Estd. 2008
Vol. 8 | No.1 |152-155| January - March | 2015
ISSN: 0974-1496 | e-ISSN: 0976-0083 | CODEN: RJCABP
http://www.rasayanjournal.com
http://www.rasayanjournal.co.in
MICROBIAL ASSAY OF NOVEL 2-S-TETRA-O-BENZOYL-DGLUCOPYRANOSYL-1-ARYL-5-HEPTA-O-BENZOYL-β-DLACTOSYL-2-ISOTHIOBIURETS
Kedar P. Pande
Mauli College of Engineering and Technology, Shegaon-444203 (Maharashtra) INDIA.
E-mail : [email protected]
ABSTRACT
The present work aims to synthesize and screen the antifungal and antibacterial activities of a series of new 2-Stetra-O-benzoyl-D-glucopyranosyl-1-aryl-5-hepta-O-benzoyl-β-D-lactosyl-2-isothiobiurets by the interaction of
hepta-O-benzoyl-β-D-lactosyl isocyanate and S-tetra-O-benzoyl-D-glucopyranosyl-1-aryl isothiocarbamides.
The identities of these newly synthesised compounds have been established on the basis of usual chemical
transformations and IR, 1HNMR and Mass spectral studies. These synthesized products were evaluated for their
antimicrobial activity against some pathogenic organisms. Some of the products displayed promising activity.
Keywords: Lactosylisocyanate, glucopyranosylisothiocarbamides, lactosylisothiobiurets, antimicrobial activity.
©2015 RASĀYAN. All rights reserved
INTRODUCTION
Carbohydrate biosynthetic pathway is often amenable to interception with synthetic unnatural
subtracts1. Such metabolic interference can block the expression of oligosaccharides or alter the
structures of sugars presented on cells. The applications of glycosidase inhibitors2-5 to agricultural and
medical fields and the prospect for new therapeutic applications are reconsidered. These chemical
approaches are contributing great insight into the countless biological functions of oligosaccharides6-8.
Hence, it was the thought of interest to synthesize these N-lactosides9 and to study their antimicrobial
activity against gram +ve and gram –ve micro-organisms with the help of cup plate agar diffusion
method. In present communication, we report the microbial assay of newly synthesised 2-S-tetra-Obenzoyl-D-glucopyranosyl-1-aryl-5-hepta-O-benzoyl-β-D-lactosyl-2-isothiobiurets.
EXPERIMENTAL
All the chemicals and solvents were obtained from commercial and purified using standard procedure
wherever required. Melting points were taken by the open capillary method and were uncorrected.
The reactions were monitored by thin layer chromatography on silica gel G plates (Merck silica- 60
F258). Optical rotations [α]D31 were measured on the Equip-Tronics EQ-800 Digital Polarimeter at
31oC in CHCl3. The structures of all the newly synthesized compounds were confirmed by IR Spectra
which recorded on Perkin-Elmer spectrum RXI FTIR Spectrometer (Range: 4000-450 cm-1). 1H
NMRwas obtained on Bruker DRX-300 NMR spectrometer operating at 300 MHz Samples were
prepared in CDCl3 with TMS as an internal reference. Mass spectra were obtained on Thermo
Finnegan LCQ Advantage max ion trap mass spectrometer.
General Procedure
Synthesis of 2-S-tetra-O-benzoyl-D-glucopyranosyl-1-aryl-5-hepta-O-benzoyl-β-D-lactosyl-2isothiobiurets (3a-g)
A 0.005M of S-tetra-O-benzoyl-D-glucopyranosyl-1-aryl isothiocarbamides(2a-g) in a 5ml of
benzene was added to a 0.005M solution of hepta-O-benzoyl-β-D-lactosyl isocyanate(1) in 15ml
benzene, the reaction mixture was reflux over boiling water bath for 5hr. After refluxing, the solvent
was distilled off and the sticky residue obtained was triturated with petroleum ether (60-80 oC) to
afford a solid (3a-g). The product was purified by chloroform petroleum ether.
MICROBIAL ASSAY OF ISOTHIOBIURETS
Kedar P. Pande
Vol. 8 | No.1 |152-155 | January - March | 2015
RESULTS AND DISCUSSIONS
Herein, we report the synthesis of various 2-S-tetra-O-benzoyl-D-glucopyranosyl-1-aryl-5hepta-O-benzoyl-β-D-lactosyl-2-isothiobiurets (3a-f) by inteaction of hepta-O-benzoyl-β-Dlactosyl isocyanate(1) and S-tetra-O-benzoyl-D-glucopyranosyl-1-aryl isothiocarbamides(2ag).
All products were crystallized from ethanol before recording the physical data. The purity of
compound was checked by TLC. The spectral analysis10-12 IR, 1HNMR and Mass spectra of
the product were observed. Optical rotation of the product was also recorded. All the
compounds have been screen for both antimicrobial and antifungal activity using cup plate
agar diffusion method13-14 by measuring the inhibition zone in mm. Amikacin (100ug/mL)
was used as a standard for antibacterial activity and Fluconazole (100ug/mL) was used as a
standard for antifungal acitivity.
OBz OBz
O
OBz
BzO
O
O
N=C=O
O
BzO
BzO
OBz
RNH
+
OBz
HN
(1)
Hepta-O-benzoyl-β
β-D-lactosyl isocyanate
OBz
C
S
(2a-g)
S-tetra-O-benzoyl-D-glucopyranosyl1-aryl isothoicarbamides
Benzene
Reflux, 5hr
O
OBz OBz
OBz
NH C
O
BzO
OBz
RNH
O
O
BzO
OBz
OBz
OBz
N
C
BzO
OBz
OBz
OBz
S
O
(3a-g)
2-S-tetra-O-benzoyl-D-glucopyranosyl-1-aryl-5-hepta-O-benzyl-β
β-D-lactosyl-2-isothiobiurets
Where, Bz = COC6H5, R = (a) phenyl, (b) o-tolyl, (c) m-tolyl, (d) p-tolyl, (e) o-Cl-phenyl, (f) m-Cl-phenyl, (g)
p-Cl-phenyl.
Scheme-1
Spectral Data
3a: White solid, yield : 60%, mp : 130-132oC, [α]31D : -50.01o (c, 0.93, CHCl3), Rf = 0.42 (Pet. Ether :
Acetone, 3:2); IR (KBr, cm-1): υ 3068 (Ar-H), 1729 (C=O), 3465 (N-H), 1653 (C=N), 1269 (C-N),
771 (C-S), 1101 & 1027 (Characteristic of Lactose); 1H NMR (CDCl3): δ 8.02-7.11 (42H, m, Ar-H),
5.21-5.15 (2H, s, NH protons), 6.28-3.89 (14H, m, lactosyl and glucosyl protons); Mass (m/z): 1825
(M+), 1053, 948, 932, 579, 531, 135. Anal.Calcd.for C103H83O27N3SCl, Requires: C, 67.72; H, 4.54; N,
2.30; S, 1.75; Found: C, 67.68; H, 4.50; N, 3.24; S, 1.71%.
3c: Yellow solid, yield : 53%, mp : 108-110oC, [α]31D : +82.9o (c, 0.93, CHCl3), Rf = 0.37 (Pet. Ether :
Acetone, 3:2); IR (KBr, cm-1): υ 3066 (Ar-H), 1730 (C=O), 3451 (N-H), 1653 (C=N), 1270 (C-N),
709 (C-S), 1069 & 1025 (Characteristic of Lactose); 1H NMR (CDCl3): δ 8.20-7.12 (44H, m, Ar-H),
5.12-5.10 (2H, s, NH protons), 6.22-3.73 (14H, m, lactosyl and glucosyl protons); Mass (m/z): 1839
(M+), 1053, 948, 932, 579, 531, 135. Anal.Calcd.for C104H85O27N3S, Requires: C, 67.86; H, 4.62; N,
2.28; S, 1.74; Found: C, 67.79; H, 4.59; N, 2.22; S, 1.71%.
3f: Light yellow solid, yield : 55%, mp : 126-127oC, [α]31D : +69.28o (c, 0.93, CHCl3), Rf = 0.48 (Pet.
Ether : Acetone, 3:2); IR (KBr, cm-1): υ 3067 (Ar-H), 1729 (C=O), 3448 (N-H), 1601 (C=N), 1270
(C-N), 772 (C-S), 1099 & 1025 (Characteristic of Lactose); 1H NMR (CDCl3): δ 8.28-7.19 (44H, m,
MICROBIAL ASSAY OF ISOTHIOBIURETS
153
Kedar P. Pande
Vol. 8 | No.1 |152-155 | January - March | 2015
Ar-H), 5.23-5.21 (2H, s, NH protons), 5.88-4.19 (14H, m, lactosyl and glucosyl protons); Mass (m/z):
1859 (M+), 1053, 948, 932, 579, 531, 135. Anal.Calcd.for C103H82O27N3SCl, Requires: C, 66.46; H,
4.40; N, 2.25; S, 1.72; Found: C, 66.40; H, 4.32; N, 2.19; S, 1.68%.
Antimicrobial Studies
All compounds were screened for antibacterial activity against Escherichia coli, Staphylococcus
aureus, Proteus vulgaris, Salmonella typhi, Klebsiella pneumoniae, Pseudomonas aeruginosa,
Bacillus subtilis in nutrient agar medium and for antifungal activity against Candida albicance and
Aspergillus niger in potato dextrose agar medium. These sterilized agar media were poured into Petri
dishes and allowed to solidify on the surface of the media, microbial suspensions were spread with the
help of sterilized triangular loop. A stainless steel cylinder of 8 mm diameter (pre-sterilized) was used
to bore the cavities. 0.1 ml portions of the test compounds in solvent were added into these wells. The
drug solution was allowed to diffuse for about an hour into the medium. The plates were incubated at
37oC for 24 hr and 30oC for 48 hr for antibacterial and antifungal activities respectively. The zone of
inhibition observed around the cups after respective incubation was measured. The results are
presented in Table-1.
Antibacterial studies of these compounds indicated that compounds 3a and 3d were found to be active
against E.coliand rest of were found to be moderately active. Compound 3a, 3b and 3g exhibited most
significant activity against S.aureusand compound 3a, 3b and 3d towards P. seudomonas. All other
compounds exhibited low to moderate activity. The results of antifungal activities are also tabulated in
Table1. Almost all compounds are most effectively active against C. albicance and A. niger.
Table-1: Antimicrobial activities of newly synthesized 2-S-tetra-O-benzoyl glucopyranosyl-1-aryl-5-hepta-Obenzoyl-β-D-lactosyl-2-isothiobiurets (3a-g)
Antibacterial**
Compound
Antifungal**
E.
S.
P.
S.
Ps.
K.
B.
C.
A.
coli
Aureus
vulgaris
typhi
aeruginosa
Pneumoniae
subtilis
albicance
niger
3a
20
22
17
16
21
19
11
20
22
3b
16
20
19
11
20
18
-
21
20
3c
15
18
-
10
12
13
20
19
21
3d
21
17
18
10
20
17
15
20
19
3e
15
-
13
-
-
12
12
18
20
3f
16
19
17
12
16
16
10
20
22
3g
17
21
16
13
19
19
-
21
23
Amikacin
25
26
23
25
24
26
24
-
-
Fluconazole
-
-
-
-
-
26
28
**zone of inhibition in mm (15 or less) resistance, (16-20 mm) moderate and (more than 20mm) sensitive.
Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Proteus vulgaris (P. vulgaris), Salmonella typhi
(S. typhi), KlebsiallaPneumoniae (K. Pneumoniae), Pseudomonas auriginosa (P. auriginosa), Bacillus subtilis
(B. subtlis), Candida albicance (C. albicance) and Aspergillusniger (A. niger).
CONCLUSION
The new lactosyl isothiobiurets exhibits promising antibacterial and antifungal activities against the
organism tested. The method adopted in this investigation is simple efficient inexpensive and is useful
in synthesizing pharmacologically important molecules.
MICROBIAL ASSAY OF ISOTHIOBIURETS
154
Kedar P. Pande
Vol. 8 | No.1 |152-155 | January - March | 2015
ACKNOWLEDGEMENT
Author is thankful to SAIF, CDRI, Lucknow for providing spectral data and Dr. C. M. Jadhao, Mauli
College of Engineering and Technology, Shegaon for providing necessary facilities.
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
N. Asano, Glycobiology, 13(10), 93(2003).
A. D. Elbein, Annu. Rev. Biochem., 56, 497(1987).
A. D. Elbein, FASEB Journal, 5(15), 3055(1991).
A. Mehta, N. Zitzmann, P. M. Rudd, T. M. Block and R.A. Dwek, FEBS Letters, 430 (12), 17
(1998).
V. N. Ingle, U. G. Upadhyay and S.T. Kharche, Indian J. Chem., 44B, 1859(2005).
T.W.Rademacher, R. B. Parekh and A.R. Dwek, Annu. Rev. BioChem., 5, 785(1998).
P. Gagneau and A. Varki, Glycobiology, 9, 747(1999).
R.C.N.Reic, S.C. Oda, M.V. De Almeida, M.C.S. Lourenco, F.R.C. Vicente, N.R.Barbosa, R.
Trevizani, P.L. C. Santos and M. Le Hyaric, J. Braz. Chem. Soc., 19(6)223(2008).
K. P. Pande and S. P. Deshmukh, DerPharmaChemica, 3(6), 28(2011).
R. M.Silverstein and F. X. Webster, “Spectrometric Identification of Organic Compounds”, 6th
ed., John Wiley and Sons, Inc, New York., 119(2011).
D. H. Williams and I. Fleming, “Spectroscopic Methods in Organic Chemistry”, 5th ed., Tata
McGraw-Hill, 53(2004).
J. R. Dyer, “Applications of Absorption Spectroscopy of Organic Compounds”, PHI Learning
Private Limited, New Delhi, 88(2010).
F.Kawangh, Analytical Microbiology, Academic press, New York, 47(1963).
British pharmacopoeia-II, Biological assay and Tests, The Stationary Office Ltd., London; A-205,
102(1998).
[RJC-1220/2015]
MICROBIAL ASSAY OF ISOTHIOBIURETS
155
Kedar P. Pande