Synthesis and anticancer effect of 3,4,5-n-alkyl-benzamides on colon carcinoma HCT- 116 cells

Article abstract of DOI:10.13005/ojc/340323

The natural phenolic gallic acid has demonstrated a significant inhibition of cell proliferation in a series of cancer cell lines, as well as induced apoptosis in HCT-116 colon cancer cells. This research aims to synthesize six compounds of 3,4,5-trihydroxy-N-alkyl-benzamide derivatives of gallic acid, and investigate its anticancer effect on colon carcinoma HCT-116 cells.Six compounds of 3,4,5-trihydroxy-N-alkyl-benzamide, namely 3,4,5-trihydroxy-N-methyl-benzamide (2); 3,4,5-trihydroxy-N-ethyl-benzamide (3); 3,4,5-trihydroxy-N-butyl-benzamide (4); 3,4,5-trihydroxy-N-sec-butyl-benzamide (5); 3,4,5-trihydroxy-N-tert-butyl-benzamide (6) and 3,4,5-trihydroxy-N-hexyl-benzamide (7), have been successfully synthesized by amidation of carboxyl group of gallic acid with six corresponding alkylamines, respectively. Furthermore, anticancer effect of these six synthesized derivatives on colon HCT-116 cells were examined by MTT assay. Data were analyzed by linear regression method to generate IC₅₀ value. The results will be compared with gallic acid as an original compound and doxorubicine as a positive control.Amidation of gallic acid with six corresponding alkylamines gave desired -N-methyl-, -N-ethyl-, -N-butyl-, -N-sec-butyl-, -N-ters-butyl-, and –N-hexyl benzamide with yield ranging from 18% to 84%. Compared to gallic acid (IC₅₀: 0.05 μM) and doxorubicine (IC₅₀: 0.001 μM), all these six synthesized derivatives showed a lower anticancer effect on colon HCT-116 cells. The strongest anticancer and inhibitory effect on HCT-116 cells has shown by 3,4,5-trihydroxy-N-hexyl benzamide (7) with IC₅₀ value of 0.07 μM. Our results suggested that 3,4,5-trihydroxy-N-hexyl benzamide (7) is a potential to be developed as a promising anti-colon cancer agent.

Full text of DOI:10.13005/ojc/340323

ISSN: 0970-020 X
CODEN: OJCHEG
2018, Vol. 34, No.(3):
Pg. 1362-1367
ORIENTAL JOURNAL OF CHEMISTRY
An International Open Free Access, Peer Reviewed Research Journal
www.orientjchem.org
Synthesis and Anticancer Effect of 3,4,5-N-Alkyl-Benzamides
on Colon Carcinoma HCT- 116 Cells
JILLY OCTARIA TAgORE CHAN1, AdE ARSIANTI2,3,*, MAURIN MARCELIA1,
STEvANO JULIO WIJOYO1, FAdILAH FAdILAH2,3, RISTA PUTRIANINgSIH2,
NORMA NUR AzIzAH3, HIROkI TANIMOTO4 and kIYOMI kAkIUCHI4
1Medical Student, Faculty of Medicine University of Indonesia, Indonesia.
2Department of Medical Chemistry, Faculty of Medicine, University of Indonesia, Indonesia.
3Drug Development Research Cluster, Indonesia Medical Education and Research Institute (IMERI),
Faculty of Medicine, University of Indonesia, Jalan Salemba Raya 6 Jakarta 10430, Indonesia, Indonesia.
4Graduate School of Materials Science, Nara Institute of Science and Technology (NAIST), 8916-5
Takayama-cho, Ikoma, Nara, Japan.
*Corresponding author E-mail: arsi_ade2002@yahoo.com
http://dx.doi.org/10.13005/ojc/340323
(Received: April 02, 2018; Accepted: May 04, 2018)
ABSTRACT
The natural phenolic gallic acid has demonstrated a significant inhibition of cell proliferation
in a series of cancer cell lines, as well as induced apoptosis in HCT-116 colon cancer cells. This
research aims to synthesize six compounds of 3,4,5-trihydroxy-N-alkyl-benzamide derivatives of
gallic acid, and investigate its anticancer effect on colon carcinoma HCT-116 cells.Six compounds of
3,4,5-trihydroxy-N-alkyl-benzamide, namely 3,4,5-trihydroxy-N-methyl-benzamide (2);
3,4,5-trihydroxy-N-ethyl-benzamide (3); 3,4,5-trihydroxy-N-butyl-benzamide (4); 3,4,5-trihydroxy-
N-sec-butyl-benzamide (5); 3,4,5-trihydroxy-N-tert-butyl-benzamide (6) and 3,4,5-trihydroxy-N-
hexyl-benzamide (7), have been successfully synthesized by amidation of carboxyl group of gallic
acid with six corresponding alkylamines, respectively. Furthermore, anticancer effect of these
six synthesized derivatives on colon HCT-116 cells were examined by MTT assay. Data were
analyzed by linear regression method to generate IC50 value. The results will be compared with
gallic acid as an original compound and doxorubicine as a positive control.Amidation of gallic acid
with six corresponding alkylamines gave desired -N-methyl-, -N-ethyl-, -N-butyl-, -N-sec-butyl-,
-N-ters-butyl-, and –N-hexyl benzamide with yield ranging from 18% to 84%. Compared to gallic
acid (IC50: 0.05 µM) and doxorubicine (IC50: 0.001 µM), all these six synthesized derivatives showed
a lower anticancer effect on colon HCT-116 cells. The strongest anticancer and inhibitory effect on
HCT-116 cells has shown by 3,4,5-trihydroxy-N-hexyl benzamide (7) with IC50 value of 0.07 µM. Our
results suggested that 3,4,5-trihydroxy-N-hexyl benzamide (7) is a potential to be developed as a
promising anti-colon cancer agent.
keywords: Synthesis, Anticancer, Gallic acid, 3,4,5-trihydroxy-N-alkyl-benzamide,
HCT-116 cells.
This is an
Open Access article licensed under a Creative Commons Attribution-Non Commercial-Share Alike
4.0 International License (https://creativecommons.org/licenses/by-nc-sa/4.0/), which permits unrestricted
Non Commercial use, distribution and reproduction in any medium, provided the original work is properly cited.

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INTROdUCTION
activity, which is attributed to its strong apoptosis-
inducing and antioxidant effects.8 Gallic acid is
When benign adenomas and polyps believed to be apoptosis- inducer through the
transform into malignancies, colorectal cancer is generation of reactive oxygen species (ROS), Ca2+
brought about.It is the 3rd most frequently diagnosed influx as well as calmodulin activation.9 Gallic acid’s
cancer and the 4th most common cause of cancer cytotoxic effects on a few cancer cell lines including
death worldwide.1 According to GLOBOCAN 2012 colon cancer have been established by multiple
from the International Agency for Research on studies.10,11 In 2016, Abd-Rabou and co-workers
Cancer (IARC), the incidence of colorectal cancer in tested 5 different concentrations of gallic acid which
people of both genders was 1,360,602 (9.7%) while were 100µM, 50 µM, 25µM, 12.5µM, 6.25µM on HCT-
the mortality in that year sum up to as high as 693,933 116 cells and all of the results showed significantly
(8.5%). In Indonesia itself, there was a reported decreased (P < 0.01) cell viability; the values
27,772 (9.3%) colorectal incidence and 18,398 being 5.93%, 6.29%, 7.63%, 8.19%, and 22.87%
(9.5%) mortality cases in the year of 2012; making respectively.12 Additionally, gallic acid has been
this type of cancer reign the 4th against other types of shown to induce apoptosis in cancer cells selectively
cancer in the aspects of both incidence and mortality without doing the same to healthy, normal cells.This
in that year.2 To determine the appropriate treatment terrific finding on the anticancer properties of gallic
acid further fuels the interests of researchers to find
ways in which the activity could be further enhanced.
Owing to the knowledge that gallic acid is a
hydrophobic substance, cell membrane permeability
is restricted. A study has concluded that an increase
in potential cytotoxicity and lipophilicity can be
done by modifying the structure of gallic acid by the
addition of lipophilic groups.13 In the present study, we
have done the synthesis of gallic acid derivatives as
anti-hepatitis C virus agents.14 Recently, we also done
in silico study of gallic acid derivatives as inhibitor of
malarial dihydrofolate reductase15, as an inhibitors of
hepatitis C virus receptor NS5B16, as an inhibitor of
Bcl-xl anti-apoptotic protein of breast cancer17, and
as an inhibitor of BRAF colon cancer18.
for colorectal cancer, stage of cancer is crucial even
though other aspects such as type of tumor, patient’s
age, and patient’s degree of health are also taken
into consideration. American Joint Committee on
Cancer (AJCC) classifies colorectal cancer into
4 distinctive stages along with a 5th “recurrent”
stage.3 Surgery is mostly implied for colorectal
cancer in its early stages; in estimation, 95% of
stage 1 and 65-80% of stage II colorectal cancers
can be cured by surgery. Stage III of colorectal
cancer is usually treated firstly by tumor resection
with administration of adjuvant chemotherapy
such as 5-fluorouracil (5-FU) and leucovorin
(LV) such as FOLFOX for 6 months afterwards
(Longo et al., 2012). Sometimes, oxaliplatin will also
be prescribed in addition to minimize the likelihood of
recurrence.4,5 Stage IV of colorectal cancer is usually
being managed by the combination of therapies
which may include surgery, chemotherapy and
radiation. Chemotherapy, albeit being a traditional
In this work, as to continue research
in developing gallic acid derivatives as an
anticancer agent, we focused on the synthesize of
3,4,5-trihydroxy-N-alkyl-benzamides 2-7 (Fig.1), and
evaluate its anticancer effect on colon HCT-116 cells
treatment for cancers, possesses undesirable by MTT cell proliferation assay.
side effects.Chemotherapy drugs namely 5-FU, LV
EXPERIMENTAL
and oxaliplatin often cause neutropenia, diarrhea,
vomiting and peripheral neuropathy.6 Therefore,
design structure of gallic acid derivative
new treatments are constantly being developed and
researched on so that patients can receive the best
treatment possible for their condition. Gallic acid
(3,4,5- trihydroxybenzoic acid) is a polyphenolic acid
Six gallic acid derivatives, compound 2-7
(Fig.1) were designed by replacing carboxyl group of
gallic acid with a series of corresponding alkylamines
to generate 3,4,5-trihydroxy-N-alkyl-benzamides 2-7.
that can be found naturally in plants such as tea Four compounds, namely derivative 2, 3, 4 and 7,
leaves, grapes, mango and gall nuts.7 Gallic acid has were designed to have a linear aliphatic carbon chain
been shown to be a potent inhibitor of fungi, bacteria in N-alkyl benzamide group, where as compound 5
and some viruses. Further more, according to and 6 were designed to have a branched alphatic
Morley et al., gallic acid displays chemopreventive carbon chain.

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ARSIANTI et al., Orient. J. Chem., Vol. 34(3), 1362-1367 (2018)
H
H
H
O
OH
O
N
O
N
O
N
HO
OH
HO
OH
HO
OH
HO
OH
OH
OH
OH
OH
3,4,5-Trihydroxy-N-
butyl-benzamide (4)
Gallic acid (1)
3,4,5-Trihydroxy-N-
methyl-benzamide (2)
3,4,5-Trihydroxy-N-
ethyl-benzamide (3)
H
N
H
H
O
O
N
O
N
HO
OH
OH
HO
OH
OH
HO
OH
OH
3,4,5-Trihydroxy-N-sec-
butyl-benzamide (5)
3,4,5-Trihydroxy-N-tert-
butyl-benzamide (6)
3,4,5-Trihydroxy-N-
hexyl-benzamide (7)
Fig. 1. Structure of gallic acid (1) and desired 3,4,5-trihydroxy-N-alkyl-benzamides 2-7
general experimental method
Science andTechnology (NAIST), Japan.The tested
All chemical reactions were performed in HCT-116 cancer cells are the culture collection
an oven-dried glassware. Chloroform and methanol of Anatomical Pathology Department, Faculty of
in technical grade were purchased from Brataco Medicine, University of Indonesia.
Indonesia chemical distributor, and were distilled
prior to use. Methylamine, ethylamine, butylamine, general procedure of amidation
sec-butylamine, tert-butyl amine, hexylamine
WSCD.hydrochloride(405.0mg,3.52mmol)
and DMF were obtained from Wako Pure and NMM (0.77 mL, 7.04 mmol) was added to a
Chemical Industries. Gallic acid, doxorubicin, solution of gallic acid (300 mg, 1.76 mmol), alkylamine
N-(3-dimethylaminopropyl)-n'-ethylcarbodiimide (3.52 mmol) and HOBt (357 mg, 2.64 mmol) in DMF
hydrochloride (WSCD.HCl), 1-Hydroxybenzotriazole (5 mL).The mixture was stirred at room temperature
(HOBt) and N-methyl morpholine (NMM) were for 24 hours. The reaction was diluted by water
purchased from Sigma-Aldrich Chemical Company. (40 mL), and extracted by EtOAc (3 x 25 mL). EtOAc
Flash column chromatography was carried out extract was collected, further washed with brine
using Merck silica gel 60. Reaction and fractions of (20 mL), dried over anhydrous MgSO4, filtered and
chromatography were analyzed by Merck precoated evaporated to give a crude extract. The extract was
silica gel 60 F254 plates.Compounds were visualized flash chromatographed on silica gel (gradient elution
by an ultraviolet lamp (254 nm) and by staining with CHCl3/CH3OH) to give desired 3,4,5-trihydroxy-N-
ninhydrin (in EtOH). 1H NMR and 13C NMR spectra alkyl-benzamide.
were recorded on JEOL JNM-ECP500 (500 MHz)
3,4,5-trihydroxy-N-methylbenzamide (2)
spectrometert with tetramethylsilane (δ 0) and
acetone (δ 2.05) as an internal standard. Mass
spectra were measured by JEOL JMS-AX 700
spectrometer. Synthesis and cytotoxicity evaluation
of 3,4,5-trihydroxy-N-alkyl-benzamides were carried
out in Departement of Medical Chemistry, Faculty
of Medicine, University of Indonesia. Structure
elucidation of the synthesized 3,4,5-trihydroxy-
N-alkyl-benzamides were conducted in Graduate
School of Materials Science, Nara Institute of
Derivative 2 (248.0 mg, 75% yield) was
obtained from flash column chromatography on
silica gel of the crude extract (gradient elution 20:1
to 4:1 CHCl3/CH3OH) as a brown oil. Rf 0.13 (5:1
CHCl3/CH3OH);1HNMR (500 MHz, acetone-d6):6.94
(2H, s); 2.80 (3H, d, J = 4.6 Hz); 13CNMR (125 MHz,
acetone-d6): 168.3 (s); 146.3 (s); 136.9 (s); 126.3
(s);107.3 (d);26.6 (q);HRMS EI+ Calcd for C8H9NO4
[M]+: 183.0532, found: 183.0529.

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ARSIANTI et al., Orient. J. Chem., Vol. 34(3), 1362-1367 (2018)
3,4,5-trihydroxy-N-ethyl benzamide (3)
3,4,5-trihydroxy-N-hexyl-benzamide (7)
Derivative 7 (357.0 mg, 80% yield) was
Derivative 3 (68.5 mg, 20% yield) was
obtained from flash column chromatography on
silica gel of the crude extract (gradient elution 25:1
to 7:1 CHCl3/CH3OH) as a brown oil. Rf 0.31 (5:1
CHCl3/CH3OH);1HNMR (500 MHz, acetone-d6):7.03
(2H, s); 3.41-3.34 (2H, m); 1.15 (3H, t, J = 7.4 Hz);
13CNMR (125 MHz, acetone-d6): 167.8 (s); 146.1
(s); 136.8 (s); 126.5 (s); 107.6 (d); 35.2 (t); 15.1 (q);
HRMS EI+ Calcd for C9H11NO4 [M]+:197.0688, found:
197.0683.
obtained from flash column chromatography on silica
gel of the crude extract (gradient elution 25:1 to 5:1
CHCl3/CH3OH) as a yellow solid.Rf 0.33 (5:1 CHCl3/
CH3OH); 1HNMR (500 MHz, acetone-d6): 6.99(2H,
s); 3.59 (2H, t, J = 5.2 Hz); 1.59-1.52 (2H, m); 1.37-
1.23 (4H, m); 0.85 (3H, t, J = 8.2 Hz); 13CNMR (125
MHz, acetone-d6): 167.6 (s); 146.0 (s); 136.7 (s);
126.7 (s); 107.5 (d); 66.9 (t); 56.0 (t); 32.3 (t); 27.3
(t); 23.2 (t); 14.2 (q); HRMS EI+ Calcd for C11H15NO4
[M]+: 253.1314, found: 253.1306.
3,4,5-trihydroxy-N-butyl benzamide (4)
Cytotoxicity Evaluation
Derivative 4 (237.0 mg, 60% yield) was
Suspension of cells (100 µL) (5000 cells.
obtained from flash column chromatography on
silica gel of the crude extract (gradient elution 25:1
to 10:1 CHCl3/CH3OH) as a dark brown solid.Rf 0.38
(5:1 CHCl3/CH3OH);1HNMR (500 MHz, acetone-d6):
7.00 (2H, s); 3.37-3.31 (2H, m); 1.59-1.50 (2H, m);
1.41-1.31 (2H, m); 0.90 (3H, t, J = 7.8 Hz); 13CNMR
(125 MHz, acetone-d6): 167.6 (s); 146.0 (s); 136.6
(s);126.8 (s);107.5 (d);40.0 (t);32.5 (t);20.7 (t);14.1
(q); HRMS EI+ Calcd for C11H15NO4 [M]+: 225.1001,
found: 225.0991.
well) of HCT-116 was dispensed to a 96-well plate.
The plate was pre-incubated for 24 h in a humidified
incubator containing 5% CO2 at 37 oC. The various
concentration of tested samples (0-200 mM) were
added into the culture media in the plate. The plate
was incubated for an appropriate lenght of time
(72 h) in the incubator. 10 µL of MTT solution was
then added to each well of the plate which was
subsequently incubated for 1-4 h in the incubator.
This mixture was then measured for absorbance
by Elisa reader at 630 nm. This procedure was
triplicated for each plate. IC50 value was determined
3,4,5-trihydroxy-N-sec-butyl-benzamide (5)
Derivative 5 (331.4 mg, 84% yield) was
obtained from flash column chromatography on using probit method by plotting the concentration of
silica gel of the crude extract (gradient elution 25:1
to 9:1 CHCl3/CH3OH) as a brown oil. Rf 0.44 (5:1
CHCl3/CH3OH);1HNMR (500 MHz, acetone-d6):6.98
(2H, s); 4.05-3.94 (1H, m); 1.63-1.47 (2H, m); 1.16
(3H, d, J = 6.9 Hz); 0.90 (3H, t, J = 8.0 Hz); 13CNMR
(125 MHz, acetone-d6): 167.1 (s); 145.9 (s); 136.6
(s); 127.0 (s); 107.5 (d); 67.1 (d); 47.6 (t); 20.6 (q);
11.0 (q); HRMS EI+ Calcd for C11H15NO4 [M]+:
225.1001, found: 225.0993.
the sample against percentage of living cells.
RESULT ANd dISCUSSION
Synthesis of gallic acid derivatives
(3,4,5-trihydroxy-N-alkyl benzamide)
Synthetic pathway of 3,4,5-trihydroxy-N-
alkyl-benzamide 2-7 is summarized in Fig. 2. As
shown, amidation of gallic acid (1) with methylamine,
ethylamine, butylamine, sec-butylamine, tert-butyl
amine, and hexylamine using a combination ofWSCD.
HCl/HOBt in the presence of basic NMMafforded the
corresponding 3,4,5-trihydroxy-N-alkyl-benzamides,
which are 3,4,5-trihydroxy-N- methyl-benzamide (2),
3,4,5-tri-hydroxy-N-ethyl-benzamide (3), 3,4,5-tri-
hydroxy-N-butyl-benzamide (4), 3,4,5-tri-hydroxy-N-
sec-butyl-benzamide(5),3,4,5-trihydroxy-N-tert-butyl-
3,4,5-trihydroxy-N-t-butyl-benzamide (6)
Derivative 6 (69.2 mg, 18% yield) was
obtained from flash column chromatography on
silica gel of the crude extract (gradient elution 25:1 to
8:1 CHCl3/CH3OH) as a dark brown oil. Rf 0.52 (5:1
CHCl3/CH3OH);1HNMR (500 MHz, acetone-d6):6.97
(2H, s);1.28 (9H, s);13CNMR (125 MHz, acetone-d6):
167.1 (s); 145.9 (s); 136.6 (s); 127.0 (s); 107.5 (d);
42.6 (s); 29.8 (q); HRMS EI+ Calcd for C11H15NO4 benzamide (6) and 3,4,5-trihydroxy-N-hexyl-benzamide
[M]+: 225.1001, found: 225.0994. (7) with a yield ranging from 18% to 84%.

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Anticancer effect of 3,4,5-trihydroxy-N-alkyl- anticancer activity of the derivatives.The lower IC50
benzamides
value the higher anticancer activity. As shown in
Anticancer activity of 3,4,5-trihydroxy-N- Table 1, compared to gallic acid (IC50: 0.05 µM)
alkyl-benzamides against colon HCT-116 cellsis and doxorubicine (IC50: 0.001 µM), all these six
displayed in Table 1. Half maximal inhibitory synthesized derivatives showed a lower anticancer
concentration (IC50) value is utilized to express effect on HCT 116 cells.
H
O
OH
O
N
R
3,4,5-trihydroxy-N-methyl-benzamide (2), R= -CH3
R-NH2
3,4,5-trihydroxy-N-ethyl-benzamide
3,4,5-trihydroxy-N-butyl-benzamide
(3), R= -CH2CH3
(4), R= -(CH2)3CH3
WSCD.HCl, HOBt, NMM
3,4,5-trihydroxy-N-sec-butyl-benzamide (5), R= -CH(CH3)CH2CH3
3,4,5-trihydroxy-N-tert-butyl-benzamide (6), R= -C(CH3)3
18-84%
HO
OH
HO
OH
3,4,5-trihydroxy-N-hexyl-benzamide
(7), R= -(CH2)5CH3
OH
Gallic acid (1)
OH
3,4,5-Trihydroxy-N-
alkyl-benzamide
Fig. 2. Synthesis of 3,4,5-trihydroxy-N-alkyl-benzamides 2-7
Except for 3,4,5-trihydroxy-N-butyl- addition of methylene (-CH2) group in 3,4,5-trihydroxy-
benzamide (4), there is a visible trend of decreasing N-ethyl-benzamide (3), increases its hydrophobicity.
IC50 value and/or increasing anticancer activity from Compared to 3,4,5-trihydroxy-N-butyl-benzamide4
3,4,5-trihydroxy-N-methyl-benzamide2 (IC50: 2.43
µM), 3,4,5-trihydroxy-N-ethyl-benzamide3 (IC50:1.64
µM), and 3,4,5-trihydroxy-N-hexyl-benzamide7(IC50:
0.07 µM). This signifies that in general, 3,4,5-
trihydroxy-N-alkyl-benzamide with longer
hydro-phobic carbon chain of –N-alkyl-benzamide
group, possess a higher anticancer activity.
(IC50:3.56 µM) that has linear aliphatic carbon chain in
-N-alkyl-benzamide group,3,4,5-trihydroxy-
N-sec-butyl-benzamide5 (IC50: 1.34 µM) and
3,4,5-trihydroxy-N-tert-butyl-benzamide 6 (IC50:0.16
µM)which have a branched aliphatic carbon chain,
showed higher anticancer activity against HCT-116
cellss. Moreover, Among all the six synthesized
3,4,5-trihydroxy–N-alkyl-benzamides, derivative
3,4,5-trihydroxy–N-hexyl-benzamide (7) with
IC50 of 0.07 µM, shows the greatest potential
to be developed as an anti- colorectal cancer
agent. Additionally, further studies examining the
anti-colorectal cancer properties of 3,4,5-trihydroxy–
N-heptyl-benzamide and 3,4,5-trihydroxy–N-octyl-
benzamide that possess a longer aliphatic carbon
chain than 3,4,5-trihydroxy–N-hexyl-benzamide (7)
needs to be conducted.
However, the fact that all six compounds
of 3,4,5-trihydroxy-N-alkyl-benzamides have higher
IC50 value than original gallic acid, might suggest
that although carboxyl group is classified as being
hydrophilic, they are crucial for anti- colorectal
cancer activity. Derivative 3,4,5-trihydroxy-N-ethyl-
benzamide (3), shows improved anticancer effect on
HCT-116 cells when it compared to 3,4,5-trihydroxy-
N-methyl-benzamide (2), which implies that an
Table 1: Anticancer activity (IC50 in µM) of 3,4,5-trihydroxy-N-alkyl-benzamides 2-7,
gallic acid (1), and doxorubicine against colon HCT-116 cells
Compound
Cytotoxicity (IC50, µM)
Gallic acid (1)
3,4,5-trihydroxy-N-methyl-benzamide (2)
3,4,5-trihydroxy-N-ethyl-benzamide
3,4,5-trihydroxy-N-butyl-benzamide
3,4,5-trihydroxy-N-sec-butyl-benzamide (5)
3,4,5-trihydroxy-N-tert-butyl-benzamide (6)
0.05
2.43
1.64
3.56
1.34
0.16
0.07
0.01
(3)
(4)
3,4,5-trihydroxy-N-hexyl-benzamide
Doxorubicine
(7)

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CONCLUSION
Directorate of Research and Public Service (DRPM)
and Faculty of Medicine, University of Indonesia
for the PITTA (Publikasi Internasional Terindeks
untuk Tugas Akhir Mahasiswa) research grant
Fiscal year 2017, and to the Graduate School
of Materials Science, Nara Institute of Science
and Technology (NAIST), Japan, for International
Research Collaboration Program (NAIST Global
Six compounds of 3,4,5-trihydroxy-N-
alkyl-benzamides have been synthesized through
amidation of gallic acid with the corresponding
alkylamines. Among six synthesized derivatives,
3,4,5-trihydroxy-N-hexyl-benzamide (7) showed the
greatest anticancer effect on HCT-116 cells.
ACkNOWLEdgEMENT
We wish to express our gratitute to Initiative Program).
REFERENCES
1.
2.
Ferlay, J.; Shin, H.R.; Bray, F.; Forman, D.; 9.
Mathers, C.;Parkin, D.M.Int.J.Cancer., 2010,
127, 2893-2917.
WHO. GLOBOCAN 2012: Estimated cancer 10.
incidence, mortality and prevalence worldwide
in 2012[Internet]. 2012[updated 2014 Feb
14; cited 2017 Aug 15]. Available from:http:// 11.
globocan.iarc.fr/Pages/fact_sheets_
population.aspx
Filipiak, K.;Hidalgo, M.;Silvan, J.M.;Fabre, B.;
Carbajo, R.J.;Pineda-Lucena, A.Food Funct.,
2014, 5, 381-389.
Esteves, M.; Siquet, C.; Gaspar, A.; Rio, V.;
Sousa, J.B.; Reis, S. Arch. Pharm. Chem.
Life. Sci., 2008, 341, 164–173.
Khaledi, H.;Alhadi, A.A.;Yehye,W.A.;Ali, H.M.;
Abdulla, M.A.;Hassandarvish P.Arch.Pharm.
Chem. Life Sci., 2011, 344, 703–709.
Abd-Rabou, A.A.; Shalby, A.B.; Ahmed, H.H.
Int. J. Pharm. Bio. Sci., 2016, 7, 584-592.
Dorothea, M.; Arsianti, A.; Liwang, F.;
Christian, H.; Panjaitan, H.P. Ann. Oncol.,
2016, 17 (7), 523-532.
3.
4.
Mishra, J.;Dromund, J.;Quazi, S.H.;Karanki, 12.
S.S.; Shaw, J.J.; Chen, B. Crit. Rev. Oncol.
Hematol., 2013, 86, 232–250.
13.
Longo, D.L.; Kasper, D.L.; Jameson, J.L.;
Fauci, A.S.; Hauser, S.L.; Loscalzo, J.
Harrison’s Principles of Internal Medicine. 14.
18thed. New York: McGraw-Hill. 2012, 537-
538.
Gramont, A.D.; Figer, A.; Seymour, M.;
Homerin, M.; Hmissi, A.; Cassidy, J. J. Clin.
Arsianti, A.;Aoki-Utsubo, C.;Fadilah;Bahtiar,
A.; Apriyanto, D.R.; Dwira, S.; Pradisty, N.A.;
Tanimoto, H.; Kakiuchi, K.; Sudarmono, P.;
Hotta, H.; Paramita, R.I.; Erlina, L. Asian J.
Pharm. Clin. Res., 2017, 10, 164-167.
Arsianti, A.; Astuty, H.; Fadilah; Bahtiar, A.;
Tanimoto, H.; Kakiuchi, K. Asian J. Pharm.
Clin. Res., 2017, 10, 330-334.
Arsianti, A.; Fadilah; Bahtiar, A.; Dwira, S.;
Apriyanto, D.R.;Paramita, R.I.Int.J.Chemtech
Res., 2017, 10, 111-117.
Paramita, R.I.; Arsianti, A.; Radji, M. Int. J.
Chemtech Res., 2017, 10, 348-355.
5.
6.
7.
8.
Oncol., 2000,18, 2938-2947.
15.
16.
17.
André, T.; Boni, C.; Mounedji-Boudiaf, L.;
Navarro, M.;Tabernero, J.; Hickish, T. N Engl.
J. Med., 2004, 350, 2343-2351.
Yoon, C.H.;Chung, S.J.;Lee, S.W.;Park,Y.B.;
Lee, S.K.;Park, M.C.Joint Bone Spine., 2013,
80, 274-279.
Silva, I.C.; Polaquini, C.R.; Regasini, L.O.;
Ferreira, H.;Pavan, F.R.Food Chem.Toxicol., 18.
2017,105, 300-307.
Humaedi, A.; Arsianti, A.; Radji M. Int. J.
Chemtech Res., 2017, 10(1), 310-315.