cells. AO
indicates live cells and PI indicates dead cells. Images were captured under fluorescence
microscope (Biorevo, BZ-9000, Keyence).
13f, 88 %
11f, 86 %
aIsolated yields of 1d-12d
AO (acridine orange)/PI (propidium iodide) staining was evaluated to
see the morphological changes associated with apoptotic mode of
cell killing. Acridine orange is used as a dye that could stain nucleic
acids (DNA and RNA) with an intact cell membrane, and propidium
iodide is also a dye which penetrates only damaged cell membrane.
The morphological changes in HeLa S3 cancer cells after the
treatment of compounds SVM-2 and SVM-4 (10 µM) are displayed in
Figure 11. Treatment of compounds SVM-2 and SVM-4 caused
dramatic changes in morphology, nuclear condensation, membrane
blebbing and irregularity in shape in cancer cells (Figure 11). AO/PI
staining studies clearly confirmed that the compounds showed
significant morphological changes and induced cell death.
Conclusions
In summary, we have developed
a highly efficient and
environmentally benign synthesis of 3-aminochromones and 3-
alkylchromones via an intramolecular hydroacylation reaction of
aldehyde-nitrile and aldehyde-activated alkyne respectively using
thaizolium catalyst in ionic liquid under microwave condition. The
DFT studies showed that the formation of 3-aminochromone was
achieved via an acylanion intermediate (INT2) which was the driving
force for the reactivity. Furthermore, a series of 3-aminochromone
derivatives was constructed by using a scaffold modification strategy.
3-Aminochromones were reacted with bromoacetylbromide and
morpholine or piperidine to obtain morpholine or piperidine
fuctionalized 3-aminochromone derivatives in moderate to good
yields through one pot protocol. In addition, we have developed a
Colony formation studies
Effect of SVM-2 and SVM-4 on colony formation was investigated in
HeLa S3 cells. As shown in Figure 12, the treatment of 10 μM of SVM-
2 and SVM-4 significantly inhibited the colony formation in HeLa S3
6 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 7 of 17
New Journal of Chemistry
Please do not adjust margins
Journal Name
ARTICLE
1
2
3
4
5
6
7
8
9
new type of three-component adduct library by reacting 3- conversion (as judged by thin layer chromatographVyi,ew3A0rti%cle Oetnhlinyel
aminochromones with 3-formyl chromones in alcohol. From this acetate/hexane), water (30 mL) was addedDtoOIt:h1e0.1r0e3a9c/tCio9nNJm0i2x6t5u0rAe
library, twenty compounds were evaluated for anticancer activity in and the precipitate was extracted with ethylacetate (320 mL) and
HeLa S3 and A549 cell lines. Among which, SVM-2, SVM-4 and SVM- washed with brine (210 mL), and dried over Na2SO4. The solvent
9 showed IC50 values of 5.18, 4.89 and 27.3 µM respectively in HeLa was evaporated and the crude product was purified by flash
S3 cells. Compound SVM-5 showed IC50 values of 13.3 and 14.2 µM chromatography on silica to obtain 3-amino-4H-chromen-4-one
in A549 and HeLa S3 cells respectively. The most potent compounds (1b)35 as pale yellow solid (Yield 92 mg, 92 %). m. p. 107−109 ℃ (lit.35
1
SVM-2 and SVM-4 produced morphological changes and controlled 108 ℃); H NMR (500 MHz, CDCl3): δ 8.26 (d, J = 8 Hz, 1H), 7.78 (s,
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
the colony forming ability in HeLa S3 cells, which indicated that these 1H), 7.63–7.60 (m, 1H), 7.42 (d, J = 8.5 Hz, 1H), 7.35 (dd, J1 = 8 Hz, J2
molecules are potential candidates for anticancer activity.
= 7.5 Hz, 1H), 3.64 (brs, 2H, -NH2).
3-Amino-8-methyl-4H-chromen-4-one (2b):35 The title compound
was prepared according to the general procedure using 2-(2-formyl-
6-methylphenoxy)acetonitrile (2a) (100 mg, 0.571 mmol, 1 equiv.), 1
(30 mg, 0.114 mmol, 0.2 equiv) and triethylamine (16 μL, 0.114 mmol,
0.2 equiv) in [bmim][BF4] (106 µL, 0.571 mmol, 1 equiv.). The product
was obtained as yellow solid (Yield 91 mg, 91 %). m. p. 127−129 ℃
(lit.35 127 ℃); 1H NMR (400 MHz, DMSO-d6): δ 8.01 (s, 1H), 7.92 (d, J
= 8 Hz, 1H), 7.55 (d, J = 7.2 Hz, 1H), 7.28 (t, J = 7.6 Hz, 1H), 4.54 (brs,
2H, -NH2), 2.41 (s, 3H).
Experimental
General methods: All the reactions were conducted in a flame or
oven dried glassware under nitrogen atmosphere with freshly
distilled dry solvents under anhydrous conditions unless otherwise
indicated. Evaporation of organic solvents was achieved by rotary
evaporation at a temperature below 40 ℃. Product purification by
flash column chromatography was accomplished using silica gel 60
(0.010–0.063 nm). Chromatograms were visualized by fluorescence
quenching with UV light at 254 nm or by staining using base solution
of potassium permanganate. Technical grade solvents were used for
chromatography and were distilled prior to use. NMR spectra were
recorded at room temperature on 300 MHz Bruker ACF 300, 400 MHz
Bruker DPX 400, 500 MHz Bruker AMX 500 or 400 MHz JEOL ECA 400
NMR spectrometer. HR-MS (ESI) spectra were recorded on a Waters
Q-Tof premierTM mass spectrometer. Single crystal X-ray analysis was
done on a Bruker X8 CCD diffractometer.
Materials: All the starting materials were purchased from
commercial suppliers and used without further purification. The
thazolium catalyst (1) was purchased from a commercial supplier and
used directly for hydroacylation reaction. All the ionic liquids used
were purchased from a commercial supplier and used without
further purification. Starting materials (1a-21a; 1e-16e) were
purified through silica gel column and perfectly dried prior to use.
Experimental procedures:
Synthesis of 1a-20a:35 In a round-bottom flask, salicylaldehyde (100
mg, 0.825 mmol, 1 equiv.) was dissolved in dry DMF (0.5 mL) and
stirred with anhydrous K2CO3 (169 mg, 1.23 mmol, 1.5 equiv.) for 15
minutes at ambient temperature under nitrogen atmosphere. After
the formation of potassium salt of salicylaldehyde,
bromoacetonitrile (125 mg, 0.98 mmol, 1.2 equiv.) was added and
stirring was continued for 18-24 h. The reaction progress was
monitored using TLC. After the completion of the reaction, the
reaction mixture was diluted with ethyl acetate (10 mL), washed with
water (3x10 mL), brine (2x5 mL), and dried over Na2SO4. The solvent
was evaporated and the crude product was purified by flash
chromatography on silica (Yield 80-98 %).
Synthesis of 3-aminochromone and 3-alkylchromone (1b-21b, 1f-
15f): Thiazolium precatalyst (1) (23 mg, 0.124 mmol, 0.2 equiv.) and
2-(2-formylphenoxy)acetonitrile35 (1a)(100 mg, 0.621 mmol, 1 equiv.)
were suspended in [bmim][BF4] (116 µL, 0.621 mmol, 1 equiv.) in a
microwave sealed tube under nitrogen atmosphere. Triethylamine
(18 µL, 0.124 mmol, 0.2 equiv.) was added via syringe to the reaction
mixture which was allowed to stir at 80 ℃ under microwave
condition for 1 h (CEM Microwave synthesizer was used) to produce
3-Amino-6-methyl-4H-chromen-4-one (3b):35 The title compound
was prepared according to the general procedure using 2-(2-formyl-
4-methylphenoxy)acetonitrile (3a) (100 mg, 0.571 mmol, 1 equiv.), 1
(30 mg, 0.114 mmol, 0.2 equiv) and triethylamine (16 μL, 0.114 mmol,
0.2 equiv) in [bmim][BF4] (106 µL, 0.571 mmol, 1 equiv.). The product
was obtained as yellow solid (Yield 87 mg, 87 %). m. p. 87−89 ℃ (lit.35
88 ℃); 1H NMR (500 MHz, CDCl3): δ 8.04 (s, 1H), 7.77 (s, 1H), 7.43 (d,
J = 8.5 Hz, 1H), 7.33 (d, J = 8.5 Hz, 1H), 3.64 (brs, 2H, -NH2), 2.46 (s,
3H).
3-Amino-6,8-di-tert-butyl-4H-chromen-4-one (4b):35 The title
compound was prepared according to the general procedure using
2-(2,4-di-tert-butyl-6-formylphenoxy)acetonitrile (4a) (100 mg,
0.366 mmol, 1 equiv.), 1 (19 mg, 0.073 mmol, 0.2 equiv) and
triethylamine (10 μL, 0.073 mmol, 0.2 equiv.) in [bmim][BF4] (68 µL,
0.366 mmol, 1 equiv.). The product was obtained as yellow solid
(Yield 86 mg, 86 %). m. p. 120−122 ℃ (lit.35 121 ℃}; H NMR (400
MHz, CDCl3): δ 8.13 (d, J = 2.0 Hz, 1H), 7.84 (s, 1H), 7.64 (d, J = 2.0 Hz,
1H), 3.43 (brs, 2H, -NH2), 1.47 (s, 9H), 1.37 (s, 9H).
1
2-Amino-1H-benzo[f]chromen-1-one (5b):35 The title compound was
prepared according to the general procedure using 2-(1-
formylnaphthalen-2-yloxy)acetonitrile (5a) (100 mg, 0.473 mmol, 1
equiv.), 1 (25 mg, 0.094 mmol, 0.2 equiv.) and triethylamine (13 μL,
0.094 mmol, 0.2 equiv) in [bmim][BF4] (88 µL, 0.473 mmol, 1 equiv.).
The product was obtained as yellow solid (Yield 83 mg, 83 %). m. p.
125−127 ℃ (lit.35 125 ℃); 1H NMR (400 MHz, CDCl3): δ 10.02 (d, J =
8.4 Hz, 1H), 8.18 (d, J = 9.2 Hz, 1H), 8.07–8.03 (m, 2H), 7.77–7.73 (m,
1H), 7.67–7.61 (m, 1H), 4.66 (brs, 2H, -NH2).
3-Amino-8-methoxy-4H-chromen-4-one (6b):35 The title compound
was prepared according to the general procedure using 2-(2-formyl-
6-methoxyphenoxy)acetonitrile (6a) (100 mg, 0.523 mmol, 1 equiv.),
1 (28 mg, 0.104 mmol, 0.2 equiv) and triethylamine (14 μL, 0.104
mmol, 0.2 equiv) in [bmim][BF4] (97 µL, 0.523 mmol, 1 equiv.). The
product was obtained as yellow solid (Yield 94 mg, 94 %). m. p.
1
130−132 ℃ (lit.35 132 ℃); H NMR (400 MHz, DMSO-d6): δ 7.99 (s,
1H), 7.60 (t, J = 4.8 Hz, 1H), 7.28 (d, J = 4.4 Hz, 1H), 4.56 (brs, 2H, -
NH2), 3.93 (s, 3H).
3-Amino-6-methoxy-4H-chromen-4-one (7b):35 The title compound
was prepared according to the general procedure using 2-(2-formyl-
3-aminochromone as
a pale-yellow precipitate. Upon 100 %
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 7
Please do not adjust margins

Please do not adjust margins
New Journal of Chemistry
Page 8 of 17
ARTICLE
Journal Name
1
2
3
4
5
6
7
8
9
4-methoxyphenoxy)acetonitrile (7a) (100 mg, 0.523 mmol, 1 equiv.), 0.2 equiv.) in [bmim][BF4] (39 µL, 0.208 mmol, 1 equiv.). The product
View Article Online
DOI: 10.1039/C9NJ02650A
1 (28 mg, 0.104 mmol, 0.2 equiv.) and triethylamine (14 μL, 0.104 was obtained as yellow solid (Yield 44 mg, 88 %). m. p. 162−164 ℃
mmol, 0.2 equiv) in [bmim][BF4] (97 µL, 0.523 mmol, 1 equiv.). The (lit.35 162 ℃); 1H NMR (400 MHz, DMSO-d6): δ 8.12 (d, J = 2.4 Hz, 1H),
product was obtained as pale yellow solid (Yield 90 mg, 90 %). m. p. 7.97 (s, 1H), 7.80 (d, J = 8.8 Hz, 1H), 7.54 (d, J = 9.2 Hz, 1H), 4.64 (brs,
1
96−98 ℃ (lit.35 96 ℃); H NMR (400 MHz, DMSO-d6): δ 7.95 (s, 1H), 2H, -NH2).
7.51 (d, J = 9.2 Hz, 1H), 7.41 (d, J = 2.8 Hz, 1H), 7.29 (dd, J1 = 9.2 Hz, J2 3-Amino-6-chloro-4H-chromen-4-one (14b):35 The title compound
= 3.2 Hz, 1H), 4.52 (brs, 2H, -NH2), 3.83 (s, 1H).
was prepared according to the general procedure using 2-(4-chloro-
3-Amino-7-methoxy-4H-chromen-4-one (8b):35 The title compound 2-formylphenoxy)acetonitrile (14a) (50 mg, 0.256 mmol, 1 equiv.), 1
was prepared according to the general procedure using 2-(2-formyl- (13 mg, 0.051 mmol, 0.2 equiv.) and triethylamine (7 μL, 0.051 mmol,
5-methoxyphenoxy)acetonitrile (8a) (100 mg, 0.523 mmol, 1 equiv.), 0.2 equiv.) in [bmim][BF4] (57 µL, 0.256 mmol, 1 equiv.). The product
1 (28 mg, 0.104 mmol, 0.2 equiv.) and triethylamine (14 μL, 0.104 was obtained as yellow solid (Yield 45.5 mg, 91 %). m. p. 174−176 ℃
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
1
mmol, 0.2 equiv.) in [bmim][BF4] (97 µL, 0.523 mmol, 1 equiv.). The (lit.35 175 ℃); H NMR (400 MHz, CDCl3): δ 8.22 (d, J = 2.4 Hz, 1H),
product was obtained as pale yellow solid (Yield 93 mg, 93 %). m. p. 7.78 (s, 1H), 7.52 (dd, J1 = 9.2 Hz, J2 = 2.8 Hz, 1H), 7.39 (d, J = 9.2 Hz,
160−162 ℃ (lit.35 160 ℃); 1H NMR (500 MHz, CDCl3): δ 8.06 (d, J = 8.9 1H), 3.67 (brs, 2H, -NH2).
Hz, 1H), 7.62 (s, 1H), 6.86 (dd, J1 = 9 Hz, J2 = 2.0 Hz, 1H), 3.82 (brs, 2H, 3-Amino-8-bromo-6-chloro-4H-chromen-4-one (15b):35 The title
-NH2), 3.81 (s, 3H).
compound was prepared according to the general procedure using
3-Amino-8-methoxy-2-methyl-4H-chromen-4-one (9b):35 The title 2-(2-bromo-4-chloro-6-formylphenoxy)acetonitrile (15a) (50 mg,
compound was prepared according to the general procedure using 0.182 mmol, 1 equiv.), 1 (10 mg, 0.036 mmol, 0.2 equiv.) and
2-(2-formyl-6-methoxyphenoxy)propanenitrile (9a) (100 mg, 0.487 triethylamine (5 μL, 0.036 mmol, 0.2 equiv.) in [bmim][BF4] (34 µL,
mmol, 1 equiv.), 1 (26 mg, 0.097 mmol, 0.2 equiv.) and triethylamine 0.182 mmol, 1 equiv.). The product was obtained as pale yellow solid
1
(13 μL, 0.097 mmol, 0.2 equiv.) in [bmim][BF4] (110 µL, 0.487 mmol, (Yield 46 mg, 92 %). m. p. 191−193 ℃ (lit.35 192 ℃); H NMR (400
1 equiv.). The product was obtained as yellow solid (Yield 80 mg, 80 MHz, DMSO-d6): δ 8.16 (d, J = 2.4 Hz, 1H), 8.08 (s, 1H), 8.01 (d, J = 2.4
%). m. p. 172−174 ℃ (lit.35 172 ℃); 1H NMR (400 MHz, CDCl3): δ 7.67 Hz, 1H), 4.77 (brs, 2H, -NH2).
(d, J = 7.2 Hz, 1H), 7.14 (t, J = 8 Hz, 1H), 6.97 (d, J = 7.2 Hz, 1H), 3.88 3-Amino-6-iodo-8-methoxy-4H-chromen-4-one (16b):35 The title
(s, 3H), 3.50 (brs, 2H, -NH2), 2.37 (s, 3H).
compound was prepared according to the general procedure using
3-Amino-8-methoxy-2-phenyl-4H-chromen-4-one (10b):35 The title 2-(2-formyl-4-iodo-6-methoxyphenoxy)acetonitrile (16a) (100 mg,
compound was prepared according to the general procedure using 0.315 mmol, 1 equiv.), 1 (66 mg, 0.063 mmol, 0.2 equiv.) and
2-(2-formyl-6-methoxyphenoxy)-2-phenylacetonitrile35 (10a) (100 triethylamine (9 μL, 0.063 mmol, 0.2 equiv.) in [bmim][BF4] (58 µL,
mg, 0.374 mmol, 1 equiv.), 1 (20 mg, 0.075 mmol, 0.2 equiv.) and 0.315 mmol, 1 equiv.). The product was obtained as yellow solid
1
triethylamine (10 μL, 0.075 mmol, 0.2 equiv,) in [bmim][BF4] (70 µL, (Yield 93 mg, 93 %). m. p. 209−211 ℃ (lit.35 209 ℃); H NMR (400
0.374 mmol, 1 equiv.). The product was obtained as pale yellow solid; MHz, DMSO-d6): δ 7.99 (s, 1H), 7.87 (d, J = 1.6 Hz, 1H), 7.48 (d, J = 1.2
1
(85 mg, 85% yield); m.p. 123−125 ℃ {lit.35 125 ℃}; H NMR (400 MHz, Hz, 1H), 4.63 (brs, 2H, -NH2), 3.92 (s, 1H).
DMSO-d6): δ 7.94 (d, J = 8.4 Hz, 2H), 7.62 (dd, J1 = 6.8 Hz, J2 = 2.4 Hz, 3-Amino-8-fluoro-4H-chromen-4-one (17b):35 The title compound
1H), 7.60–7.56 (m, 2H), 7.51–7.49 (m, 1H), 7.35–7.33 (m, 2H), 4.73 was prepared according to the general procedure using 2-(2-fluoro-
(brs, 2H, -NH2), 3.95 (s, 3H).
6-formylphenoxy)acetonitrile (17a) (30 mg, 0.167 mmol, 1 equiv.), 1
3-Amino-6-nitro-4H-chromen-4-one (11b):35 The title compound (9 mg, 0.033 mmol, 0.2 equiv.) and triethylamine (5 μL, 0.033 mmol,
was prepared according to the general procedure using 2-(2-formyl- 0.2 equiv.) in [bmim][BF4] (31 µL, 0.167 mmol, 1 equiv.). The product
4-nitrophenoxy)acetonitrile (11a) (100 mg, 0.485 mmol, 1 equiv.), 1 was obtained as yellow solid (Yield 27 mg, 92 %). m. p. 163−165 ℃
(26 mg, 0.097 mmol, 0.2 equiv.) and triethylamine (13 μL, 0.097 (lit.35 165 ℃); 1H NMR (500 MHz, CDCl3): δ 8.01 (td, J1 = 8 Hz, J2 = 1.2
mmol, 0.2 equiv.) in [bmim][BF4] (90 µL, 0.485 mmol, 1 equiv.). The Hz, 1H), 7.81 (s, 1H), 7.38 (ddd, J1 = 10.4 Hz, J2 = 8 Hz, J3 = 1.2 Hz, 1H),
product was obtained as yellow solid (Yield 78 mg, 78 %). m. p. 7.28 (dd, J1 = 9 Hz, J2 = 4.4 Hz, 1H), 3.69 (brs, 2H, -NH2).
168−170 ℃ (lit.35 168 ℃); 1H NMR (400 MHz, DMSO-d6): δ 8.76 (d, J 3-Amino-6,8-difluoro-4H-chromen-4-one
(18b):35
The
title
= 2 Hz, 1H), 8.40 (dd, J1 = 9.2 Hz, J2 = 2 Hz, 1H), 8.01 (s, 1H), 7.78 (d, J compound was prepared according to the general procedure using
= 9.2 Hz, 1H), 4.81 (brs, 2H, -NH2).
3-Amino-6,8-dichloro-4H-chromen-4-one
2-(2,4-difluoro-6-formylphenoxy)acetonitrile (18a) (20 mg, 0.101
title mmol, 1 equiv.), 1 (5 mg, 0.02 mmol, 0.2 equiv.) and triethylamine (3
(12b):35
The
compound was prepared according to the general procedure using μL, 0.02 mmol, 0.2 equiv.) in [bmim][BF4] (19 µL, 0.101 mmol, 1
2-(2, 4-dichloro-6-formylphenoxy)acetonitrile (12a) (100 mg, 0.434 equiv.). The product was obtained as yellow solid (Yield 17 mg, 89 %).
mmol, 1 equiv.), 1 (23 mg, 0.086 mmol, 0.2 equiv.) and triethylamine m. p. 150−152 ℃ (lit.35 149 ℃); 1H NMR (400 MHz, DMSO-d6): δ 8.05
(12 μL, 0.086 mmol, 0.2 equiv.) in [bmim][BF4] (81 µL, 0.434 mmol, 1 (s, 1H), 7.81 (ddd, J1 = 10.8 Hz, J2 = 8.4 Hz, J3 = 2.8 Hz, 1H), 7.57 (td, J1
equiv.). The product was obtained as yellow solid (Yield 87 mg, 87 %). = 8.4 Hz, J2 = 2 Hz, 1H), 4.73 (brs, 2H, -NH2).
m. p. 180−182 ℃ (lit.35 182 ℃); 1H NMR (400 MHz, DMSO-d6): δ 8.07 3-Amino-7-fluoro-4H-chromen-4-one (19b):35 The title compound
(s, 1H), 8.01 (d, J = 2.4 Hz, 1H), 7.94 (d, J = 2.4 Hz, 1H), 4.77 (brs, 2H, was prepared according to the general procedure using 2-(5-fluoro-
-NH2).
2-formylphenoxy)acetonitrile (19a) (100 mg, 0.558 mmol, 1 equiv.),
3-Amino-6-bromo-4H-chromen-4-one (13b):35 The title compound 1 (30 mg, 0.111 mmol, 0.2 equiv.) and triethylamine (15 μL, 0.111
was prepared according to the general procedure using 2-(4-bromo- mmol, 0.2 equiv.) in [bmim][BF4] (104 µL, 0.558 mmol, 1 equiv.). The
2-formylphenoxy)acetonitrile (13a) (50 mg, 0.208 mmol, 1 equiv.), 1 product was obtained as yellow solid (Yield 88 mg, 88 %). m. p.
(11 mg, 0.041 mmol, 0.2 equiv.) and triethylamine (6 μL, 0.041 mmol, 172−174 ℃ (lit.35 172 ℃); 1H NMR (400 MHz, CDCl3): δ 8.25 (dd, J1=
8 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 9 of 17
New Journal of Chemistry
Please do not adjust margins
Journal Name
ARTICLE
1
2
3
4
5
6
7
8
9
9.6 Hz, J2 = 6.4 Hz, 1H), 7.74 (s, 1H), 7.11–7.06 (m, 2H), 3.64 (brs, 2H, (Yield 95 mg, 95 %). m. p. 73−75 ℃ (lit.34 75 ℃); 1H NMR (400 MHz,
View Article Online
DOI: 10.1039/C9NJ02650A
-NH2).
CDCl3): δ 7.98 (s, 1H), 7.91 (s, 1H), 7.45 (d, J = 8.8 Hz, 1H), 7.33 (d, J =
3-Amino-6-fluoro-2-methyl-4H-chromen-4-one (20b):35 The title 8.4 Hz, 1H), 4.17 (q, J = 7.2 Hz, 2H), 3.46 (s, 2H), 2.43 (s, 3H), 1.26 (t,
compound was prepared according to the general procedure using J = 7.2 Hz, 3H).
2-(5-fluoro-2-formylphenoxy)propanenitrile (20a) (60 mg, 0.31 mmol, Ethyl-2-(6,8-di-tert-butyl-4-oxo-4H-chromen-3-yl)acetate (5f):34 The
1 equiv.), 1 (16 mg, 0.062 mmol, 0.2 equiv.) and triethylamine (8 μL, title compound was prepared according to the general procedure
0.062 mmol, 0.2 equiv.) in [bmim][BF4] (58 µL, 0.31 mmol, 1 equiv.). using ethyl-4-(2,4-di-tert-butyl-6-formylphenoxy)but-2-ynoate (5e)
The product was obtained as pale yellow solid (Yield 48 mg, 81 %). (100 mg, 0.290 mmol, 1 equiv.), 1 (15 mg, 0.058 mmol, 0.2 equiv.)
m. p. 165−167 ℃ (lit.35 165 ℃); 1H NMR (400 MHz, DMSO-d6): δ 7.66 and triethylamine (8 μL, 0.058 mmol, 0.2 equiv.) in [bmim][BF4] (54
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
(dd, J1 = 8.8 Hz, J2 = 3.2 Hz, 1H), 7.61 (dd, J1 = 9.2 Hz, J2 = 4.8 Hz, 1H), µL, 0.290 mmol, 1 equiv.). The product was obtained as pale white
7.54 (dt, J1 = 8 Hz, J2 = 2.8 Hz, 1H), 4.41 (brs, 2H, -NH2), 2.39 (s, 1H).
3-Amino-6-(trifluoromethoxy)-4H-chromen-4-one (21b):35 The title MHz, CDCl3): δ 8.11 (d, J = 2 Hz, 1H), 8.01 (s, 1H), 7.69 (d, J = 1.6 Hz,
compound was prepared according to the general procedure using 1H), 4.20 (q, J = 7.2 Hz, 2H), 3.48 (s, 2H), 1.48 (s, 9H), 1.36 (s, 9H), 1.30
2-(2-formyl-4-(trifluoromethoxy)phenoxy)acetonitrile (21a) (50 mg, (t, J = 7.2 Hz, 3H).
1
solid (Yield 85 mg, 85 %); m. p. 92−94 ℃ (lit.34 92 ℃); H NMR (400
0.204 mmol, 1 equiv.), 1 (10 mg, 0.04 mmol, 0.2 equiv.) and Ethyl-2-(7-methoxy-4-oxo-4H-chromen-3-yl)acetate (6f):34 The title
triethylamine (6 μL, 0.04 mmol, 0.2 equiv.) in [bmim][BF4] (38 µL, compound was prepared according to the general procedure using
0.204 mmol, 1 equiv.). The product was obtained as yellow solid ethyl-4-(2-formyl-5-methoxyphenoxy)but-2-ynoate (6e) (100 mg,
1
(Yield 43 mg, 87 %). m. p. 107−109 ℃ (lit.35 108 ℃); H NMR (400 0.381 mmol, 1 equiv.), 1 (20 mg, 0.076 mmol, 0.2 equiv.) and
MHz, DMSO-d6): δ 8.02 (s, 1H), 7.92 (s, 1H), 7.75 (d, J = 9.2 Hz, 1H), triethylamine (11 μL, 0.076 mmol, 0.2 equiv.) in [bmim][BF4] (71 µL,
7.72 (dd, J1 = 9.2 Hz, J2 = 1.1 Hz, 1H), 4.67 (brs, 2H, -NH2).
0.381 mmol, 1 equiv.). The product was obtained as a pale white solid
Ethyl-2-(4-oxo-4H-chromen-3-yl)acetate (1f): 34 The title compound (Yield 94 mg, 94 %). m. p. 103−104 ℃ (lit.34 104 ℃); H NMR (400
was prepared according to the general procedure using ethyl-4-(2- MHz, CDCl3): δ 8.10 (d, J = 8.8 Hz, 1H), 7.86 (s, 1H), 6.95 (dd, J1 = 8.8
formylphenoxy)but-2-ynoate (1e) (100 mg, 0.431 mmol, 1 equiv.), 1 Hz, J2 = 1.6 Hz, 1H), 6.81 (s, 1H), 4.17 (q, J = 7.2 Hz, 2H), 3.88 (s, 3H),
(23 mg, 0.086 mmol, 0.2 equiv.) and triethylamine (12 μL, 0.086 3.44 (s, 2H), 1.26 (t, J = 7.2 Hz, 3H).
1
mmol, 0.2 equiv) in [bmim][BF4] (97 µL, 0.431 mmol, 1 equiv.). The Ethyl-2-(6-methoxy-4-oxo-4H-chromen-3-yl)acetate (7f):34 The title
product was obtained as a pale white solid (Yield 93 mg, 93 %). m. p. compound was prepared according to the general procedure using
78−80 ℃ (lit.34 80 ℃); 1H NMR (300 MHz, CDCl3): δ 8.21 (dd, J1 = 8.1 ethyl-4-(2-formyl-4-methoxyphenoxy)but-2-ynoate (7e) (100 mg,
Hz, J2 = 1.8 Hz, 1H), 7.94 (s, 1H), 7.65 (ddd, J1 = 8.7 Hz, J2 = 7.2 Hz, J3 0.381 mmol, 1 equiv.), 1 (20 mg, 0.076 mmol, 0.2 equiv.) and
= 1.8 Hz, 1H), 7.44 (d, J = 8.7 Hz, 1H), 7.41–7.38 (m, 1H), 4.18 (q, J = triethylamine (11 μL, 0.076 mmol, 0.2 equiv.) in [bmim][BF4] (71 µL,
7.2 Hz, 2H), 3.47 (s, 2H), 1.27 (t, J = 7.2 Hz, 3H).
0.381 mmol, 1 equiv.). The product was obtained as a pale white solid
Ethyl-2-(8-methyl-4-oxo-4H-chromen-3-yl)acetate (2f):34 The title (Yield 90 mg, 90 %); m. p. 94−96 ℃ (lit.34 95 ℃); 1H NMR (400 MHz,
compound was prepared according to the general procedure using CDCl3): δ 7.93 (s, 1H), 7.57 (d, J = 3.2 Hz, 1H), 7.39 (d, J = 9.2 Hz, 1H),
ethyl-4-(2-formyl-6-methylphenoxy)but-2-ynoate (2e) (100 mg, 7.27–7.24 (m, 1H), 4.19 (q, J = 7.2 Hz, 2H), 3.88 (s, 3H), 3.48 (s, 2H),
0.406 mmol, 1 equiv.), 1 (22 mg, 0.081 mmol, 0.2 equiv.) and 1.28 (t, J = 7.2 Hz, 3H).
triethylamine (11 μL, 0.081 mmol, 0.2 equiv.) in [bmim][BF4] (91 µL, Ethyl-2-(8-methoxy-4-oxo-4H-chromen-3-yl)acetate (8f):34 The title
0.406 mmol, 1 equiv.). The product was obtained as pale white solid compound was prepared according to the general procedure using
(Yield 96 mg, 96 %). m. p. 80−82 ℃ (lit.34 82 ℃); 1H NMR (400 MHz, ethyl-4-(2-formyl-6-methoxyphenoxy)but-2-ynoate (8e) (100 mg,
CDCl3): δ 8.06 (d, J = 7.6 Hz, 1H), 7.99 (s, 1H), 7.50 (d, J = 6.8 Hz, 1H), 0.381 mmol, 1 equiv.), 1 (20 mg, 0.076 mmol, 0.2 equiv.) and
7.30 (d, J = 8 Hz, 1H), 4.19 (q, J = 6.8 Hz, 2H), 3.48 (s, 2H), 2.47 (s, 3H), triethylamine (11 μL, 0.076 mmol, 0.2 equiv.) in [bmim][BF4] (71 µL,
1.28 (t, J = 6.8 Hz, 3H).
0.381 mmol, 1 equiv.). The product was obtained as a pale white solid
Ethyl-2-(1-oxo-1H-benzo[f]chromen-2-yl)acetate (3f):34 The title (Yield 88 mg, 88 %). m. p. 125−127 ℃ (lit.34 127 ℃); H NMR (400
compound was prepared according to the general procedure using MHz, CDCl3): δ 7.99 (s, 1H), 7.77 (dd, J1 = 8 Hz, J2 = 1.2 Hz, 1H), 7.31
ethyl-4-(1-formylnaphthalen-2-yloxy)but-2-ynoate (3e) (100 mg, (t, J = 8 Hz, 1H), 7.16 (dd, J1 = 8 Hz, J2 = 1.2 Hz, 1H), 4.17 (q, J = 7.2 Hz,
0.354 mmol, 1 equiv.), 1 (19 mg, 0.070 mmol, 0.2 equiv.) and 2H), 3.99 (s, 3H), 3.47 (s, 2H), 1.26 (t, J = 7.2 Hz, 3H).
1
triethylamine (10 μL, 0.070 mmol, 0.2 equiv) in [bmim][BF4] (75 µL, Ethyl-4-(3-(2-ethoxy-2-oxoethyl)-4-oxo-4H-chromen-8-yloxy)but-2-
0.354 mmol, 1 equiv.). The product was obtained as a pale white solid ynoate (9f):34 The title compound was prepared according to the
1
(Yield 94 mg, 94 %). m. p. 114−116 ℃ (lit.34 116 ℃); H NMR (400 general
procedure
using
diethyl-4,4'-(3-formyl-1,2-
MHz, CDCl3): δ 10.03 (d, J = 8.8 Hz, 1H), 8.06 (d, J = 8.8 Hz, 1H), 8.00 phenylene)bis(oxy)dibut-2-ynoate (9e) (100 mg, 0.279 mmol, 1
(s, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.73 (dd, , J1 = 8.4 Hz, J2 = 1.2 Hz, 1H), equiv.), 1 (15 mg, 0.056 mmol, 0.2 equiv.) and triethylamine (8 μL,
7.62–7.58 (m, 1H), 7.48 (d, J = 8.8 Hz, 1H), 4.22 (q, J = 7.2 Hz, 2H), 0.056 mmol, 0.2 equiv.) in [bmim][BF4] (52 µL, 0.279 mmol, 1 equiv.).
3.54 (s, 2H), 1.29 (t, J = 7.2 Hz, 3H).
The product was obtained as a pale white solid (Yield 89 mg, 89 %).
Ethyl-2-(6-methyl-4-oxo-4H-chromen-3-yl)acetate (4f):34 The title m. p. 113−115 ℃ (lit.34 114 ℃); 1H NMR (400 MHz, CDCl3): δ 7.99 (s,
compound was prepared according to the general procedure using 1H), 7.87 (dd, J1 = 8 Hz, J2 = 1.2 Hz, 1H), 7.34 (d, J = 8 Hz, 1H), 7.28
ethyl-4-(2-formyl-4-methylphenoxy)but-2-ynoate (4e) (100 mg, (dd, J1 = 8 Hz, J2 = 1.2 Hz, 1H), 4.99 (s, 2H), 4.24 (q, J = 7.2 Hz, 2H),
0.406 mmol, 1 equiv.), 1 (22 mg, 0.081 mmol, 0.2 equiv.) and 4.19 (q, J = 7.2 Hz, 2H), 3.48 (s, 2H), 1.30 (t, J = 7.2 Hz, 2H), 1.28 (t, J
triethylamine (11 μL, 0.081 mmol, 0.2 equiv.) in [bmim][BF4] (91 µL, = 7.2 Hz, 2H).
0.406 mmol, 1 equiv.). The product was obtained as a pale white solid
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 9
Please do not adjust margins

Please do not adjust margins
New Journal of Chemistry
Page 10 of 17
ARTICLE
Journal Name
1
2
3
4
5
6
7
8
9
Ethyl-2-(6,8-dichloro-4-oxo-4H-chromen-3-yl)acetate (10f):34 The 1H), 7.66 (t, J = 7.2 Hz, 1H), 7.58 (t, J = 7.2 Hz, 1H), 7.50V–ie7w.4A4rti(cmle ,O3nHlin)e,
title compound was prepared according to the general procedure 7.39 (t, J = 7.6 Hz, 1H), 4.15 (s, 2H).
DOI: 10.1039/C9NJ02650A
using ethyl-4-(2,4-dichloro-6-formylphenoxy)but-2-ynoate (10e) Synthesis
(100 mg, 0.322 mmol, 1 equiv.), 1 (17 mg, 0.066 mmol, 0.2 equiv.) benzaldehyde (16e): Step 1. In a round-bottom flask, salicylaldehyde
and triethylamine (9 μL, 0.066 mmol, 0.2 equiv.) in [bmim][BF4] (62 tethered with the propargyl group 2-(prop-2-yn-1-
of
2-((4-(adamantan-2-yl)-4-oxobut-2-yn-1-yl)oxy)
µL, 0.322 mmol, 1 equiv.). The product was obtained as a pale white yloxy)benzaldehyde (300 mg, 1.87 mmol, 1 equiv.) was dissolved in
1
solid (Yield 86 mg, 86 %). m. p. 103−105 ℃ (lit.34 105 ℃); H NMR dry ethanol (10 mL) under nitrogen atmosphere. Triethyl
(400 MHz, CDCl3): δ 8.07 (d, J = 2.4 Hz, 1H), 8.01 (s, 1H), 7.70 (d, J = orthoformate (0.60 mL, 3.20 mmol, 1.7 equiv.) and PPTS (5 mg, 0.02
2.4 Hz, 1H), 4.18 (q, J = 7.2 Hz, 2H), 3.47 (s, 2H), 1.26 (t, J = 7.2 Hz, mmol, 0.01 equiv.) were added and the resulting solution was
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
3H).
refluxed for 3 h. The progress of the reaction was monitored using
Ethyl-2-(6-chloro-4-oxo-4H-chromen-3-yl)acetate (11f):34 The title TLC. Upon completion, the reaction mixture was quenched with few
compound was prepared according to the general procedure using drops of Et3N (0.05 equiv.) and concentrated. The oil was diluted with
ethyl-4-(4-chloro-2-formylphenoxy)but-2-ynoate (11e) (100 mg, ethyl acetate (50 mL), washed with 10 % NaHCO3 (2×20 mL), followed
0.375 mmol, 1 equiv.), 1 (20 mg, 0.075 mmol, 0.2 equiv.) and by brine (2×10 mL) and then dried over anhydrous Na2SO4. The
triethylamine (10 μL, 0.075 mmol, 0.2 equiv.) in [bmim][BF4] (70 µL, solvent was evaporated and the residue was dried under vacuum.
0.375 mmol, 1 equiv.). The product was obtained as a pale white solid The obtained protected salicylaldehyde derivative was used for the
(Yield 86 mg, 86 %). m. p. 87−89 ℃ (lit.34 88 ℃); 1H NMR (400 MHz, next step.
CDCl3): 8.17 (d, J = 2.4 Hz, 1H), 7.94 (s, 1H), 7.60 (dd, J1 = 8.8 Hz, J2 = Step 2. In a round-bottom flask, under nitrogen atmosphere, the
2.4 Hz, 1H), 7.41 (d, J = 8.8 Hz, 1H), 4.18 (q, J = 7.2 Hz, 2H), 3.47 (s, acetal protected salicylaldehyde derivative (1-(diethoxymethyl)-2-
2H), 1.27 (t, J = 7.2 Hz, 3H).
(prop-2-yn-1-yloxy)benzene from step 1) (439 mg, 1.88 mmol, 1
Ethyl-2-(7-fluoro-4-oxo-4H-chromen-3-yl)acetate (12f):34 The title equiv.) was dissolved in dry THF (10 mL). The solution was stirred at –
compound was prepared according to the general procedure using 78 ℃ (dry ice-acetone bath) for 10 min, butyl lithium (2 M in
ethyl-4-(5-fluoro-2-formylphenoxy)but-2-ynoate (12e) (100 mg, 0.4 cyclohexane) (1.40 mL, 2.82 mmol, 1.5 equiv.) was slowly added to
mmol, 1 equiv.), 1 (21 mg, 0.08 mmol, 0.2 equiv.) and triethylamine the flask over 10 min and stirred for another 30 min at the same
(11 μL, 0.08 mmol, 0.2 equiv.) in [bmim][BF4] (74 µL, 0.4 mmol, 1 temperature. 1-Adamantanecarbonylchloride in dry THF (631 mg,
equiv.). The product was obtained as a pale white solid (Yield 90 mg, 3.20 mmol, 1.7 equiv.) was slowly added to the reaction mixture and
90 %). m. p. 101−103 ℃ (lit.34 102 ℃); 1H NMR (400 MHz, CDCl3): 8.22 stirring was continued for another 30-60 min. The progress of the
(dd, J1 = 9.6 Hz, J1 = 6.4 Hz, 1H), 7.91 (s, 1H), 7.13 (d, J = 6.4 Hz, 2H), reaction was monitored using TLC. Upon completion, the reaction
4.18 (q, J = 7.2 Hz, 2H), 3.45 (s, 2H), 1.27 (t, J = 7.2 Hz, 3H).
mixture was allowed to warm to ambient temperature and quenched
(E)-Ethyl-2-(2,2-dimethyl-4-oxochroman-3-ylidene)acetate (13f):34 with saturated NH4Cl (20 mL). Then the reaction mixture was diluted
The title compound was prepared according to the general with ethyl acetate (50 mL), washed with water (3×50 mL), followed
procedure using ethyl-4-(2-formylphenoxy)-4-methylpent-2-ynoate by brine (2×10 mL), and then dried over Na2SO4. The solvent was
(13e) (50 mg, 0.192 mmol, 1 equiv.), 1 (10 mg, 0.038 mmol, 0.2 equiv.) evaporated and the crude product was dissolved in CHCl3 (20 mL),
and triethylamine (5 μL, 0.038 mmol, 0.2 equiv.) in [bmim][BF4] (35 then water (10 mL) and 50 % TFA (3 mL) were added, and stirred the
µL, 0.192 mmol, 1 equiv.). The product was obtained as yellow biphasic mixture for 15 to 30 min. The progress of deprotection was
viscous oil (Yield 44 mg, 88 %). 1H NMR (400 MHz, CDCl3): δ 7.90 (d, J monitored by using TLC. Upon completion, the reaction mixture was
= 7.6 Hz, 1H), 7.48 (t, J = 7.6 Hz, 1H), 7.01 (t, J = 7.6 Hz, 1H), 6.91 (d, J diluted with ethyl acetate (50 mL), washed with aqueous NaHCO3
= 8.4 Hz, 1H), 6.19 (s, 1H), 4.31 (q, J = 7.2 Hz, 2H), 1.62 (s, 6H), 1.32 (2×50 mL), water (3×50 mL), followed by brine (2×10 mL), and then
(t, J = 7.2 Hz, 3H).
dried over anhydrous Na2SO4. The solvent was evaporated and the
3-(2-Oxopropyl)-4H-chromen-4-one (14f):34 The title compound was crude product was purified by flash column chromatography to
prepared according to the general procedure using 2-(4-oxopent-2- obtain 16e as a viscous oil (Yield 480 mg, 79 %). 1H NMR (400 MHz,
ynyloxy)benzaldehyde (14e) (100 mg, 0.495 mmol, 1 equiv.), 1 (26 CDCl3): δ 10.46 (s, 1H), 7.83 (dd, J = 7.6 Hz, 1.6 Hz, 1H), 7.54–7.50
mg, 0.099 mmol, 0.2 equiv.) and triethylamine (14 μL, 0.099 mmol, (m,1H), 7.10–7.03 (m, 2H), 4.89 (s, 2H), 1.98 (s, 1H), 1.86 (s, 2H), 1.78
0.2 equiv.) in [bmim][BF4] (92 µL, 0.495 mmol, 1 equiv.). The product (d, 2.16 Hz, 3H), 1.68 (q, 12.0 Hz, 4H), 1.57 (s, 4H), 1.37 (d, 11.4 Hz,
was obtained as a pale white solid (Yield 84 mg, 84 %). m. p. 96−98 1H); 13C{1H} NMR (100 MHz, CDCl3): δ 189.6, 174.4, 159.7, 135.6,
℃ (lit.34 97 ℃); 1H NMR (400 MHz, CDCl3): δ 8.20 (d, J = 7.6 Hz, 1H), 128.4, 125.5, 121.5, 113.6, 83.4, 80.0, 72.8, 56.5, 42.1, 41.4, 38.6,
7.88 (s, 1H), 7.67 (t, J = 7.8 Hz, 1H), 7.45 (d, J = 8.8 Hz, 1H), 7.40 (t, J 38.5, 36.5, 36.4, 36.1, 28.1, 27.8.
= 7.6 Hz, 1H), 3.56 (s, 2H), 2.33 (s, 3H).
Amine functionalization of 3-aminochromones for active drug
3-(3,3-Dimethyl-2-oxobutyl)-4H-chromen-4-one (15f):34 The title candidates (1c-12c): Stepwise process; Procedure A: In a round-
compound was prepared according to the general procedure using bottom flask, under nitrogen atmosphere, 3-aminochromone 1b
2-((4-oxo-4-phenylbut-2-yn-1-yl)oxy)benzaldehyde (15e) (100 mg, (200 mg, 1.24 mmol, 1 equiv.) was dissolved in acetonitrile (5 mL)
0.378 mmol, 1 equiv.), 1 (20 mg, 0.075 mmol, 0.2 equiv.) and and stirred the reaction mixture with Et3N (346 μL, 2.48 mmol, 2
triethylamine (10 μL, 0.075 mmol, 0.2 equiv.) in [bmim][BF4] (70 µL, equiv.) at 0 ℃. Bromoacetylbromide (118 μL, 1.36 mmol, 1.1 equiv.)
0.378 mmol, 1 equiv.). The product was obtained as a pale white solid was added via syringe over 5 min at 0 ℃ and the reaction mixture
(Yield 81 mg, 81 %). m. p. 131−133 ℃ (lit.34 132 ℃); H NMR (400 was allowed to stir for another 30 min at room temperature. After
MHz, CDCl3): δ 8.22 (d, J = 8 Hz, 1H), 8.09 (d, J = 8 Hz, 2H), 7.97 (s, the formation of amide bond (judged by TLC), morpholine (321 μL,
1
3.726 mmol, 3 equiv.) was added and stirring was continued for 12
10 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 11 of 17
New Journal of Chemistry
Please do not adjust margins
Journal Name
ARTICLE
1
2
3
4
5
h. The progress of the reaction was monitored using TLC. After the 2-Morpholino-N-(1-oxo-1H-benzo[f]chromen-2-yl)acetamide (4c):
View Article Online
completion of the reaction, the reaction mixture was quenched with The title compound was prepared according DtoOpI:r1o0c.e10d3u9r/eCB9NuJs0in2g65202A-
water (50 mL) and extracted with ethyl acetate (3x30 mL). The ((1-formylnaphthalen-2-yl)oxy)acetonitrile (5a) (100 mg, 0.473
combined organic layer was washed with brine (210 mL) and dried mmol), triethylamine (132 µL, 0.947 mmol,
2
equiv.),
6
over anhydrous Na2SO4. The solvent was evaporated and the crude bromoacetylbromide (45 μL, 0.521 mmol, 1.1 equiv.) and morpholine
product was purified by flash chromatography on silica to obtain 2- (122 µL, 1.421 mmol, 3 equiv.). The product was obtained as dirty
morpholino-N-(4-oxo-4H-chromen-3-yl)acetamide (7c) as a white white solid (Yield 100 mg, 63 %); m. p. 155–157 ℃;1H NMR (500 MHz,
solid (over two steps) (Yield 232 mg, 65 %). m. p. 150−152 ℃ {lit.35 CDCl3): δ 10.00 (d, J = 8.3 Hz, 2H), 9.42 (s, 1H), 8.06 (d, J = 8.9 Hz, 1H),
150 ℃}; 1H NMR (500 MHz, CDCl3): δ 9.72 (s, 1H, -NH), 9.31 (s, 1H), 7.91 (d, J = 7.6 Hz, 1H), 7.78 (t, J = 7.3 Hz, 1H), 7.64 (t, J = 7.0 Hz, 1H),
8.19 (d, J = 7.9 Hz, 1H), 7.63 (t, J = 7.5 Hz, 1H), 7.45 (d, J = 8.4 Hz, 1H), 7.51 (d, J = 8.9 Hz, 1H), 3.89 (s, 4H), 3.23 (s, 2H), 2.66 (s, 4H); 13C {1H}
7.35 (t, J = 7.5 Hz, 1H), 3.77 (t, J = 4 Hz, 4H), 3.13 (s, 2H), 2.57 (t, J = 4 NMR (126 MHz, CDCl3): δ 172.7, 168.9, 157.1, 142.6, 135.7, 130.5,
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Hz, 4H).
130.2, 129.4, 128.4, 126.9, 126.7, 125.9, 117.8, 115.5, 67.1, 62.3,
One pot process; Procedure B: After the completion of microwave 53.8. FT-IR (KBr): vmax 3267, 2918, 2816, 1683, 1643, 1600, 1514,
reaction for the synthesis of 1b using 1a (100 mg, 0.621 mmol), the 1116, 829 cm-1; HR-MS (ESI) m/z [M+H]+: calcd. for C19H19N2O4:
microwave tube was kept at 0 ℃, triethylamine (141 µL, 1.242 mmol, 339.1345, found: 339.1346.
2 equiv.) followed by bromoacetylbromide (59 µL, 0.683 mmol, 1.1 N-(6-Methoxy-4-oxo-4H-chromen-3-yl)-2-morpholinoacetamide
equiv.) were added using syringe and stirred the reaction mixture for (5c): The title compound was prepared according to procedure B
30 min at room temperature. After the formation of amide bond using 2-(2-formyl-4-methoxyphenoxy)acetonitrile (7a) (100 mg,
(judged by TLC), morpholine (160 µL, 1.863 mmol, 3 equiv.) was 0.523 mmol), triethylamine (145 µL, 1.047 mmol, 2 equiv.),
added and stirring was continued for 12 h. The progress of the bromoacetylbromide (50 μL, 0.575 mmol, 1.1 equiv.) and morpholine
reaction was monitored using TLC. After the completion of the (135 µL, 1.57 mmol, 3 equiv.). The product was obtained as pale
1
reaction, the reaction mixture was quenched with water (50 mL) and yellow solid (Yield 111 mg, 67 %); m. p. 170–172 ℃; H NMR (300
extracted with ethyl acetate (3x30 mL). The combined organic layer MHz, CDCl3): δ 9.83 (s, 1H), 9.36 (s, 1H), 7.57 (d, J = 3.0 Hz, 1H), 7.45
was washed with brine (210 mL) and dried over anhydrous Na2SO4. (d, J = 9.2 Hz, 1H), 7.29 (dd, J1 = 9.2 Hz, J2 = 3 Hz, 1H), 3.89 (s, 3H),
The solvent was evaporated and the crude product was purified by 3.84 (t, J = 4.6 Hz, 4H), 3.18 (s, 2H), 2.63 (t, J = 4.6 Hz, 4H); 13C{1H}
flash chromatography on silica to obtain 2-morpholino-N-(4-oxo-4H- NMR (75 MHz, CDCl3): δ 171.6, 168.7, 156.7, 150.8, 144.9, 124.5,
chromen-3-yl)acetamide (7c) as a white solid (over three steps) 123.1, 122.7, 119.9, 104.1, 67.1, 62.1, 55.9, 53.8; FT-IR (KBr): vmax
(Yield 107 mg, 60 %).
3278, 2959, 2854, 1680, 1631, 1587, 1491, 1113, 868 cm-1; HR-MS
N-(6-Chloro-4-oxo-4H-chromen-3-yl)-2-morpholinoacetamide (2c): (ESI) m/z [M+H]+: calcd. for C16H19N2O5: 319.1294, found: 319.1294.
The title compound was prepared according to procedure B using 2- N-(8-Methoxy-4-oxo-4H-chromen-3-yl)-2-morpholinoacetamide
(4-chloro-2-formylphenoxy)acetonitrile (14a) (100 mg, 0.512 mmol), (6c): The title compound was prepared according to procedure B
triethylamine (142 µL, 1.02 mmol, 2 equiv.), bromoacetylbromide (49 using 2-(2-formyl-3-methoxyphenoxy)acetonitrile (6a) (100 mg,
μL, 0.564 mmol, 1.1 equiv.) and morpholine (132 µL, 1.53 mmol, 3 0.523 mmol), triethylamine (145 µL, 1.047 mmol, 2 equiv.),
equiv.). The product was obtained as pale white solid (Yield 107 mg, bromoacetylbromide (50 μL, 0.575 mmol, 1.1 equiv.) and morpholine
1
65 %). m. p. 215–217 ℃; H NMR (400 MHz, CDCl3): δ 9.78 (s, 1H), (135 µL, 1.57 mmol, 3 equiv.). The product was obtained as pale
1
9.38 (s, 1H), 8.21 (d, J = 2.5 Hz, 1H), 7.63 (dd, J1 = 9 Hz, J2 = 2.5 Hz, yellow solid (Yield 116 mg, 70 %). m. p. 145–147 ℃; H NMR (400
1H), 7.48, (d, J = 9 Hz, 1H), 3.83 (t, J = 4.5 Hz, 4H), 3.18 (s, 2H), 2.62 MHz, CDCl3): δ 9.71 (s, 1H), 9.35 (s, 1H), 7.70 (d, J = 8.2 Hz, 1H), 7.24
(t, J = 4.5 Hz, 4H); 13C{1H} NMR (100 MHz, CDCl3): δ 170.9, 168.8, (t, J = 8.0 Hz, 1H), 7.09 (d, J = 8.0 Hz, 1H), 3.92 (s, 3H), 3.75 (t, J = 4.5
154.1, 145.2, 134.2, 130.9, 125.0, 124.1, 123.1, 120.3, 67.0, 62.1, Hz, 4H), 3.10 (s, 2H), 2.55 (t, J = 4.3 Hz, 4H); 13C{1H} NMR (100 MHz,
53.8. FT-IR (KBr): vmax 3285, 2855, 1685, 1631, 1606, 1531, 1111, 831 CDCl3): δ 171.7, 168.7, 148.9, 146.3, 144.8, 124.6, 124.1, 123.1, 116.1,
cm-1; HR-MS (ESI) m/z [M+H]+: calcd. for C15H16N2O4Cl: 323.0720, 114.0, 67.0, 62.1, 56.3, 53.8; FT-IR (KBr): vmax 3307, 1689, 1573,1392,
found: 323.0719.
1060, 812 cm-1; HR-MS (ESI) m/z [M+H]+: calcd. for C16H19N2O5:
N-(6-Fluoro-4-oxo-4H-chromen-3-yl)-2-morpholinoacetamide (3c): 319.1294, found: 319.1296.
The title compound was prepared according to procedure B using 2- N-(6,8-Dichloro-4-oxo-4H-chromen-3-yl)-2-morpholinoacetamide
(4-fluoro-2-formylphenoxy)acetonitrile (100 mg, 0.558 mmol), (8c) & (1c, HCl salt): The title compound was prepared according to
triethylamine (155 µL, 1.117 mmol, 2 equiv.), bromoacetylbromide procedure B using 2-(2,4-dichloro-6-formylphenoxy)acetonitrile (12a)
(53 μL, 0.614 mmol, 1.1 equiv.) and morpholine (144 µL, 1.675 mmol, (100 mg, 0.434 mmol), triethylamine (121 µL, 0.869 mmol, 2 equiv.),
3 equiv.). The product was obtained as dirty white solid (Yield 104 mg, bromoacetylbromide (41 μL, 0.478 mmol, 1.1 equiv.) and morpholine
1
61 %). m. p. 202–204 ℃; H NMR (400 MHz, CDCl3): δ 9.77 (s, 1H), (112 µL, 1.304 mmol, 3 equiv.). The product was obtained as dirty
9.39 (s, 1H), 7.87 (dd, J1 = 8.1 Hz, J2 = 3 Hz, 1H), 7.53 (dd, J1 = 9.2 Hz, white solid (Yield 107 mg, 69 %); m. p. 170–172 ℃; 1H NMR (400 MHz,
J2 = 4.2 Hz, 1H), 7.45–7.40 (m, 1H), 3.83 (t, J = 4.5 Hz, 4H), 3.18 (s, CDCl3): δ 9.77 (s, 1H), 9.47 (s, 1H), 8.12 (d, J = 2.3 Hz, 1H), 7.75 (d, J =
2H), 2.63 (d, J = 4.3 Hz, 4H); 13C{1H} NMR (100 MHz, CDCl3): δ 168.8, 2.3 Hz, 1H), 3.84 (t, J = 4.4 Hz, 4H), 3.20 (s, 2H), 2.64 (t, J = 4.4 Hz, 4H);
1
4
159.4 (d, JCF = 241 Hz), 152.1, 145.3, 123.6, 123.0 (d, JCF = 7.3 Hz), 13C{1H} NMR (100 MHz, CDCl3): δ 170.4, 168.9, 150.0, 145.1, 134.0,
122.6, 122.3, 120.7 (d, 3JCF = 8.0 Hz), 110.3 (d, 2JCF = 23.6 Hz) 67.0, 130.5, 124.8, 124.3, 123.8, 123.7, 67.0, 62.1, 53.8; FT-IR (KBr): vmax
62.1, 53.8; FT-IR (KBr): vmax 3244, 3059, 2961, 2859, 2821, 1690, 1636, 3337, 1694, 1628, 1599, 1495, 1182, 866 cm-1. HR-MS (ESI) m/z
1581, 1485, 1111, 833 cm-1; HR-MS (ESI) m/z [M+H]+: calcd. for [M+H]+: calcd. for C15H15N2O4Cl2: 357.0345, found: 357.0342.
C15H16N2O4F: 307.1094, found: 307.1093.
Compound 8c (50 mg, 0.14 mmol) was dissolved in ethanol (0.2 ml)
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 11
Please do not adjust margins

Please do not adjust margins
New Journal of Chemistry
Page 12 of 17
ARTICLE
Journal Name
1
2
3
4
5
6
7
8
9
and added 2 M HCl solution in diethyl ether (2 mL) and stirred the using 2-(2-formyl-3-methoxyphenoxy)acetonitrile (6a) (100 mg,
View Article Online
reaction mixture for 10 min. After slow evaporation with needle 0.523 mmol), triethylamine (145 µL, 1.D04O7I: 10m.1m03o9l,/C92NJe0q2u6i5v0.)A,
pressure of nitrogen gas followed by high vacuum drying yielded bromoacetylbromide (50 μL, 0.575 mmol, 1.1 equiv.) and piperidine
quantitative amount of HCl salt (1c). The product was obtained as (154 µL, 1.57 mmol, 3 equiv.). The product was obtained as pale
1
white solid (Yield 50 mg, 91 %); m. p. 107–109 ℃; 1H NMR (500 MHz, yellow solid (Yield 102 mg, 62 %). m. p. 130–132 ℃; H NMR (400
DMSO-d6 & 3-drops of CD3OD): δ 9.44 (s, 1H), 8.29 (s, 1H), 8.15 (s, 1H), MHz, CDCl3): δ 9.94 (s, 1H), 9.43 (s, 1H), 7.79 (d, J = 8.1 Hz, 1H), 7.32–
4.15 (bs, 2H), 4.05 (bs, 2H), 3.41 (s, 2H), 3.13 (s, 2H). 13C{1H} (126 MHz, 7.25 (m, 1H), 7.15 (d, J = 7.9 Hz, 1H), 3.98 (s, 3H), 3.09 (s, 2H), 2.52 (s,
DMSO-d6 & 3-drops of CD3OD): δ 170.3, 164.1, 150.0, 148.1, 134.3, 4H), 1.70 (p, J = 5.4Hz, 4H), 1.47 (d, J = 4.7Hz, 2H); 13C{1H} NMR (100
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
130.1, 124.5, 124.4, 124.1, 123.8, 63.4, 57.2, 52.3.
MHz, CDCl3): δ 171.8, 169.9, 148.9, 146.3, 144.8, 124.5, 124.3, 123.3,
N-(8-(Cyanomethoxy)-4-oxo-4H-chromen-3-yl)-2-morpholinoaceta- 116.5, 113.9, 62.5, 56.3, 55.0, 26.2, 23.6; HR-MS (ESI) m/z [M+H]+:
mide (9c): The title compound was prepared according to procedure calcd. for C17H21N2O4: 317.1501, found: 317.1501.
B using 2,2'-((3-formyl-1,2-phenylene)bis(oxy))diacetonitrile (100 mg, N-(6-Fluoro-4-oxo-4H-chromen-3-yl)-2-(piperidin-1-yl)acetamide
0.465 mmol), triethylamine (129 µL, 0.93 mmol,
bromoacetylbromide (44 μL, 0.511 mmol, 1.1 equiv.) and morpholine using 2-(4-fluoro-2-formylphenoxy)acetonitrile (100 mg, 0.558
(120 µL, 1.395 mmol, 3 equiv.). The product was obtained as pale mmol), triethylamine (155 µL, 1.117 mmol,
yellow solid (Yield 104 mg, 66 %). m. p. 159–161 ℃; H NMR (300 bromoacetylbromide (53 μL, 0.614 mmol, 1.1 equiv.) and piperidine
MHz, CDCl3): δ 9.78 (s, 1H), 9.41 (d, J = 3.5 Hz, 1H), 7.95 (dd, J1 = 6.8 (144 µL, 1.675 mmol, 3 equiv.). The product was obtained as pale
2 equiv.), (12c): The title compound was prepared according to procedure B
2
equiv.),
1
1
Hz, J2 = 2.8 Hz, 1H), 7.39–7.33 (m, 2H), 4.95 (s, 2H), 3.81 (t, J = 4.6 Hz, yellow solid (Yield 108 mg, 64 %). m. p. 141–143 ℃; H NMR (400
4H), 3.17 (s, 2H), 2.61 (t, J = 4.5 Hz, 4H); 13C{1H} NMR (75 MHz, CDCl3): MHz, CDCl3): δ 9.92 (s, 1H) 9.40 (s, 1H), 7.86 (dd, J1 = 8.2 Hz, J2 = 3 Hz,
δ 171.2, 168.8, 146.7, 145.5, 144.6, 124.5, 124.3, 123.6, 119.9, 117.9, 1H), 7.51 (dd, J1 = 9.2Hz, J2 = 4.2Hz, 1H), 7.43–7.38 (m, 1H), 3.10 (s,
114.2, 66.9, 62.0, 55.0, 53.7; FT-IR (KBr): vmax 3298, 2989, 2843, 1686, 2H), 2.53 (s, 4H), 1.70 (p, J = 5.6Hz, 4H), 1.48 (d, J = 5.1 Hz, 2H); 13C{1H}
1627, 1582, 1454, 1109, 868 cm-1; HR-MS (ESI) m/z [M+H]+: calcd. for NMR (100 MHz, CDCl3): δ 170.0, 159.1 (d, JCF = 245.5 Hz), 152.1,
C17H18N3O5: 344.1246, found: 344.1244.
Synthesis of 2,2'-((3-formyl-1,2-phenylene)bis(oxy))diacetonitrile: Hz), 110.3 (d, JCF = 23.7 Hz), 62.5, 55.0, 26.2, 23.6; FT-IR (KBr): vmax
1
4
3
145.2, 123.8, 123.2 (d, JCF = 7.9 Hz), 122.4, 122.2, 120.7 (d, JCF = 8
2
In
a
round-bottom flask, under nitrogen atmosphere, 2,3- 3294, 2936, 2855, 1692, 1643, 1582, 1483, 1159, 827 cm-1; HR-MS
dihydroxybenzaldehyde (200 mg, 1.449 mmol, 1 equiv.) was stirred (ESI) m/z [M+H]+: calcd. for C16H18N2O3F: 305.1301, found: 305.1302.
with anhydrous K2CO3 (599 mg, 4.347 mmol, 3 equiv.) in dry DMF (1 Procedure for three component addition reaction; Procedure C: 3-
mL) for 15 min at room temperature. After formation of yellow aminochromone (1b) (100 mg, 0.621 mmol, 1 equiv.) was dissolved
precipitate, bromoacetonitrile (240 µL, 3.478 mmol, 2.4 equiv.) was in methanol (5 mL) and reacted with 3-formylchromone (108 mg,
added and stirring was continued for 24 h. The progress of the 0.621 mmol, 1 equiv.) at 80 ℃ for about 2 h. The three component
reaction was monitored using TLC. After the completion of the adduct was precipitated upon cooling to room temperature. The
reaction, the reaction mixture was quenched with water (50 mL), obtained solid was filtered off, washed with cold methanol (3×3 mL)
extracted with ethyl acetate (320 mL), washed with brine (25 mL), and then vacuum dried to obtain pure form of (Z)-3-(((2-methoxy-4-
and dried over anhydrous Na2SO4. The solvent was evaporated and oxochroman-3-ylidene)methyl)amino)-4H-chromen-4-one (1d) as
the crude product was purified by column chromatography on silica pale yellow solid (Yield 147 mg, 68 %); m. p. 244–246 ℃; 1H NMR (500
1
to obtain white solid (Yield 274 mg, 88 %). m. p. 84–86 ℃; H NMR MHz, DMSO-d6): δ 11.57 (d, J = 12.9 Hz, 1H), 8.84 (s, 1H), 8.18 (d, J =
(400 MHz, CDCl3): δ 10.35 (s, 1H), 7.60 (dd, J1 = 9.3 Hz, J2 = 4.1 Hz, 7.8 Hz, 1H), 7.95 (d, J = 12.9 Hz, 1H), 7.86 (dd, J1 = 17.3 Hz, J2 = 8.0
1H), 7.34–7.30 (m, 2H), 4.93 (s, 2H), 4.89 (s, 2H); 13C{1H} NMR (100 Hz, 2H), 7.72 (d, J = 8.5 Hz, 1H), 7.54 (q, J = 8.0 Hz, 2H), 7.16 (t, J = 7.4
MHz, CDCl3): δ 188.3, 149.4, 148.6, 130.7, 126.4, 123.6, 120.7, 114.7, Hz, 1H), 7.11 (d, J = 8.1 Hz, 1H), 5.77 (s, 1H), 3.40 (s, 3H);13C{1H} NMR
114.3, 58.7, 54.9.
N-(7-Methoxy-4-oxo-4H-chromen-3-yl)-2-morpholinoacetamide
(10c): The title compound was prepared according to procedure B 107.1, 101.6, 55.4; FT-IR (KBr): vmax 3442, 2933, 1643, 1471, 1205 cm-
using 2-(2-formyl-5-methoxyphenoxy)acetonitrile (8a) (100 mg, 1; HR-MS (ESI) m/z [M+H]+: calcd. for C20H16NO5: 350.1028, found:
0.523 mmol), triethylamine (145 µL, 1.047 mmol, 2 equiv.), 350.1034. (Note: 13C{1H} NMR experiments of compounds 1d-12d
bromoacetylbromide (50 μL, 0.575 mmol, 1.1 equiv.) and morpholine suffered solubility issues during the sample preparation in both CDCl3
(135 µL, 1.57 mmol, 3 equiv.). The product was obtained as pale and DMSO-d6. The maximum solubility of the compounds was found
(126 MHz, DMSO-d6): δ 181.3, 177.6, 156.6, 149.1, 147.5 (2C), 144.1,
142.8, 141.6, 136.0, 126.2, 122.8 (2C), 122.7, 120.0, 118.8, 115.4,
1
yellow solid (Yield 109 mg, 66 %). m. p. 145–147 ℃; H NMR (400 to be 4 mg/0.5 mL of the solvent. In some cases, a mixture of solvents
MHz, CDCl3): δ 9.76 (s, 1H), 9.26 (s, 1H), 8.10 (dd, J1 = 8.9Hz, J2 = 2.1Hz, such as CDCl3 and 3 drops of CD3OD was necessary to dissolve the
1H), 6.96 (dd, J1 = 9.2Hz, J2 = 2.3Hz, 1H), 6.84 (d, J = 2.3 Hz, 1H), 3.88 compounds on slight pre-heating of the NMR tube. The additional
(d, J = 1.8Hz, 3H), 3.81 (t, J = 3.7 Hz, 4H), 3.15 (s, 2H), 2.60 (s, 4H); signals are believed to be obtained due to the diastereomeric nature
13C{1H} NMR (125 MHz, CDCl3): δ 171.8, 168.8, 149.0, 146.4, 144.9, of the compounds and partial decomposition of the compound due
124.7, 124.1, 123.2, 116.5, 114.1, 67.0, 62.1, 56.4, 53.8; FT-IR (KBr): to heat.
vmax 3273, 3132, 2972, 2853, 1672, 1631, 1524, 1440, 1115, 864 cm- (Z)-3-(((2-Ethoxy-4-oxochroman-3-ylidene)methyl)amino)-4H-
1; HR-MS (ESI) m/z [M+H]+: calcd. for C16H19N2O5: 319.1294, found: chromen-4-one (2d): The title compound was prepared according to
319.1292.
procedure C using 1b (100 mg, 0.621 mmol), ethanol (5 mL) and 3-
N-(8-Methoxy-4-oxo-4H-chromen-3-yl)-2-(piperidin-1-yl)acetamide formylchromone (108 mg, 0.621 mmol, 1 equiv.). The product was
(11c): The title compound was prepared according to procedure B obtained as yellow solid (Yield 175 mg, 78 %); m. p. 256–258 ℃; FT-
12 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 13 of 17
New Journal of Chemistry
Please do not adjust margins
Journal Name
ARTICLE
1
2
3
4
IR (KBr): vmax 3445, 2967, 1644, 1471, 1207 cm-1. 1H NMR (400 MHz, mmol, 1 equiv.). The product was obtained as pale yellVoiwewsAortliicdle(OYnielinlde
1
DMSO-d6): δ 11.55 (d, J = 12.8 Hz, 1H), 8.85 (s, 1H), 8.19–8.17 (m, 150 mg, 73 %); m. p. 316–318 ℃; H NMR D(4O0I:010M.1H0z3,9/DCM9NSJO0-2d665):0Aδ
1H), 7.95 (d, J = 13.2 Hz, 1H), 7.87–7.83 (m, 2H), 7.73 (d, J = 8.4 Hz, 11.51 (dd, J1 = 13 Hz, J2 = 2.3 Hz, 1H), 8.82 (d, J = 3.0 Hz, 1H), 7.89 (dd,
1H), 7.56–7.52 (m, 2H), 7.16–7.07 (m, 2H), 5.87 (s, 1H), 3.80–3.64 (m, J1 = 13.0 Hz, J2 = 3.0 Hz, 1H), 7.71–7.56 (m, 2H), 7.50–7.38 (m, 2H),
2H), 1.10 (t, J = 6.8 Hz, 3H); 13C{1H} NMR: (100 MHz, DMSO-d6) δ 180.9, 7.33 (dd, J1 = 8.4 Hz, J2 = 2.4 Hz, 1H), 6.98 (dd, J1 = 8.2 Hz, J1 = 3.2 Hz,
171.30, 156.0, 155.9, 144.38, 143.07, 135.0, 134.9, 126.40, 126.20, 1H), 5.70 (d, J = 3.4 Hz, 1H), 3.94 (d, J = 3.4 Hz, 3H), 3.34 (d, J = 3.6 Hz,
125.9, 125.7, 123.0, 122.4, 122.3, 119.1, 118.4, 104.9, 100.4, 63.5, 3H), 2.29 (d, J = 2.7 Hz, 3H); 13C{1H} NMR (126 MHz, CDCl3 & 3 drops
15.4.; HR-MS (ESI) m/z [M+H]+: calcd. for C21H18NO5: 364.1185, found: of CD3OD): δ 187.9, 181.0, 160.1, 152.7, 147.9, 136.9, 134.5, 131.3,
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
364.1181.
130.7, 125.2, 124.2, 123.0, 122.7, 116.7, 115.8, 113.4, 103.9, 103.9,
(Z)-3-(((2-Isopropoxy-4-oxochroman-3-ylidene)methyl)amino)-4H-
55.4, 54.3, 19.5; FT-IR (KBr): vmax 3424, 2937, 1652, 1488, 1218, 1070
chromen-4-one (3d): The title compound was prepared according to cm-1; HR-MS (ESI) m/z [M+H]+: calcd. for C22H20NO6: 394.1291, found:
procedure C using 1b (100 mg, 0.621 mmol), isopropanol (5 mL) and 394.1286.
3-formylchromone (108.8 mg, 0.621 mmol, 1 equiv.). The product (Z)-3-(((2-Ethoxy-4-oxochroman-3-ylidene)methyl)amino)-8-
was obtained as yellow solid (Yield 159 mg, 68 %). m. p. 244–246 ℃; methoxy-4H-chromen-4-one (7d): The title compound was prepared
1H NMR (400 MHz, DMSO-d6): δ 11.52 (d, J = 13.2 Hz, 1H), 8.85 (s, according to procedure C using 6b (100 mg, 0.523 mmol), ethanol (5
1H), 8.18 (dd, J1 = 6.8 Hz, J2 = 1.2 Hz, 1H), 7.92–7.83 (m, 3H), 7.73 (d, mL) and 3-formylchromone (91 mg, 0.523 mmol, 1 equiv.). The
J = 8.4 Hz, 1H), 7.55–7.52 (m, 2H), 7.15–7.11 (t, J = 7.2 Hz, 1H), 7.05 product was obtained as pale yellow solid (Yield 184 mg, 75 %); m. p.
1
(d, 8.4 Hz, 1H), 5.96 (s, 1H), 4.13–4.07 (m, 1H), 1.19 (d, J = 6 Hz, 3H), 230–232 ℃; H NMR (400 MHz, DMSO-d6): δ 11.52 (d, J = 13.0 Hz,
1.03 (d, J = 6.4 Hz, 3H); 13C{1H} NMR (100 MHz, DMSO-d6): δ 181.1, 1H), 8.83 (s, 1H), 7.91 (d, J = 12.9 Hz, 1H), 7.83 (dd, J1 = 7.8 Hz, J2 = 1.2
171.3, 156.2, 155.9, 144.0, 143.0, 135.0, 134.8, 126.4, 126.2, 125.9, Hz, 1H), 7.66 (dd, J1 = 7.2 Hz, J2 = 2.1 Hz, 1H), 7.58–7.47 (m, 1H), 7.42
125.7, 123.0 (2C), 122.2, 119.1, 118.5, 105.3, 99.1, 70.1, 23.6, 22.5; (q, J = 7.7 Hz, 2H), 7.28–6.98 (m, 3H), 5.85 (s, 1H), 3.94 (s, 3H), 3.77–
FT-IR (KBr): vmax 3448, 2966, 1644, 1472, 1212 cm-1; HR-MS (ESI) m/z 3.50 (m, 2H), 1.06 (t, J = 7.0 Hz, 3H); 13C{1H} NMR (126 MHz, CDCl3 &
[M+H]+: calcd. for C22H20NO5: 378.1341, found: 378.1341.
3 drops of CD3OD): δ 188.7, 181.7, 161.0, 155.9, 148.8, 143.7, 143.5,
(Z)-3-(((2-Ethoxy-6-methyl-4-oxochroman-3-ylidene)methyl)amino) 141.6, 134.9, 134.5, 126.4, 125.1, 124.7, 122.2, 117.8, 116.8, 114.3,
-4H-chromen-4-one (4d): The title compound was prepared 104.6, 101.5, 56.3, 55.3, 14.0. FT-IR (KBr): vmax 3446, 2974, 1651,
according to procedure C using 1b (100 mg, 0.621 mmol), ethanol (5 1468, 1205, 1019 cm-1; HR-MS (ESI) m/z [M+H]+: calcd. for C22H20NO6:
mL) and 3-formyl-6-methylchromone (116 mg, 0.621 mmol, 1 equiv.). 394.1291, found: 394.1310.
The product was obtained as pale yellow solid (Yield 170 mg, 72 %). (Z)-3-(((2-Ethoxy-6-methyl-4-oxochroman-3-ylidene)methyl)amino)
m. p. 260–262 ℃; 1H NMR (400 MHz, DMSO-d6): δ 11.52 (d, J = 12.8 -8-methoxy-4H-chromen-4-one (8d): The title compound was
Hz, 1H), 8.85 (s, 1H), 8.18 (dd, J1 = 6.4 Hz, J2 = 1.6 Hz, 1H), 7.93–7.83 prepared according to procedure C using 6b (100 mg, 0.523 mmol),
(m, 2H), 7.73 (d, J = 8.4 Hz, 1H), 7.65 (d, J = 2.0 Hz, 1H), 7.56–7.52 (m, ethanol (5 mL) and 3-formyl-6-methylchromone (98 mg, 0.523 mmol,
1H), 7.34 (dd, J1 = 6.4 Hz, J2 = 2.0 Hz, 1H), 6.97 (d, J = 8.4 Hz, 1H), 5.83 1 equiv.). The product was obtained as pale yellow solid (Yield 165
(s, 1H), 3.78–3.62 (m, 2H), 2.32 (s, 3H), 1.08 (t, J = 7.2 Hz, 3H); 13C{1H} mg, 65 %); m. p. 290–292 ℃; 1H NMR (400 MHz, DMSO-d6): δ 11.49
NMR (75 MHz, DMSO-d6): δ 181.1, 171.3, 155.8, 154.0, 144.1, 142.9, (d, J = 12.9 Hz, 1H), 8.80 (s, 1H), 7.87 (d, J = 12.9 Hz, 1H), 7.75–7.54
135.8, 134.8, 131.3, 126.4, 126.0, 125.7, 125.5, 122.7, 122.2, 119.1, (m, 2H), 7.48–7.35 (m, 2H), 7.31 (dd, J1 = 8.4 Hz, J2 = 2.2 Hz, 1H), 7.02–
118.3, 105.1, 100.3, 63.4, 20.6, 15.4; FT-IR (KBr): vmax 3449, 2959, 6.83 (m, 1H), 5.80 (s, 1H), 3.93 (s, 3H), 3.76–3.54 (m, 2H), 2.28 (s, 3H),
1648, 1467, 1216 cm-1; HR-MS (ESI) m/z [M+H]+: calcd. for C22H20NO5: 1.05 (t, J = 7.1 Hz, 3H); 13C{1H} NMR (126 MHz, CDCl3): δ 189.1, 176.5,
378.1341, found: 378.1339.
(Z)-8-Methoxy-3-(((2-methoxy-4-oxochroman-3-ylidene)methyl)
160.5, 154.5, 148.7, 143.5, 137.5, 135.9, 125.5, 124.9, 124.8, 123.8,
123.0, 120.2, 118.7, 118.3, 118.1, 116.7, 112.8, 56.3, 56.3, 29.7, 21.0;
amino)-4H-chromen-4-one (5d): The title compound was prepared FT-IR (KBr): vmax 3444, 2970, 1640, 1487, 1212, 1066 cm-1; HR-MS (ESI)
according to procedure C using 6b (100 mg, 0.523 mmol), methanol m/z [M+H]+: calcd. for C23H22NO6: 408.1447, found: 408.1459.
(5 mL) and 3-formylchromone (91 mg, 0.523 mmol, 1 equiv.). The (Z)-7-Methoxy-3-(((2-methoxy-4-oxochroman-3-ylidene)methyl)
product was obtained as pale yellow solid (Yield 181 mg, 77 %). m. p. amino)-4H-chromen-4-one (9d): The title compound was prepared
270–272 ℃;1H NMR (400 MHz, DMSO-d6): δ 11.53 (d, J = 13.0 Hz, 1H), according to procedure C using 8b (100 mg, 0.523 mmol), methanol
8.83 (s, 1H), 7.92 (d, J = 13 Hz, 1H), 7.83 (dd, J1 = 7.9, J2 = 1.6 Hz, 1H), (5 mL) and 3-formylchromone (91 mg, 0.523 mmol, 1 equiv.). The
7.64 (ddd, J1 = 12.8 Hz, J2 = 7.4 Hz, J3 = 2.1 Hz, 1H), 7.60–7.48 (m, 2H), product was obtained as pale yellow solid (Yield 170 mg, 72 %); m. p.
1
7.48–7.36 (m, 3H), 7.18–7.02 (m, 1H), 5.74 (s, 1H), 3.93 (d, J = 6.9 Hz, 282–284 ℃; H NMR (400 MHz, DMSO-d6): δ 11.64–11.30 (m, 1H),
3H), 3.36 (s, 3H); 13C{1H} NMR (126 MHz, CDCl3 & 3-drops of CD3OD): 8.75 (s, 2H), 8.03 (d, J = 9.0 Hz, 1H), 7.86 (ddd, J1 = 10.4 Hz, J2 = 9.2 Hz,
δ 187.8, 180.8, 160.1, 154.8, 147.8, 137.2, 134.0, 133.6, 125.4, 124.2, J3 = 3.2 Hz, 2H), 7.59–7.43 (m, 1H), 7.20 (d, J = 2.3 Hz, 1H), 7.17–6.98
123.0, 121.2, 116.9, 115.8, 115.4, 113.4, 112.1, 105.1, 100.6, 55.4, (m, 2H), 5.83 (s, 1H), 3.88 (s, 3H), 3.32 (s, 3H); 13C{1H} NMR (126 MHz,
54.4; FT-IR (KBr): vmax 3446, 2970, 1640, 1487, 1212, 1018 cm-1; HR- CDCl3): δ 188.7, 176.1, 160.5, 137.2, 136.9, 135.9 (2C), 131.1, 127.8,
MS (ESI) m/z [M+H]+: calcd. for C21H18NO6: 380.1134, found: 127.5, 127.0, 125.5 (2C), 118.6, 118.3, 116.2, 114.3, 100.0, 99.7, 55.7,
380.1132.
55.7; HR-MS (ESI) m/z [M+H]+: calcd. for C21H18NO6: 380.1134, found:
380.1124.
(Z)-8-Methoxy-3-(((2-methoxy-6-methyl-4-oxochroman-3-ylidene)
methyl)amino)-4H-chromen-4-one (6d): The title compound was (Z)-3-(((2-Ethoxy-4-oxochroman-3-ylidene)methyl)amino)-7-
prepared according to procedure C using 6b (100 mg, 0.523 mmol), methoxy-4H-chromen-4-one (10d): The title compound was
methanol (5 mL) and 3-formyl-6-methylchromone (98 mg, 0.523 prepared according to procedure C using 8b (100 mg, 0.523 mmol),
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 13
Please do not adjust margins

Please do not adjust margins
New Journal of Chemistry
Page 14 of 17
ARTICLE
Journal Name
1
2
3
ethanol (5 mL) and 3-formylchromone (91 mg, 0.523 mmol, 1 equiv.). 96-well plates (2×104/plate) and incubated with fresh DMEM at 37
View Article Online
The product was obtained as pale yellow solid (Yield 174 mg, 71 %). ℃ under 5% CO2 and 95% air for 24 h. AfteDr OthI:e10c.e1l0ls39w/Ce9reNJw0a2s6h5e0Ad
m. p. 292–294 ℃; 1H NMR (400 MHz, DMSO-d6): δ 11.48 (d, J = 13.0 twice with PBS (phosphate buffered saline), the medium was
Hz, 1H), 8.74 (s, 1H), 8.03 (d, J = 9.0 Hz, 1H), 7.89 (d, J = 13.0 Hz, 1H), changed to serially diluted test samples in DMEM (100 μL) with a
7.83 (dd, J1 = 7.8 Hz, J2 = 1.7 Hz, 1H), 7.51 (ddd, J1 = 8.2 Hz, J2 = 7.3 Hz, control and blank in each test plate. After 24 h of treatment, the cells
J3 = 1.8 Hz, 1H), 7.19 (d, J = 2.4 Hz, 1H), 7.16–7.00 (m, 3H), 5.83 (s, were washed twice with PBS and incubated with 100 μL of DMEM
1H), 3.88 (s, 3H), 3.79–3.55 (m, 2H), 1.06 (t, J = 7.1 Hz, 3H); 13C{1H} containing MTT solution (5 mg/mL). Then, the cells were incubated
NMR (126 MHz, CDCl3 & few drops of CD3OD): δ 181.6, 171.4, 164.4, for 3-4 h at 37 ℃ and DMSO (100 μL) was added to each well.
157.7, 155.7, 143.8, 141.4, 135.0, 134.4, 127.1, 126.1, 125.9, 122.6, Addition of DMSO gave purple colored precipitate due to the
122.1, 117.7, 115.2, 104.3, 101.4, 100.0, 55.7, 55.1, 13.6; HR-MS (ESI) formation of formazan in live cells. After 3-4 h of incubation at 37 ℃,
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
m/z [M+H]+: calcd. for C22H20NO6: 394.1291, found: 394.1299.
it formed purple colored precipitate due to the formation of
formazan in live cells. For quantifying the precipitate, the absorbance
(Z)-7-Methoxy-3-(((2-methoxy-6-methyl-4-oxochroman-3-ylidene)
methyl)amino)-4H-chromen-4-one (11d): The title compound was at 570 nm was measured (BioRad).56 Cell viability was calculated from
prepared according to procedure C using 8b (100 mg, 0.523 mmol), the mean values of data from three wells by using the following
methanol (5 mL) and 3-formyl-6-methylchromone (98 mg, 0.523 equation:
mmol, 1 equiv.). The product was obtained as pale yellow solid (Yield Cell viability (%) = [Abs (Test sample) – Abs (Blank)]/ [Abs (Control) – Abs (Blank)
186 mg, 76 %). m. p. 266–268 ℃; H NMR (400 MHz, DMSO-d6): δ x 100
]
1
11.46 (d, J = 13.0 Hz, 1H), 8.72 (s, 1H), 8.03 (d, J = 9.0 Hz, 1H), 7.86 (d, Fluorescence study
J = 13.1 Hz, 1H), 7.61 (d, J = 1.3 Hz, 1H), 7.39–7.25 (m, 1H), 7.18 (d, J 1×105 HeLa S3 cancer cells were seeded in 30 mm plates; cells were
= 2.4 Hz, 1H), 7.08 (dd, J1 = 9.0 Hz, J2 = 2.4 Hz, 1H), 7.03–6.91 (m, 1H), treated with compounds SVM-2 (50 μM) and SVM-4 (10 μM), and
5.68 (s, 1H), 3.88 (s, 3H), 3.38 (s, 3H), 2.68 (s, 3H); 13C{1H} NMR (126 incubated for 24 h. After treatment, cells were stained with AO/PI
MHz, CDCl3): δ 188.7, 176.1, 163.5, 160.5, 157.9, 154.5, 137.2, 135.9, (10 µM) solution for 15 min at dark condition. The images were
127.0, 125.5, 125.0, 120.2, 118.3, 116.2, 114.6, 114.3,101.1, 99.7, captured under fluorescence microscope.56-58
55.8, 55.7, 29.7, 21.0. HR-MS (ESI) m/z [M+H]+: calcd. for C22H20NO6: Colony formation study
394.1291, found: 394.1279.
Freshly prepared HeLa S3 cells (2×103/well) were plated in 24-well
(Z)-3-(((2-Ethoxy-6-methyl-4-oxochroman-3-ylidene)methyl) amino)- plates and incubated for overnight under condition (5 % of CO2 and
7-methoxy-4H-chromen-4-one (12d): The title compound was 95 % of air) for cell attachment. Next day, HeLa S3 cells were treated
prepared according to procedure C using 8b (100 mg, 0.523 mmol), with different concentrations of SVM-2 (1, 5 and 10 μM) and SVM-4
ethanol (5 mL) and 3-formyl-6-methylchromone (98 mg, 0.523 mmol, (2.5, 5 and 10 μM) for 24 h, and washed twice with PBS. Then, cells
1 equiv.). The product was obtained as pale yellow solid (Yield 175 were incubated with freshly prepared DMEM for 10 days to grow
mg, 69 %). m. p. 244–246 ℃; 1H NMR (400 MHz, DMSO-d6): δ 11.45 colonies. After 10 days of incubation, cells were washed with distilled
(d, J = 13.0 Hz, 1H), 8.74 (s, 1H), 8.03 (d, J = 9.0 Hz, 1H), 7.86 (d, J = H2O and incubated with crystal violet solution for 15-20 min at room
13.0 Hz, 1H), 7.62 (d, J = 1.8 Hz, 1H), 7.31 (dd, J = 8.3, 2.3 Hz, 1H), 7.20 temperature. Cells were then washed with distilled H2O and dried.
(d, J = 2.4 Hz, 1H), 7.08 (dd, J1 = 9.0 Hz, J2 = 2.4 Hz, 1H), 6.94 (d, J = 8.3 The images were captured and colony area was calculated by ImageJ
Hz, 1H), 5.79 (s, 1H), 3.88 (s, 3H), 3.81–3.53 (m, 2H), 2.29 (s, 3H), 1.05 software.
(t, J = 7.1 Hz, 3H); 13C{1H} NMR (126 MHz, DMSO-d6): δ 188.7, 176.0,
172.9, 163.5, 160.5, 154.5, 137.2, 136.9, 135.9, 127.0, 125.5, 125.0,
120.2, 118.3, 118.1, 117.7, 116.2, 114.32, 100.0, 55.8, 55.7, 21.0,
20.5. HR-MS (ESI) m/z [M+H]+: calcd. for C23H22NO6: 408.1447, found:
Conflicts of interest
There are no conflicts to declare
408.1459.
Computational studies
All the calculations were carried out using the Gaussian09 quantum
Acknowledgements
chemical program.48 Geometry optimization of the reactants,
S. V. thanks the Department of Science and Technology, New Delhi
intermediates and transition states were performed using the
for DST-INSPIRE faculty award (DST/INSPIRE/04/2015/000328). We
B3LYP51-55 functional in conjunction with the basis set 6-31G**.
thank Dr. Liu Xuewei group, CBC, SPMS, Nanyang Technological
Frequency calculations have been done to confirm that the
University, Singapore for their constant support for spectral analysis.
optimized structures were minima on the potential energy surface.
For the stable structures, no imaginary frequency was observed. For
the transition states, one imaginary frequency on the reaction path
was observed. Further intrinsic reaction coordinate (IRC) energy
profiles have been calculated to confirm that the tracked transition
states (TS) were the correct ones.
Anticancer activity
HeLa S3 and A549 cancer cells were cultured and maintained in
standard DMEM (Dulbecco’s modified eagle medium) with 10 % fetal
bovine serum. The cancer cells were stored at 37 ℃ under humidified
condition (5 % of CO2 and 95 % of air). Briefly, cells were seeded in
Notes and references
1
2
3
M. N. Hopkinson, C. Richter, M. Schedler and F. Glorius,
Nature 2014, 510, 485–496.
X. -Y. Chen, S. Li, F. Vetica, M. Kumar and D. Enders, iScience
2018, 2, 1–26.
a) D. M. Flanigan, F. Romanov-Michailidis, N. A. White and T.
Rovis, Chem. Rev. 2015, 115, 9307–9387. b) Y. Wang, L. -B. Qu
and D. Wei, Chem. Asian J. 2019, 14, 293300. c) Y. Wang, Q.-Y.
Wu, T. -H. Lai, K. -J. Zheng, L. B. Qu and D. Wei, Catal. Sci.
Technol. 2019, 9, 465476. d) X. Li, Y. Wang, Y. Wang, M. Tang,
14 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 15 of 17
New Journal of Chemistry
Please do not adjust margins
Journal Name
ARTICLE
1
2
3
4
5
6
7
8
9
L. -B. Qu, Z. Li and D. Wei, J. Org. Chem. 2018, 83, 85438555. 26 M. L. D. Wang, J. Du, R. Y. Chen and D. Q. Yu, ChiVni.ewChAretimcle.OLnelitnte.
338360. f) Y. Wang, L. -B. Qu and D. Wei, Chem. Asian J., 2019, 27 T. Sawada, S. Hashimoto, S. Tohma, YD. ONIi:s1h0i.o10k3a9,/TC.9NNJa0g2a6i5,0TA.
e) Y. Wang, D. Wei and W. Zhang, ChemCatChem, 2018, 10,
2000, 11, 129–130.
14, 293300. g) Y. Wang, Q. -Y. Wu, T. -H. Lai, K. -J. Zheng, L.
-B. Qu and D. Wei, Catal. Sci. Technol., 2019, 9, 465476. h) Q.
Sato, K. Ito, T. Inoue, M. Iwata and K. Yamamoto,
Immunopharmacol. 2000, 49, 285–294.
Shi, Y. Wang, Y. Wang, L. -B. Qu, Y. Qiao and D. Wei, Org. Chem. 28 T. Inaba, K. Tanaka, R. Takeno, H. Nagaki, C. Yoshida and
Front., 2018, 5, 27392748. i) Y. Wang, Y. Qiao, D. Wei and M.
Tang, Org. Chem. Front., 2017, 4, 19871998. j) Y. Qiao, D. Wei 29 C. Beudot, M. P. De Méo, D. Dauzonne, R. Elias, M. Laget, H.
Takano, S. Chem. Pharm. Bull. 2000, 48, 131–139.
and J. Chang, J. Org. Chem., 2015, 80, 8619–8630.
R. S. Menon, A. T. Biju and V. Nair, Chem. Soc. Rev. 2015, 44,
5040–5052.
J. Reis, A. Gaspar, N. Milhazes and F. Borges, J. Med. Chem.
2017, 60, 7941–7957.
C. F. M. Silva, D. C. G. A. Pinto and A. M. S. Silva,
ChemMedChem 2016, 11, 2252–2260.
M. J. Matos, S. Vazquez-Rodriguez, E. Uriarte, L. Santana and
F. Borges, Expert Opin. Ther. Patents 2015, 25, 1285–1304.
R. S. Keri, S. Budagumpi, R. K. Pai and R. G. Balakrishna, Eur. J.
Med.Chem. 2014, 78, 340–374.
Guiraud, G. Balansard and G. Duménil, Mutat. Res. 1998, 417,
141–153.
30 Y. Aikawa, N. Tanuma, T. Shin, S. Makino, K. Tanaka and Y.
Matsumoto, J. Neuroimmunol. 1998, 89, 35–42.
31 D. Dauzonne, B. Folléas, L. Martinez and G. G. Chabot, Eur. J.
Med. Chem. 1997, 32, 71–82.
32 a) T. Patonay and R. V. Hoffman, J. Org. Chem. 1995, 60, 2368–
2377. b) C. W. Robinson, C. S. Rye, N. E. A. Chessum and K.
Jones, Org. Biomol. Chem. 2015, 13, 7402–7407. c) J. Reis, F.
Cagide, D. Chavarria, T. Silva, C. Fernandes, A. Gaspar, E.
Uriarte, F. Remião, S. Alcaro, F. Ortuso and F. Borges, J. Med.
Chem. 2016, 59, 5879–5893.
33 J. F. Geissler, J. L. Roesel, T. Meyer, U. P. Trinks, P. Traxler and
N. B. Lydon, Cancer Res. 1992, 52, 4492–4498.
34 S. Vedachalam, Q.-L. Wong, B. Maji, J. Zeng, J. Ma and X. -W.
Liu, Adv. Synth. Catal. 2011, 353, 219–225.
35 S. Vedachalam, J. Zeng, B. K. Gorityala, M. Antonio and X. -W.
Liu, Org. Lett. 2010, 12, 352–355.
36 B. Maji, S. Vedachalan, X. Ge, S. Cai and X. -W. Liu, J. Org. Chem.
2011, 76, 3016–3023.
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
4
5
6
7
8
9
A. Gaspar, M. J. Matos, J. Garrido, E. Uriarte and F. Borges,
Chem. Rev. 2014, 114, 4960–4992.
10 H. A. Tawfik, E. F. Ewies and W. S. El-Hamouly, Int. J. Res.
Pharm. Chem. 2014, 4, 1046–1085.
11 G. A. Reynolds, J. A. Van Allan and A. K. Seidel, J. Heterocyclic
Chem. 1979, 16, 369–370.
12 B. Bauvois, M. -L. Puiffe, J. -B. Bongui, S. Paillat, C. Monneret
and D. Dauzonne, J. Med. Chem. 2003, 46, 3900–3913.
13 J. Morris, D. G. Wishka, A. H. Lin, W. R. Humphrey, A. L. Wiltse,
R. B. Gammill, T. M. Judge, S. N. Bisaha and N. L. Olds, J. Med. 37 S. Vedachalam, S. M. Tan, H. P. Teo, S. Cai and X. -W. Liu, Org.
Chem. 1993, 36, 2026–2032. Lett. 2012, 14, 174–177.
14 M. Zhao, J. -L. Liu, H. -F. Liu, J. Chen and L. Zhou, Org. Lett. 38 S. Vedachalam, N. Murugesh, P. Chakraborty, R. Karvembu
2018, 20, 2676–2679. and X.-W. Liu, New J. Chem. 2018, 42, 1832–1839.
15 M. Fridén-Saxin, N. Pemberton, K. da Silva Andersson, C. 39 C. Dai, J. Zhang, C. Huang and Z. Lei, Chem. Rev. 2017, 117,
Dyrager, A. Friberg, M. Grøtli and K. Luthman, J. Org. Chem.
2009, 74, 2755–2759.
6929–6983.
40 M. A. P. Martins, C. P. Frizzo, D. N. Moreira, N. Zanatta and H.
G. Bonacorso, Chem. Rev. 2008, 108, 2015–2050.
41 T. Welton, Biophys. Rev. 2018, 10, 691–706.
16 J. Wu, C. Zhao and J. Wang, J. Am. Chem. Soc. 2016, 138,
4706–4709.
17 D. Janssen-Müller, M. Fleige, D. Schlüns, M. Wollenburg, C. G. 42 R. L. Vekariya, J. Mol. Liq. 2017, 227, 44–60.
Daniliuc, J. Neugebauer and F. Glorius, ACS Catal. 2016, 6, 43 K. Dong, X. Liu, H. Dong, X. Zhang and S. Zhang, Chem. Rev.
5735–5739.
2017, 117, 6636–6695.
18 M. Zhao, H. Yang, M. -M. Li, J. Chen and L. Zhou, Org. Lett. 44 Suresh and J. S. Sandhu, Green Chem. Lett. Rev. 2011, 4, 289–
2014, 16, 2904–2907. 310.
19 M. Schedler, D. -S. Wang and F. Glorius, Angew. Chem., Int. 45 A. Sharma, P. Appukkuttan and E. Van der Eycken, Chem.
Ed. 2013, 52, 2585–2589. Commun. 2012, 48, 1623–1637.
20 a) H. U. Vora and T. Rovis, Aldrichimica Acta 2011, 44, 3–11. 46 V. Polshettiwar and R. S. Varma, Acc. Chem. Res. 2008, 41,
b) M. Christmann, Angew. Chem., Int. Ed. 2005, 44, 2632– 629–639.
2634. c) S. P. Lathrop and T. Rovis, Chem. Sci. 2013, 4, 1668– 47 D. Dallinger and C. O. Kappe, Chem. Rev. 2007, 107, 2563–
1673. d) Z. Rafiński, A. Kozakiewicz and K. Rafińska, ACS Catal.
2591.
2014, 4, 1404–1408. e) M. -Q. Jia, C. Liu and S. -L. You, J. Org. 48 See the supporting information for more details.
Chem. 2012, 77, 10996–11001. T. Ema, Y. Nanjo, S. Shiratori, 49 G. Kalaiarasi, G. Aswini, S. Rex Jeya Rajkumar, S. Dharani, V.
Y. Terao and R. Kimura, Org. Lett. 2016, 18, 5764–5767.
M. Lynch and R. Prabhakaran, Appl Organometal Chem. 2018,
32, e4466.
21 a) A. T. Biju, N. Kuhl and F. Glorius, Acc. Chem. Res. 2011, 44,
1182–1195. b) A. T. Biju and F. Glorius, Angew. Chem., Int. Ed. 50 M. K. M. Subarkhan and R. Ramesh, Inorg. Chem. Front. 2016,
2010, 49, 9761–9764. c) X. Bugaut, F. Liu and F. Glorius, J. Am.
3, 1245–1255.
Chem.Soc. 2011, 133, 8130–8133. d) I. Piel, M. Steinmetz, K. 51 A. D. Becke, J. Chem. Phys., 1993, 98, 5648–5652.
Hirano, R. Fröhlich, S. Grimme and F. Glorius, Angew. Chem., 52 A. D. Becke, J. Chem. Phys., 1993, 98, 1372–1377.
Int. Ed. 2011, 50, 4983–4987.
53 C. Lee, W. Yang and R. G. Parr, Phys. Rev. B, 1988, 37, 785–
22 C. Quiney, D. Dauzonne, C. Kern, J. -D. Fourneron, J. -C. Izard,
789.
R. M. Mohammad, J. -P. Kolb and C. Billard, Leukemia Res. 54 A. D. Becke, Phys. Rev. A, 1988, 38, 3098–3100.
2004, 28, 851–861.
55 L. R. Domingo, R. J. Zaragozá, J. A. Saéz and M. Arnó,
23 K. Tanaka, T. Yamamoto, Y. Aikawa, K. Kizawa, K. Muramoto,
Molecules 2012, 17, 1335–1341.
H. Matsuno and A. Muraguchi, Rheumatology 2003, 42, 1365– 56 C. Balachandran, J. Haribabu, K. Jeyalakshmi, N. S. P.
1371.
Bhuvanesh, R. Karvembu, N. Emi and S. Awale, J. Inorg.
Biochem. 2018, 182, 208–221.
57 J. Haribabu, G. Sabapathi, M. M. Tamizh, C. Balachandran, N.
S. P. Bhuvanesh, P. Venuvanalingam, R. Karvembu,
Organometallics 2018, 37, 1242–1257.
24 K. Kuriyama, C. Higuchi, K. Tanaka, H. Yoshikawa and K. Itoh,
Biochem. Biophys. Res. Commun. 2002, 299, 903–909.
25 Y. Aikawa, M. Yamamoto, T. Yamamoto, K. Morimoto and K.
Tanaka, Inflammation Res. 2002, 51, 188–194.
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 15
Please do not adjust margins

Please do not adjust margins
New Journal of Chemistry
Page 16 of 17
ARTICLE
Journal Name
1
2
3
4
5
6
7
58 J. Haribabu, M. M. Tamizh, C. Balachandran, Y. Arun, N. S. P.
Bhuvanesh, A. Endo and R. Karvembu, New J. Chem. 2018, 42,
10818–10832.
View Article Online
DOI: 10.1039/C9NJ02650A
59 13C{H} NMR experiments were carried out using a mixture of
solvents such as CDCl3 and 3 drops of CD3OD with slight pre-
heating of the NMR tube.
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
16 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 17 of 17
New Journal of Chemistry
1
2
3
4
View Article Online
DOI: 10.1039/C9NJ02650A
5
6
Table of Content:
7
8
9
NHC catalyzed green synthesis of functionalized chromones: DFT
mechanistic insights and in vitro activities in cancer cells
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Nithya Murugesh,a Jebiti Haribabu,a Krishnamoorthy Arumugam,a Chandrasekar Balachandran,b Rajagopal
Swaathy,a Shin Aoki,b, c Anandaram Sreekanth,*a Ramasamy Karvembu*a and Seenuvasan Vedachalam*a
A simple green protocol for the synthesis of 3-aminochromone derivatives using NHC catalyzed intramolecular hydroacylation reaction
was developed. Further functional 3-aminochromes was subjected to anticancer activites.