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A new synthetic approach and biological evaluation of novel phenothiazines

bearing tert-butyl group
Amit R. Trivedi a; Arif B. Siddiqui a; Dipti K. Dodiya a; Manish J. Soalnki a; Viresh H. Shah a
a
Chemical Research Laboratory, Department of Chemistry, Saurashtra University, Rajkot, India

First Published on: 18 August 2009

To cite this Article Trivedi, Amit R., Siddiqui, Arif B., Dodiya, Dipti K., Soalnki, Manish J. and Shah, Viresh H.(2009)'A new synthetic
approach and biological evaluation of novel phenothiazines bearing tert-butyl group',Journal of Sulfur Chemistry,99999:1,
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 Journal of Sulfur Chemistry
iFirst, 2009, 1–7

A new synthetic approach and biological evaluation of novel

phenothiazines bearing tert-butyl group
Amit R. Trivedi, Arif B. Siddiqui, Dipti K. Dodiya, Manish J. Soalnki and Viresh H. Shah*
Chemical Research Laboratory, Department of Chemistry, Saurashtra University, Rajkot 360005, India
Downloaded By: [Cranfield University] At: 11:35 18 August 2009

(Received 16 October 2007; final version received 27 June 2009 )

A new and facile synthetic route is proposed for the synthesis of some novel phenothiazines (5a–5g) based
on the reaction of 2-amino substituted benzenethiols with p-tert-butyl phenol in good yield. The newly
synthesized compounds were characterized by IR, 1 H NMR and mass spectral studies. Their antimicrobial
activities against three strains of bacteria: Bacillus subtilis, Bacillus megaterium, Escherichia coli, and two
strains of fungi: Aspergillus niger and Aspergillus oryzae, were investigated.

Keywords: phenothiazine; p-tert-butyl phenol; 2-amino benzenethiols; oxidative cyclization;

antimicrobial activity

1. Introduction

During the past five decades, studies devoted to the phenothiazine class of compounds (1–4) have
been stimulated by the discovery of the pharmacodynamic properties of a number of derivatives in
which a variety of amino alkyl side chains are connected to the nitrogen atom of the heterocyclic
unit. Consequently, drugs incorporating a phenothiazine ring system (5–7) have played a crucial
role in medicinal chemistry and have occupied a place of choice in the arsenal of pharmaceuti-
cal drugs, owing to their antimicrobial (8), tranquilizing (9), anti-inflammatory (10), antimalarial
(11), antipsychotropic (12), antitubercular (13, 14) and antitumor (15–17) properties, and for their
ability to stimulate the penetration of anticancer agents via blood-brain barrier.
A slight variation in the substitution pattern in the phenothiazine nucleus causes a marked dif-
ference in activity, and therefore phenothiazines with varied substituents are being synthesized
and tested for activities in search of better medicinal agents. Moreover, in spite of its very simple
hydrocarbon structure, the tert-butyl group has attracted much attention from organic chemists
for many decades due to its unique biomedical applications and significant contribution in the
enhancement of bioactivities (18–24). Keeping in mind the biomedical applications, as a contin-
uation of our previous work (25) and with a view to further assess the pharmacological profile
of this class of compounds, we thought it worthwhile to synthesize some new congeners of phe-
nothiazines 5a–5g by incorporating the phenothiazine nucleus and tert-butyl group in a single

*Corresponding author. Email: [email protected]

ISSN 1741-5993 print/ISSN 1741-6000 online

© 2009 Taylor & Francis
DOI: 10.1080/17415990903173511
http://www.informaworld.com
 2 A.R. Trivedi et al.

molecular framework with a potential spectrum of bio-responses. The antimicrobial activities of

the newly synthesized compounds 5a–5g against three strains of bacteria: Bacillus subtilis, Bacil-
lus megaterium and Escherichia coli, and two strains of fungi: Aspergillus niger and Aspergillus
oryzae, were investigated.

2. Chemistry

To the best of our knowledge, there are no previous reports in the literature for phenothiazines
having a tert-butyl group. A literature survey reveals general synthetic strategies towards phe-
nothiazines, such as thionation of diphenylamines (26), reaction of aminothiols with halonitro
or dihalogenobenzene derivatives (27), cyclization of diphenyl sulfides (28), condensation of
aminothiols with quinone or 1,3-dione (29), cyclization of 2-substituted diphenylamines (30),
etc., which require long reaction times or include multi-step synthesis. In contrast, for the same re-
Downloaded By: [Cranfield University] At: 11:35 18 August 2009

action performed under our conditions, the reaction time was markedly reduced and improvement
in yields was also observed. The work up is simple and does not require any tedious procedures.
Herein we report a novel, efficient, single-step synthesis of novel phenothiazines bearing a
tert-butyl group (5a–5g). The synthetic pathway for the preparation of the title compounds 5a–5g
is depicted in Scheme 1.
The ring system 5a–5g was obtained via oxidative cyclization (31) by merely heating a mixture
of p-tert-butyl phenol (1) and appropriately substituted 2-mercapto aniline (2a–5g) in dimethyl
sulfoxide at 140 ◦ C for 45 min. Since the 2-mercapto anilines (2a–g) readily oxidize to bis
(o-aminophenyl)disulfide (3) under these reaction conditions, the reaction is considered to pro-
ceed via the intermediate 4, which is readily cyclized by the scission of the sulfur–sulfur bond
(32, 33) upon the attack by the nucleophilic enaminone system. The possible mechanism of the
reaction is proposed in Scheme 2.
The IR spectra of 5a–5g exhibit a sharp peak in the region 3300–3350 cm−1 due to −NH
stretching vibrations, and a sharp peak in the range of 600–700 cm−1 , suggesting the presence of
C−S−C linkage. Phenothiazines 5d and 5g having a chlorine atom show a peak in the range of
720–780 cm−1 . In phenothiazines 2c and 2f, two peaks appear in the range of 1050–1075 cm−1 and
1200–1275 cm−1 due to asymmetric and symmetric vibrations of C−O−C linkage. The 1 H NMR
spectra of phenothiazines 5a–5g exhibit a broad signal in the region 9.01–9.92 δ due to an NH
proton, while aromatic protons appear as a multiplet between 7.11–8.91 δ. In all phenothiazines
5a–5g, a sharp singlet of 9H was observed in the range of 1.28–1.45 δ due to the presence of a
tert-butyl group.

R1 R1
H
OH H2N (CH3)2SO N
H3C + H3C
HS R2 140 ºC S R2
H3C H3C
CH3 CH3
1 (2a–g) (5a–g)

R1 R2
2a, 5a H H
2b, 5b CH3 H
2c, 5c OCH3 H
2d, 5d Cl H
2e, 5e H CH3
2f, 5f H OCH3
2g, 5g H Cl

Scheme 1.
 Journal of Sulfur Chemistry 3

R1
OH H2N R1
(CH3)2SO
+ H2N
H3C
140 ºC
HS R2
H3C S R2
CH3
2
1 (2a–g) 3

R1 R1
H H
N N
H3C H3C
S R2 S R2
H3C H3C
CH3 S R2 CH3
(5a–g)

H2N
R1
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Scheme 2.

3. Biological activity

Compounds 5a–5g were screened in vitro for their antimicrobial activities against three strains
of bacteria: B. subtilis, B. megaterium, E. coli, and two strains of fungi: A. niger and A. oryzae,
by the agar-diffusion technique (34). A 1 mg/ml solution in dimethylformamide was used. The
bacteria and fungi were maintained on nutrient agar and Czapek’s-Dox agar media, respectively.
DMF showed no inhibition zones. The agar media were incubated with different microorganism
cultures. After 24 h of incubation at 30 ◦ C for bacteria and 48 h of incubation at 28 ◦ C for fungi,
the diameter of the inhibition zone (mm) was measured (Table 1). Ampicillin, chloramphenicol
and fluconazole were purchased from an Indian market and used in a concentrations of 25 mg/ml
as references for antibacterial and antifungal activities. The results depicted in Table 1 revealed
that phenothiazines 5a, 5b, 5e, 5g and 5h showed a very high activity with respect to the used ref-
erences, while phenothiazines 5c and 5f showed comparable activity. On the other hand, nearly all

Table 1. Inhibition zone (mean diameter of inhibition, in mm) as a criterion of antibacterial and antifungal
activities of the newly synthesized phenothiazine derivatives.

Inhibition zone
Gram-positive bacteria Gram-negative bacteria Fungi
B. subtilis B. megaterium E. coli A. niger A. oryzae

Compound
5a 69 56 56 69 68
5b 41 46 59 56 72
5c 36 42 29 62 65
5d 53 61 69 53 69
5e 44 57 57 71 77
5f 32 35 26 72 63
5g 60 69 55 63 76
Reference drugs
Ampicillin 40 29 26 33 –
Chloramphenicol 30 55 48 35 –
Fluconazole – – – 22 16
 4 A.R. Trivedi et al.

of the prepared compounds exhibited an interesting high antifungal activity against the reference
chemotherapeutics.

4. Conclusion

To sum up, we have developed a concise and efficient approach for the synthesis of some novel
phenothiazines bearing a tert-butyl group. We also believe that the procedural simplicity, efficiency
and the easy accessibility of the reaction partners should be rewarded by giving access to a wide
array of heterocyclic frameworks equipped with a pendant phenothiazine unit. Further work is
under progress to design new synthetic methodologies for phenothiazines, and for screening these
compounds for their biological activities.
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5. Experimental

Melting points were determined in open capillary tubes and are uncorrected. Formation of the
compounds was routinely checked by TLC on silica gel-G plates of 0.5-mm thickness, and spots
were located by iodine. 1 H NMR spectra were determined in a CDCl3 solution at 300 MHz on a
Bruker spectrometer. IR spectra were recorded using a Shimadzu 8400 spectrometer in KBr (γ in
cm−1 ). Elemental analysis of the newly synthesized compounds was carried out on a Carlo Erba
1108 analyzer.

5.1. General experimental procedure

A mixture of p-tert-butyl phenol 1 (0.01 mol), respective 2-mercapto aniline 2a–2g (0.01 mol) in
dimethyl sulfoxide (10 ml) was heated with stirring at 140–145 ◦ C form 45 min. The crystalline
product 5a–5g separated on cooling was filtered and recrystallized from methanol.

5.2. 7-Tert-butyl-10H-phenothiazine (5a)

Yield 78%, m.p. 177 ◦ C; IR (KBr) cm−1 : 3334 (NH), 1328 (C−N), 653 (C−S−C); 1 H NMR
(300 MHz, CDCl3 ) δ: 1.43 (s, 9H, tert-Bu), 7.21–8.40 (m, 7H, Ar−H), 9.11 (s, 1H, NH); MS:
m/z 255; Anal. calcd. for C16 H17 NS: C, 75.25; H, 6.71; N 5.48. Found: C, 75.29; H, 6.73;
N, 5.54%.

5.3. 7-Tert-butyl-1-methyl-10H-phenothiazine (5b)

Yield 83%, m.p. 204 ◦ C; IR (KBr) cm−1 : 3335 (NH), 1332 (C−N), 655 (C−S−C); 1 H NMR
(300 MHz, CDCl3 ) δ: 1.32 (s, 9H, tert-butyl), 2.46 (s, 3H, CH3 ), 7.43–8.21 (m,6H, Ar−H), 9.01
(s, 1H, NH); MS: m/z 269; Anal. calcd. for C17 H19 NS: C, 75.79; H, 7.11; N, 5.20. Found: C,
75.83; H, 7.16; N, 5.25%.

5.4. 7-Tert-butyl-1-methoxy-10H-phenothiazine (5c)

Yield 74%, m.p. 118 ◦ C; IR (KBr) cm−1 : 3338 (NH), 1330 (C−N), 650 (C−S−C); 1 H NMR
(300 MHz, CDCl3 ) δ: 1.45 (s, 9H, tert-butyl), 3.67 (s, 3H, OCH3 ), 7.80–8.91 (m, 6H, Ar−H),
9.73 (s, 1H, NH); MS: m/z 285; Anal. calcd. for C17 H19 NOS: C, 71.54; H, 6.71; N, 4.91. Found:
C, 71.5; H, 6.77; N, 4.88%.
 Journal of Sulfur Chemistry 5

5.5. 7-Tert-butyl-1-chloro-10H-phenothiazine (5d)

Yield 85%, m.p. 167 ◦ C; IR (KBr) cm−1 : 3332 (NH), 1325 (C−N), 648 (C−S−C); 1 H NMR
(300 MHz, CDCl3 ) δ: 1.38 (s, 9H, tert-butyl), 7.11–8.36 (m, 6H, Ar−H), 9.28 (s, 1H, NH); Mass
(m/z): 290; Anal. calcd. for C16 H16 N5 SCl: C, 66.31; H, 5.56; N, 4.83. Found: C, 66.36; H, 5.53;
N, 4.88%.

5.6. 3-Tert-butyl-7-methyl-10H-phenothiazine (5e)

Yield 82%, m.p. 137 ◦ C; IR (KBr) cm−1 : 3337 (NH), 1327 (C−N), 645 (C−S−C); 1 H NMR
(300 MHz, CDCl3 ) δ: 1.40 (s, 9H, tert-butyl), 2.52 (s, 3H, CH3 ), 7.37–8.44 (m, 6H, Ar−H), 9.18
(s, 1H, NH); MS: m/z 269; Anal. calcd. for C17 H19 NS: C, 75.79; H, 7.11; N, 5.20. Found: C,
75.74; H, 7.08; N, 5.16%.
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5.7. 3-Methoxy-7-tert-butyl-10H-phenothiazine (5f)

Yield 72%, m.p. 97 ◦ C; IR (KBr) cm−1 : 3330 (NH), 1325 (C−N), 657 (C−S−C); 1 H NMR
(300 MHz, CDCl3 ) δ: 1.28 (s, 9H, tert-butyl), 3.64 (s, 3H, OCH3 ), 7.21–8.68 (m, 6H, Ar−H),
9.92 (s, 1H, NH); MS: m/z 285; Anal. calcd. for C17 H19 NOS: C, 71.54; H, 6.71; N, 4.91. Found:
C, 71.58; H, 6.75; N, 4.98%.

5.8. 3-Chloro-7-tert-butyl-10H-phenothiazine (5g)

Yield 68%, m.p. 153 ◦ C; IR (KBr) cm−1 : 3333 (NH), 1330 (C−N), 658 (C−S−C); 1 H NMR
(300 MHz, CDCl3 ) δ: 1.34 (s, 9H, tert-butyl), 7.12–8.52 (m, 6H, Ar−H), 9.29 (s, 1H, NH); MS:
m/z 290; Anal. calcd. for C16 H16 NSCl: C, 66.31; H, 5.56; N, 4.83. Found: C, 66.28; H, 5.50;
N, 4.78%.

Acknowledgements
The authors thank the Professor and Head, Department of Chemistry, Saurashtra University, Rajkot for providing research
facilities. Authors are thankful to CDRI Lucknow for spectral and analytical data.

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 Journal of Sulfur Chemistry 7

Amit R. Trivedi, Arif

B. Siddiqui, Dipti K.
R1 R1
H Dodiya, Manish J.
OH H2N N Soalnki and Viresh
(CH3)2SO
H3C + H3C
H. Shah
HS R2 140 ºC S R2
H3C H3C A new synthetic
CH3 CH3
1 (2a–g) (5a–g) approach and
biological evaluation
of novel
phenothiazines
bearing tert-butyl
group

1–6
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