Isolation of several kinds of bacterial pyrogen was attempted by paper ionophoresis. Pyrogenic substances were found in portions coloring with bromophenol blue and ninhydrin, and in a fluorescent portion.
Several kinds of microörganisms which are liable to be occluded in glucose solution were cultured in synthetic medium. Filtrate of such a culture medium was extracted and condensed by various methods and pyrogenic portion was isolated by paper ionophoresis. It was found that the substances which act as pyrogen varied by the method of extraction, sometimes the substance being a protein, at others, non-protein fluorescent substance, or substances giving positive ninhydrin reaction.
Gas flow meter fitted with an injection needle as a semi-variable capillary tube described herein is shown in Fig. 1. The simplicity for the exchange of capillary tube and compactness of its whole structure offer many advantages in carbon and hydrogen determinations; e.g. wide range of flow rate, saving of the space of apparatus, and easy handlings, in addition to rapid and accurate reading. The suitable dimensions of capillary tube are to be determined by the nomograph shown in Fig. 3, which was calibrated from Poiseuille's equation.
Inhibition of the growth of tubercle bacilli, in vitro, was effectively shown in 400, 000-800, 000 dilution of sodium p-aminosalicylate, 40, 000 dilution of 4-acetylamino-benzaldehyde thiosemicarbazone, and in 12, 800, 000 dilution of isonicotinic acid hydrazide, all used as the control, while 2, 4-dihydroxy-5-hexylbenzalisonicotinic acid hydrazone, one among the 64 compounds tested, effectively prevented growth in 2, 560, 000 dilution. The esters of p-aminosalicylic acid, reported in Part VI of this series, were again test-ed and some of them were found to possess powerful growth inhibitory action, as the following effectively prevented growth at respective dilutions: Phenyl ester at 5, 120, 000-12, 800, 000, o-tolyl ester at 2, 560, 000, p-tolyl ester at 2, 560, 000-6, 400, 000, m-tolyl ester at 1, 280, 000-6, 400, 000, o-methoxyphenyl ester at 6, 400, 000, and p-methoxyphenyl ester at 6, 400, 000-12, 800, 000. Other compounds showed only a weak growth-preventive activity.
In order to prepare the derivative of decarbonylmethylmatrinamine, its acylation was attempted by the application of tosyl chloride, 4-nitrobenzoyl chloride, and 3, 4-dinitrobenzoyl chloride, but acylated compounds could not be obtained in a crystalline state. Application of acetic anhydride yielded a monoacetate of needles, m. p. 123.5-124°, and the application of α-chlorocinchonic chloride gave the acid amide of needles, m.p. 162-164°. Application of sodium butoxide to the acid amide resulted in the formation of α-butoxycinchonic acid decarbonylmethylmatrinamide, which possesses a structure similar to Nupercaine but failed to show any striking local anesthetic action. Reduction of methyl methylmatrinate with lithium aluminum hydride gives the corresponding primary alcohol, i.e. methylmatrine alcohol as prisms of m.p. 82-83°, [α]D12: -35.52°, which gives a monomethiodide of needles, m.p. 177-178°, and a phenylurethane of prisms, m.p. 269-272°. Treatment of this methylmatrine alcohol with thionyl chloride in chloroform yields the corresponding chloride which, in a free state, transits to the quaternary ammonium chloride. This quaternary salt was found to be identical with matridine methochloride from which it was realized that the quaternary nitrogen in matridine methiodide is the tertiary amine-nitrogen that was newly formed by the reduction of the lactim ring in matrine. Boiling of methylmatrine alcohol with diphen-ylmethyl bromide in toluene results in the formation of a corresponding diphenylmethyl ether of prisms, m.p. 68-70°, which gives a monomethiodide of needles, m.p. 185-186°, picrate of needles, m.p. 220-222° (decomp.), a perchlorate of cubic crystals, m.p. 178-181° (decomp.), and a hydriodide of m.p. 217-219° (decomp.). The structure of this ether is similar to that of Benadryl but no antihistamine activity was found to be shown by it.
Cleavage reactions of ethers by the Grignard reagent were applied to several kinds of methoxydiphenyl ether derivatives and the ease or difficulty of the reaction and conditions of the reaction were comparatively examined (Table I).
A new high-frequency titration apparatus with a circuit shown in Fig. 1 was made, the feature being the arrangement of L1, L2 and L3 as shown in Fig. 2. L1 and L2 are combined oscillating circuits, each wound in a reverse direction so that the action of one would work in the reverse direction in the other and the difference is magnified to a fairly large degree. There is also no necessity of shielding each other, structure is extremely simple, sensitivity is great, and the stability is large, so that the response curve covers a wide range of concentrations, as shown in Fig. 6. The stability is better when the characteristics of the vacuum tubes V1 and V2 are uniform, one example of which is shown in Table II. Commercial alternate current can be used without any stabilization apparatus. Both the distance between the condenser plates inserted in the cell and the solution, and the plate area, closely affect the sensibility and possible concentration range to be measured (Table I). In the present apparatus, body effect is practically nil. In a high concentration range, the existing apparatus in which the sample was inserted into the coil gave response curves as shown in Fig. 8 and a similar curve (Fig. 9) is obtained by the present apparatus. This is because, in such a high concentration, the dielectric constant of the solution changes as shown in Fig. 11 (Huckel's theory confirmed by Hasted). Application of this to equation (1) in the text results in the decrease of C2, i.e. increase of Gp. Both the linearity and reproducibility are good. Some titration curves are shown in Fig. 12.
Preparation of coriaria dilactone by the condensation of 1, 1-dimethylindane and bromoisovaleryl chloride was attempted but only one of its isomers was obtained. Examination was also made of the synthetic method for 1, 1-dimethylindane and the structure of by-products obtained during the course of this synthesis.
In order to confirm the assumed structural formula of coriaria dilactone, derived from coriamyrtin by aromatization with hydriodic acid and oxidation, methylmagnesium iodide was applied to benzene-1, 2, 3, 4-tetracarboxylic acid dianhydride and the objective lactone was obtained. Since its potash fusion yielded phthalic acid, it was clarified that coriaria dilactone was different from the formula assumed earlier and that the gem-dimethyl groups were found to be present in a symmetrical position. These results have confirmed the fact that both coriaria lactone and coriaric acid are isomers of the structural formulae assumed previously.
Various properties of coriamyrtin were found to be similar to those of picrotoxinin and, therefore, their comparative examination was carried out. A structural formula of coriamyrtin was assumed on the basis of the formula given for picrotoxinine by Conroy by which the carbon skeleton formed a part of steroidal skeleton as in picrotoxinin and the reductive action of coriamyrtin was assumed to be due to its acetal group. Comparison of the infrared absorption spectra of coriamyrtin and its derivatives with those of picrotoxinin and its derivatives endorsed the assumed structure for coriamyrtin.
Bromination of butylsulfonylacetone (I) in glacial acetic acid yielded a monobromo compound, m.p. 119° (from alcohol), when the reaction mixture was allowed to stand for about 0.5-1.0 hour after discoloration of bromine. No depression of the melting point was observed on admixture with 1-butylsulfonyl-3-bromopropanone (II), obtained by the dry hydrogen bromide decomposition of the diazoketone (VIII) formed from butylsulfonylacetic chloride (VII) and diazomethane. The oily substance formed by the reduction of (II) with zinc dust in glacial acetic acid gives the same 2, 4-dinitrophenylhydrazone as that of m.p. 140° of (I). Condensation of (II) and thiourea gave 2-amino-4-butyl-sulfonylmethylthiazole (III). 1-Butylmercapto-3-chloro-2-propanone obtained from butylmercaptoacetic chloride (IX) through diazoketone (X) gave 2-amino-4-butylmercapto-methylthiazole (XII) by condensation with thiourea, and its acetate (XIII), m.p. 85°, is different from that of the isomer, m.p. 96-98°. Admixture of (IV), obtained by the oxidation of (XIII), with the acetate of (III) showed no depression, but the admixture with the isomer of the same melting point, m.p. 138°, i.e. 2-acetamino-4-methyl-5-butylsulfonylthiazole, showed depression. Reaction of 2-amino-5-bromothiazole (XIV) and alkylmercaptane, under the same conditions as those in the case of 2-amino-4-methyl-5-bromothiazole, gave the objective 5-alkylmercapto (XVI) and 5-alkylsulfonyl (XVII) derivatives, but these could not be detected by the method from butylmercapto-acetaldehyde diethyl acetal.
The reaction of 2-acetamino-4-methyl-5-bromothiazole (I) and sodium butoxide failed to give the objective 2-acetamino-4-methyl-5-butoxythiazole (II) under the various conditions tested and only a substance of unknown structure, melting at over 350°, was obtained. Bromination of butoxyacetone (V) followed by reaction with thiourea gave a minute amount of 2-aminothiazole derivative whose acetate melted at 116-117°. The isomer of (II), i. e. 2-acetamino-4-butoxymethylthiazole (XIII) was obtained by the acetylation of 2-amino-4-butoxymethylthiazole (XII), formed in a good yield by the condensation of thiourea and 1-butoxy-3-chloro-2-propanone (XI) obtained by the hydro-chloric acid decomposition of the diazoketone (X). Attempted preparation of 2-amino-5-butoxythiazole [Fig. 2, (III)] by the bromination of butoxyacetaldehyde diethyl acetal (I) followed by condensation with thiourea failed to provide a crystalline product, although the formation of 2-aminothiazole derivatives was proved by the diazo color reaction. The antibacterial activity of the thiazole derivatives thereby prepared are shown in Table I, and the ultraviolet absorption maxima of the thiazole derivatives reported in the present and the preceding papers are shown in Table II.
Periodic acid was applied to o-, m- and p-toluidine, m- and p-nitraniline, o-, m-, and p-aminobenzoic acid, aniline, monoethylaniline and its hydrochloride, dimethylaniline, and diphenylamine, and it was found that all these compounds, with the exception of nitranilines, absorbed the said oxidant at the same reaction rate as those of α-glycols. Moreover, the periodical progress of these reactions were found to be closely similar to the portion of periodical curve above 5 moles of absorption of periodic acid by arylamine-N-glucoside from which it was realized that the oxidation of arylamine-N-glucoside by above 5 moles of periodic acid was not due to formic acid and formaldehyde but by the action of liberated arylamine itself.
Reaction of 4-nitroquinoline 1-oxide and SH compounds was examined and it was found that this quinoline compound reacted with cysteine or thioglycollic acid to form 4-cysteinylquinoline 1-oxide and 4-thioglycollylquinoline 1-oxide.
2-Mercaptothiazolinone-(5), an isomer of rhodanine which is tentatively designated as isorhodanine, and its 4-arylidene derivatives are slightly less stable than rhodanine but their photographic emulsion effect is approximately equal to that of rhodanine. Addition of a small amount of isorhodanine and the derivatives results in the acceleration of fogging but their large addition results in the appearance of desensitizing and fog inhibiting actions. Hydrolysis of 4-arylideneisorhodanine with methanolic potash gives 5-substituted 4-carboxythiazolidine whose SH or =S group at 2-position is harmful in that it accelerates fogging. Difference of the substituent at 5-position was found not to give much influence on the emulsion effect. Antibacterial action of the compounds of this series was tested and p-nitrobenzylideneisorhodanine was found to be effective against various bacteria in 104 dilution.
Derivation of the SH group in 2-mercapto-4-carboxythiazolidine 5-substituted compound to methylmercapto group with diazomethane results in the loss of the photographic emulsion effect. It was found that its substitution to the hydroxyl group by treatment with alkaline hydrogen peroxide or the removal of the methylmercapto group with aluminum amalgam results in the appearance of a fog inhibiting effect. The imino group in the 3-position of the thiazolidine nucleus is the action point of the fog inhibiting effect and its acetylation makes it ineffective. The carboxyl group in the 4-position is connected with fog inhibition that its esterification results in the loss of such action. Carbamide group was found to be effective in fog inhibition.
High-frequency papyrography was carried out with the instrument shown in Fig. 1, whose electric source was stabilized by the use of a battery so as to make the measurement of each substance more sensitive. Various experimental conditions for the use of this instrument were found, especially for inorganic compounds. In order to attain sensibility, the filter paper should be dried perfectly by storing in a vacuum desiccator. For a qualitative purpose, however, the filter paper need not be so perfectly dried as the sensibility of substances is higher on a moist paper although the degree of sensibility is not constant.
Antibacterial activities were tested with 2-propylmercapto-, 2-butylmercapto-, 2-allylmercapto-, 2-thiophenyl-, 2-p-thiocresyl-, 2-p-chlorothiophenyl-, 2-methylamino-, benzylamino-, 2-anilino-3-chloro-, 2-anilino-3-methylmercapto-, 2, 3-di(thiophenyl)-, 2, 3-di-(p-thiocresyl)-, 2, 3-dichloro-, and 2-chloro-3-hydroxy-1, 4-naphthoquinone, and results are shown in Tables I and II. There were no compounds that showed better results than 2-methylmercapto-and 2, 3-di(methylmercapto)-1, 4-naphthoquinone, reported in the previous paper, but dichloro- and allylmercapto-1, 4-naphthoquinone were found to possess a fair degree of antibacterial power. 2, 3-Di(thiophenyl)-1, 4-naphthoquinone gives two kinds of crystals according to the conditions of recrystallization, one of purple leaflets, and the other of orange red needles, m.p. 148°. The former was found to transit to the latter without melting at 100-105° to orange red crystals that melt at 148°.
The two crystal forms of 2, 3-dithiophenyl-1, 4-naphthoquinone, the purple form and orange red form, were studied to find what kind of isomers they were. Molecular weight determination showed both to be a monomer, and the visible absorption spectra (Fig. 1) were also identical. Infrared absorption spectra (Fig. 2) failed to indicate any specific structural characteristics but such was indicated by the electron diffraction analysis. It is, therefore, assumed that the foregoing phenomenon is a monotropic dimorphism and a similar phenomenon was found to exist in structurally similar 2-p-thiocresy1-3-thiophenyl- and 2-p-thiocresyl-3-p-chlorothiophenyl-1, 4-naphthoquinone. Infrared absorption spectra of 1, 4-naphthoquinone, 2, 3-dimethylmercapto-, and 2-thiophenyl-1, 4-naphthoquinone were determined.
Synthesis of isonicotinic acid hydrazide (INAH) from citric acid was examined and its industrial application was perfected by the successful establishment of an advantageous process of preparing phosphoryl bromide and halogenation of citrazinic acid at an ordinary pressure (cf. Table II). Phosphoryl bromide was obtained in an 80% yield by the oxidation of phosphorus tribromide with potassium chlorate. Application of phosphoryl bromide to citrazinic acid, at ordinary pressure, yielded 2, 6-dibromoisonicotinic acid in a 70% yield.
Unsymmetric trinuclear and unsymmetric Hamer-type dyes were synthesized from the intermediate of binuclear trimethine, possessing an anilino group in the meso-position and with two different nuclei, such as 1, 1′-diethyl-meso-anilino-4, 2′-trimethylenequinocyanine dihalide, by the application of a cycloammonium salt different from that constituting the foregoing nuclei, such as 2-methylbenzothiazole ethiodide, in the presence of anhydrous potassium acetate, in acetic anhydride.
Hydrogenation of 2-methyl-4-amino-5-cyanopyrimidine under various conditions was observed and it was found that under some conditions a large amounts of secondary amine and Schiff's base are formed as by-products. In the presence of ammonia, the present reaction was found to constitute the relationship of reactions (1) and (2) >> reaction (3), while in its absence, the relationship was (1) > (3). It was also found that the foregoing by-products were formed through reaction (3) and that the formation ratio of these products were dependent on the temperature.
Reëxamination was carried out on the synthesis of dehydroacetic acid by the liquid condensation of ethyl acetoacetate, and it was found that silver oxide, barium oxide, and magnesium oxide catalyst also gave a good yield besides the usual catalyst of sodium bicarbonate. From the pyrolytic decomposition of dehydroacetic acid, especially in the case of using sodium bicarbonate, it was also observed that the dehydroacetic acid formed underwent pyrolytic decomposition at a reaction temperature above 190°. When the reaction temperature was kept below 190°, the theoretical yield against the consumed ester was increased although there was an increase in the recovery of the ester. This tendency was also observed in the case of magnesium oxide. Use of ion exchange resin, such as Diaion K (sodium form), Diaion A (hydroxyl form), and Amberlite IR-120 (sodium form), as a catalyst resulted in milder reactions and theoretical yield reached approximately 50%. Other catalysts give a large amount of tarry residue so that the ion exchange resins may possibly have specific catalytic effect.
Examination was made of the synthetic method for dehydroacetic acid by the vaporphase condensation of ethyl acetoacetate. The ester was passed through a reaction tube filled with pumice stone, heated to about 300°, at a comparatively slow space velocity, and the distillate thereby obtained was purified by fractional distillation. It was found that the best reaction temperature between 200 and 300° was at 300°, and that the formation of dehydroacetic acid was nil when sodium hydroxide was added as a catalyst to the pumice filling. Addition of magnesium oxide, zinc oxide, ferric oxide, aluminum oxide, reduced iron, or Amberlite IR-120 (sodium form) tended to cause the reaction to proceed in the direction of ester cracking and the results were much poorer than by the single use of pumice stone. The cracked gas contained ethylene and carbon dioxide. Use of activated carbon or silica gel in place of pumice stone resulted in vigorous decomposition of the ester and no formation of dehydroacetic acid was detected. It was also clarified that the increase of reaction surface by the change in the length of pumice layer resulted in the betterment of theoretical yield and reaction rate and that the deposition of carbon and other matter on pumice by its repeated use extremely injured the reaction. It was assumed from these experimental results that pumice stone is comparatively inactive to ester cracking and gives a good influence on the formation of dehy-droacetic acid.
Thermal analysis by moist-melting method was carried out on 16 kinds of two-component system composed of aromatic heterocyclic sulfa drugs, such as sulfapyridine, sulfathiazole, sulfadiazine, sulfamerazine, and sulfisoxazole, as one component, and acid amides, acid hydrazides, and phenols, as the other component, from which the ability of these compounds to form a molecular compound was examined. It was found that only sulfamerazine-nicotinamide and sulfamerazine-p-nitrophenol systems formed molecular compounds that undergo thermal decomposition while other two-component systems failed to form molecular compounds.
Thermal analysis by moist-melting method was carried out on 18 kinds of two-component system composed of barbituric acid derivatives, such as barbital, ethylhexabital, methylhexabital, and isoamylethylbarbituric acid, as one component, and aminopyrine, p-nitrophenol, and succinimide, as the other component. It was found that only three, barbital-resorcinol, ethylhexabital-aminopyrine, and isoamylethylbarbituric acid-aminopyrine systems formed molecular compounds.
Thermal analysis by the moist-melting method was carried out on 13 kinds of two-component system composed of several pyridine-carboxylic acid derivatives, such as nicotinamide and isonicotinic acid hydrazide, as one component, and aminopyrine, p-nitrophenol, and succinimide, which possess CO, NO2, OH, and NH radicals that form hydrogen bond comparatively easily, as the other component. Seven of these systems were found to form molecular compounds. p-Nitrophenol was found to form molecular compounds with all the pyridine-carboxylic acid derivatives tested, while aminopyrine failed to bond with any of them. Succinimide was found to form molecular compounds with nicotinamide and isonicotinic acid hydrazide, but not with the others.
High frequency titration of organic bases and amino acids in non-aqueous solution was carried out and good results were obtained. The apparatus used was the one described in the previous report, the solvent used was a mixture of benzene, methanol, and glacial acetic acid, and the titration was made with 0.1N acetic acid solution of HClO4. Titration could be carried out with methanol content of 20-45%. Glacial acetic acid was used in 2-10cc. amount when it was needed for dissolving a compound, glacial acetic acid containing 1-9% of water being satisfactory, but its use is not essential. The use of over 99.8% glacial acetic acid was found to cause acetylation that the foregoing hydrous acid seemed to give a better result. The titration was possible when the dissociation constant of a base in aqueous solution is approximately 10-10, but not under 10-12. The apparent dissociation constants of the bases of amino acids are generally under 10-12, but they were satisfactorily titrated. Amino acids that are insoluble in methanol or hydrous acetic acid were dissolved in a known excess of HClO4 normal solution and back-titrated with 0.1N glacial acetic acid solution of Na2CO3 (present as CH3COONa). The amount of samples used was 4-130mg., and the concentration of the solution to be titrated was 3.6×10-4 to 2×10-2 moles. The present method, which uses a large amount of methanol or a slightly hydrated glacial acetic acid, seems to be inconsistent with the existing method of titrating in non-aqueous solutions but it bases the change in conductivity and is different from the usual methods which depend on hydrogen ion activity.
Diazo compounds react with malonic acid ester in weak acid medium, in the presence of cuprous salts, to form phenylmalonic acid derivatives. The yield is comparatively good when reaction is carried out on solid diazonium salts in an anhydrous state.
1) Alcohols: As high boiling alcohols, β-phenylethyl, benzyl, and α-phenylethyl alcohol, and benzhydrol were reacted with formamide, and it was found that three forms of reaction occured to give respective product of a formate, formylamine, and ether. In other words, the difference in chemical structure appeared in the reaction so that formylamine and ether were chiefly formed from benzhydrol while the formate formation was more likely to occur from β-phenylethyl alcohol. 2) Ketoximes: Reaction of acetophenoxime, benzophenoxime, acetoxime, and cyclohexanone oxime with formamide resulted in the reductive activity of formamide to give one product of corresponding formylamines. 3) Aromatic Nitroso Compounds: Reaction of nitrosobenzene, 4-nitrosophenetole, and 4-nitrosodimethylaniline with formamide chiefly yielded the corresponding azoxybenzene compounds as the product. However, the nitroso compounds that can easily take the quinoid form were found to prevent the formation of azoxybenzene compound as 4-nitrosophenol, 1-nitroso-2-naphthol, and 1-nitroso-4-naphthol only formed black, amorphous substance of unknown structure.
A large amount of l-pinocarvone, b.p5 68-69°, nD20: 1.4942, d416: 0.9829, [α]D: -55.86°, in addition to d-α-pinene, b.p. 153.5-155.5°, nD: 1.4670, d414.5: 0.8610, [α]D: +44.92°, was found in the essential oil from wild Chenopodium ambrosioides L.
Manufacture of thiophene by the Philipps method and its modified Hachihame method was examined. It was found that by the addition of sand for dry distillation of sodium succinate and phosphorus trisulfide, the yield of thiophene was increased and made industriallization possible. On the other hand, the Krause method for the manufacture of 2-bromothiophene was examined and the increse in the yield to 76% was found possible by the addition of ether as a solvent.
Ethyl crotonate was chosen as the α, β-unsaturated carboxylic acid ester, and the Grignard reaction of 2-bromothiophene and the ester was carried out. In either 1:1 or 1:2 molar ratio, the product was 2-(1′-thienyl)-propylthienone-1′, and the unsaturated ketone (I) and saturated ester (II), the intermediates of the reaction, could not be obtained.
Digenic acid, m.p. 251° (decomp.), [α]D24: -14.8°±0.5° (c=1.01, H2O), C10H17O5N, and digeneaside (sodium mannosidoglycerate), m.p. 261° (decomp.), [α]D10: +94.0°±0.5° (c=3.00, H2O), C9H15O9Na⋅H2O, were isolated from Digenea simplex Aq. The acid was found to have an anthelmintic action.