Journal of pharmacobio-dynamics最新文献

In vitro effects of 3-formylamino-7-methylsulfonylamino-6-phenoxy-4H-1-benzopyran-4-on e (T-614), a novel antiinflammatory compound, on the production of interleukin-1 (IL-1) and/or interleukin-6 (IL-6) by human monocytes and the THP-1 cells of a human monocytic cell line, were examined. T-614 inhibited the release of immunoreactive IL-1 beta from these cells stimulated with lipopolysaccharides (LPS) in a dose-dependent manner (0.3-30 micrograms/ml). The release of IL-6 from THP-1 cells, as determined by the assays for its hepatocyte-stimulating activities and immunoreactivities, was inhibited by T-614 with the IC50 values of 2.0 and 6.6 micrograms/ml, respectively. Northern blotting analysis using LPS-stimulated THP-1 cells indicated that the inhibitory effect of T-614 on IL-1 beta production is caused by the suppression of IL-1 beta mRNA expression. The inhibition of cytokine production by T-614 may provide an important insight into the additional mechanisms contributing to its antiinflammatory activities.

Disulfide bridges in fibrinogen (Fbg) were cleaved by cis-diamminedichloroplatinum(II) (cis-DDP). Incubation with 120 molar excess of cis-DDP at pH 7.4 and 37 degrees C in the presence of EDTA resulted in cleavage of seven disulfide bridges out of 29. In the presence of calcium, however, the number of cleavages were reduced to four. The result indicates that calcium, which has three high affinity sites on Fbg, protects disulfide bridges from the rupture. Thrombin clottability was examined by turbidity measurement. The cis-DDP treated Fbg was shown to give a decreased fiber thickness. As the cleavage proceeded, the clotting ability of Fbg decreased.

The pharmacokinetics of zonisamide (ZNS) and the effects of phenobarbital (PB), valproic acid (VPA), carbamazepine (CBZ) and phenytoin (PHT) on ZNS kinetics were investigated in rats. The effects of other antiepileptics on the serum protein binding, erythrocyte distribution and metabolism of ZNS were also studied in vitro to elucidate the mechanism of pharmacokinetic interaction of ZNS. ZNS showed a linear disposition kinetics after oral administration of ZNS within the dose examined. Moreover, the pharmacokinetic behaviors of ZNS were not altered after multiple dosing. The decreased t1/2 value of ZNS by PB or CBZ pretreatment and the increased Vd/F value of ZNS by VPA pretreatment were observed, although it showed no marked effect of PHT on ZNS kinetics. The enhanced metabolism of ZNS was observed by PB or CBZ pretreatment from an in vitro metabolism study. The serum protein binding and erythrocyte distribution of ZNS showed no significant change in the presence of other antiepileptics in vitro. These results indicate that the decreased t1/2 value of ZNS is attributable to the enzyme inducing effect of PB or CBZ, and that neither protein binding nor erythrocyte distribution of ZNS could be the reason for the increased Vd/F value of ZNS by VPA coadministration.

An enzyme immunoassay (EIA) for assessment of cell activation and proliferation was developed by the use of monoclonal antibodies (MoAb) directed against a ubiquitous growth-associated antigen, gp125. Test cells distributed in a microtest plate were labeled with anti-rat gp125 MoAb, B3, for rat cells or anti-human gp125 MoAb, HBJ127, for human cells and subsequently with rabbit anti-mouse immunoglobulins. The cell-bound antibody was assessed by a colorimetric enzyme assay using horse-radish peroxidase-modified protein A and 2,2'-azinobis(3-ethylbenzthiazoline sulfonic acid). To terminate the reaction and to make the reaction mixture transparent, sodium dodecyl sulfate (SDS) was added to the mixture. The intensity of the developed color was measured by use of a multiwell scanning spectrometer. The titration curves obtained by the EIA for cells were practically similar to those obtained by the conventional 3H-thymidine (Tdr) uptake method, and the appropriate cell number to be used for the assay was indicated to be 3 to 50 x 10(3) cells per well. This method was applicable not only for quantitation of cell number of growing cells but also for measuring mitogenic responses of lymphocytes as revealed by the data obtained from Con A stimulation of lymphocytes. These results indicate that this new EIA method using anti-gp125 antibodies is useful for quantitative assays of cell growth and cell activation in the research fields of oncology and immunology.

The influence of para substituent of phenol on the property of phospholipid dependence of uridine diphosphate-glucuronyltransferase (UDPGT) was studied using hepatic microsomes of the rats. para Substituents used were p-ethyl, p-tert-butyl and p-phenyl. Glucuronidations of phenol and its derivatives were inhibited with each other. When phospholipids of the microsomes were removed with phospholipase A2, Vmax and Km decreased in the order phenol, p-ethylphenol, p-tert-butylphenol, p-phenylphenol, indicating that this change by delipidation increased with van der Waals volume (Vw) of para substituent. Arrhenius plots of glucuronidation for phenol and p-ethylphenol exhibited a change in slope, responding to thermotropic property of the membrane labelled with 1,6-diphenyl-1,3,5-hexatriene (DPH). In contrast, p-tert-butyl and p-phenyl substituents showed linear Arrhenius plots. The activation energy (Ea) for glucuronidation of p-phenylphenol was increased by delipidation, whereas Ea of other substrates did not show any remarkable change.

Constant exposure of mastocytoma P-815 cells to adenosine 3',5'-cyclic decylphosphoramidate (1), which is permeable to the cell membrane and resistant to the action of phosphodiesterase, caused a dose-dependent (1 to 50 microM) inhibition in the synthesis of DNA and cell proliferation. Pretreating the cells with compound 1 (20 microM, 4 h) caused considerable inhibition of the incorporation of [3H]thymidine ([3H]TdR) into [3H]deoxythymidine 5'-triphosphate ([3H]dTTP) and that of [14C]hypoxanthine into nucleic acid, but not the synthesis of [14C]dTTP from [U-14C]aspartate. These results indicate that compound 1 preferentially inhibits the salvage synthesis of intracellular nucleotides and nucleic acids. Thymidine kinase, a key enzyme in salvage synthesis of nucleotides, was almost undetectable in cells pretreated with compound 1 at 20 microM for 4 h or at 5 microM for 15 h. On the other hand, compound 1 activated partially purified cAMP-dependent protein kinase A from bovine heart. Judging from these observations, it is likely that compound 1 readily permeates the cell membrane, activates cAMP-dependent protein kinase, then inhibits the salvage synthesis of nucleotides and nucleic acids by inhibiting thymidine kinase, which results in the inhibition of cell growth.

The characteristics of calcium antagonism and vascular effect of 2-methoxyethyl(E)-3-phenyl-2-propen-1-yl(+/-)-1,4-dihydro-2,6-dime thyl-4-(3- nitrophenyl)pyridine-3,5-dicarboxylate (FRC-8653) were investigated. FRC-8653 inhibited an increase in intracellular free calcium concentration during membrane depolarization in PC12 cells. FRC-8653 also inhibited the specific binding of 3H-nitrendipine to cardiac membranes, in a similar manner to nifedipine and nicardipine. FRC-8653 inhibited KCl- and CaCl-induced contractions in isolated rabbit aorta, but failed to affect norepinephrine-induced contraction. The vasorelaxing effect of FRC-8653 in rabbit aorta developed more slowly than those of nifedipine and nicardipine. In ex vivo experiment, the inhibitory effect of orally administered FRC-8653 against KCl-contraction in rat aorta lasted longer than that of nifedipine. These findings suggest that FRC-8653 dilates blood vessels by blocking calcium influx via dihydropyridine-sensitive, voltage-dependent calcium channels and that the vascular effects are slow in development and long in duration.

The interaction of pranoprofen, pranoprofen glucuronide and pranoprofen methylester with human serum albumin (HSA), was investigated by equilibrium dialysis and spectroscopic techniques. The binding affinities of pranoprofen glucuronide and pranoprofen methylester to HSA were found to be almost the same, although they were remarkably small as compared to that of the parent compound, pranoprofen. Pranoprofen and pranoprofen methylester showed stereoselective affinities to HSA. It was found from the competitive displacement experiments using the fluorescent probes that the specific binding site for pranoprofen was site II, the diazepam site, and that the binding sites of pranoprofen glucuronide and pranoprofen methylester were site I, the warfarin site. In addition, from the binding data with modified HSA, it seemed that tyrosine-411 was specifically involved in the pranoprofen binding. The absorption spectral changes which accompanied the binding of pranoprofen and pranoprofen methylester to HSA or detergents implied that the HSA binding site of pranoprofen consisted of a cationic site on the surface of the albumin molecule with a hydrophobic region to accommodate the aromatic ring and that the binding site for pranoprofen methylester seemed to occupy a wide hydrophobic area. These limited data indicated differences in the location and microenvironments of binding sites for pranoprofen and its glucuronide on the HSA molecule.

The H2-receptor antagonists, cimetidine, ranitidine, famotidine and nizatidine, were tested for their effect on the isolated smooth muscle strips from the rabbit stomach fundus and sigmoid colon. These H2-receptor antagonists were found to possess a concentration-dependent contractile effect on the above smooth muscle preparations and the order of potency was: ranitidine = nizatidine > famotidine > cimetidine. In addition, the smooth muscle preparations from the sigmoid colon were significantly more sensitive to the above compounds than the smooth muscle preparations from the stomach fundus.

Glucocorticoids inhibit IgE antibody-mediated passive cutaneous anaphylaxis (PCA) and chemical mediator-induced cutaneous reactions elicited in the mouse ear. In the present study, we investigated the effect of actinomycin D, a protein synthesis inhibitor, on dexamethasone-caused inhibition of PCA and histamine-induced cutaneous reaction in the mouse ear. Tyrosine aminotransferase (TAT) activity in the liver, which was estimated as an index for protein synthesis, significantly increased by the administration of hydrocortisone, prednisolone and dexamethasone. Significant increase in TAT activity was observed from 2 h after glucocorticoid administration and peaked at 4 h, and declined gradually thereafter. Cycloheximide even at high doses of 100 and 300 mg/kg failed to affect the increase in TAT activity by dexamethasone. On the contrary, actinomycin D at doses of 1 and 10 mg/kg abrogated the TAT activity increase by dexamethasone almost completely. Treatment with 1 mg/kg of actinomycin D, however, failed to affect the inhibition of PCA and histamine-induced cutaneous reaction by dexamethasone. These results suggest that glucocorticoids exhibit their inhibitory action of PCA and chemical mediator-induced cutaneous reactions in mice through a mechanism resistant to actinomycin D treatment.