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Design, synthesis of novel N prenylated indole-3-carbazones and evaluation

of in vitro cytotoxity and 5-LOX inhibition activities

Article  in  Arabian Journal of Chemistry · February 2015

DOI: 10.1016/j.arabjc.2015.02.006

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Accepted Manuscript

Design, synthesis of novel N prenylated indole-3-carbazones and evaluation of

in vitro cytotoxity and 5-LOX inhibition activities

Praveen Choppara, Y.V. Prasad, C.V. Rao, K. Hari Krishna, G. Trimoorthulu,

G.U. Maheswara Rao, J.V. Rao, M.S. Bethu, Y.L.N. Murthy

PII: S1878-5352(15)00040-4
DOI: http://dx.doi.org/10.1016/j.arabjc.2015.02.006
Reference: ARABJC 1576

To appear in: Arabian Journal of Chemistry

Received Date: 23 August 2014

Accepted Date: 11 February 2015

Please cite this article as: P. Choppara, Y.V. Prasad, C.V. Rao, K. Hari Krishna, G. Trimoorthulu, G.U. Maheswara
Rao, J.V. Rao, M.S. Bethu, Y.L.N. Murthy, Design, synthesis of novel N prenylated indole-3-carbazones and
evaluation of in vitro cytotoxity and 5-LOX inhibition activities, Arabian Journal of Chemistry (2015), doi: http://
dx.doi.org/10.1016/j.arabjc.2015.02.006

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Design, synthesis of novel N prenylated indole-3-carbazones and evaluation of in vitro cytotoxity and 5-
LOX inhibition activities

Praveen Chopparaa, Y.V. Prasada, C.V. Raob, K. Hari Krishnab, G. Trimoorthulub, G.U. Maheswara
Raob, J.V. Raoc, M.S. Bethuc, Y.L.N. Murthya*
a
Department of Organic Chemistry, Foods, Drugs & Water, Andhra University, Visakhapatnam, 530 003, India
b
Resaerch & Development Centre, Laila impex, Vijayawada, 520 007, India
c
Biology Division, Indian Institute of Chemical Technology, Hyderabad 500 007, (AP), India.
*Corresponding author: Tel.: +919849229804; fax: +91891-2713813. E-mail: murthyyln@gmail.com

Abstract

A series of novel N-1 and C-3 substituted indole derivatives (5a-f) were designed, synthesized and
evaluated for their cytotoxic properties, viz brine shrimp lethality bioassay (BSLB) besides 5-lipoxygenase (5-
LOX) inhibitory activities through in vitro assays. Structure Activity Relation (SAR) studies showed that
compound 5d with an LC50 of 6.49 µM and 5c with an IC50 of 33.69 µM were found to be interesting for
cytotoxicity and 5-LOX inhibitory activity respectively.

Key words
Brine shrimp lethality bioassay, Cytotoxicity, 5-Lipoxygenase inhibition, N prenylated indole-3- carbazones.

1. Introduction

Indole derivatives are an important class of heterocyclic compounds with a wide range of biological
activities (Bandini and Eichholzer, 2009). Indole is a substructural element of many natural products which is an
important pharmacophore moiety of a large number of molecules with significant biological activities (a,b,c
Diana et al., 2011; a,b Carbone et al., 2014) and is widely used as a scaffold in agricultural and medicinal
chemistry. In particular, N-1 and C-3-substituted indole derivatives have been found to play an important role in
many biologically active compounds especially with anti inflammatory (Hall et al., 2008; Singh et al., 2008),
anti cancer (Madadi et al., 2014; Singh et al., 2008; Mashayekhi et al., 2013), anti nociceptive (Adam et al.,
2010; Moir et al., 2010) and anti psychotic activity (Madadi et al., 2013). On the other hand, carbazones
(semicarbazones, thiosemicarbazones) are compounds of considerable interest because of their important
chemical properties and potentially beneficial biological activities.

Literature survey reveals that carbazone analogues attain a wide range of biological activities.
Thiosemicarbazones appear to be a structural class with anti-pox virus activity (Katz et al., 1987; Katz et al
1975; Rao et al., 1966). Methisazone (I) plays an important role as prophylactic agent against several viral
diseases (Sethi., 2002). Thiosemicarbazone is the important pharmacophore in the therapy and prophylaxis of
mycobacteria infections and can represent a template for the development of novel antimycobacterial drugs like
SRI-224 (II), SRI-286 (III) (Bermudez et al., 2003; Dover et al., 2007). oxazolyl thiosemicarbazones (IV)
(Sriram et al., 2006) and some S-alkylisothiosemicarbazones (V) (Cocco et al., 2002; Logu et al., 2005).
Thiacetazone (VI) is a thiosemicarbazone antimicrobial that has been widely used for the treatment of Multi
Drug Resistant-Tuberculosis (MDR-TB) in many developing countries (Houston and Fanning., 1994).
Thiosemicarbazide analogues possess a wide range of biological activities including anti convulsant (Tripathi et
al., 2012), anti microbial (Zhong et al., 2011), anti viral (Garcial et al., 2003), anti trypanosomal (Moreira et al.,
2014), and mushroom tyrosinase inhibitors (Yi, W et al., 2011). On the other hand, Semicarbazide analogues
exhibits anti convalsant (Rajak et al., 2013), anti tubercular (Sriram et al., 2004), anti trypanosomal (Cerecetto et
al., 2000), anti inflammatory (Vieira et al., 2012), anti amnesic, cognition enhancing and anti cholinesterase
(Sinha and Shrivastava., 2013) and anti cancer activity (Qi et al., 2013).

S
NH2
CH3
N NH H H
N NH2 N NH2
N N
O S S
N MeO N
CH3

I II III

H
N N NH2
H N NH2 CH3 N
N N R Ar N
O N S
H S O N
S R H
R1

IV V VI

Thus the importance of indole and carbazone nuclei in medicinal chemistry prompted us to synthesize novel N
prenylated indole carrying carbazones at C-3 position and in order to extend the boundaries of pharmacological
properties of indole and carbazone moieties, the target compounds were screened for cytotoxic (BSLB) and 5-
LOX inhibitory activities.

2. Results and Discussion

2.1 Chemistry
Due to various biological and pharmacological activities associated with indole and carbazone skeletons, it is
worthwhile to incorporate the two moieties in a single molecular frame with an assumption to obtain more
potent biologically active compounds viz., 1-(5-substituted-1-(3-methylbut-2-enyl)-1H-indole-3-yl)methylene)
carbazides (5a-f). The target molecules have been accomplished by modifying first at N-H position and later at
third position (C-3) of indole. The modifications mainly include replacement of hydrogen at N-H by prenylation
(2-methyl but-2-ene) and formation of carbazones at C-3 with semicarbazide/thiosemicarbazide to increasing its
pharmacological efficacy.
The target molecules (5a-f) were achieved by Vilsmeier–Haack reaction of 5-substituted indoles (1a-c) yields 5-
substituted 3-formyl indoles (2a-c), the modification at N-H position was brought by the reaction with Prenyl
bromide in presence of NaH, DMF at 0 oC for 15 min yields N-prenylated 3-formyl indoles (3a-c). The
formation of carbazones at C-3 was accompanied by the reaction of compounds (3a-c) with
semicarbazide/thiosemicarbazide (4a,b). The synthetic path involved in synthesis of 1-(5-substituted-1-(3-
methylbut-2-enyl)- 1H-indole-3-yl)methylene) carbazides (5a-f) was presented in Scheme- 1. All the
synthesized compounds were well characterized by advanced spectroscopic techniques.

CHO CHO
R R Br R
DMF, POCl3

N NaOH, H2O N
H NaH, DMF N
H
0oC, 15 min
1a-c 2a-c 3a-c

H
N NH2.HCl
H2N
X 4 a,b
R X
1a, 2a, 3a H - KOH, EtOH
1b, 2b, 3b Br - reflux
1c, 2c, 3c CN -
4a - O
4b - S
5a H O X
5b H S
5c Br O CH=N-NH-C-NH2
5d Br S R
5e CN O
5f CN S N

5a-f

Scheme-1. Synthetic scheme for the synthesis of 1-(5-substituted-1-(3-methylbut-2-enyl)- 1H-indole-3-

yl)methylene) carbazides (5a-f)

2.2 Bio evaluation

2.2.1 In vitro cytotoxic activity by Brine Shrimp Lethal Bioassay (BSLB)

Brine shrimp Lethal Bioassay (BSLB) (Meyer et al., 1982), i.e. toxicity to the Artemia salina (brine shrimp)
nauplii was performed and has been used for establishing of fungal toxins, plant extract toxicity, heavy metals,
cyano bacteria toxins, pesticides, cytotoxicity testing of dental materials (Carballo et al., 2002), natural and
synthetic organic compounds (Choudhary and Thomsen, 2001). Toxicity to A. salina has a good correlation with
anti tumour, pesticidal (Mc Laughlin, 1991), anti trypanosomal activities (Zani et al., 1995), rodent and human
acute oral toxicity data. The brine shrimp larvae respond similarly to the corresponding mammalian systems
(Solis et al., 1993). Since the DNA-dependent RNA polymerases of A. salina have been shown to be similar to
the mammalian type (Birndorf et al., 1975). This test is not only used for predicting cytotoxicity, but also it is
used to predict anti tumour, anti bacterial and pesticidal activities (Sanchez et al., 1993). The brine shrimp
lethality method is widely used in the bioassay for identification of the bioactive compounds (Venkateswara Rao
et al., 2007; Olowa and Nuneza, 2013). The bioassay determines lethal concentrations of newly synthesized 1-
(5-substituted-1-(3-methylbut-2-enyl)-1H-indole-3-yl)methylene) carbazides (5a-f) in brine medium adopting
the microplate assay developed by Meyer et al., 1982. The Median lethal concentration (LC50) values of the
target compounds were compared with the LC50 value of the positive control, Podophyllotoxin. The LC50 values
(µM) for the test compounds (5a-f) obtained from BSLB were tabulated in Table-1.
Table-1. In vitro cytotoxicity of title compounds (5a-f) by Brine Shrimp Lethal Bioassay.

Logarithmic Regression 95% Confidence

*
equation Fiducial limits
Entry Compound LC50 ± SE (µM) Relative
Y = Ybar + b (x - xbar) activity
LCL UCL

1 5a 477.14±44.66 Y=-0.61+2.66x 114.72 149.79 70.39

2 5b 315.95±16.91 Y=0.42+2.34x 81.45 105.66 49.55
3 5c 206.04±11.57 Y=0.71+2.31x 64.79 79.21 39.29
4 5d 6.49±0.30 Y=3.59+3.74x 2.158 2.599 1.29
5 5e 274.02±11.08 Y=-0.83+3.05x 74.99 87.86 44.32
6 5d 205.27±9.06 Y=-0.11+2.83x 58.39 69.42 34.88

7 Standard† 4.42±0.28 Y=4.10+3.40x 1.63 2.02 1.00

48 h old nauplii of brine shrimp (A. salina) were treated with different concentrations of indole carbazides (5a-f) for 24 h and lethal
concentration was investigated through probit analysis.*Regression equation Y = Ybar +b (X- xbar) or (Ybar– b* xbar) + b* x). Ybar =
weighted avg. of working probit values; xbar = avg. of log10 (concentration); b = slope of the line; X = x scale concentration in Log).
†Podophyllotoxin used as positive control. Three replicates for each concentration and the control (without test compound), were tested for
lethal bio-efficacy and the mean data was used to calculate the probit regression equation. *LC50 is defined as the concentration of extract
that can cause 50% mortality in the exposed population.

The results from Table-1 show that compound 5d exhibit good cytotoxity with LC50 of 6.49 µM and its
relative active (1.29) was close-at-hand to Podophyllotoxin (1.00). Among the semicarbazides (5a,5c,5e),
unsubstituted indole analogue (5a) was found to be weak than the substituted analogues (5c and 5e), bromo
analogue (5c) was interesting candidate with LC50 of 206.04 µM followed by cyano analogue (5e) with LC50
274.02 µM and their relative activities were 39.29 and 44.32 respectively. The results of thiosemicarbazides
(5b,5d,5f) showed that bromo analogue (5d) was found to be interesting with LC50 of 6.49 µM then cyano
analogue (5f) with 205.27 µM and unsubstituted indole analogue (5b) was weak with LC50 of 315.95 µM. In
conclusion, among the synthesised compounds, unsubstituted analogues (5a,5b) showed poor activity than the
corresponding substituted analogues (5c-f).

2.2.2 5-lipoxygenase inhibition assay

Lipoxygenases (LOXs), which are widely distributed in both the plant and animal kingdoms, belong to
a class of non-heme iron-containing enzymes which catalyze the hydroperoxidation reaction of fatty acids to
peroxides (Peters-Golden and Henderson, 2007). 5-Lipoxygenase (5-LOX) is the key enzyme in the
biosynthesis of leukotrienes (LTs) through catalyzing the initial two steps in conversion of arachidonic acid to
LTs (Samuelson, 1983). LTB4 and the cysteinyl-leukotrienes are potent constrictors of human airways, and
have powerful proinflammatory properties (Funk, 2001). Inhibition of 5-LOX may lead to the development of
new therapeutic treatments for pathologies such as asthma, allergies, and other inflammatory disorders
(Samuelson, 1983., Young, 1999., Drazen, 1998). Recent studies have implicated a role of 5-LOX products
involved in a number of other diseases, including cancer (Romano and Claria, 2003), atherosclerosis (Spanbroek
and Habenicht, 2003), stroke (Helgadottir et al., 2004) and osteoporosis (Werz and Steinhilber, 2005)
Therapeutic potential of 5-LOX inhibition has been widely highlighted in recent years (Balkan and Berk, 2003.,
Funk et al., 2005., Yoshimura et al., 2005). Literature survey reveals N-1 and C-3 substituted indoles are
identified as 5-LOX inhibitors (Prasher et al., 2014., Singh and pooja, 2013., Zheng et al., 2007). Based on the
literature survey the novel 1-(5-substituted-1-(3-methylbut-2-enyl)-1H-indole-3-yl) methylene) carbazides (5a-f)
were screened for their inhibitory properties against 5-LOX enzyme employing the assay described by
Reddanna et al., 1990. 5-LOX inhibition of the tested compounds was determined by measuring the Inhibition
Concentration; IC50 (IC50 represents the concentration of a drug that is required for 50% inhibition) expressed in
µM. The results for test compounds (5a-f) were compared with the positive control Curcumin. The IC50 values
for the test compounds (5a-f) obtained from 5-LOX inhibition assay were tabulated in Table-2.

Table-2. IC50 values obtained from in vitro 5-Lipoxygenase inhibition assay for the compounds (5a-f)

Entry Compound IC50 (µM)

1 5a >100

2 5b >100

3 5c 33.69

4 5d 36.65

5 5e >100

6 5f >100

7 Standard* 27.58

IC50 represents the concentration of a drug that is required for 50% inhibition expressed in µM.
* Curcumin as positive control.

The results obtained from the Table-2 shows that the unsubstituted (5a,5b) and cyano (5e,5f) analogues were
weak with an IC50 value of >100 µM and bromo analogues (5c, 5d) were the engrossing compounds compared
with the standard “curcumin” with an IC50 of 33.69 and 36.65 µM respectively.

2.2.3 Structure –Activity Relationship Study (SARS)

In 1-(5-substituted-1-(3-methylbut-2-enyl)-1H-indole-3-yl)methylene) carbazides (5a-f), substituent
plays an important role to make difference in the activity. From the results of Brine Shrimp Lethal Bioassay as
shown in Table-1, compound 5d (Table-1 entry-4) is more potent among the series, in which bromine is
present on the 5th position of indole and thiosemicarbazone at 3rd position. From the results of 5-LOX inhibition
assay as shown in Table-2, compound 5c (Table-2 entry-3) is more potent 5-LOX inhibitor. In compound 5c
bromine is present on the 5th position of indole and semicarbazone at 3rd position.
Bio evaluation assays reveal that among the target compounds (5a-f), compounds 5c and 5d exhibit
potent activity. In both the compounds 5c and 5d bromine is present on the 5th position of indole, The 3rd
position was occupied by semicarbazone and thiosemicarbazone in compounds 5c and 5d respectively. The
SAR studies of novel 1-(5-substituted-1-(3-methylbut-2-enyl)-1H-indole-3-yl)methylene) carbazides (5a-f), it
was observed that the presence of hydrophobic substituent (bromine) at 5th position of indole moiety make the
compounds 5c and 5d more potent than unsubstituted analogues (5a,5b) and cyano analogues (5e,5f) which lack
hydrophobic substituent.

3. Experimental
3.1 General
All chemical reagents were obtained from Sigma Aldrich and were used without further purification.
Melting points were determined in open capillaries and are uncorrected. Infrared (IR) spectra were recorded
using FT-IR Bruker Alpha spectrometer, ESI-Mass spectra were recorded on Finnigan Matt Mass spectrometer
and NMR (1H and 13C) spectra were recorded with a Bruker Ascend-400 and Jeol JNM EX-90 spectrometer.

3.2 General procedure for the synthesis of substituted 1H-indole-3-carbaldehyde (2a-c):

To a solution of substituted indoles (1a-c) (42.6 mmol) in dry DMF (187.4 mmol) in an ice-salt bath
and POCl3 (47.1 mmol) is subsequently added with stirring over a period of 30 min. After completion of
addition, raise the temperature to 40 oC and stir the syrup for 1.5 h at that temperature. At the end of the reaction
(as indicated by TLC) 25 gms crushed ice was added to the reaction mixture. The obtained solution is
transferred into 250 mL RB flask, add NaOH (470 mmol) dissolved in 50 mL water with constant stirring and
the resultant suspension is heated rapidly to the boiling point and allowed to cool to room temperature, after
which it is placed in refrigerator overnight. The precipitate was filtered off, washed thrice with 100 mL water,
yielding substituted 1H-indole-3-carbaldehyde (2a-c).

3.2.1 1H-indole-3-carbaldehyde (2a): (James P.N and Snyder, H.R., 1959)

Yield: 92%; Mp: 196-198 oC. 1H NMR (DMSO-d6, 400 MHz): δ 7.14 (t, J = 7.2 Hz, 1H), 7.22 (t, J = 7.2 Hz,
1H), 7.34 (d, J = 8 Hz, 1H), 7.52 (s, 1H), 7.62 (d, J = 8 Hz, 1H), 8.12 (s, 1H), 9.52 (s, 1H). 13C NMR (DMSO-
d6, 22.4 MHz): δ 111.4, 118.0, 119.4, 120.5, 122.4, 127.7, 131.8, 137.2, 182.7. ESI-MS: m/z 146.20 [M+H]+. IR
(KBr, cm−1): 1224, 1632, 3442. Anal. Calcd. for C9H7NO: C, 74.47, H, 4.86, N, 9.65 %; Found: C, 74.44, H,
4.91, N, 9.64 %

3.2.2 5-bromo-1H-indole-3-carbaldehyde (2b):

Yield: 90%; Mp: 192 oC. 1H NMR (DMSO-d6, 400 MHz): δ 7.34 (d, J = 8.8 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H),
7.75 (s, 1H), 8.25 (s, 1H), 8.32 (s, 1H), 9.94 (s, 1H). 13C NMR (DMSO-d6, 22.4 MHz): δ 113.0, 114.8, 117.3,
123.1, 125.6, 135.2, 136.7, 144.4, 183.9. ESI-MS: m/z 245.95 [M+Na]+; 247.95 [M+Na+2]+ . IR (KBr, cm−1):
1229, 1643, 3312. Anal. Calcd. for C9H6BrNO: C, 48.25, H, 2.70, N, 6.25 %; Found: C, 48.22, H, 2.76, N, 6.24
%

3.2.3 3-formyl-1H-indole-5-carbonitrile (2c):

Yield: 84%; Mp: 220-224 oC. 1H NMR (DMSO-d6, 400 MHz): δ 7.27 (d, J = 8.4 Hz, 1H), 7.41 (d, J = 8.4 Hz,
1H), 7.69 (s, 1H), 8.21 (s, 1H), 8.43 (s, 1H), 10.23 (s, 1H). 13C NMR (DMSO-d6, 100 MHz): δ 103.4, 111.3,
113.6, 116.1, 117.2, 123.8, 126.1, 134.8, 137.1, 139.8, 185.8. ESI-MS: m/z 171. [M+H]+. IR (KBr, cm−1): 1221,
1650, 3345. Anal. Calcd. for C10H6N2O: C, 70.58, H, 3.55, N, 16.46 %; Found: C, 70.60, H, 3.52, N, 16.45 %
3.3 General procedure for the synthesis of substituted 1-(3-methylbut-2-enyl)- 1H-indole-3-carbaldehyde (3a-c):
To a solution of substituted 1H-indole-3-carbaldehyde (2a-c) (2.20 mmol) in dry DMF (5 mL) was
added NaH (2.64 mmol, 60% oil dispersion) and the resulting mixture was stirred for 10 min in an ice bath. 3,3-
dimethylallyl bromide (2.20 mmol) was added and the resulting mixture stirred for 15 min at 0 oC. The mixture
was diluted with EtOAc (20 mL), washed five times with distilled water (50 mL). The organic layer was dried
over anhydrous Na2SO4, filtered and the solvent was removed under reduced pressure to get crude residue which
was purified by silica gel column chromatography (100-200 mesh) using 8:2 (Hexane:EtOAc) as eluents
affording 3a-c.

3.3.1 1-(3-methylbut-2-enyl)-1H-indole-3-carbaldehyde (3a):

Yield: 87%; Mp: 79-81 oC. 1H NMR (CDCl3, 400 MHz): δ 1.85 (s, 3H), 1.86 (s, 3H), 4.74 (d, J = 7.2 Hz, 2H),
5.44 (t, J = 7.2 Hz, 1H), 7.28-7.41 (m, 4H), 7.76 (s, 1H), 9.99 (s, 1H). 13C NMR (CDCl3, 22.4 MHz): δ 17.9,
25.4, 44.6, 110.0, 117.7, 117.9, 121.8, 122.7, 123.6, 125.5, 137.3, 137.5, 138.7, 184.3, ESI-MS: m/z 214.20
[M+], IR (KBr, cm−1): 1161, 1640. Anal. Calcd. for C14H15NO: C, 78.84, H, 7.09, N, 6.57 %; Found: C, 78.80,
H, 7.16, N, 6.55 %

3.3.2 5-bromo-1-(3-methylbut-2-enyl)-1H-indole-3-carbaldehyde (3b):

Yield: 89%; Mp: 95-98 oC. 1H NMR (CDCl3, 400 MHz): δ 1.85 (s, 3H), 1.86 (s, 3H), 4.71 (d, J =7.2 Hz, 2H),
5.42 (t, J =7.2 Hz, 1H), 7.25 (d, J = 9.6 Hz, 1H), 7.43 (dd, J = 2, 8.8 Hz, 1H), 7.73 (s, 1H), 8.47 (d, J = 2 Hz,
1H), 9.96 (s, 1H). 13C NMR (CDCl3, 22.4 MHz): δ 12.7, 20.1, 39.4, 106.0, 110.8, 111.7, 119.0, 121.1, 121.4,
130.3, 132.6, 132.8, 133.8, 178.6, ESI-MS: m/z 292.03 (M)+; 294.03 (M+2)+; 314.01 (M+Na)+; 316.01
[M+Na+2]+. IR (KBr, cm−1): 1162, 1654. Anal. Calcd. for C14H14BrNO: C, 57.55, H, 4.83, N, 4.79 %; Found: C,
57.53, H, 4.90, N, 4.77 %

3.3.3 3-formyl-1-(3-methylbut-2-enyl)-1H-indole-5-carbonitrile (3c):

Yield: 79%; Mp: 90-92 oC 1H NMR (CDCl3, 400 MHz): δ 1.73 (s, 3H), 1.75 (s, 3H), 4.68 (d, J =7.2 Hz, 2H),
5.44 (t, J =7.2 Hz, 1H), 7.27 (d, J = 8.8 Hz, 1H), 7.51 (d, J = 2, 8.4 Hz, 1H), 7.81 (s, 1H), 8.32 (d, J = 2 Hz, 1H),
10.03 (s, 1H). 13C NMR (CDCl3, 100 MHz): δ 17.7, 25.8, 40.2, 109.1, 111.4, 112.5, 116.7, 120.3, 122.2, 122.6,
129.5, 132.2, 133.0, 133.7, 179.1, ESI-MS: m/z 239 (M+H)+ ; IR (KBr, cm−1): 1134, 1649. Anal. Calcd. for
C15H14N2O: C, 75.61, H, 5.92, N, 11.76 %; Found: C, 75.62, H, 5.93, N, 11.77 %

3.4 General procedure for the synthesis of 1-(5-substituted-1-(3-methylbut-2-enyl)- 1H-indole-3 yl)methylene)

semicarbazide (5a,5c,5e)
To the solution of substituted 1-(3-methylbut-2-enyl)-1H-indole-3-carbaldehyde (3a-c) (3 mmol) and
KOH (4 mmol) in 10 mL absolute ethanol, solution of semicarbazide hydrochloride (4a) (4 mmol) in absolute
ethanol and water (9:1mL) was added slowly and the reaction mixture was refluxed for 1.5 h. Removal of the
solvent in vacuo furnished a thick liquid which produced precipitate on addition of water. The precipitate was
filtered off, washed with hot water, and dried to afford as a solid, which was purified by silicagel column
chromatography (100-200 mesh) using 6:4 (Hexane:EtOAc) as eluents to afford target compounds 5a, 5c and 5e
3.4.1 1-((1-(3-methylbut-2-enyl)- 1H-indol-3-yl)methylene) semicarbazide (5a):
Yield: 79%; Mp: 148 oC. 1H NMR (DMSO-d6, 400 MHz): δ 1.70 (s, 3H), 1.81 (s, 3H), 4.76 (d, J = 7.2 Hz, 2H),
13
5.32 (t, J = 7.2 Hz, 1H), 6.25 (s, 2H), 7.34-7.45 (m, 4H), 7.79 (s, 1H), 8.09 (s, 1H), 8.94 (s, 1H). C NMR
(DMSO-d6, 22.4 MHz): δ 17.8, 25.4, 44.1, 109.8, 111.0, 118.8, 120.8, 121.8, 122.7, 125.2, 130.7, 137.0, 137.3,
138.8, 158.7. ESI-MS: m/z 271.29 [M+H] +. IR (KBr, cm−1): 1165, 1694, 3148, 3469. Anal. Calcd. for
C15H18N4O: C, 66.64, H, 6.71, N, 20.73 %; Found: C, 66.63, H, 6.75, N, 20.71 %

3.4.2 1-(5-bromo-1-(3-methylbut-2-enyl)-1H-indole-3-yl) methylene) semicarbazide (5c):

Yield: 82%; Mp: 188-190 oC. 1H NMR (DMSO-d6, 400 MHz): δ 1.71 (s, 3H), 1.81 (s, 3H), 4.76 (d, J = 6.8 Hz,
2H), 5.34 (t, J =7.2 Hz, 1H), 6.25 (s, 2H), 7.34 (d, J = 8.8 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H), 7.75 (s, 1H), 8.02
13
(s, 1H), 8.25 (s, 1H), 9.94 (s, 1H). C NMR (DMSO-d6, 100 MHz): δ 18.3, 25.8, 44.3, 111.0, 112.8, 113.7,
119.9, 124.2, 125.4, 126.6, 133.0, 135.8, 137.0, 137.9, 157.2. ESI-MS: m/z 349.06 [M+]; 351.06 [M+2]+; 387.02
[M+K]+; 389.02 [M+K+2]+. IR (KBr, cm−1): 1196, 1623, 3142, 3473. Anal. Calcd. for C15H17BrN4O: C, 51.59,
H, 4.91, N, 16.04 %; Found C, 51.57, H, 4.99, N, 16.01 %

3.4.3 1-(5-cyano-1-(3-methylbut-2-enyl)-1H-indole-3-yl) methylene) semicarbazide (5e):

Yield: 84%; Mp: 178-180 oC. 1H NMR (DMSO-d6, 400 MHz): δ 1.69 (s, 3H), 1.74 (s, 3H), 4.69 (d, J = 7.2 Hz,
2H), 5.41 (t, J =7.2 Hz, 1H), 6.51 (s, 2H), 7.21 (d, J = 8.4 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.75 (s, 1H), 8.19
(s, 1H), 8.43 (s, 1H), 10.37 (s, 1H). 13C NMR (DMSO-d6, 100 MHz): δ 17.7, 26.9, 43.9, 105.1, 111.3, 113.2,
113.7, 116.9, 120.1, 125.1, 125.9, 132.6, 136.9, 137.3, 138.8, 160.1. ESI-MS: m/z 296 [M+H]+. IR (KBr, cm−1):
1254, 1642, 3135, 3489. Anal. Calcd. for C16H17N5O: C, 65.07, H, 5.80, N, 23.71 %; Found C, 65.08, H, 5.81,
N, 23.70 %

3.5 General procedure for the synthesis of 1-(5-substituted-1-(3-methylbut-2-enyl)- 1H-indole-3 yl)methylene)

thiosemicarbazide (5b,5d,5f)
To the solution of substituted 1-(3-methylbut-2-enyl)- 1H-indole-3-carbaldehyde (3a-c) (3 mmol) and
KOH (3 mmol) in 10mL absolute ethanol, solution of thiosemicarbazide (4b) (4 mmol) in absolute ethanol and
water (9:1mL) was added slowly, and the reaction mixture was refluxed for 1 h. Removal of the solvent in
vacuo furnished a thick liquid which produced precipitate on addition of water. The precipitate was filtered off,
washed with excess of hot water and dried to afford crude products, which was purified by silicagel column
chromatography (100-200 mesh) using 6:4 (Hexane:EtOAc) as eluents to afford target compounds 5b, 5d and
5f.

3.5.1 1-((1-(3-methylbut-2-enyl)- 1H-indol-3-yl)methylene)thiosemicarbazide (5b):

Yield: 85%; Mp: 162 oC. 1H NMR (DMSO-d6, 400 MHz): δ 1.75 (s, 3H), 1.78 (s, 3H), 4.51(d, J = 7.2 Hz, 2H),
13
5.26 (t, J = 7.2 Hz 1H), 5.90 (s, 2H), 7.20-7.29 (m, 4H), 7.78 (s, 1H), 8.04 (s, 1H), 9.99 (s, 1H). C NMR
(DMSO-d6, 22.4 MHz): δ 17.9, 25.5, 44.0, 109.9, 110.6, 118.6, 121.1, 121.8, 122.8, 125.0, 131.8, 137.0, 137.5,
140.8, 177.0. ESI-MS: m/z 287.25 [M]+. IR (KBr, cm−1): 1161, 1613, 3148, 3413. Anal. Calcd. for C15H18N4S:
C, 62.91, H, 6.33, N, 19.56 %; Found: C, 62.88, H, 6.39, N, 19.55 %
3.5.2 1-(5-bromo-1-(3-methylbut-2-enyl)-1H-indole-3-yl) methylene) thiosemicarbazide (5d):
Yield: 83%; Mp: 220 oC. 1H NMR (DMSO-d6, 400 MHz): 1.71 (s, 3H), 1.81 (s, 3H), 4.76(d, J = 6.8 Hz, 2H),
5.32 (t, J = 1.2 Hz, 1H), 6.26 (s, 2H), 7.34 (d, J = 8.4 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H), 7.75 (s, 1H), 8.02 (s,
1H), 8.25 (s, 1H), 9.95 (s, 1H). 13C NMR (DMSO-d6, 22.4 MHz): δ 18.4, 26.0, 44.2, 110.9, 112.7, 114.2, 120.0,
124.3, 124.5, 126.7, 133.1, 135.5, 137.5, 138.0, 179.3. ESI-MS: m/z 365.04 (M+); 367.04 (M+2)+; 387.03
(M+Na)+; 389.02 [M+Na+2]+. IR (KBr, cm−1): 1187, 1621, 3160, 3387. Anal. Calcd. for C15H17BrN4S: C,
49.32, H, 4.69, N, 15.34 %; Found: C, 49.31, H, 4.72, N, 15.31 %

3.5.3 1-(5-cyano-1-(3-methylbut-2-enyl)-1H-indole-3-yl) methylene) thiosemicarbazide (5f):

Yield: 91%; Mp: 192-194 oC. 1H NMR (DMSO-d6, 400 MHz): 1.75 (s, 3H), 1.79 (s, 3H), 4.75 (d, J = 7.2 Hz,
2H), 5.21 (t, J = 1.6 Hz, 1H), 6.84 (s, 2H), 7.29 (d, J = 8.0 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.72 (s, 1H), 8.14
(s, 1H), 8.46 (s, 1H), 10.41 (s, 1H). 13C NMR (DMSO-d6, 22.4 MHz): δ 17.2, 26.4, 45.1, 105.2, 111.1, 112.9,
114.7, 115.7, 119.5, 125.6, 126.2, 133.7, 134.8, 136.3, 138.7, 180.1. ESI-MS: m/z 312 (M+H)+; IR (KBr, cm−1):
1192, 1626, 3178, 3421. Anal. Calcd. for C16H17N5S: C, 61.71, H, 5.50, N, 22.49 %; Found: C, 61.72, H, 5.51,
N, 22.48 %

3.6 Procedure for in vitro cytotoxic activity of 1-(5-substituted-1-(3-methylbut-2-enyl)-1H-indole-3-yl)

methylene) carbazides (5a-f) by Brine Shrimp Lethal Bioassay (BSLB)
Brine Shrimp Lethal Bioassay was used according to method of Meyer et al., 1982. The lethality
experiments were conducted in a 12 well micro plate, each well consisting of 5 mL artificial sea water with
required test concentration. The derivatives were dissolved in dimethylsulphoxide (DMSO) and diluted with
artificial sea salt water to achieve the required test concentrations, and maintained final concentration of DMSO,
which did not exceed 0.05%. The control experiments were also performed simultaneously with an addition of
carrier solvent alone (without sample). Ten numbers of 48 h old nauplii were added into each well and exposed
for 24 h to different test concentrations under illumination with a minimum of three replicates. A drop of dry
east suspension (3mg in 5 mL artificial sea water) was added as food to each well. The plates were then
examined under magnification (10x) and the number of dead nauplii in each well was counted. Percent
mortalities were corrected for natural mortality in controls using Abbots formula (Abbot, 1925),i.e., corrected
percent mortality = (PI – C/100 – C)*100, where PI denotes the observed mortality rate, and C is natural
mortality. The median lethal concentration (LC50 for 24 h value) for each assay was calculated by taking
average of three experiments using statistical software, BioStat-2008 based on the Finney Probit analysis
(Finney, 1953). The results for test compounds were compared with the positive control Podophyllotoxin. All
samples were done in triplicate. The LC50 values for the test compounds (5a-f) obtained from BSLB were
expressed in µM.

3.7 Procedure for 5-lipoxygenase inhibition assay for 1-(5-substituted-1-(3-methylbut-2-enyl)-1H-indole-3-yl)

methylene) carbazides (5a-f)
5-Lipoxygenase inhibition assay mixture contained 2.97 mL of 50 mmol phosphate buffer pH at 6.3. 5 µL
of 80 mmol Linoleic acid and sufficient amount of potato 5-lipoxygenase enzyme. The reaction was started by
the addition of substrate (Linoleic acid) and the increase in UV absorption at 234 nm was followed at 25 oC. The
activities were measured at 2 minutes. The reaction was linear during this time period. In the inhibition studies,
the activities were measured in presence of various concentrations of target molecules (5a-f). All the assays
were performed in triplicate. The IC50 values for the test compounds (5a-f) obtained from 5-LOX inhibition
assay were expressed in µM.
4. Conclusion
In conclusion, we report a novel series of six N prenylated indole-3-carbazones, in which five compounds
were interesting in cytotoxic and two compounds in 5-LOX inhibition assay.

Acknowledgment
We are grateful to DRDO, New Delhi for providing financial assistance through the project
ERIP/ER/1003916/M/01/1354 and UGC, New Delhi for the award of UGC-BSR JRF to the author P.C.

REFERENCES
Abbot, W.B., 1925. A method of computing the effectiveness of an insecticide. J. Econ. Entomol. 18, 265-267
Adam, J., Cairns, J., Caulfield, W., Cowley, P., Cumming, I., Easson, M., Edwards, D., Ferguson, M., Goodwin,
R., Jeremiah, F., Kiyoi, T., Mistry, A., Moir, E., Morphy, R., Tierney, J., York, M., Baker, J., Cottney, J.,
Houghton, A., Westwood, P., Walker, G., 2010. Design, synthesis, and structure–activity relationships of
indole-3-carboxamides as novel water soluble cannabinoid CB1 receptor agonists. Med. Chem. Commun.
1, 54-60.
Balkan, A.; Berk, B., 2003. Novel Approaches to Anti-Inflammatory Agents as Therapeutics for Asthma. Curr.
Med. Chem. Anti-Inflamm. Anti-Allergy Agents. 2, 9-18.
Bandini, M., Eichholzer, A., 2009. Catalytic functionalization of indoles in a new dimension. Angew. Chem.
Int. Ed. 48, 9608-9644.
Bermudez, L.E., Reynolds, R., Kolonoski, P., Aralar, P., Inderlied, C.B., Young, L.S., 2003. Thiosemicarbazole
(Thiacetazone-Like) compound with activity against Mycobacterium avium in mice. Antimicrob. Agents
Chemother. 47, 2685-2687.
Birndorf, H.C., Alossio, J.D. Bagshaw, J.C., 1975. DNA-dependent RNA-polymerases from Artemia embryos:
Characterization of polymerases I and II from nauplius. Dev. Biol. 45, 34.
Carballo, J.L., Hernandez-Inda, Z.L., Perez, P., Garcia-Gravalos, M.D., 2002. A comparison between two brine
shrimp assays to detect in vitro cytotoxicity in marine natural products. BMC Bio- technol. 2, 17
Carbone, A., Pennati, M., Barraja, P., Montalbano, A., Parrino, B., Spanò, V., Lopergolo, A., Sbarra, S., Doldi,
V., Zaffaroni, N., Cirrincione, G., Diana P., 2014. Synthesis and antiproliferative activity of substituted
3[2(1Hindol3yl)1,3thiazol4yl]1Hpyrrolo[3,2b] piridine, marine alkaloid nortopsentin analogues. Curr. Med.
Chem. 21, 1654-1666. (b) Carbone, A., Parrino, B., Barraja, P., Spano, V., Cirrincione, G., Diana, P.,
Maier, A. , Kelter, G. , Fiebig, H.H., 2013. Synthesis and antiproliferative activity of 2,5bis( 3′Indolyl)
pyrroles, analogues of the marine alkaloid nortopsentin. Marine Drugs. 11, 643-654.
Cerecetto, H., Maio, R.D., Gonzalez, M., Risso, M., Sagrera, G., Seoane, G., Denicola, A., Peluffo, G., Quijano,
C., Stoppani, A.O.M., Paulino, M., Olea-Azar, C., Basombrio, M.A., 2000. Synthesis and antitrypanosomal
evaluation of E-isomers of 5-nitro-2-furaldehyde and 5-nitrothiophene-2-carboxaldehyde semicarbazone
derivatives. Structure–activity relationships. Eur. J. Med. Chem. 35, 343-350.
Choudhary, M.I., Thomsen, W.J., 2001. Bioassay techniques for drug development. Harwood Academic
Publishers 9.
Cocco, M.T., Congiu, C., Onnis, V., Pellerano, M.L., De Logu, A., 2002. Synthesis and antimycobacterial
activity of new S-alkylisothiosemicarbazone derivatives. Bioorg. Med. Chem. 10, 501-506.
Diana, P., Carbone, A., Barraja, P., Montalbano, A., Parrino, B., Lopergolo, A., Pennati, M., Zaffaroni, N.,
Cirrincione, G., 2011. Synthesis and antitumor activity of 3(2Phenyl1,3thiazol4yl)1Hindoles and
3(2Phenyl1,3thiazol4yl)1H7azaindoles. Chem Med Chem. 6, 1300-1309. (b) Diana, P., Stagno, A., Barraja,
P., Montalbano, A., Carbone, A., Parrino, B., Cirrincione, G., 2011. Synthesis of the new ring system
pyrrolizino[2,3b] indol4(5H)one. Tetrahedron 67, 3374-3379. (c) Diana, P., Stagno, A., Barraja, P.,
Carbone, A., Parrino, B., Dall'acqua, F., Vedaldi, D., Salvador, A., Brun, P., Castagliuolo, I.,; Issinger, O.G.
Cirrincione, G., 2011. Synthesis of Triazenoazaindoles: a new class of triazenes with antitumor activity.
Chem Med Chem 6(7), 1291-1299.
Drazen, J., 1998. Clinical pharmacology of leukotriene receptor antagonists and 5-Lipoxygenase inhibitors. Am.
J. Resp. Crit. Care., 157, S233-S237.
Dover, L.G., Alahari, A., Gratraud, P., Gomes, J.M., Bhowruth, V., Reynolds, R.C., Besra, G.S., Kremer, L.,
2007. EthA, a common activator of thiocarbamide-containing drugs acting on different mycobacterial
targets. Antimicrob. Agents Chemother. 51, 1055-1063.
Finney, D.J., 1971. Probit Analysis, Cambridge University press, London. pp. 68-72.
Funk, C.D., 2005. Leukotriene modifiers as potential therapeutics for cardiovascular disease. Nat. Rev.
Drug Discov. 4, 664-672.
Funk, C.D., 2001. Prostaglandins and Leukotrienes: Advances in Eicosanoid Biology. Science. 294, 1871-
1875.
Garcial, C.C., Brousse, B.N., Carlucci1, M.J., Moglioni, A.G., Alho, M.M., Moltrasio, G.Y., Accorso, N.B.D.,
Damonte, E.B., 2003. Inhibitory effect of thiosemicarbazone derivatives on Junin virus replication in vitro.
Antivir. Chem. Chemoth. 14, 99-105.
Hall, A., Billinton, A., Brown, S.H., Chowdhury, A., Giblin, G.M.P., Goldsmith, P., Hurst, D.N., Naylor, A.,
Patel, S., Scoccitti, T., Theobald, P.J., 2008. Discovery of a novel indole series of EP1 receptor antagonists
by scaffold hopping. Bioorg. Med. Chem. Lett. 18, 2684-2690.
Helgadottir, A., Manolescu, A., Thorleifsson, G., Gretarsdottir, S., Jonsdottir, H., Thorsteinsdottir, U., Samani,
N.J., Gudmundsson, G., Grant, S.F., Thorgeirsson, G., Sveinbjornsdottir, S., Valdimarsson, E.M.,
Matthiasson, S.E., Johannsson, H., Gudmundsdottir, O., Gurney, M.E., Sainz, J., Thorhallsdottir, M.,
Andresdottir, M., Frigge, M.L., Topol, E.J., Kong, A., Gudnason, V., Hakonarson, H., Gulcher, J.R.,
Stefansson, K. 2004. The gene encoding 5-lipoxygenase activating protein confers risk of myocardial
infarction and stroke. Nat. Genet. 36, 233.
Houston, S., Fanning, A., 1994. Current and Potential Treatment of Tuberculosis. Drugs. 48, 689-708.
James P.N., Snyder, H.R., 1959. Org. Synth. 39, 30.
Katz, E., 1987. Thiosemicarbazones: inhibition of the growth of pox viruses and requirement for the growth of
an isatin-beta-thiosemicarbazone dependent mutant. Rev. Clin. Basic Pharm. 6, 119-130.
Katz, E., Margalith, E., Winer, B., 1975. Isolation and characterization of a γ-Thiochromanone-4-
thiosemicarbazone-resistant mutant of vaccinia virus. Antimicrob. Agents Chemother. 7, 85-90.
Krishnarjuna, A.V., Rao, T.V.N., Sundara raju, D., Vanisree, M., Tsay, H.S., Subbaraju, G.V., 2006. Biological
screening of medicinal plants collected from Eastern Ghats of India using Artemia salina. Int. J. Appl. Sci.
Eng. 4, 115-125.
Logu, A.D., Saddi, M., Onnis, V., Sanna, C., Congiu, C., Borgna, R., Cocco, M.T., 2005. In vitro
antimycobacterial activity of newly synthesised S-alkylisothiosemicarbazone derivatives and synergistic
interactions in combination with rifamycins against Mycobacterium avium. Int. J. Antimicrob. Agents. 26,
28-32.
Madadi, N.R., Penthala, N.R., Brents, L.K., Ford, B.M. Prather, P.L., Crooks, P.A., 2013. Evaluation of (Z)-2-
((1-benzyl-1H-indol-3-yl)methylene)-quinuclidin-3-one analogues as novel, high affinity ligands for CB1
and CB2 cannabinoid receptors. Bioorg. Med. Chem. Lett. 23, 2019-2021.
Madadi, N.R., Penthala, N.R., Janganati, V., Crooks, P.A., 2014. Synthesis and anti-proliferative activity of
aromatic substituted 5-((1-benzyl-1H-indol-3-yl)methylene)-1,3-dimethylpyrimidine-2,4,6(1H,3H,5H)-
trione analogs against human tumor cell lines. Bioorg. Med. Chem. Lett. 24, 601-603.
Mashayekhi, V., Tehrani, K.H.M.E., Azerang, P., Sardari, S., Kobarfard, F., 2013. Synthesis, antimycobacterial
and anticancer activity of novel indole-based thiosemicarbazones. Arch. Pharm. Res. DOI 10.1007/s12272-
013-0242-z
Mc laughlin, J.L., 1991. Crown-gall tumors in potato discs and brine shrimp lethality: two simple bioassays for
higher plant screening and fractionation, in: K. Hostettmann (Ed.), Methods in Plant Biochemistry,
Academic Press, London,
Meyer, B.N., Ferrigni, N.R., Putnam, J.E., Jacobsen, L.B., Nichols, D.E., Mc Laughlin, J.L., 1982. Brine
Shrimp: A convenient general bioassay for active plant constituents. Planta Med. 45, 31-34.
Moir, E.M., Yoshiizumi, K., Cairns, J., Cowley, P., Ferguson, M., Jeremiah, F., Kiyoi, T., Morphy, R., Tierney,
J., Wishart, G., York, M., Baker, J., Cottney, J.E., Houghton, A.K., McPhail, P., Osprey, A., Walker, G.,
Adam, J.A., 2010. Design, synthesis, and structure–activity relationship study of bicyclic piperazine
analogs of indole-3-carboxamides as novel cannabinoid CB1 receptor agonists. Bioorg. Med. Chem. Lett.
20, 7327-7330.
Moreira, D.R.M. De Oliveira, A.D.T., Gomes, P.A.T.M., de Simone, C.A., Villela, F.S., Ferreira, R.S., De
Silva, A.C., Santos, T.A.R., De Castro, M.C.A.B., Pereira, V.R.A., Leite, A.C.L., 2014. Conformational
restriction of aryl thiosemicarbazones produces potent and selective anti-Trypanosoma cruzi compounds
which induce apoptotic parasite death. Eur. J. Med. Chem. 75, 467-478.
Olowa L.F., Nuneza O.M., 2013. Brine shrimp lethality assay of the ethanolic extracts of three selected species
of medicinal plants from Iligan city, Philippines. Int. Res. J. Biological Sci. 2, 74-77
Peters-Golden, M., Henderson, W.R., 2007. Mechanisms of Disease: Leukotrienes. New. Engl. J. Med., 357,
1841-1854.
Prasher, P., Pooja, Singh, P., 2014. Lead modification: Amino acid appended indoles as highly effective
5-LOX inhibitors. Bioorg. Med. Chem. 22, 1642-1648.
Qi, B., Mi, B., Zhai, X., Xu, Z., Zhang, X., Tian, Z., Gong, P., 2013. Discovery and optimization of novel 4-
phenoxy-6,7-disubstituted quinolines possessing semicarbazones as c-Met kinase inhibitors. Bioorg. Med.
Chem. 21, 5246-5260.
Rajak, H., Thakur, B.S., Singh, A. Raghuvanshi, K., Sah, A.K., Veerasamy, R., Sharma, P.C., Pawar, R.S.,
Kharya, M.D., 2013. Novel limonene and citral based 2,5-disubstituted-1,3,4-oxadiazoles: A natural
product coupled approach to semicarbazones for antiepileptic activity. Bioorg. Med. Chem. lett. 23, 864-
868.
Rao, A.R., McKendrick, G.D., Velayudhan, L., Kamalakshi, K., 1966. Assessment of an isothiazole
thiosemicarbazone in the prophylaxis of contacts of variola major. Lancet. 287, 1072-1074.
Rao, J.V., Kavitha, P., Jakka, N.M., Sridhar, V., Usman, P.K., 2007. Toxicity of organophosphates on
morphology and locomotor behavior in Brine shrimp, Artemia salina. Arch. Environ. Contam. Toxicol.
53, 227–232
Reddanna, P., Whelan, J., Maddipati, K.R., Reddy, C.C., 1990. Purification of arachidonate 5-lipoxygenase
from potato tubers. Methods Enzymol. 187, 268-277.
Romano, M., Claria, J., 2003. Cyclooxygenase-2 and 5-lipoxygenase converging functions on cell proliferation
and tumor angiogenesis: implications for cancer therapy. Faseb J. 17, 1986-1985.
Samuelson, B., 1983. Leukotrienes: mediators of immediate hypersensitivity reactions and inflammation.
Science. 220, 568-575.
Sanchez, C., Gupta, M., Vasquez, M., de Noriega, Montenegro, G., 1993. Bioessay with brine Artemia to
predict antibacterial and pharmacologic activity. Rev. Med. Panama. 18, 62-69.
Sethi, M.L., 2002. Antiviral agents and protease inhibitors. In Principles of Medicinal Chemistry, Foye, W. O.,
Lemke, T.L., Williams, D.A., Eds., Williams and Wilkins: Baltimore. p 952.
Singh, P., Mittal, A., Bhardwaj, A., Kaurb, S., Kumar, S., 2008. 1-Toluene-sulfonyl-3-[(3′-hydroxy-5′-
substituted)-γ-butyrolactone]-indoles: Synthesis, COX-2 inhibition and anti-cancer activities. Bioorg. Med.
Chem. Lett. 18, 85-89.
Singh, P., Pooja., 2013. N-1, C-3 substituted indoles as 5-LOX inhibitors—In vitro enzyme immunoaasay, mass
spectral and molecular docking investigations. Bioorg. Med. Chem. Lett. 23, 1433-1437.
Sinha, S.K., Shrivastava, S.K., 2013. Synthesis, evaluation and molecular dynamics study of some new 4-
aminopyridine semicarbazones as an antiamnesic and cognition enhancing agents. Bioorg. Med. Chem. 21,
5451-5460.
Solis, P.N., Wright, C.W., Anderson, M.M., Gupta, M.P., Phillipson, J.D., 1993. A microwell cytotoxicity assay
using Artemia salina (Brine Shrimp). Planta Med. 59, 250-252.
Spanbroek, R., Habenicht, A.J. 2003. The potential role of antileukotriene drugs in atherosclerosis. Drug News
Perspect. 16, 485-489.
Sriram, D., Yogeeswari, P., Thirumurugan, R., 2004. Antituberculous activity of some aryl semicarbazone
derivatives. Bioorg. Med. Chem. lett. 14, 3923-3924.
Sriram, D., Yogeeswari, P., Thirumurugan, R., Pavana, R.K., 2006. Discovery of New Antitubercular Oxazolyl
Thiosemicarbazones. J. Med. Chem. 49, 3448-3450.
Tripathi, L., Kumar, P., Singh, R., Stables, J.P., 2012. Design, synthesis and anticonvulsant evaluation of novel
N-(4-substituted phenyl)-2-[4-(substituted) benzylidene]-hydrazinecarbothio amides. Eur. J. Med. Chem.
47, 153-166.
 Vieira, R.P., Lessa, J.A., Ferreira, W.C., Costa, F.B., Bastos, L.F.S., Rocha, W.R., Coelho, M.M., Beraldo, H.,
2012. Influence of susceptibility to hydrolysis and hydrophobicity of arylsemicarbazones on their anti-
nociceptive and anti-inflammatory activities. Eur. J. Med. Chem. 50, 140-148.
Werz, O., Steinhilber, D., 2005. Development of 5-lipoxygenase inhibitors—lessons from cellular enzyme
regulation. Biochem. Pharmacol. 70, 327-333.
Yi, W., Dubois, C., Yahiaoui, S., Haudecoeur, R., Belle, C., Song, H., Hardré, R., Réglier, M., Boumendjel, A.,
2011. Refinement of arylthiosemicarbazone pharmacophore in inhibition of mushroom tyrosinase. Eur. J.
Med. Chem. 46, 4330-4335
Yoshimura, R., Matsuyama, M., Kuratsukuri, K., Tsuchida, K., Takemoto, Y., Nakatani, T., 2005. A novel
approach to anticancer therapies for prostate cancer: lipoxygenase as a new target in the treatment of
prostate cancer. Drugs Future. 30, 351-358.
Young, R.N., 1999. Inhibitors of 5-lipoxygenase: a therapeutic potential yet to be fully realized? Eur. J. Med.
Chem. 34, 671-685.
Zani, C.L., Chaves, P.G., Queiroz, R., Mendes, N.M., De Oliveira, A.B., Cardoso, J.E., Anjos, M.G., Grandi, T.
S.M., 1995. Brine shrimp lethality assay as a pre-screening system for anti-Trypanosoma cruzi activity.
Phytomedicine. 2, 47-50.
Zheng, M., Zheng, M., Ye, D., Deng, Y., Qiu, S., Luo, X., Chen, K., Liua, H., Jiang, H., 2007. Indole
derivatives as potent inhibitors of 5-lipoxygenase: Design, synthesis, biological evaluation, and molecular
modelling. Bioorg. Med. Chem. Lett. 17, 2414-2470.
Zhong, Z., Aotegen, B., Xu, H., 2011. The influence of the different inductivity of acetyl phenyl–
thiosemicarbazone–chitosan on antimicrobial activities. Int. J. Biol. Macromol. 48, 713-719.

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