Received 14 March 1999; received in revised form 23 April 1999; accepted 17 May 1999
Abstract
Isatin, its 5-chloro and 5-bromo derivatives have been reacted with N-[4-(49-chlorophenyl)thiazol-2-yl] thiosemicarbazide to form
Schiff bases and the N-Mannich bases of these compounds were synthesized by reacting them with formaldehyde and three secondary
amines. Their chemical structures have been confirmed by means of IR, 1 H-NMR data and by elemental analysis. Investigation of
antimicrobial activity of compounds was done by agar dilution method against 28 pathogenic bacteria, 8 pathogenic fungi and anti-HIV
activity against replication of HIV-1 (IIIB) in MT-4 cells. Among the compounds tested 1-[N,N-dimethylaminomethyl]-5-bromo
isatin-3-h19-[40-(p-chlorophenyl) thiazol-20-yl] thio semicarbazonej 10 showed the most favourable antimicrobial activity. 1999
Elsevier Science B.V. All rights reserved.
Keywords: Isatin; Thiazole; Thiosemicarbazone; Schiff bases; Mannich bases; Antimicrobial
1. Introduction
Isatin (Indolin-2,3-dione) derivatives are reported to
show variety of biological activities like antibacterial
(Daisley and Shah, 1984) antifungal (Piscopo et al., 1987)
and anti-HIV (Pandeya et al., 1998) activities. Thiazoles
have been reported to possess antibacterial (Agarwal et al.,
1997), antifungal (Sup et al., 1995) and anti-HIV (Maass et
al., 1993) activities. It is also reported that isatin-bthiosemicarbazones have shown antimicrobial activity
(Teitz et al., 1994). Further amino derivative of pyridine
2-carboxaldehyde thiosemicarbazone have been found to
be shown antitumor activity (Liu et al., 1992). Recently
cytotoxic activities of Mannich bases of chalcones (Dimmock et al., 1998) have been reported. In view of the
antimicrobial property of the above pharmacophores, it
was envisaged that the combined effect of all the entities
will result in increased antimicrobial activity. Thus the
2. Experimental
2.1. Chemistry
The melting points were determined by using ThomasHoover melting point apparatus and are uncorrected.
Spectroscopic data were recorded on the following instruments IR, Jasco infrared spectrometer, Jeol FX 90Q FTNMR spectrometer (90 MHz). Microanalyses were performed by the microanalytical unit Central Drug Research
Institute, India.
0928-0987 / 99 / $ see front matter 1999 Elsevier Science B.V. All rights reserved.
PII: S0928-0987( 99 )00038-X
26
2.1.4. Synthesis of 1 -(morpholino methyl) isatin-3 -h l9 [40 -( p-chlorophenyl) thiazol-20 -yl] thiosemicarbazonej
(6)
A slurry consisting of the S-1 (0.005 mol), tetrahydrofuran (5 ml) and 37% formalin (2 ml) was made. To this
morpholine (0.005 mol) was added dropwise, with cooling
and shaking. The reaction mixture was allowed to stand at
room temperature for 1 h with occasional shaking after
which it was warmed on a steam bath for 15 min. At the
27
end of the period the contents were cooled and the product
obtained was recrystallized from chloroformpetroleum
ether. Yield 90.4%; m.p. 1318C; IR (KBr): 1615 cm 21
(C5N), 2850 cm 21 (CH 2 ); 1 H-NMR (CDCl 3 ) d (ppm):
2.6 (t, 4H, CH 2 NCH 2 ), 3.7 (t, 4H, CH 2 OCH 2 ) 4.45
(s, 2H, CH 2 ), 6.3 (s, 1H, H-5), 6.97.8 (m, 7H, Ar-H),
9.45 (s, 2H, 23-NH-, D 2 O exchangeable); Anal.
(C 23 H 21 O 2 N 6 S 2 C1) C, H, N.
Table 1
Physical constants of the synthesized compounds
Code
R1
Yield
(%)
M.P.
8C
Mol. formula
Elemental analysis
calculated / found
%C
%H
%N
52.1
52.3
48.1
48.0
43.8
43.9
53.5
53.7
2.8
2.9
2.4
2.5
2.2
2.2
4.0
4.2
16.9
17.0
15.6
15.9
14.2
14.2
17.8
17.6
94.6
125
C 18 H 12 ON 5 S 2 C1
Cl
86.7
208
C 18 H 11 ON 5 S 2 C1 2
Br
91.2
128
C 18 H 11 ON 5 S 2 ClBr
H-
CH 2 -N(CH 3 ) 2
82.8
186
C 21 H 19 ON 6 S 2 C1
91.3
122
C 24 H 23 ON 6 S 2 C1
56.3
56.4
4.5
4.6
16.4
16.4
90.4
131
C 23 H 21 O 2 N 6 S 2 C1
Cl
89.0
268
C 21 H 18 ON 6 S 2 C1 2
53.7
53.8
49.8
49.1
4.0
4.1
3.5
3.5
16.3
16.4
16.6
16.8
Cl
90.6
224
C 24 H 22 ON 6 S 2 C1
52.7
52.8
4.0
4.1
15.3
15.4
Cl
90.4
220
C 23 H 20 0 2 N 6 S 2 C1 2
10
Br
80.0
197
C 21 H 18 ON 6 S 2 ClBr
50.4
50.3
45.8
45.8
3.6
3.7
3.2
3.3
15.3
15.2
15.2
15.3
11
Br
72.0
207
C 24 H 22 ON 6 S 2 ClBr
48.8
48.5
3.7
3.7
14.2
14.2
12
Br
89.5
191
C 23 H 20 O 2 N 6 S 2 C1Br
46.6
46.6
3.3
3.4
14.2
14.0
-CH 2 -N(CH 3 ) 2
-CH 2 -N(CH 3 ) 2
28
Table 2
Antibacterial activity of the compounds MICs in mg / mL a
Microorganisms / Drugs
1. Salmonella typhimurium
2. Vibrio parahaemolyticus
3. Salmonella paratyphi B
4. Edwardsiella tarda
5. Vibrio cholerae 0139
6. Staphylococcus aureus (G1ve)
7. Escherchia coli NCTC 10418
8. Vibrio cholerae non-01
9. Enterococcus faecalis (G1ve)
10. Salmonella typhi
11. Pseudomonas aeruginosa
12. Klebsiella pneumoniae
13. Staphylococcus albus (G1ve)
14. Salmonella enteritidis
15. Aeromonas hydrophile
16. Vibrio cholerae-01
17. Bacillus subtilis (G1ve)
18. Shigella sonnei
19. Shigella boydii
20. Plesiomonas shigelloides
21. Proteus rettgeri
22. Shigella flexnari
23. Proteus vulgaris
24. Enterobacter
25. Morgonella morgonii
26. Citrobacter ferundii
27. Proteus morgonii
28. Salmonella paratyphi B
625
78.1
625
625
312.5
1250
625
39.1
4.9
312.5
2500
625
625
312.5
156.2
312.5
312.5
156.2
312.5
312.5
156.2
312.5
312.5
156.2
156.2
156.2
312.5
156.2
625
156.2
312.5
312.5
312.5
1250
312.5
39.1
9.8
156.2
1250
312.5
312.5
312.5
156.2
156.2
156.2
78.1
156.2
78.1
156.2
312.5
78.12
156.2
312.5
156.2
312.5
78.1
312.5
78.1
312.5
312.5
78.1
1250
312.5
39.1
9.8
156.2
1250
625
312.5
156.2
156.2
78.1
156.2
156.2
156.2
156.2
156.2
78.1
156.2
156.2
78.1
78.1
156.2
19.5
156.2
19.5
78.1
156.2
39.1
625
312.5
39.1
2.4
156.2
1250
312.5
625
156.2
39.1
78.1
156.2
78.1
156.2
78.1
39.1
156.2
156.2
156.2
78.1
39.1
156.2
39.1
312.5
39.1
156.2
156.2
78.1
1250
156.2
19.5
2.4
156.2
1250
312.5
625
156.2
78.1
78.1
156.2
156.2
156.2
312.5
39.1
156.2
78.1
156.2
156.2
39.1
312.5
156.2
312.5
78.1
312.5
312.5
156.2
1250
312.5
39.1
4.9
78.1
1250
625
625
312.5
78.1
156.2
78.1
156.2
312.5
156.2
78.1
312.5
156.2
156.2
156.2
78.1
312.5
156.2
312.5
39.1
78.1
156.2
312.5
312.5
78.1
19.5
2.4
78.1
625
156.2
312.5
78.1
156.2
39.1
156.2
78.1
78.1
78.1
39.1
78.1
78.1
39.1
78.1
156.2
78.1
78.1
29
Table 3
Antibacterial activity of the compounds MICs in mg / mL a
Microorganisms / Drugs
10
11
12
Suiphamethoxazole
Trimethoprim
1. Salmonella typhimurium
2. Vibrio parahaemolyticus
3. Salmonella paratyphi B
4. Edwardsiella tarda
5. Vibrio cholerae 0139
6. Staphylococcus aureus (G1ve)
7. Escherchia coli NCTC 10418
8. Vibrio cholerae non-01
9. Enterococcus faecalis (G1ve)
10. Salmonella typhi
11. Pseudomonas aeruginosa
12. Klebsiella pneumoniae
13. Staphylococcus albus (G1ve)
14. Salmonella enteritidis
15. Aeromonas hydrophile
16. Vibrio cholerae-01
17. Bacillus subtilis (G1ve)
18. Shigella sonnei
19. Shigella boydii
20. Plesiomonas shigelloides
21. Proteus rettgeri
22. Shigella flexnari
23. Proteus vulgaris
24. Enterobacter
25. Morgonella morgonii
26. Citrobacter ferundii
27. Proteus morgonii
28. Salmonella paratyphi B
312.5
78.1
312.5
156.2
78.1
625
156.2
39.1
9.8
156.2
625
156.2
156.2
156.2
78.1
39.1
156.2
39.1
78.1
39.1
39.1
156.2
78.1
156.2
312.5
78.1
156.2
78.1
312.5
156.2
156.2
156.2
156.2
625
312.5
39.1
9.8
156.2
625
312.5
156.2
312.5
156.2
78.1
78.1
78.1
156.2
156.2
39.1
312.5
156.2
78.1
156.2
78.1
156.2
156.2
78.1
39.1
39.1
78.1
78.1
312.5
312.5
9.8
1.2
156.2
625
625
78.1
78.1
78.1
39.1
78.1
19.5
78.1
156.2
156.2
9.8
39.1
39.1
19.5
39.1
78.1
19.5
156.2
78.1
156.2
312.5
39.1
625
312.5
39.1
4.9
78.1
625
312.5
156.2
156.2
78.1
78.1
156.2
78.1
156.2
156.2
78.1
39.1
78.1
39.1
78.1
78.1
78.1
78.1
156.2
39.1
156.2
156.2
156.2
625
156.2
39.1
4.9
156.2
1250
625
156.2
156.2
78.1
156.2
78.1
156.2
156.2
156.2
156.2
78.1
39.1
156.2
19.5
78.1
78.1
39.1
5000
1250
5000
5000
.5000
5000
2500
312.5
5000
2500
78.12
2500
2500
2500
2500
5000
5000
2500
2500
5000
2500
2500
2500
1250
2500
5000
5000
2500
5000
2.4
9.8
312.5
39.1
.5000
19.5
1.2
78.1
4.9
5000
5000
.5000
4.9
1250
5000
5000
9.8
9.8
4.9
2500
156.2
156.2
156.2
156.2
19.5
156.2
156.2
30
Table 4
Antifungal activity of the compounds MICs in mg / mL
Drugs /
Microorganism
Cryptococcus
neoformans
Microsporum
audouinii
Trichophyton
mentagrophytes
Epidermophyton
floccosum
Microsporum
gypsum
Histoplasma
capsulatum
Candida
albicans
Aspergillus
niger
1
2
3
4
5
6
7
8
9
10
11
12
Clotrimazole
4.9
2.4
4.9
4.9
4.9
2.4
2.4
4.9
4.9
2.4
4.9
9.8
2.4
4.9
2.4
2.4
4.9
2.4
1.2
2.4
4.9
2.4
2.4
9.8
4.9
4.9
9.8
4.9
1.2
4.9
4.9
2.4
2.4
4.9
2.4
2.4
2.4
9.8
2.4
1.2
1.2
2.4
1.2
2.4
2.4
1.2
4.9
2.4
2.4
1.2
1.2
2.4
2.4
1.2
1.2
1.2
1.2
1.2
1.2
1.2
2.4
1.2
1.2
1.2
2.4
39.l
19.5
19.5
78.1
39.1
39.1
19.5
19.5
19.5
19.5
39.1
78.1
19.5
156.2
78.1
39.1
78.1
156.5
156.5
78.2
78.1
78.1
78.1
78.1
78.1
0.3
39.1
19.5
9.8
39.1
39.1
9.8
19.5
9.8
9.8
9.8
9.8
9.8
2.4
EC 50 a , mg / mL
CC 50 b , mg / mL
SI c
1
2
3
4
5
6
7
8
9
10
11
12
.12
.10
.10
.54
.57
.19
.10
.12
.12
.29
.21
.14
12.1
10.1
10.4
53.8
57.2
19.4
10.3
11.8
11.7
29.2
20.7
13.7
,1
,1
,1
,1
,1
,1
,1
,1
,1
,1
,1
,1
hydrophobic group is necessary (Teitz et al., 1994) whereas in our case, we have a big p-chlorophenyl thiazolyl
group at thiosemicarbazide end.
Acknowledgements
The authors would like to express their gratitude and
thanks to the Head, Dept. of Pharmaceutics and Dept. of
Microbiology, Banaras Hindu University for providing
necessary facilities for this research work.
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