The products from the positive Voges-Proskauer reaction namely, the reactions of 1-phenylpropane-1, 2-dione (PPD) with 1, 1-pentamethyleneguanidine (PMG), and those from the negative Voges-Proskauer reaction, namely the reactions of benzil with PMG, PPD with benzamidine, and of benzil with benzamidine were isolated, and the relation between the structural difference in these products and their color developments was elucidated. The reaction of PMG with PPD afforded 4-hydroxy-4-methyl-5-phenyl-2-piperidino-4H-imidazole [Ic], which is known as the intermediate in the Voges-Proskauer reaction. The reactions of PMG with benzil, benzamidine with PPD, and benzamidine with benzil afforded 2, 4, 4/2, 5, 5-trisubstituted-2-imidazolin-4/5-one-s [II, IV, V] and 4-hydroxy-4, 5-diphenyl-2-piperidino-4H-imidazole [III], respectively. It was found that II, IV, and V were formed through the benzilic acid rearrangement, while the intramolecular rearrangement was not involved in the formation of III. II, IV, and V did not react to give any color development in the presence of alkali, but III which had a similar skeleton to that of Ic gave II with an intramolecular rearrangement. Since Ic gave the pigments (A, B, and C), whose structures were already elucidated, by the release of formaldehyde from 4-hydroxy and 4-methyl groups of 4H-imidazole or the reaction with 1-naphthol in the presence of alkali, this reaction process was found to be of importance for exhibiting the positive Voges-Proskauer reaction. A tautomerism was found between IV and V by examination of their IR spectra.
Rat liver preparation metabolized ο-chloro-α-(tert-butylaminomethyl)-benzyl alcohol hydrochloride (C-78) to 4-hydroxy-C-78 (II), 3-hydroxy-C-78 (III), 1-(ο-chlorophenyl)-1, 2-ethanediol (V), and ο-chloromandelic acid (VI), as in in vivo experiments. Further, two metabolites, 5-hydroxy-C-78 (VII) and 1-(ο-chlorophenyl)-2-aminoethanol (VIII) were newly obtained from in vitro system. However, 4-hydroxy-5-methoxy-C-78, a major metabolite in vivo, was not detected. The microsomal fraction had all the activity for forming the metabolites in the presence of NADPH-generating system. Two types of the reaction by the microsomal fraction, such as ring hydroxylation (major) and cleavage of the side chain (minor), were differently affected by inhibitors, and this fact suggested that both reactions were catalyzed by different enzyme systems.
From the ether extract of Cnidium officinale MAKINO (Umbelliferae ; Japanese name "Senkyu"), six phthalides were isolated by column and preparative thin-layer chromatography. One of them was a new phthalide and named senkyunolide. The structure of senkyunolide was determined by correlation with the known butylphthalide as (3S)-3-butyl-4, 5-dihydrophthalide (VI). The mass spectroscopy was found to be the most satisfactory method for structural analysis of phthalides and showed a definate systematic fragmentations starting from the cleavages at the allylic position.
The metabolic fate of ο-chloro-α-(tert-butylaminomethyl)-benzyl alcohol hydrochloride (C-78) in the rat was studied by oral administration of 14C-C-78, and the bronchodilating activities of the metabolites were also examined. The major metabolites in the urine, bile, and feces were unchanged C-78 (I) and 4-hydroxy- (II), 3-hydroxy- (III), and 4-hydroxy-5-methoxy-C-78 (IV), and their corresponding conjugates. As the minor metabolites, ο-chlorobenzoic acid, ο-chloromandelic acid, and 1-(ο-Chlorophenyl)-1, 2-ethanediol were detected by gas chromatography. These results suggested that the major metabolic pathway of C-78 in the rat is hydroxylation of the benzene ring. Although IV had no activity, II had a stronger bronchodilating activity than C-78, and III was as effective as C-78, suggesting the participation of the metabolites in the bronchodilating action and durability of C-78.
The stability against separation of oil phase from 25% liquid petrolatum-water emulsions stabilized with 0.5% mixed surfactants of sorbitan-fatty acid monoesters and polyoxyethylene sorbitan-fatty acid monoesters was evaluated by centrifugation. Reproducibility of the centrifugal test was very good. Result of the test after aging was closely related to the induction period of oil phase separation and the quantity of separated oil phase after a long-time storage of the emulsions. Change of the specific interfacial area with time revealed that the stabilizing effect is better for a single hydrophilic surfactant with maximum hydrophile-lypophile balance (HLB) than for any mixed surfactant in the relation of HLB value with stability. It thus appears that the so-called "optimum HLB value" determines the best dispersibility, because the specific interfacial area immediately after emulsification shows a maximum value at the "optimum HLB value" while the best stabilizing effect is not attained.
The concentrations of surfactant and oil, emulsifying method, and interfacial tention were examined as the possible factors affecting the stability of liquid petrolatum-water emulsions stabilized with mixed surfactants of sorbitan monolaurate and polyoxyethylene sorbitan monolaurate in its relation to hydrophile-lypophile balance (HLB) value. The so-called "optimum HLB value" is obtained by the water phase separation test. This is closely related to the particle size and flow properties, but has little or no relation to the properties of interfacial film. On the other hand, oil phase separation test can be considered to evaluate the coalescence of oil droplets in relation to the properties of interfacial film. Change in the surfactant composition affected oil phase separation in relation to the interfacial tention. The conventional screening method for emulsifying agants by the HLB concept was criticized on the basis of the present investigation.
Reaction of 1-nitro-2, 2-bis(methylthio)ethylene with amines gave the corresponding monoamine and diamine derivatives which were nitroenamines. Heterocyclic compounds, pyrrole, 1, 2, 3, 6-tetrahydropyrimidine, thiazole, and 2(1H)-pyridone derivatives, were synthesized by using the reaction of these nitroenamines.
Effect of sodium copper chlorophyllin (SCC) on the lipid peroxidation in rat liver homogenate was examined. Formation of lipid peroxides was estimated by the thiobarbituric acid (TBA) method, and SCC was found to inhibit the increases of TBA value caused by both ferrous ion and L-ascorbic acid. The ferrous ion and ascorbic acid-stimulated lipid peroxidation in liver homogenates was also expressed as the increase in the oxygen consumption of liver homogenates. SCC repressed the acceleration of oxygen consumption of liver homogenates by both ferrous ion and ascorbic acid. The antioxidative effect of SCC was thought to be attributed neither to free cupric ion nor to pro-oxidative action of SCC on ascorbic acid.
The quality estimation of Ch' an Su (Senso) based on the chromatographic determination of three major bufadienolides (resibufogenin, cinobufagin, and bufalin) was investigated. Reliable methods established by the use of gas-liquid chromatography, high performance-liquid chromatography, and thin-layer densitometry were found to be routinely applicable for bufadienolide determination of "Rokushingan, " a commercial drug containing.
Eye-lens capsule has one layer of epithelium inside its anterior part and this epithelium was found to be a negatively charged layer from measurement of the membrane potential of epithelium-capsule.2) Theoretical equations were derived for the membrane potential and the membrane permeability coefficient of the membrane, composed of a positively charged membrane and a negatively charged membrane. The membrane composed of a capsule, which was positively charged, and a cellulose membrane, which was negatively charged, were considered as a model membrane of epithelium-capsule and the theoretical equations were applied to this composed membrane. These theoretical values were in good agreement with the observed values when the parameter values of the membrane charge density θ* and the transport number of anion t*- measured for each component membrane were used.
Some pharmacological actions related to analgesic and anti-inflammatory activities of 5-aminomethylsalicylic acid (AMS) were investigated. (1) Analgesic activity of AMS (100-200 mg/kg, s.c.) was about twice as potent as that of salicylamide (200 mg/kg, s.c.) and the effect persisted for about 150 min. (2) AMS (100 mg/kg, s.c.) inhibited carrageenin-induced paw edema by approximately 36% and the activity was the same as that of sodium salicylate (100 mg/kg, s.c.). (3) AMS (200 mg/kg, s.c.) increased the excretion of uric acid and the activity was slightly more potent than phenylbutazone (30 mg/kg, p.o.). (4) AMS (10-5 M) inhibited the serum plasmin activity. (5) AMS (200 mg/kg, s.c.) slightly enhanced the increased vascular permeability induced by histamine. (6) AMS did not affect the effluent volume of the perfused ear vessel in the rabbit. (7) AMS (100-200 mg/kg, s.c.) slightly increased the blood sugar level in the rabbit.
The mechanism of anti-inflammatory action of 5-aminomethylsalicylic acid (AMS) was investigated. (1) AMS as well as salicylate showed more significant inhibition of the leucocyte migration, in normal rats than in adrenalectomized rats. AMS (100 mg/kg, i.p.) inhibited it by 50.4% in normal rats and 28.7% in adrenalectomized rats. (2) AMS decreased the ascorbic acid level in the adrenals by 22.8 and 52.3% at the dose of 100 and 200 mg/kg, i.p., respectively. (3) Although a single treatment with 100 mg/kg, i.p., was ineffective, 7-day administration of AMS (50 mg/kg, i.p.) increased significantly the rate of corticosterone release from isolated rat adrenals (49.0%) and its blood level (44.0%). (4) Oxygen uptake and glucose consumption by isolated rabbit leucocytes were markedly increased with AMS (15 mg/dl), while these effects were not observed in the system with an addition of E.coli. (5) AMS (2.4×10-5 M) inhibited markedly the respiration and coupled phosphorylation in the cyclophorase system from brain and liver. (6) With 7-day administration of AMS (50 mg/kg, i.p.), the hemogram of rats was not affected. (7) AMS inhibited the heat coagulation of bovine serum albumin by 6.8 and 13.6% at the concentration of 15 and 30 mg/dl, respectively.
A practical method for the quantitative determination of primary and secondary amines by gas chromatography was developed. Amines were readily converted into their N-n-amyloxycarbonyl derivatives by a simple procedure involving n-amyloxycarbonylation with n-amyl chloroformate in aqueous alkaline medium, and the resulting derivatives were extracted with isopropyl ether. The derivatives thus obtained have good gas chromatographic properties and are stable under normal laboratory conditions. The reproducibility of response was found to be good for derivatives carried through the entire chemical and chromatographic procedure, and the calibration graphs were linear.
Partial synthesis of dihydroburnamicine (IV) and naturally occurring 2-acylindole alkaloid, burnamicine (I), from hirsutine (IIIb) and geissoschizine methyl ether (II), respectively, is described. By the first route, 7-acetoxy-7H-hirsutine methiodide (VIb) was converted to 2-acylindole derivative (VII), whose conversion into IV has already been reported, 4 but the conversion of II into the corresponding 2-acylindole derivative (IX) was unsuccessful by this route. In the other route, ethyl chloroformate as a reagent for C/D ring cleavage of indole alkaloids was used. Dihydrocorynantheine (IIIa) and IIIb reacted with ethyl chloroformate to give the urethane derivatives (XIIa, b), which were converted to IV by several steps but the conversion of methane derivative (XX) prepared from II into the desired 2-acylindole derivative (XXI) was not successful. Reaction of hirsutinol (XXIV), derived from IIIb with ethyl chloroformate, gave the urethane derivative (XXV), which was also converted to IV. By the same route, geissoschizol (XXVIII), derived from II, was converted to I. This partial synthesis of I revealed its absolute configuration and geometry of the ethylidene group. It was very interesting that I and IV having the same absolute configuration at C-15 exhibited nearly opposite circular dichroism curves.
N-Nitrosopyrrolidine (I), N-nitroso-2-pyrrolidone (II), and N-nitroso-5-methyl-2-pyrrolidone (III) were prepared, and fed in drinking water to male Sprague-Dawley rats for 365 days to compare the effects of their chemical consitution on the carcinogenicity of these compounds. In the animals surviving beyond 276 days, I induced hepatocellular carcinoma in 15 of 18 animals (83%), II induced colon carcinoma in 1 of 15 animals, and III did not induce carcinoma in any of 16 animals. Obviously I, II, and III have entirely different pattern of carcinogenicity. LD50 (oral, measured by Litchfield-Wilcoxon method in male ddY mice) increased in the order of II, III, and I, and the mutagenic activity ("Rec-assay, " using B.subtilis mutant, H-17 and M-45) increased in the order of I, III, and II. These orders have little relation to their carcinogenicity.
Several peptides, H-Arg-Pro-Lys-Thr-OH (I), H-Thr-Arg-Pro-Lys-OH (II), H-Thr-Arg-Pro-Arg-OH (III), H-Thr-Lys-Pro-Lys-OH (IV), H-Thr-Lys-Gly-Arg-OH (V), H-Thr-Lys-Pro-Orn-OH (VI), H-Thr-Orn-Pro-Arg-OH (VII), and H-Thr-Gly-Lys-Pro-Arg-OH (VIII), related to tuftsin were synthesized. I exhibited a phagocytosis-stimulating activity, to the same degree as tuftsin.
A mixture of cholesteryl chloride and cholesteryl nonanoate shows a liquid crystalline state, termed compensated nematic (S0), in a limited range of composition (60-70 molar% of cholesteryl chloride). The compensated nematic state transformed to cholesteric liquid crystal by the addition of cetyl alcohol, but the addition of anthracene did not induce such a change.