Physical constants, i.e. surface tension, partial specific volume, reduced viscosity, and refractive index, were measured with a mixture of 10 kind of solubilizing agent and water. Mixed solutions of having only hydrophilic group, ethylene glycol, diethylene glycol, triethylene glycol with water did not show a break in the foregoing physical properties and these stubstances were considered not to undergo association. On the other hand, substances having both hydrophobic and hydrophilic groups, such as cellosolve, butylcellosolve, carbitol, butylcarbitol, hexylene glycol, dipropylene glycol and diethylin with water showed a break in physical properties, suggesting that association occurred in these mixtures at a certain concentration of these substances, resulting in a sudden change in their internal structure. A mixed solutions of butylcellosolve, butylcarbitol, hexylene glycol, and diethylin with water showed a clear break point in physical properties suggesting that they are highly associated in the solution.
Perhydrobenzo [c] quinolizine derivatives (VI and VII) and benzo [c] quinolizidine derivative (IV) were synthesized from quinaldine by the routes shown in Charts 3 and 4, involving diethyl β-(2-quinolyl) ethylmalonate intermediates (XI). In connection with stereoisomer of perhydrobenzo [c] quinolizine (V), stereostructure of their parent skeleton were examined.
A peptide-like substance was isolated from pseudoglobulin fraction of the exudative inflammatory skin by carboxymethylcellulose. This substance showed an accelerating effect on the permeability of the venule wall, while it had no influence on the leucocyte migration. Further, contraction of the rat uterus was not induced by this permeability substance in vitro, unlike bradykinin. The interrelation between some anti-inflammatory drugs and the permeability substance was also examined by dye leakage method using pontamine sky blue in rat skin.
4, 5-Diphenylpyrimidine (DPP) was obtained on heating deoxybenzoin (DB) with formamide and POCl3.1) Several deoxybenzoins to generalize this preparative method, and following results were obtained. None from 4, 4'-di (CH3O) DB (I), 3, 4-3', 4'-bis (OCH2O)-DB (II), and 3'-(NH2)-4-(CH3O) DB (XIXc) ; 4-(4-CH3O)-5-(4-NO2) DPP (V) from 4-(CH3O)-4'-(NO2) DB (III), 4-(4-NH2)-5-(4-CH3O) DPP (VI) from 4'-(CH3O)-4-(NO2) DB (IV), 5-(4-NH2) DPP (XIa) from 4'-(NO2) DB (Xa) ; 5-(3-NO2) DPP (XIb) from 3'-(NO2) DB (Xb) ; 4-(4-CH3O)-5-(3-NH2) DPP (XIc) from 3'-(NO2)-4-(CH3O) DB (Xc) ; 6, 8-dimethoxy-3-phenylisoquinoline (XIII) from 3', 5'-di(CH3O) DB (XII) ; 5-(3, 4-di (CH3O) DPP (XVI) with some 6, 7-dimethoxy-3-phenylisoquinoline (XV) from 3', 4'-di(CH3O) DB (XIV) ; 5-(3-CH3O)- DPP (XIII), from 3'-(CH3O) DB (XVII) ; 5-(3-NH2) DPP (XXa) from 3'-(NH2) DB (XIXa) ; 5-(3-HO) DPP (XXb) from 3'-(HO) DB (XIXb) ; 4-(4-CH3O)-5-(3-HO) DPP (XXd) from 3'-(HO)-4-(CH3O) DB (XIXd). From these results, the following will be concluded : (1) To form isoquinolines, at least two +R groups (e.g. CH3O) must be substituted in the meta and other positions of benzyl in DB. When there is only one substituent at the meta-position of the benzyl group, even though more effective as NH2 or OH, isoquinoline ring-closure does not take place but the corresponding pyrimidines are obtained. (2) NO2 substituent is often reduced to NH2 in this procedure. (3) The presence of +R substituent at the para-position of the benzoyl group of DB deactivates vates this reaction and -R substituent accelerates it.
An evidence was provided that a kind of a complex is formed by interaction between sodium dehydroacetate with flavin in aqueous solution from the result of physicochemical examination proving the following four points. In the presence of sodium dehydroacetate, (1) quenching of fluorescence characteristic to flavin occurs, (2) a change occurs in the ultraviolet absorption spectrum of flavin, the absorption maximum at 445-450 mμ shifing to a longer wavelength region, with decrease in its intensity, (3) photodecomposition of flavin is markedly inhibited, and (4) solubility of sparingly soluble riboflavin increases markedly. Dissociation constant of flavin-sodium dehydroacetate complex was measured by the fluorescence method, absorption spectroscopy, and by the solubilization method and the value was found to be approximately 2 to 6×10-2 M/L (pH 5.8-8.6, 20°). The site of the dehydroacetic acid molecule interacting with flavin seems to be the enolic hydroxyl in 4-position of its pyrone ring.
N-Substituted 3-glucosyloxy-2-methyl-4-pyridones were synthesized from natural maltol glucoside (III) and primary amines. Aliphatic amines reacted readily with (III) in aqueous methanol under reflux, while aromatic amines required the addition of potassium hydroxide as a catalyst. The aglycones, N-substituted 3-hydroxy-2-methyl-4-pyridones, were obtained by acid hydrolysis of the above pyridone glucosides.
In order to examine the dispersibility of sulfisoxazole, sulfadimethoxine, and N1-acetyl-sulfisoxazole in aqueous sodium lauryl sulfate (SLS)-sucrose solutions, the apparent median redius (γmed) obtained from the particle size distribution as determined by the sedimentation balance and sedimentation volume were measured at sucrose concentrations of 0-30% and SLS concentrations of 1- 8×10-3 mole/liter, and the effect of SLS and sucrose on the dispersibility of those sulfonamides was investigated. It was thereby recognized that both γmed and sedimentation volume markedly decreased until the SLS concentration reachec 4×10<-3> mole/liter, beyond which they were almost constant. This SLS concentration was the same as that at the maximum point of ζ-potential. From these results, it was concluded that the dispersive sedimentation begins at the SLS concentration of 4×10-3 mole/liter, and that the addition of sucrose has no effect on this concentration. Correlation of dispersibility with the amount of SLS adsorbed on the sulfonamides was examined. It was found that the adsorbed amount of SLS required for dispersive sedimentation of the three sulfonamides was 47, 51, and 50% of the saturated amount adsorbed, respectively.
Hexaethylditin (II) reacted with several halomethylbenzenes to give triethyltin halide (V) and the corresponding dimerization products ; 63% bibenzyl (III) from benzyl bromide (Ib), 19.3% trans-stilbene (VIII) from benzal halide (VI), 92.5% cis- and trans-α, α'-dichlorostilbene (XI, XII) from benzotrichloride (IX), and 83.2% tetraphenylethylene (XV) from diphenyldichloromethane (XIV). It has been proved that the stilbene derivatives are formed in this reaction, by way of the dehalogenation reaction of vic-dihalides which were initialy produced as an intermediate. Reaction of DDT (XX) with II, however, mainly afforded DDD (XXI), and trans-p, p'-dichlorostilbene (XXIII) was obtained from the reaction of XXI with II.
Ammoxidation of 2-furaldehyde was carried out using multi-staged reactor of gas-liquid counter-flow type. This reaction was examined in methanolic solution at 30°, in the presence of cupric chloride and sodium methoxide. Mass balance equations in steady state, (19)-(22), were obtained based on the rate equations (5)-(8), previously proposed. The values calculated from these equations agreed well with experimental results, Therefore, the equations (5)-(8) can be utilized for designing any reactor to carry out this reaction.
Ammoxidation of 2-furaldehyde by using oxygen gas as an oxidant was examined in methanolic solution at 30°, in the presence of cupric chloride and sodium methoxide catalysts, and the kinetics of the reaction was studied. The presence of furimido-methyl ester in addition to 2-furonitirile was confirmed as main oxidation products. The ratio of the yield of nitrile to imido ester depends on the concentration of sodium methoxide. It became clear from kinetic analysis that the overall reaction proceeds according to the following scheme : [numerical formula] The reaction order and the rate constant of each step were determined. It was found that k1, k2, and k-2 are affected by the concentration of catalysts, and that the activity of cupric chloride decreases during reaction. The rate of reaction of 2-furaldehyde and the rate of formation of furfurylidenimine, 2-furonitrile, and furimidomethyl ester were determined as the equations (23), (24), (6) and (12), respectively.
Methylindole dithiocarboxylate derivatives were synthesized by the application of sodium hydride and carbon disulfide on indole derivatives, followed by treatment with dimethyl sulfate. Substitution reactions were carried out between methylthio group in these compounds and various amines to obtain various thioamide derivatives. Pyridazino-[4, 5-b] indole derivatives were synthesized by the use of these reactions.
A method for separatory determination of xylanase activity was investigated and a procedure was established as follows. Two ml of 1.0% carboxymethylxylan solution (250 μeq/g), 1 ml of 0.1M acetate buffer solution (pH 4.5), and 1 ml of enzyme solution in a 25 ml Nesler tube was incubated at 37°. After 30 min, xylose formed in the reaction mixture was determined by the Tauber -Kleiner's method (exo-xylanase activity). By using the Somogyi-Nelson's method instead of Tauber -Kleiner's method, free reducing sugar in the reaction mixture was determined (total xylanase activity). At the same time, a blank test was performed by replacing active enzyme solution with heat-inactivated enzyme solution and the amount of reducing sugar in a blank test was subtracted from that of the sample. Exo-xylanase activity subtracted from total xylanase activity was defined as endo-xylanase activity. One unit of xylanase activity was defined as the amount of enzyme which will liberated 1 mg of xylose under the above condition.
Appropriate working conditions have been studied to separately determine α-chymotrypsin and trypsin in a preparation which is used as an anti-inflamatory enzyme. Soybean trypsin inhibitor (STI) was used as the inhibitor. Trypsin activity in the preparation was completely inhibited by 8×10-3% (w/v) of STI in the reaction system. α-Chymotrypsin activity was partially inhibited and the value of remaining activity was 54.9±3.5%. Thus, it was possible to determine α-chymotrypsin and trypsin activity separately with 8×10-3% of STI. The reaction between trypsin and STI was nearly stoichiometric even in the presence of casein, a substrate. The determination was little affected by most of diluents and binders.
Qualitative and quantitative determinations were made on free amino acids contained in 75 kinds of domestic lichens, using an automatic amino acid analyzer (Hitachi Model KLA-2) and the total nitrogen was determined by the semimicro-Kjeldahl method. Correlation among the genus and species of lichens with the kind and content of amino acids and total nitrogen was examined. Some difference was observed even among the same genus and there seemed to be hardly any correlation. The kind of amino acids contained in individuals was 19 at the most, while only five kinds were present in some. As far as the 75 kinds of lichens examined in the present series of work were concerned, total nitrogen was less than 1% in 53 kinds, 1-2% in 12 kinds, 2 -3% and 3-4% in four each, and more than 4% in one. The total nitrosen content of Peltigera variolosa (MASS, ) GYEL. was the highest among all the lichens examined with 4.52%, but its free amino acid content rather low. This is probably due to the present of nitrogenous compounds other than amino acids. Of these 75 kinds of lichens, four kinds of Lobaria genus and three kinds of Sticta genus, both of Stictaceae family, had larger content of total nitrogen than other genus. It was of interest that taurine was found in 11 kinds of lichens including Alectoria sulcata YLN., Cetraria pseudocomplicata ASAHINA lad Cladonia aggregata (SW.) ACH. All Of the 75 kinds of lichens examined were found to have a few Ninhydrin-positive substances which could not be identified.
Oxidation of 2, 3-diphenyl-7-hydroxyindole (VI) with Fremy's salt, [ON (SO3K)2] gave 4, 7-dihydro-2, 3-diphenylindole-4, 7-dione (X), mp 226-227°, (yield, 20%), 6, 7-dihydro-2, 3-diphenylindole-6, 7-dione (XII), mp>280°, (yield, 61%). By the same reaction, its N-methyl derivative (VII) afforded 6, 7-dihydro-2, 3-diphenyl-1-methylindole-6, 7-dione (XXI), mp 251-253°, (yield, 38.5%), as a sole product.
The structural confirmations of the migration product of phenyl group in benzo [b] thiophene derivatives from C3 to C2-position during the following reactions are shown. a) Cyclization of 2-(2, 5-dimethoxyphenylthio) acetophenone (X) with polyphosphoric acid. b) Demethylation of 7-methoxy-3-phenylbenzo [b] thiophene (IV) with anhydrous aluminium chloride. Oxidation of 7-hydroxy-3-phenylbenzo [b] thiophene (VI) with Fremy's salt gives 4, 7-dihydro-3-phenylbenzo [b] thiophene-4, 7-dione (VII), mp 103-104° (yield 35.3%), and 6, 7-dihydro-3-phenylbenzo [b] thiophene-6, 7-dione (VIII), mp 158-159° (yield 40.7%). On the other hand, the same reaction of 7-hydroxy-2-phenylbenzo [b] thiophene (XIV) yields 4, 7-dihydro-2-phenylbenzo [b] thiophene-4, 7-dione (XV), mp 174-175° (yield 31.5%), as a sole product. Examination of the oxidation products from the nine 7-hydroxyheterocyclic compounds with Fremy's salt (Table I) shows that the presence of a phenyl group at C3-position is effective in producing o-quinone derivative. This phenomenon may be explained by the steric hindrance of a C3-phenyl group to the formation of a key intermediate (XXb) which is the important step towards formation of p-quinone.
Examination of the nuclear magnetic resonance (NMR) and the spin system of 12 compounds of heterocyclic quinone (Table I) showed that the quinoid proton signal of such a p-quinone appears as an A2 singlet or AB quartet, while that of an o-quinone appears as an AX type. Application of this observation to the oxidation products of 4-hydroxy-2-phenylbenzoxazole (II) and 1-hydroxydibenzo-p-dioxin (III) with Fremy's salt leads to the assignment of the p-quinone structures to them. Nuclear magnetic resonance spectra of 16 kinds of heterocyclic compounds containing several methoxy groups (Table II) indicated that the C4-methoxy signal of the compound bearing a phenyl group at C-3 shifted to a higher magnetic field by the anisotropic effect of the phenyl group at C-3.
The inhibitory effect of 1, 2-bis(2-benzimidazolyl) ethane and 1, 2-bis (2-benzimidazolyl)-ethanol derivatives on poliovirus and adenovirus replication was investigated by the tube method. Among the compounds tested, 1, 2-bis (5-(or 6-) chloro-2-benzimidazolyl) ethane (III), 1, 2-bis(5-(or 6-) methoxy-2-benzimidazolyl) ethane (V), 1, 2-bis (5-(or 6-) methyl-2-benzimidazolyl) ethanol (VII), and 1, 2-bis (5-(or 6-) methoxy-2-benzimidazolyl) ethanol (IX) exerted a clear inhibition of the cytophathic effect of poliovirus. In the concentration of 10-3-10-5 mole, V and IX showed complete inhibition of the plaque formation by poliovirus, and indicated that the inhibition index, which is expressed as the logarithmic difference of TCID50/ml in treated and untreated, was about 7.0. This inhibitory effect is greater than that of 2-(α-hydroxybenzyl) benzimidazole. These compounds also show almost complete block of the reproduction mechanism in intracellular viral growth of poliovirus.
Solubility and hydrolysis rate of 1-monolaurin in aqueous solutions were measured at 25°, 29.5°, and 35°. The concentration of 1-monolaurin was determined by colorimetry of glycerol formed from hydrolysed 1-monolaurin. Hydrolysis progressed very slowly and reversibly, until water was saturated with lauric acid. As a result of calculation from the temperature dependence of solubility, rate constants of forward and backward reactions, and equilibrium constant of hydrolysis, it has been found that heat of solution is ΔHs=4.04 kcal/mole, activation energies of forward and backward reactions are Ea=14.7 and E'a=10.6 kcal/mole, respectively, and enthalpy change of hydrolysis is ΔH=4.1 kcal/mole.
Effect of inorganic electrolytes on the interaction between Sephadex gels and solutes was examined in respect to gel filtration of several food dyes, oligo- and poly-peptides. Elution volume (Ve) of all the food dyes investigated and certain peptides involving lysine-vasopressin showed an increase to some appreciable extent according to lyotropic series of the anions (NCS-<Cl-<SO42-) in eluates. The linear relationships were obtained in the plots of the Ve values of the dyes on Sephadex G-25 and G-50 against swelling characteristics of the gels ; namely, effective diffusion coefficient of media in xerogels and ζ-potential of the gel particles in the electrolyte solutions. Because the diffusion coefficients of these dyes and peptides in these electrolyte solutions were smaller than those in deionized water solutions by porous disk method, it was suggested that the increase of the Ve values described above was mainly due to the enhancement of the aromatic sorption accompanied by a suppression of the charge effect in the presence of electrolytes. As a practical application of the effect of electrolytes on aromatic sorption, oxytocin and lysine-vasopressin were effectively separated on Sephadex G-50 column by the use of 0.1 F K2SO4 as an eluate, based on the difference in the location of a phenylalanyl residue in the latter peptide.
A simple apparatus is described for use in one-dimensional ascending paper chromato-graphy. A large glass tube of 4 cm in internal diameter and 45 cm in height is stoppered with a cork stopper which is provided, on the inner side, with five to six each of stainless steel staples, plastic rings, and paper clips, all parallel to each other. Strips of filter paper (2×40 cm) are held by paper clips, one to each clip, and a small glass rod is placed at the lower part of each paper strip, the end of which is immersed in the developing solvent. Eight of these glass tubes can be stood in a circle on a round holder of 22 cm in diameter. When compared with the existing appartus, this simple apparatus has the following advantages. (a) The cost is very low, (b) both the space and volume of solvent are small, (c) filter paper strips are kept apart, (d) there are no parts than can be damaged, (e) no special procedures are necessary for the operation or measurement of Rf, and (f) ascending speed of the solvent is slightly faster.
4-Oxo-1-piperidinecarboxylic acid esters (IV) were prepared by the reaction of 1-benzyl-4-piperidone (Ia) with alkoxycarbonyl chlorides (III) in 70-95% yields, but in the reaction of 1-methyl-4-piperidone (Ib) with ethoxycarbonyl chloride (IIIa), the yield of ethyl 4-oxo-1-piperidinecarboxylate (IVa) was 28%.
Neutral substances in plants of the Genus Corydalis (C. pallida var. tenuis YATABE, C. incisa PERS., and C. platycarpa MAKINO) were examined and two kinds of aliphatic alcohol, compd. A, mp 82.5-83°, and compd. B, mp 89-89.5°, were isolated as common components. Compd. A was found to be identical with nonacosan-10-ol, C19H39CH(OH)C9H19, from its mass spectrum and some degradation reactions of its ketoxime. Compd. B was identified with hentriacontan-1-ol, C31H63OH, from its infrared spectrum and mass spectrum of its trimethylsilyl ether.