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+ ATPase inhibitor. For this case the attack of glutathione takes place at the C-2 of the benzimidazole. For a possible mechanism see Weidolf, L.; Karlsson, K.-E.; Nilsson, I. Drug Metab. Dispos. 1992, 20, 262.
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0002384106
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6. Replacement of the 1,3,4-oxadiazole with other heterocycles or replacement of the sulfoxide group of 1,3,4-oxadiazole with a carbonyl or a sulfone resulted in compounds that were inactive in the HUVEC assay. 7. For 2-mercapto-1,3,4-oxadiazole synthesis see: (a) Hoggarth, E. J. Chem. Soc. 1952, 4811.
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0011962213
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note
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8. (a) The ELISA for the determination of presence's of ELAM-1 and VCAM-1 is as follows: 96-well plates were coated with 2% gelatin (1 h). The excess gelatin was aspirated from the wells. HUVE cells were seeded at 20,000 cells/well in a 200 μL volume. The test compounds were dissolved in DMSO at a stock concentration of 50 mM. When the monolayer reached confluence (24-28h after seeding), the test compounds were added to the wells at a final concentrations of 30 μM, 10 μM, 3 μM, and 1 μM. Each compound was added in duplicate. The plates were incubated for 4 h at 37 °C. Then, IL-1 β was added at a final concentration of 2 ng/mL and the plates were incubated for 4 h at 37 °C. The supernatant was removed from the plates by decanting and the wells were washed 3X with 1% BSA-Dulbecco's PBS. Primary antibody [7A9 for ELAM-1 (Otsuka Pharm.) and 2G7 for VCAM-1 (Otsuka Pharm.)] at a concentration of 2.5 μg/mL in a 50 μL volume was added to each well. The plates were incubated at 4 °C for 45 min to 1 h. The wells were washed 3X with 1% BSA Dulbecco's PBS. Secondary antibody (F (Ab')2 fragment biotinylated goat antimouse IgG-IgM (Jackson ImmunoResearch) at 50 μL/well at a final dilution of 1:1,000 was added and incubated for 45 min to 1 h at 4 °C. The wells were washed 3X with 1% BSA-Dulbecco's PBS. Streptavidin-peroxidase (Sigma) at 50 μL/well with a final dilution of 1:30,000 was added to each well and the plates were incubated at 4°C for 30 min. Substrate solution (OPD tablets, citric acid buffer and hydrogen peroxide solution) was added at a concentration of 100 μL/well. The plates were incubated for 15 min at room temperature on plate shaker and the reaction was stopped with 12.5% sulfuric acid. Plates were read for absorbance at 492 nm and the reading for the duplicate averaged.
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17
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0011921128
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note
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9. (a) Morphological changes to the monolayer was determined by light microscopy. Toxicity was determined by crystal violet and MTT staining assays in the presence of the compounds at a 30 μM dose.
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18
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0000129949
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10. (a) The 2-chloromethyl-3,5-dimethyl-4-methoxypyridine starting material can be prepared according to the procedure outlined in GB patent 2134523 and references ourtlined in Brändström, A.; Lindberg, P.; Bergman, N.; Alminger, T.; Ankner, K.; Junggren, U.; Lamm, B.; Nordberg, P.; Erickson, M.; Grundevik, I.; Hagin, I.; Hoffmann, K.; Johansson, S.; Larssom, S.; Lofberg, I.; Ohlson, K.; Björn, P.; Skånberg, I.; Tekenbergs-Hjelte, L. Acta Chem. Scand. 1989, 43, 536.
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Brändström, A.1
Lindberg, P.2
Bergman, N.3
Alminger, T.4
Ankner, K.5
Junggren, U.6
Lamm, B.7
Nordberg, P.8
Erickson, M.9
Grundevik, I.10
Hagin, I.11
Hoffmann, K.12
Johansson, S.13
Larssom, S.14
Lofberg, I.15
Ohlson, K.16
Björn, P.17
Skånberg, I.18
Tekenbergs-Hjelte, L.19
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19
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0028290402
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(b) 9-chloro-4-methoxy-6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridine precursor was prepared according to the procedure of Yamada, S.-I.; Goto, T.; Narita, S.-I. Chem. Pharm. Bull. 1994, 42, 718.
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Goto, T.2
Narita, S.-I.3
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0011981106
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note
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11. A selective alkylation (nucleophilic attack on the C-2 of the 1,3,4-oxadiazole) of a receptor or an enzyme in the signal transduction pathway is suspected. This may explain the decrease in activity for those compounds that appear to be more stable to attack at C-2 of the oxadiazole.
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0003009329
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La du, B. W., Mandel, H. G., Way, E. L., Eds.; Robert E. Krieger: Florida
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Mazel, P.1
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0026652811
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Each compound was tested on a group of seven mice
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13. Procedure conditions: C57BL/6 mice were shaven and an IV solution injection of 100 μL of BSA at 10 mg/mL in saline was administered. This was followed immediately by an ID 25 μL injection of anti-BSA (0.5 mg/mL solution) along with the test compound or vehicle on a predetermined site on the backside of the mice. Four hours post injection, the animals were sacrificed and the skins removed and assayed for myeloperoxidase (MPO) activity as a measure of neutrophils, according to the procedure of Mulligan, M. S.; Ward, P. A. J. Imm. 1992, 149, 331. Each compound was tested on a group of seven mice.
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Mulligan, M.S.1
Ward, P.A.2
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