Combined effect of aminopyrine with phenopyrazone (1, 4-diphenyl-3, 5-pyrazolidinedione) at the weight ratio of 1 : 1 on analgesic, antipyretic and anti-inflammatory activities was examined by oral administration in experimental animals. ED50 values (mg/kg) of aminopyrine, phenopyrazone, and the combination were 117.2 (n=85), more than 640, and 126.2 (n=85), respectively, for suppressing the phenylquinone-induced writhing in mice, 13.6 (n=56), more than 640, and 10.2 (n=56), respectively, for reducing the yeast-induced fever in rats, and 182.3 (n=128), 574.0 (n=47), and 216.4 (n=78), respectively, for inhibiting the carrageenin-induced edema in rats. Their LD50 values in mice were 522.6 (n=110), more than 3200, and 848.5 (n=110) mg/kg, p.o., respectively. Anti-writhing activity of aminopyrine in the mice pretreated with phenopyrazone daily for 5 days was significantly better than that in the mice pretreated with a vehicle 24 hr after the last administration, and the plasma level of phenopyrazone at the time was almost nil. The anti-writhing activity of aminopyrine (60 mg/kg) was significantly potentiated by the addition of 60 mg/kg of phenopyrazone but not with 6 mg/kg. The parallel shift of the dose-response curve for aminopyrine to the left was significant by the addition of a fixed dose (60 mg/kg) of phenopyrazone to each dose of aminopyrine. These results suggest that analgesic and antipyretic activities of aminopyrine were potentiated by the combined use with phenopyrazone at the weight ratio of 1 : 1 without any potentiating effect on acute lethal toxicity, though the anti-inflammatory activity tended to be potentiated, and the synergism between the drugs appeared to be due to the raised sensitivity of the acting site.
The interaction between alkanes with various chain length (solute) and liquid crystal substance, cholesteryl myristate (ChM : crystal→smectic→cholesteric→isotropic liquid), was studied by gas-liquid chromatography (GLPC), its stationary phase being composed of ChM and solid support, Chromosorb W. With variations in the gas flow-rate, ChM content in the stationary phase, and column temperature, the specific retention volume of each solute, Vg, was measured and its standard free energy, enthalpy, and entropy of solution in ChM, Δsoln G2&ohbar;, Δsoln H2&ohbar;, Δsoln S2&ohbar;, were calculated. The gas flow-rate had a slight effect on Vg of the stationary phase with a large ChM content, although this effect could be nullified by the extrapolation of Vg to zero flow-rate. Logarithm of Vg or Δsoln G2&ohbar; depends almost linearly on the ChM content. From the analysis of Δsoln G2&ohbar; vs. temperature relationship, the effect of ChM content on Δsoln G2&ohbar; was found to be due to the contribution of Δsoln S2&ohbar;, i.e., destruction of the liquid crystal structure by solute molecules.
In order to estimate the impact strength of tablets reasonably, load impact tests and static load tests were carried out using the Konsistometer. The impact force for 50% tablet breaking (F50) and the static load for 50% tablet breaking (L50) were obtained from the values of the impact acceleration of the load or the static load, and were compared with the tablet hardness which is regarded as the static compression breaking strength. Deformation of tablets was also examined from the viscoelastic considerations. From the results of these experiments, the following conclusions were obtained. 1) The values of F50 and L50 were directly proportional to the values of hardness. This means that each of these three indices is similar measure of the compression breaking strength, but the values of L50 were 5-6 times greater than those of F50. 2) The breaking deformation (Db) of lactose/starch tablets was approximately constant, regardless of the compaction pressure for tableting or the hardness of tablets. On the other hand, the Db values of crystalline cellulose tablets increased with decreasing compaction pressure. 3) WEO was directly proportional to the hardness for each kind of tablets. Crystalline cellulose tablets which have higher values of Db and total work for tablets breaking deformation (Wb) gave higher values of WEO than those for lactose/starch tablets.
Through the observation of chemical shifts in nuclear magnetic resonance spectra, formation constants of 1 : 1 metal complexes with nucleosides were found in dimethyl sulfoxide at room temperature. The complex formation constants of CdCl2-adenine and CdCl2-benzyladenine are 1.6-1.7 times larger than the values for the HgCl2 complexes of adenine and benzyladenine. On the other hand, the formation constant of HgCl2-6-mercaptopurine is larger than the value of CdCl2-6-mercaptopurine. 6-Methylmercaptopurine did not complex with HgCl2 or CdCl2. In nucleosides, the complex formation constant with HgCl2 is in the order of 4-thiouridine, cyclocytidine, and cytidine. The complex formation with HgCl2 or CdCl2 was not observed with nucleoside analogs prepared from 5, 6-diamino-1, 3-dimethyluracil with D-glucose or D-galactose.
The binding properties of prazepam, desalkylprazepam, and diazepam to human serum albumin (HSA) were studied. Protein-bound drug and unbound drug were separated by Sephadex gel filtration. Binding ability of the test drugs to HSA decreased in the order of diazepam, desalkylprazepam, and prazepam. HSA was found to have just one class of binding site for prazepam, but had two different binding sites for diazepam and desalkylprazepam. Binding of the primary site for diazepam and desalkylprazepam was high, whereas that of the secondary site was low. The reason why the intact prazepam level in plasma was very low after its oral administration was discussed from the viewpoint of the binding properties of prazepam to HSA.
The metabolic fate of Prazepam (PZ), (7-chloro-1-cyclopropylmethyl-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepin-2-one) was studied in rats. After oral or intravenous administration of 5-14C-PZ, the radioactivity was detected in various tissues, indicating extensive uptake of this compound. A higher level of the radioactivity was seen in the liver, small intestine, stomach, adrenal, and kidneys. After oral administration, the radioactivity level in the brain was nearly comparable to that in the blood. In the case of intravenous administration, however, the level in the brain was several times higher than that in the blood. In each route of administration, fecal excretion mostly resulting from biliary excretion was the major elimination route of this drug. A major metabolite in the urine was 4'-hydroxydesalkylprazepam (4'-HDPZ) sulfate which was also found in the bile. Other major metabolites in the bile were 3-hydroxyprazepam (HPZ) glucuronide and metabolite A which was present as free and conjugated forms. Most of the biliary metabolites were found in the feces as a free form with the unchanged drug. Metabolite A was isolated from the bile and identified as 3'-hydroxydesalkylprazepam (3'-HDPZ). Major metabolites in the plasma, liver, and kidneys were desalkylprazepam (DPZ), oxazepam (OX), 4'-HDPZ, and its sulfate. In the brain, DPZ and OX were the major metabolites, but a small amount of the unchanged drug and HPZ was also detected. These results suggest that the main reactions in the PZ metabolism are C3-hydroxylation to HPZ and N-dealkylation to DPZ followed by aromatic hydroxylation to 3'- and 4'-HDPZ or C3-hydroxylation to OX. These metabolic patterns were not altered after a repeated administration. The relationship between the pharmacological activity and metabolism of this drug is also discussed.
Metabolism of prazepam (PZ), diazepam (DZ), and desalkylprazepam (DPZ) by rat liver microsomes was studied. When PZ concentration was low, PZ was mainly metabolized to DPZ and oxazepam (OX), whereas DPZ and 3-hydroxy-PZ were the major metabolites when PZ concentration was high. As minor metabolites, 4'-hydroxy-DPZ (HDPZ) and 4'-hydroxy-OX (HOX) were also formed from PZ. In the case of DZ, 3-hydroxy-DZ (HDZ) was the major metabolite regardless of DZ concentration, and the amount of demethyl-DZ was much less than that of HDZ. OX, HDPZ, and HOX were also detected but their amount was extremely small. In the case of DPZ, OX was the major metabolite regardless of DPZ concentration. HDPZ and HOX were also detected in trace quantities. The liver microsomal enzyme activities for PZ, DZ, and DPZ were studied. The total metabolite formation activity for PZ was almost equal to that for DZ, but it was twice as active as that for DPZ. The C(3)-hydroxylation activity for DZ was approximately four times the activity for PZ or DPZ. The N(1)-dealkylation activity for PZ was four times as active as that for DZ. The C(4')-hydroxylation activity for PZ was almost equal to that for DPZ but was six times as active as that for DZ. The repeated administration of 10 mg/kg/day of PZ for consecutive 28 days did not have any significant effect on the liver microsomal enzyme activities for PZ.
The activities of Cl- ion (aCl-) of six phenothiazine hydrochlorides (PTZ·HCl) in aqueous solutions were obtained by EMF measurements of Ag-AgCl electrode. Above a concentration specific for each PTZ·HCl, the chemical was considerably adsorbed on to the surface of the electrode with increase in the concentration. The EMF obtained was not equal to the intrinsic EMF corresponding to aCl- in the bulk solution. It was necessary to eliminate the adsorption effect by an appropriate method. The critical micelle concentration (cmc) was determined by means of the crooked points on the plot of log aCl- vs. PTZ·HCl concentration. The crooked point of each PTZ·HCl aqueous solution corresponded to the break point on the plot of pH vs. PTZ·HCl concentration. It was interpreted that the aggregation of PTZH+ ion was accompanied with hydrolysis reaction, PTZH++H2O⇄PTZ+(H3O)+. Taking the hydrolysis into account, the abnormality of aCl- in the region just above cmc was thermodynamically explained.
Ferric hydroxide was peptized by pouring the precipitate into aqueous solutions of various amino acids and boiling for 8 hr. The degree of peptization was evaluated by the concentration of ferric hydroxide in the sol thus formed. For this purpose, the concentration was determined by chelatometry after decomposition of colloidal particles into ferric ions. The adsorption of hydrogen ions on ferric hydroxide particles in amino acid solutions was measured as well. In the case of glycine, L-α-alanine, DL-α-aminobutyric acid, L-valine, and L-isoleucine, the addition of a small amount of hydrochloric acid increased the degree of peptization. However, no peptization took place in the case of L-arginine and L-lysine hydrochlorides, and L-methionine. In spite of no difference between glycine and L-α-alanine in the amount of hydrogen ion adsorbed on ferric hydroxide particles, the degree of peptization and stability of sol were higher in L-α-alanine than in glycine. It was found that L-α-alanine is one of the most effective peptizer of ferric hydroxide among amino acids, and it was concluded that the adsorption of amino acids except L-methionine promotes peptization.
Effect of NADPH on lipid peroxidation in microsomes with the soluble fraction of rat liver was investigated. NADPH was found to inhibit lipid peroxidation in 9000×g supernatant fraction when over a certain amount of the fraction was added to the reaction mixture, even in the presence of Fe2+ or Fe2+-ADP which is known to activate the microsomallipid peroxidation induced by NADPH. In addition, the soluble fraction-stimulated lipid peroxidation in microsomes was found to be effectively inhibited by NADPH in the presence of a certain amount of the soluble fraction. NADPH extended the duration of depressing the lipid peroxidation in 9000×g supernatant fraction followed by a rise in the formation of lipid peroxides. When Zn2+, an inhibitor of glutathione reductase, was added to 9000×g supernatant fraction or the soluble fraction-microsomes system, lipid peroxide formation was not depressed by the addition of NADPH. Inversely, the addition of GSSG to the soluble fraction-microsome system was found to enhance the antiperoxidative effect of NADPH. These results indicate that a factor, which participates in the inhibitory action of NADPH on the microsomal lipid peroxidation in the presence of the soluble fraction, may be a glutathione reductase system present in the soluble fraction. Therefore, it is thought that NADPH does not only stimulate the microsomal lipid peroxidation, but also acts as an inhibitory factor on the lipid peroxide formation in microsomes together with the soluble fraction.
Action of parotin on the circulating leukocytes of a rabbit was examined and the following facts were found. 1) Intravenous injection of parotin in rabbits resulted in the leukocytosis-promoting activity in the serum 4-6 hr later. 2) The leukocytosis-promoting factor in the rabbit serum after administration of parotin is inactivated by heat treatment at 100°for 5 min, but is a stable, non-dialyzable substance after heat treatment at 56°for 30 min or at 37°for 60 min. 3) The leukocytosis-promoting factor was isolated from the serum after parotin administration, using ammonium sulfate fractionation, DEAE-cellulose column chromatography, gel filtration over Sephadex G-200, isoelectric focusing with carrier ampholyte, and preparative slab disc electrophoresis. It was found that this factor has a molecular weight of ca. 160000 by gel filtration over Sephadex G-200 and disc electrophoresis, and has an isoelectric point at pH 5.02. The factor is not stained by basic fuchsin. From sodium dodecyl sulfate treatment, this factor was considered to be formed from two subunits with a molecular weight of ca. 80000. This factor was found to be immunochemically different from parotin.
In order to study the photostability of phylloquinone (K) and menaquinone-4 (MK-4) in injection and infusion solutions, high-speed liquid chromatographic determination was examined. Determination of K and MK-4 was made on a Permaphase ODS column with mobile phases of dioxane-H2O (68 : 32) and methanol-H2O (89 : 11), respectively for K and MK-4. Internal standards used were α-heptadecanyl naphthoate for K and α-pentadecanyl naphthoate for MK-4. K and MK-4 in the injection and infusion solution were stable when stored in the darkness, but decomposed by photo-irradiation according to the pseudofirst order reaction to yield phyllochromenol and menachromenol respectively as main decomposition products. These decompositions occurred by irradiation with the light of 350-500 nm region, with the maxima around 430 nm. Photodegradation of K and MK-4 was inhibited by excluding a light in the 350-500 nm region.
Sedimentation volume (V2/V1), yield value (F), and differential viscosity (df/dg), obtained from the flow curve, were studied by changing the concentration of sodium chondroitin sulfate (Ch-Na) and/or simple salt in the concentrated system of aqueous kaolinite suspension. The above parameters decreased with increasing concentration of Ch-Na or simple salt by breaking the card-house structure of kaolinite (dispersing effect), but these parameters increased in the region of a high concentration of simple salt, showing aggregation between faces and/or edges of kaolinite by electrostatic shielding effect of the added salt. On the other hand, relative viscosity (ηrel) decreased with the concentration of simple salt or Ch-Na because the hydrodynamic radius of secondary particle became smaller with the concentration of addend, and the structure of secondary particle was more compact than card-hause structure at high concentrations of simple salt. The mean diameter (dm) of secondary particles, measured by the sedimentation balance in a dilute suspension, also decreased with the concentration of addend, as did F, df/dg, and V2/V1. It was concluded that the dispersing effect corresponds to making small particles in dilute suspension and to breaking the card-house structure in the concentrated suspension. The effective dispersing concentration of Ch-Na increased with the concentration of kaolinite because dispersing effect appears owing to its adsorption onto kaolinite but, in the case of simple salt, the concentration dependence was small because the ionic strength of the medium is effective for dispersing kaolinite particles by shielding the electrostatic attractive force between the face and edge of kaolinite particles.
Three new compounds, (±)-car-3-ene-2, 5-dione (II), 3, 4, 5-trimethoxytoluene (VI), and 2, 3, 5-trimethoxytoluene (VII), were isolated from the essential oil of the subterranean part of Asiasarum heterotropoides (Aristolochiaceae). Structures of these substances were determined from chemical and spectroscopic evidences. Orcinol aromatics, most of which have been reported as metabolic product of molds, were confirmed to be synthesized in higher plants. It is an interesting fact that, in the case of Asiasarum sieboldii, the proportion of these primitive compounds is larger in the earlier growing period of the individual.
The degree of swelling of bovine eye-lens capsule was measured in solutions of various pH and ionic strength, and the effect of swelling on the membrane permeability was examined. It was found that, when ionic strength of solution was low, capsule swelled both in a more acidic region than the isoelectric point and in a more alkaline region than the isoionic point of collagen, which is the main component of the capsule. The swelling is, therefore, due to repulsion among electric charges of the collagen. The void ratio, the apparent membrane constant, and the membrane permeability of capsule increased in propotion to swelling of the capsule. On the other hand, when ionic strength of the solution was high, the capsule was dehydrated and shrank. Because of the decrease of free water in the capsule by dehydration, the membrane permeability decreased and, in the case of an electrolyte, the transport number was affected in the manner that the transport number of a cation increased and that of an anion decreased. This resulted because the frictional resistance of the matrix of dehydrated capsule was smaller for cations than for anions since the radius of cations was smaller than that of anions.
Knoevenagel condensation of o-nitrobenzaldehydes (Ia-d) with ethyl 2, 4-dioxopentanoate (II) gave 3-acetyl-2, 4-dihydroxy-4-(o-nitrophenyl) crotonic acid lactones (IVa-d), which take the open-form, 3-(o-nitrobenzylidene)-2, 4-dioxopentanoic acids (IV'a-d), in a polar solvent. Reductive cyclization of IVa-d in the presence of 5% Pd-C as a catalyst gave α-hydroxy-2-methyl-3-quinolineacetic acid 1-oxides (Va-d) in a good yield. These were easily derived to esters (VIa-c, VII, and VIIIa), α-keto esters (IXa-c), α-hydroxyimino esters (Xa-c), and aldehydes (XIa-c). The reaction of VIa with acetyl chloride followed by deoxygenation with phosphorus trichloride afforded methyl α-acetoxy-2-methyl-3-quinolineacetate (XVII), which was hydrolyzed to α-hydroxy acid (XVIII). Reduction of Xa with zinc dust in acetic acid and acetic anhydride gave α-acetamido ester (XIX), which was hydrolyzed to α-quinaldylglycine (XX). The reaction of VIa with acetic anhydride gave methyl α-acetoxy-2-acetoxymethyl-3-quinolineacetate (XXI), which was oxidized to the N→oxide derivative (XXIII). Hydrolyses of XXI and XXIII gave δ-lactones (XXII, XXIV). The reaction of IVa-d with diazomethane in ether gave 3-acetyl-4-hydroxy-2-methoxy-4-(o-nitrophenyl) crotonic acid lactones (XXVIa-d), the structure of which was determined from UV spectra. Similar reductive cyclization of XXVIa-d in alcohols (R3OH) gave alkyl (R3) 1, 4-dihydro-1-hydroxy-α-methoxy-2-methyl-4-oxo-3-quinolineacetates (XXVIIa-d, XXVIIIa-XXXa) in a fairly good yield. The mechanism of the formation of quinoline N-oxide derivatives and 1, 4-dihydro-4-quinolones was discussed. Finally for biological tests, resolution of Va with cinchonidine was carried out, and its effect on Aspergillus flavus and soy beans was tested.
The acid catalyzed cyclization reaction with formaldehyde of the secondary amides possessing a hydroxyl group at α, β, or γ-position was attempted. New oxazolidinone derivatives (1 to 7), 1, 3-oxazinone derivatives (8 to 12), and 1, 3-oxazepinone derivatives (13 to 16) were obtained with the formation of 1, 3, 5-dioxazepinones (Va and Vb) as the by-product of 1 and 2, respectively.