S. Ghosh et al. / Bioorg. Med. Chem. Lett. 19 (2009) 386–389
389
reduced pressure and the crude product was purified over SiO2 using hexane-
EtOAc (2:1) as eluant to furnish pure compound 5a–e.
13. (a) Schurr, P. E.; Schultz, J. R.; Parkinson, T. M. Lipids 1972, 7, 68; (b) Deeg, R.;
Ziegehorn, J. Clin. Chem 1983, 29, 1798; (c) Batra, S.; Bhaduri, A. P.; Joshi, B. S.;
Roy, R.; Khanna, A. K.; Chander, R. Bioorg. Med. Chem. 2001, 9, 3093.
14. Buccolo, G.; David, H. Clin. Chem. 1973, 19, 476.
Research Fellowship. A.K.M. thanks DST, New Delhi for Ramanna
fellowship.
Supplementary data
15. Zilversmit, D. B.; Davis, A. K.; Memphis, B. S.; Tenn, J. L. Clin. Med. 1985, 35, 831.
16. (a) Wing, D. R.; Robinson, D. F. Biochem. J. 1968, 109, 841; (b) Mosinger, F. J.
Lipid. Res. 1965, 6, 157.
Supplementary data associated with this article can be found, in
17. Lipid lowering activity: Adult male Charles Foster rats (200 225 g) bred in the
animal house of the institute were used for the lipid lowering activity. Rats
were divided in control, triton induced, triton plus compounds and Gemfibrozil
(100 mg/Kg) treated groups containing six rats in each. Hyperlipidemia was
developed by administration of Triton WR-1339 (Sigma chemical co., St. Louis,
USA) at a dose of 400 mg/Kg body wt. intraperitoneally to animals of all groups
except the control. Compounds 4a–k and 5a–e were macerated with gum
acacia (0.2% w/v), suspended in water and fed simultaneously with triton at a
dose of 100 mg/Kg po to the animals of treated groups. Animals of the control
and triton group without treatment with test compounds were given same
amount of gum acacia suspension (vehicle). After 18 h of treatment (50 mg/kg
b. wt.) 1.0 mL blood was withdrawn from retro-orbital sinus using glass
capillary in EDTA coated eppendorf tube (3.0 mg/mL blood). The blood was
centrifuged (at 2500 g) at 4 °C for 10 minand the plasma was separated. Plasma
was diluted with normal saline (ratio 1:3) and used for analysis of total
cholesterol (TC), phospholipids (PL), triglycerides (Tg) by standard procedures.
18. Antioxidant activity (generation of free radicals): Super oxide anions (O2À) were
generated enzymatically by xanthine (160 mM), xanthine oxidase (0.04 U), and
References and notes
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nitroblue tetrazolium (320
lM) in absence or presence of compounds 4a–k
and 5a–e (100 g/mL) in 100 mM phosphate buffer (pH 8.2). Fractions were
l
sonicated well in phosphate buffer before use. The reaction mixtures were
incubated at 37 °C and after 30 min the reaction was stopped by adding 0.5 mL
glacial acetic acid. The amount of formazone formed was calculated
spectrophotometrically. In another set of experiment effect of compounds on
the generation of hydroxyl radical (OHÀ) was also studied by non-enzymatic
reactants. Briefly, OHÀ were generated in a non-enzymatic system comprising
deoxy ribose (2.8 mM), FeSO4Á7H2O (2 mM), sodium ascorbate (2.0 mM) and
H2O2 (2.8 mM) in 50 mM KH2PO4 buffer (pH 7.4) to a final volume of 2.5 mL.
The above reaction mixtures in the absence or presence of test compounds
(100 lg/mL and 200 lg/mL) were incubated at 37 °C for 90 min. The test
9. Jose Camarasa, M.; Fernandez-Resa, P.; Garcia Lopez, M. T.; De Las Heras, F. G.;
Mendez-Castrillon, P. P.; Felix, A. S. Synthesis 1984, 509–510.
10. Preparation of N-(2,3,4,6-tetra-O-acetyl-b-D-glucopyranosyl) thiosemicarbazide
compounds were also studied for their inhibitory action against microsomal
lipid peroxidation in vitro by non-enzymatic inducer. Reference tubes and
reagents blanks were also run simultaneously. Malondialdehyde (MDA)
contents in both experimental and reference tubes were estimated
spectrophotometrically by thiobarbituric acid as mentioned above.
(3): To a solution of compound 2 (5 g, 12.85 mmol) in CH2Cl2 (100 mL) was
added hydrazine monohydrate (1.2 g, 24 mmol) and the reaction mixture was
allowed to stir at room temperature for 2 h. The solvents were removed under
reduced pressure and the crude product was purified over SiO2 using hexane-
EtOAc (2:1) as eluant to furnish pure compound 3 (5.1 g, 94%).
Alloprinol, Mannitol and
a-tocopherol were used as standard drugs for
superoxide, hydroxylations and microsomal lipid peroxidation. All
experimental data were analyzed using Student’s t-test. Oxidized LDL was
compared with the test compounds treated oxidized LDL. The generation of
oxygen free radicals were compared in the presence and absence of test
compounds. The hyperlipidemic group was compared with control and
hyperlipidemic plus drug treated groups P < 0.05 was considered to be
significant.
11. Typical experimental condition for the preparation of compounds (4a–k): To a
solution of compound 3 (500 mg, 1.2 mmol) in 2-propanol (5 mL) was added
appropriate aldehyde or ketone (1.4 mmol) followed by 5 drops of AcOH and
the reaction mixture was allowed to stir at 80 °C for appropriate time (Table 1).
The solvents were removed under reduced pressure and the crude product was
purified over SiO2 using hexane-EtOAc (2:1) as eluant to furnish pure
compound 4a–k.
12. Typical experimental condition for the preparation of compounds (5a–e): To a
solution of compound 2 (500 mg, 1.28 mmol) in anhydrous CH2Cl2 (5 mL) was
added appropriate aryl hydrazine (1.1 mmol) and the reaction mixture was
allowed to stir at room temperature for 2 h. The solvents were removed under
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Ghosh, Samir
