1) Pheophorbide a, which is one of the decomposition products from chlorophyll, is widely distributed in foodstuffs, e.g. in salted vegetables and abalone viscera. This compounds is known to cause photosensitivity in human and animals. We have studied the mechanism of this phenomenon and shown that in the presence of pheophorbide a, visible light irradiation leads to the production of singlet oxygen. This, in turn, results in cell damage by oxydation of lipids, especially in the cell membranes. Further, we discovered that some kinds of vitamins display a preventive action against the photosensitivity caused by pheophorbide a. 2) It is well known that iodine dificiency caused edemic goiter in human subjects. On the other hand, many goitrogenic substances were found in various vegetables. We have studied the goitrogenic activity of soybean, and found that goitrogenic factors are some glucosides, saponine and isoflavonoides such as genistin and daidzin. In the process of this study, hyperplastic goiters were observed frequently in rats when they were given soybean under conditions of iodine deficiency for as long a term as 6 to 12 months. However, enlargement of thyroid and malignancy was completely inhibited when a small amount of iodine was added to the diet. The study was undertaken to determine the involvement of intestinal bacteria in the induction of goitrogens from soybean glucosides using germ-free mice. It was found that germ-free mice were more sensitive to iodine deficiency than conventional mice, and it was suggested that iodine metabolism was modified significantly with intestinal flora.
Reaction of 1-aminopyridinium and 1-aminoquinolinium mesitylene-sulfonates with ketenethioacetals [bis(methylthio)methylenemalonate (2a), 2, 2-bis(methylthio)-1-nitroethylene (8), and 2, 2-bis(methylthio)-2-sulfonylethylenes (10a, b)] gave N-substituted (1-pyridinio)amides (3a-d), (1-quinolinio)amide (5), pyrazolo[1, 5-α]pyridines (9a-f, 11a-g) and pyrazolo[1, 5-α]quinolines (12a-d) in good yield. 1-Aminoquinolinium mesitylene-sulfonate also reacted with 1, 2-dicyano-2-methylthio-1-(phenylsulfonyl)-ethylene (13) to give 2, 3-dicyanopyrazolo[1, 5-α]quinoline (14) in good yield.
Anti-ulcerogenic activities of the 70% methanol extracts of Panax ginseng C.A. MEYER were examined in rats. The 70% methanol extracts were found to show inhibitory effects on the Shay-ulcer in pylorus-ligated rats, serotonin- and endotoxin-induced ulcer. Furthermore, in order to examine the mechanism of the 70% methanol extracts for the serotonin- and endotoxin-induced ulcer, the effects of the 70% methanol extracts on the gastric mucosal blood flow in rats were investigated. As a result, the 70% methanol extracts inhibited the decrease of gastric mucosal blood flow induced by serotonin and endotoxin.
Solvent system optimization for digitalis glycosides was examined by silica gel normalphase liquid chromatography. 2-Propanol was added to a diluent, n-hexane, for increasing the solubility of water which was found to be a fine modifier for reducing peak tailing. In addition, ethanol was incorporated into the carrier solvent as a strong component to control the retentivity. As a systematically designed four component phase system, n-hexane-2-propanol-ethanol-water (52 : 20 : 25 : 3 v/v) was applied to the high performance liquid chromatographic analysis of digoxin in commercially available tablet preparations, using the clean-up procedure. The phase system reported here showed preferable selectivity for the simultaneous resolution of three digitalis glycosides.
The mechanism of resistance of Serratia marcescens to chlorhexidine was studied. Two strains of S. marcescens were isolated from the patients at Shinshu University Hospital. One of these strains was resistant to chlorhexidine and the other was sensitive. Minimum bactericidal concentration (MBC) of the resistant strain ((R1) was 4 mg/ml, whereas that of sensitive one (S1) was 0.08 mg/ml. Effects of chlorhexidine on the biosyntheses of macromolecules were comparatively examined on the S1 and R1 strains. The biosyntheses of protein, deoxyribonucleic acid, ribonucleic acid, and lipid in vivo were apparently inhibited by chlorhexidine in the S1 strain within 30s after the addition of chlorhexidine, while those of the R1 strain were not inhibited. Adsorption and permeation of chlorhexidine to the R1 and S1 strain were examined by using [14C]-chlorhexidine. The percent adsorptions of radioactive chlorhexidine onto the intact cells of the R1 and the S1 strain were about 74% and 78%, respectively. The percent distributions of radioactivity in the cell envelope of the R 1 and S 1 strain were about 29% and 42% to the total radioactivity incorporated into the cytoplasm of the R1 and the S1 strain were about 3% and 7%, respectively. These data suggested that membraneous permeability of bacteria played an important role in resistant mechanism.
In order to develop new cholelitholytic agents, sarcosine conjugated chenodeoxycholic acid (SCDC) and ursodeoxycholic acid (SUDC) were synthesized. The effect of oral administration of SCDC and SUDC on the formation of gallstones was studied in a mouse model of cholesterol cholelithiasis. Groups were fed a lithogenic diet with or without the sarcosine-conjugated bile acids for 12 weeks. Cholesterol gallstones formed in 100% of the animals fed a lithogenic diet, 75% of those fed 0.03% of SCDC and SUDC, and 35% and 15% of those fed 0.15% of SCDC and SUDC, respectively. Acute toxicity studies show that the LD50 of SCDC and SUDC for the mice administered orally were 3750 mg/kg and >10000 mg/kg, respectively. Intestinal absorption studies using bile fistula rats show that more than 90% of both the sarcosine-conjugated bile acids administered intraduodenally were excreted in the bile within 24 h. No changes of the administered bile acids were seen during the absorption from the intestine, the passage of the liver, and the excretion into the bile. Studies of enzymatic hydrolysis with cholyglycine hydrolase of SCDC and SUDC show that the sarcosine-conjugated bile acids were not hydrolysed at all within 120 min, while glycine-conjugated bile acids were hydrolysed completely.
In order to develop the aqueous infusion packed in a polypropylene (PP) container, the properties of the PP containers, packed with distilled water and a physiological saline, and then stored under several conditions, were examined concerning the following items. 1. Physicochemical properties of the container. 2. Components soluble in the liquid inside the container. 3. Components of the head space gas in the container. Under every storage condition, it was found that the pp container met the requirements of Plastic Container for Aqueous Infusion, General Test, Pharmacopoeia of Japan (JP) and any changes in the physicochemical properties were not observed. No aldehyde, ketone, and carbonyl compounds derived from the oxidative degradation of PP were detected in the inside liquids. The number of insoluble matters was not more than that in the regulation of Infusions, General Rules for Preparations, JP and did not increase at every storage conditions. The saturated and unsaturated hydrocarbons, of which the number of carbon was 4 to 7, having largest quantity among these hydrocarbons, turned out to be about 120 ppb. It is clarified that these hydrocarbons result from the partial pyrolysis of pp through the heat treatment of the container.
A new type of suppository with a hollow cavity into which drugs, in the form of powder, liquid, or solid could be placed was constructed. The advantage of the use of this hollow-type suppository is that interactions between drug and the oily-base suppository materials can be essentially eliminated. Brilliant blue FCF (BB) was selected as a model drug since the rate of dissolution could be observed visually. New type suppositories and conventional type suppositories were constructed with four different oily-base materials. The dissolution rates of BB in vitro from new-type could then be compared with those of conventional-type suppositories made from the same base material. It was found that with conventional construction, the rates differed considerably depending on the base material. With the new type no difference was found between the four materials tested. Similar results were obtained if the BB was added to the suppository as a fine powder, aqueous solution or macrogol mixture. Plasma concentration of BB were measured in rabbits after the rectal administration of the suppositories. These data on bioavailability showed that the hollow-type suppositories were less influenced by kinds of the base material than were the conventional types.
Dissolution behavior from solid dispersion system obtained from nifedipine and enteric coating agent (hydroxypropylmethylcellulosephthalate and copolymer based on methacrylic acid and methacrylic acid methyl ester) were investigated. The X-ray diffraction data suggested that nifedipine was present in its amorphous form in these systems. Dissolved amounts of nifedipine from the system in JP X 1st fluid (pH 1.2) were poorer than that of nifedipine crystalline. But the dissolution rate of nifedipine from the system, especially nifedipine-hydroxypropylmethylcellulosephthalate, was very rapid and the system showed supersaturated concentration in JP X 2nd fluid (pH 6.8). Dissolution behavior, that is dissolution rate and amount of nifedipine dissolved in the system was better in the case of the use of hydroxypropylmethylcellulosephthalate as carrier than that using copolymer based on methacrylic acid and methacrylic acid methyl ester. The above reason was discussed from the inhibitory effect of these polymers on the crystallization of nifedipine.
The coalescence of droplets was studied to elucidate the process of demulsification by using the apparatus based on Cockbain and McRoberts method. Hydrocarbons were used as oil phase and their droplets in water containing SDS were formed by rapid injection from capillary. The processes of coalescence were studied by the height of droplet column. The coalescence processes of the emulsion formed by the same samples were studied. The correlations between the coalescence of droplets and that of emulsions were found to be the same behaviour. Furthermore the droplets of coconut oil in the gum acacia solutions were also examined.
A computer-assisted structure-activity study using pattern recognition methods was applied to handle a set of 153 aromatic amines containing 37 carcinogens and 116 noncarcinogens. All pharmacological data were obtained from rat experimental animals and their liver active site. A set of 48 descriptors are generated by the ADAPT system program which was specially developed for structure-activity studies. These 48 descriptors are divided into 6 small subsets. Each subset contains 40 descriptors. Important descriptors of each subset that could support to dichotomize the aromatic amines into carcinogen and noncarcinogen are selected by the variance method. These selected descriptors are not influenced by any perturbed factors caused by statistics or pattern recognition methods. Most of these important descriptors are related to molecular sizes and shapes. Various pattern recognition methods are applied to classify the aromatic amine data set mentioned above. The linear learning machine, Bayesian quadratic discriminant and the iterative least-squares linear discriminant development routine obtained good classification values. the prediction value by the linear learning machine using the leave two out method achieved 90.1% as the highest value.
The effects of Shosaiko-to and its constituent crude drugs on the inflammatory responses were investigated in rats. The intraperitoneal administration of Shosaiko-to produced a dose-dependent inhibitory effect on the carrageenin-induced paw edema in rats. The suppression of the carrageenin-induced paw edema by Shosaiko-to decreased significantly in adrenalectomized rats. The elevations of serum and adrenal corticosterone levels were observed in rats treated with Shosaiko-to. Bupleuri Radix, Scutellariae Radix, Zingiberis Rhizoma and Ginseng Radix at an intraperitoneal dose of 100 mg/kg suppressed significantly carrageenin-induced paw edema, but Glycyrrhizae Radix, Zizyphi Inermi Fructus and Pinelliae Tuber did not. The active principles of Shosaiko-to producing anti-inflammatory effects were absorbed on Amberlite XAD-2 and eluted with ethanol. These results suggest that Shosaiko-to suppressed the carrageenin paw edema and that the anti-inflammatory action was partly due to an increase in adrenal function.
The present experiments were carried out to investigate the mechanisms of anti-inflammatory action of Shosaiko-to. The intraperitoneal administration of Shosaiko-to to rats produced increases in corticosterone level in the serum, adrenocorticotropine (ACTH) level in the plasma and cyclic adenosine monophosphate contents in the pituitary and adrenal gland. The elevations of serum corticosterone and plasma ACTH level induced by Shosaiko-to were inhibited by the treatment with dexamethasone (2.5 mg/kg, i.p.). Basal and ACTH-induced secretions of corticosterone from adrenal slices were not influenced by Shosaiko-to in vitro. Although Shosaiko-to did not increase plasma ACTH level in adrenalectomized rats, the action of Shosaiko-to to increase plasma ACTH level was observed in adrenalectomized rats which was administered corticosterone to maintain normal corticosterone level in the serum. Shosaiko-to produced a marked inhibition in exudate volume and leukocyte number in carrageenin-induced pleurisy of rats. Also chemotaxis of rat polymorphonuclear leukocytes and histamine release from rat peritoneal mast cells were suppressed by Shosaiko-to in vitro. Of the constituent crude drugs, Bupleuri Radix, Scutellariae Radix, Glycyrrhizae Radix and Ginseng Radix induced a marked elevation of serum corticosterone level. The histamine release from rat peritoneal mast cells was suppressed by Bupleuri Radix and Glycyrrhizae Radix. These results suggest that Shosaiko-to may produce an antiinflammatory effect by at least two mechanisms in rats. One is a stimulating action on the pituitary-adrenocortical function, and the other is direct inhibitions of histamine release from rat peritoneal mast cells and chemotaxis of polymorphonuclear leukocytes.
From the root of Scutellaria baicalensis GEORGI, three new flavonoids, II, III and IV, were isolated, together with 5, 7, 2'-trihydroxyflavone (I) and baicalein-7-O-β-D-glucopyranoside (V). The structures of II, III and IV were established as 5, 7, 2'-trihydroxy-8, 6'-dimethoxyflavone, 5, 7, 2', 5'-tetrahydroxy-8, 6'-dimethoxyflavone and 5, 2', 5'-trihydroxy-6, 7, 8-trimethoxyflavone, respectively.
From the root of Scutellaria baicalensis GEORGI, six new flavonoids (II-VII) were isolated, together with 5, 7, 2'-trihydroxy-8-methoxyflavone (I). The structures of II-VII were established as shown in Chart 1.
In order to investigate the constituents of Osmanthus species, the components of the leaves of Osmanthus fragrans LOUR. var. aurantiacus MAKINO were examined. Ligustroside, 10-hydroxyligustroside, oleuropein, 10-hydroxyoleuropein, 10-acetoxyligustroside, 10-acetoxyoleuropein, acteoside, phillyrin, (+)-pinoresinol-β-D-glucopyranoside, apigenin, luteolin, quercetin, apigenin 7-O-β-D-glucoside, luteolin 7-O-β-D-glucoside, p-hydroxyphenethyl alcohol, 3, 4-dihydroxyphenethyl alcohol, p-hydroxyphenethyl β-D-glucoside, 3, 4-dihydroxyphenethyl β-D-glucoside, p-coumaric acid, ferulic acid, caffeic acid, oleanolic acid, ursolic acid and β-sitosterol were isolated from the methanolic extract.
A new and sensitive method for the fluorometric determination of homovanillic acid using o-dianisidine-peroxidase system was established. Under the optimum conditions, the calibration curve was linear from 0.5μg/ml to 20μg/ml (0.5-20μg/tube) of homovanillic acid, and coefficients of variation was below 5 percents. Salicylic acid, glucose, urea, p-hydroxyphenylacetic acid, and homogentisic acid did not interfer with the value of homovanillic acid, but the coexistence of uric acid, dopamine, and serotonine caused clearly decreases in the homovanillic acid value by the present method.
The disintegration time of a tablet using hydroxypropylcellulose (HPC) as a binder increased under the condition of high humidity. The mechanism of the increase of disintegration time was studied. The penetration rate of disintegration medium into the tablet decreased with an increase in moisture content of the tablet. Two reasons for this phenomena were considered. The first is a change in the pore size distribution of the tablet by absorption of moisture. The second is a change of the properties of HPC in the pore of the tablet. From the following two results, the second reason was considered to be main one. 1) The pore size distribution of the tablet was not changed under the condition of high humidity. 2) The disintegration time increased by moisture absorption was decreased reversibly by drying.