Research J. Pharm. and Tech.

13(12): December 2020

ISSN 0974-3618 (Print) www.rjptonline.org

0974-360X (Online)

RESEARCH ARTICLE

Synthesis and Biological Screening of some Novel 3-Subsituted Indole

Derivatives
Raju Kundavaram1*, M. Gayathri Devi1, B. Siva Kumar2, T. Archana3, M. Gowri Manoja4,
Dr. J. Naga Sai Kumar5
1
Assistant Professor, Pharmaceutical Chemistry, Karnataka College of Pharmacy, Bangalore.
2
Creative Educational Society College of Pharmacy, Chinna Tekuru, Andhra Pradesh.
3
Assistant Professor, Pharmaceutics, Karnataka College of Pharmacy, Bangalore.
4
Assistant Professor, Pharmaceutical Analysis and Quality Assurance, Anurag Group of Institutions, Hyderabad.
5
Assistant Professor, Pharmacy Practice, Karnataka College of Pharmacy, Bangalore.
*Corresponding Author E-mail:

ABSTRACT:
In the present work Indole and substituted glycolic acid have been chosen as starting materials. These are broad
class of chemical compounds, with many important pharmacological properties. Because of ease of reaction with
electrophiles, indole is widely used in various pharmaceutical preparations. The present investigation was
concerned with synthesis of some novel 3-substituted indole derivatives with objective of discovery novel and
potent analgesic, anti-inflammatory and anti-oxidant agents. These compounds were synthesized by cyclisation
of indole-3-acetic acid with different aromatic amines. The structures of new compounds were confirmed by FT
IR, 1HNMR and Mass. These compounds were tested in-vivo for analgesic, anti-inflammatory activates and
biochemical studies antioxidant activity. The statistical analysis was carried out by ANOVA. All the compounds
tested (NHR-I-NHR-VII) showed analgesic, anti-inflammatory and antioxidant activities. Among seven
compounds three compounds (NHR-I, NHR-II, NHR-IV) showed significant analgesic, anti-inflammatory and
antioxidant activities. Compounds NHR-II and NHR-V showed 74.38% and 70.68% inhibition of paw oedema in
anti-inflammatory activity compared to 75.28% inhibition of standard Indomethacin. Compounds NHR-I and
NHR-II showed 68.84% and 73.30 % inhibition against acetic acid induced writings’ in analgesic activity
compared to 75.23% inhibition of standard Diclofenac. Compound NHR-II showed 71.6% inhibition of DPPH
radical in Anti-oxidant activity compared to 84% inhibition of standard Ascorbic acid. The results showed that
incorporation of appropriately substituted aromatic amines at third position of Indole nucleus can afford good
analgesic, anti-inflammatory and antioxidant with reduced side effects of compounds.

KEYWORDS: Indole, glycolic acid, Indomethacin, analgesic activity, anti-inflammatory activity.

INTRODUCTION: Indole chemistry began to develop with the study of the

INDOLES dye indigo found in many natural products such as the
Indole is an aromatic, heterocyclic, organic compound. It indole alkaloids, fungal metabolites and marine natural
has a bicyclic structure, consisting of a six-membered products which is benzopyrrole in which the benzene
benzene ring fused to a five-membered pyrrole ring at and pyrrole rings are fused through the 2- and 3-
2,3 – position. The name indole is portmanteau of the positions of the pyrrole nucleus. The IUPAC name of the
words indigo and oleum, since indole was first isolated indole is 1H-benzo (b) pyrrole, it is being the b-face
by treatment of the indigo dye with oleum. benzo-fused isomer.

Indole is a solid at room temperature, can be produced

by bacteria as a degradation product of the amino acid
Received on 24.12.2019 Modified on 05.03.2020 tryptophan. It also occurs in coal tar. Indoles are basic in
Accepted on 06.05.2020 © RJPT All right reserved nature and form salts with acids (Morrin Acheson et al.,
Research J. Pharm. and Tech. 2020; 13(12):5787-5792. 2009)1,2. Pandeya et al. synthesized a series of p-nitro
DOI: 10.5958/0974-360X.2020.01009.4
phenyl substituted semicarbazones. The newly
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synthesized derivatives were screened for their potent analgesic, anti-inflammatory and anti-oxidant
anticonvulsant activities against maximal electroshock activities with least possible unwanted effects.
(MES), subcutaneous pentylenetetrazole (scPTZ) and
subcutaneous strychnine (scSTY) tests. (Pandeya et al., MATERIAL AND METHODS:
1999)3. Gerard A. Pinna et al. investigated various new Research Design:
derivatives and structural analogues of N-(1-ethyl-2- To start the work, initially literature survey has been
pyrrolidinylmethyl)-4,5-dihydro-1H-benzo [g] indole-3- done. After literature survey, a general scheme was
carboxamide, a representative term of a series of 2- prepared as follows.
aminomethyl pyrrolidinyl derived 4,5-dihydrobenzo[g]
indole carboxamides with good D2-like affinity, and O
evaluated for their ability to bind to dopamine D2-like
HO
receptors in vitro. From these studies, compound (2- N
H OH

chloro-N-(1-ethyl-2-pyrrolidinylmethyl)-5, 6-dihydro- 1H-indole glycolic acid

4H benzo [6, 7] cyclohepta [b] pyrrole-3-carboxamide),

was found to possess a potent affinity for D2-like KOH,Water
receptors. (Gerard A. Pinna et al., 2002)4. Istvan Borza et 250°c 18 hr
al., synthesized a novel series of indole-2-carboxamide
derivatives and identified as NR2B selective NMDA CH2COOK

receptor antagonists. The influence of the number and

position of OH groups on the indole skeleton as well as
the substitution of the piperidine ring on the biological N
H
activity of the compounds was studied. From the results potassium 2-(1H-indol-3-yl)acetate

the compound was found to be more potent among the

all derivatives. (Istvan Borza et al., 2003.)5. HCl

Able, E et al., synthesized some istatin and indole CH2COOH

oximes and evaluated for their anti-hypertensive activity.

(Able, E et al., 2003)6, VitoBoido et al., reported A set of
ten 2-phenyl-3-(quinolizidin-1-yl)-5-substituted indoles N
H
was prepared through the Fischer cyclization of lupin 2-(1H-indol-3-yl)acetic acid
and epilupinylphenylketone4-substituted phenyl R

hydrazones. Compounds were tested for antiaggregating

activity on human platelets activated by adenosine NH2

diphosphate (ADP), collagen and adrenaline. (VitoBoido

DCC,Ethylacetate
et al.,2004)7. Girolamo Cirrincione et al., reported the O
stirr 24 hr
R
Eight derivatives of the new ring system [1,2,3,5]
tetrazino [5,4-a] indole-4-one 7, were synthesised in N
H
good yields by reaction of 2-diazoindoles with alkyl or
aryl isocyanates. Compounds 7 were screened for their
activity against human tumour cell lines. (Girolamo
N
Cirrincione et al., 2005)8. H

Importance of the proposed investigation: Indole and substituted glycolic acid have been chosen as
Indoles found to have analgesic, anti-inflammatory, starting materials. The formations of the final products
antioxidant activities which are potent molecules and were monitored by TLC. The completed products
hence indole derivatives can effectively treat these showed significant color under UV chamber. All the
diseases. The starting materials in the present work are compounds prepared were purified by recrystallization
Indole and substituted glycolic acid which are broad with suitable solvents. Structures of the synthesized
class of chemical compounds, with many important compounds have been characterized and confirmed by
pharmacological properties. It is widely used in various IR, NMR and Mass techniques.
pharmaceutical preparations. Therefore, various efforts
have been made to produce new compounds with Chemicals and Instruments:
different properties and least possible unwanted effects. The chemicals used in this project was obtained from SD
The present investigation mainly concerned with Fine chemicals, the instruments such as FT-IR was of
synthesis and characterization of some novel 3- Shimadzu make, NMR of Joel D-3000, and UV
substituted Indole derivatives and evaluated for their Spectrophotometer of Labo Med Inc 2602.

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Table No. 1: Summary of the synthesized compounds

S. No. Compound Molecular formula Molecular Colour Solubility Melting point % of
code weight Yield
1 NHR-I C16H15N2O 250 Light buff Chloroform, DMSO 126-128°C 78
2 NHR-II C15H13N2OCl 284.5 Buff Chloroform, 80-82°C 80
Ethanol
3 NHR-III C17H16N2o2 280 Dark brown Chloroform, DMSO 79
4 NHR-IV C17H16N2O 264 Brick red Chloroform, acetone 166-167°C 83
5 NHR-V C22H18N4O 354 Dark yellow Chloroform, 130-132°C 80
acetone. DMSO

RESULTS:
Synthetic methodology (table-1). 2. Compound: N-(3-chlorophenyl)-2-(1H indole-3-yl)
acetamide
Spectral results: The synthesized compounds were Code: NHR-II
characterized by IR, NMR and Mass spectral studies IR
spectra were recorded on SHIMADZU as KBr disc and
Mull technique (γ cm-1).1H-NMR spectra were recorded
on Bruker AC 80 MHz Spectrophotometer using CDCl3
as an internal standard (chemical shift in δ, ppm).Mass
spectra were recorded on an Electron impact Mass
spectrometer at 70 eV using direct insertion probe. The
molecular weight was calculated in positive mode by
standard addition method.

1. Compound: 2-(1H-indol-3-yl)-N-phenylacetamide
Code: NHR-I

Fig. 2: FT-IR, 1H NMR, Mass Spectra of Compound NHR-I

3. Compound: 2-(1H-indole-3-yl)-N-(3-
methoxyphenyl) acetamide
Code: NHR-III

Fig. 1: FT-IR, 1H NMR, Mass Spectra of Compound NHR-I

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Fig. 3: FT-IR 1H NMR Spectra of Compound NHR-III

Table No. 2: IR and 1H-NMR and Mass spectral data of compound NHR-I, II, III
Data of Compound NHR-I
1
S. FTIR H-NMR Mass
No. Vibrational moiety Wave number, Chemical shift Nature of proton Data of positive Molecular
γ (cm-1) (δ, ppm) addition method weight
1. Indole N-H stretch 3432 3.928 2H, s, CH2 (M + BP) = 250 250
2. N-H stretch of RCONHR1 3276 7.0-7.6 10H, s, of Aromatic
ring
3. C-H stretch in aromatic ring 2854 8.2 1H, br-s, NH
4. C-H stretch in aliphatic ring 2930 8.4 1H, br-s, NH
5. C=O stretch in RCONHR1 1654
Data of Compound NHR-II
1
S. FTIR H-NMR Mass
No. Vibrational moiety Wave number, Chemical shift Nature of proton Data of positive Molecular
γ (cm-1) (δ, ppm) addition method weight
1. Indole N-H stretch 3372 3.928 2H, s, CH2 Aliphatic (M + BP) = 250 250
ring
2. Br N-H stretch of RCONHR1 3314 7.0-7.6 9H, m, of Aromatic
ring
3. C-H stretch in aromatic ring 2854 8.1 1H, br-s, NH
4. C-H stretch in aliphatic ring 2930 8.3 1H, br-s, NH
5. C=O stretch in RCONHR1 1654
Data of Compound NHR-III
1
S. FTIR H-NMR Mass
No. Vibrational moiety Wave number, Chemical shift Nature of proton Data of positive Molecular
γ (cm-1) (δ, ppm) addition method weight
1. Indole N-H stretch 3413 3.89 3H, s, CH2 Aliphatic (M +) = 281 281
ring Base peak 277
2. Br N-H stretch of RCONHR1 3294 3.75 3H, s, of OCH3
3. C-H stretch in aromatic ring 2854 8.1 1H, br-s, NH
4. C-H stretch in aliphatic ring 2929 8.3 1H, br-s, NH
5. C=O stretch in RCONHR1 1666 6.60 1H,m of Aromatic ring

Pharmacological Studies:
Analgesic and anti-inflammatory activities of
synthesised compounds were evaluated by in vivo
methods.

Analgesic Activity:
The analgesic activity of the compounds was evaluated
by acetic acid induced writhing model and Eddy’s hot
plate methods.

Results of Analgesic Activity using acetic acid

induced writhing in Mice:
Analgesic effect of synthesized derivatives was Fig. 4: Bar diagram showing Analgesic activity results
measured in acetic acid induced writhing test. Data are
expressed as the Mean ± SEM (n=5), one-way analysis
of variance (ANOVA) followed by multiple comparison
method. It is represented in the Fig. 4
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Results of Analgesic Activity using Eddy’s Hot-Plate “3-substituted some novel indole derivatives” were
Test in Mice: synthesized in two steps facile procedure. The reaction
involves interaction of indole, Glycolic acid and
Table No. 3: Eddy’s Hot-Plate Test substituted anilines. Seven derivatives were synthesized,
S. Compound Response in minutes characterized and evaluated for analgesic, anti-
No. Group 30min 60min 90min
1 Control 2±0.29 3.83±0.8 3±0.52
inflammatory and antioxidant activities. The purity of
2 Standard 6±0.45 ns
7±1.16** 9.16±0.76** the title compounds were tested in the precoated TLC
(Diclofenac) plates, structural characterization was performed by IR,
3 NHR-I 3.33±0.5ns 6.6±0.25* 5.83±0.7** 1H NMR and Mass. All the three derivatives were
4 NHR –II 3.33±0.4** 6±0.2** 6.83±0.3** screened for their analgesic, anti-inflammatory and
5 NHR –III 4.66±0.5*** 6.1±0.4** 8.33±0.33**
antioxidant activities. Synthesized derivatives were
evaluated for analgesic activity in this activity,
Effects of the all compounds on the latency time of mice
Diclofenac was used as standard drug.
exposed to the hot plate test were studied. All values are
expressed as Mean ± Sem n=5, one way analysis of
ANALAGESIC ACTIVITY:
variance (ANOVA) followed by multiple comparison
Acetic acid induced writhing model:
Dunnett’s test *P<0.005, **P<0.01, and***P<0.001 as
From the results the synthesized derivatives screened at
compared to control group. Some of the values are said
20 mg/kg body weight NHR-I has shown 68.8% of
to be NS (no significant)
inhibition which is more potent when compare to control
and less potent than standard (72.5%) due to
Anti-Inflammatory Activity:
unsubstituted aniline. The result of NHR-II revealed that
The anti-inflammatory activity of the compounds was
73.30% of inhibition which is less potent than standard
evaluated in Carrageenan-induced paw method.
Diclofenac may be presence of Chlorine group at Meta
position might be favoured more potent analgesic
Results of Anti-inflammatory Activity in Mice using
activity.
Carrageenan-induced paw oedema model in mice:

Table No. 4: Anti-inflammatory activity (Percentage inhibition of

NHR-III has given 61.6% inhibition which is less potent
paw volume) than all other derivatives and standard due to presence of
S. Compound % Inhibition of Paw oedema Time in electron releasing group like Methoxy at Meta position.
No. Group minutes
30min 60min 90min 180 min Eddy’s hot plate method:
1 Standard 77 79 83 75.28
2 NHR-I 56.74 55.61 48.03 62.99
All derivatives which are screen by Acetic acid induced
3 NHR –II 74.38 72.70 73.51 74.38 model where also screened for their Analgesic activity
4 NHR –III 41.97 26.03 21.05 62.73 using Eddy’s hot plate method at different time intervals
(30, 60, and 90). The results at various reaction time of
all the derivatives. As the reaction time increase the time
taken for the jumping also increases. Among all the
novel 3-substituted indole derivatives NHR-III have
shown potent more activity when compared to control
and almost equipotent to that of standard. This may due
to presence of electron donating group Methoxy of
NHR-III at Meta position.

ANTI-INFLAMMATORYACTIVITY:
Carrageenan-induced paw oedema model:
From the results of synthesized derivatives screened at
20 mg/kg body weight, NHR-I shown percentage of
inhibition 62.9 which is shown to be more potent than
Fig. 5: Graph showing Anti-inflammatory activity results control and less potent than standard that of 75.28% may
be unsubstituted aniline. NHR-II revealed 74.38% of
DISCUSSION: inhibition which shows that it is equipotent to that of
The wide literature survey kept in view for the present standard Indomethicin (75.28%) may be presence of
research work, and it has been carried out to synthesize Chlorine group at meta position. NHR-III has given
the 3-substituted indole acetamide and their evaluation 62.73% of inhibition which is more potent.
for the analgesic, anti-inflammatory anti-oxidant
activities.

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SUMMARY: With great pleasure I acknowledge my sincere thanks to

Indole is a common component of fragrances and the honorable chairman, honorable secretary and founder of
precursor to many pharmaceuticals. The biological Karnataka Educational Trust Prof. Basawaraj Ramnal
importance of Indole heterocyclic directly associated and our Director Dr K. Ramesh for granting us the
with their pharmacological and medical potential, Indole permission to carry out the research work. I express my
extremely attractive and rewarding research targets and heartfelt thanks to Miss. Julisa Suwail, and Mrs. M.
has motivated countless researchers to study their Gayathri Devi for their support to complete this work.
synthesis and pharmacological properties. Indole posses’
wide range pharmacological activities like analgesic, REFERENCES:
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methodology and applications". J. Chem. Soc. Perkin Trans. 2000
inflammatory antioxidant etc, due to its pharmacological
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activities inspired us to synthesize novel 3-substituted 2. Cacchi, S.; Fabrizi, G. (2005). "Synthesis and Functionalization of
Indole derivatives and screen for their anti-inflammatory Indoles Through Palladium-catalyzed Reactions". Chem. Rev. 105
analgesic and antioxidant activities respectively. The (7): 2873–2920.
3. Morrin Acheson R. An Introduction to the Chemistry of
structures of novel compounds were confirmed by FT-
heterocyclic compounds. A Wiley Interscience Publication, 2009,
IR, 1HNMR and Mass. These compounds (NHR-I-NHR- 3rd edition, 189-195.
III) were tested in-vivo for analgesic, anti-inflammatory 4. S. N. Pandeya, I. Ponnilavarasan, A. Pandey, R. Lakhan and J. P.
activates and biochemical studies antioxidant activity. Stables, Evaluation of p-nitrophenyl substituted semicarbazones for
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Compounds NHR-I and NHR-II showed 68.84% and
5. Gerard A. Pinna,a, Maria A. Pirisi, Giorgio Chelucci, Jean M.
73.30 % inhibition against acetic acid induced writings’ Mussinu, Gabriele Murineddu, Giovanni Loriga, Paolo S. D
in analgesic activity compared to 75.23% inhibition of Aquila and Gino Serra. Synthesis and D2-Like Binding Affinity of
standard Diclofenac. Compound NHR-II has showed New Derivatives of N-(1-Ethyl-2-pyrrolidinylmethyl)-4,5-dihydro-
1H-benzo[g]indole-3-carboxamide and Related 4H-[1]
71.6% inhibition of DPPH radical in Anti-oxidant
Benzothiopyrano [4,3-b] pyrrole and 5,6-Dihydro-4H-benzo[6,7]
activity compared to 84% inhibition of standard cyclohepta [b] pyrrole-3-carboxamide Analogues. Bioorganic and
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Agai, Gabor Tarkanyi, Csilla Horvath, Gizella Barta-Szalai, Eva
position of Indole nucleus can afford good analgesic,
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RESEARCH: 8. Marcella Ercoli, Lorenzo Mina , Caterina Canu Boido,Vito Boido,
All the title compounds (NHR-I to NHR-III) were Fabio Sparatore, Ugo Armani, Antonietta Piana 2-Phenyl-3-
(quinolizidin-1-yl)-5-substituted indoles as platelet antiaggregating
synthesized and characterized by analytical data and
agents IL FARMACO 2004, 59, 101–109.
screened for their analgesic, anti-inflammatory and
antioxidant activates. Among all the derivatives, the
compound NHR-II N-(3-chlorophenyl)-2-(1H indole-3-
yl) acetamide) was found to be more potent analgesic
when compared with all other derivatives and less potent
than standard in acetic acid induced writhing model. The
compounds NHR-III and has shown more potent
analgesic activity when compare to other derivatives and
less potent than standard in Eddy’s hot plate method.
The derivatives NHR-II N-(3-chlorophenyl)-2-(1H
indole-3-yl) acetamide) was found to be more potent
when compared with all other derivatives and less potent
than standard in the Carrageenan-induced paw oedema
model.

ACKNOWLEDGEMENTS:
I am very grateful to my parents who have struggled
their present days for the wealth of my tomorrow, who
shows their blessings, love and encouragement in every
activity of me which gave strength and power to achieve
my goals, even in hard times.

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