1) In order to compare the anthelmintic effect of coumarin and dihydrocoumarin derivatives, 7 kinds of methyl derivatives of octahydrocoumarin were prepared and their action against hog ascaris was tested. 2) The compounds prepared and tested were octahydrocoumarin, and 3-methyl-, 4-methyl-, 5-methyl-, 6-methyl-, 7-methyl-, and 8-methyl-octohydrocoumarins. Of these, all except the first are newly synthesized compounds. 3) The octahydrocoumarin derivatives showed less effect than the corresponding coumarin derivatives, whether the living time or time elapsed until appearance of motionless state was taken as the standard of potency judgement. The effect of the octahydrocoumarin derivatives varied compared to the dihydrocoumarin derivatives. 4) The effect of octahydrocoumarin derivatives was also varied in accordance with the position of the methyl group but the order of potency was not in parallel with that of coumarin or dihydrocoumarins. 5) The strongest effect was found in 3-methyloctahydrocoumarin. Since 4-methyloctahydrocoumarin effected curling motion in ascaris, the importance of the methyl group in the lactone ring of santonin molecule is pointed out by the analogy of structure of this compound with that of santonin.
1) Although coumarin derivatives do not react with carbonyl reagents, thiocoumarin derivatives undergo condensation with them. In order to find the role of the lactone ring in coumarin derivatives on their anthelmintic action by comparison of the anthelmintic actions of these derivatives, 10 kinds of thiocoumarin derivatives were prepared and their anthelmintic action was tested. 2) Compounds tested were thiocoumarin, and 3-methyl-, 4-methyl-*, 5-methyl-*, 6-methyl-*, 7-methy-, 8-methyl-*, 4, 7-dimethyl-, 3, 4, 7-trimethyl-, and 3-ethyl*-thiocoumarin (those with * are new compounds). 3) Whatever is taken as the standard for anthelmintic action, thiocoumarin derivatives in general had weaker effect than the corresponding coumarin derivatives and the order of efficacy of methylcoumarin and methylthiocoumarin derivatives was not parallel. 4) The effect of 3-alkylthiocoumarins tended to decrease with the lengthening of the alkyl group. 5) If the spasmodic and paralytic actions of santonin, the chief cause of its anthelmintic action, are to be attributed to the ketone group and the lactone ring in its molecule, thiocoumarin derivatives should have stronger anthelmintic action than coumarin derivatives. Since this was found to be reverse of the case by experiments, it was assumed that coumarin derivatives should also possess some amount of ketonic properties.
If it may be assumed that the spasmodic and paralytic actions that are the chief causes of the anthelmintic effect of santonin are due to the ketone group and the lactone ring in its molecule, thiocoumarin derivatives which possess ketonic properties should have stronger paralytic action than the coumarin derivatives not possessing such ketonic property. However, such was found to be the reverse of the case and, therefore, some ketonic property was assumed to exist in coumarin derivatives. In order to prove this assumption, solvent effect against absorption spectrum of coumarin was studied. The absorption band of coumarin in the shorter wave length range shifted towards the visible range by the increase of the dipolar character of the solvent, while the absorption band in the longer wave length range shifted towards the ultraviolet region. Since these characteristics agree with the regularity of solvent effect observed in α, β-unsaturated ketones, it is known that there is some ketonic property in coumarin. Therefore, the decrease of paralytic action in dihydro- and octahydro-coumarins compared to coumarin derivatives may be attributed to the loss of the ketonic property. It follows, further, that the ketone group in the santonin molecule is important for the paralytic and spasmodic actions which are the chief cause of the anthelmintic action of santonin.
Addition of 1% NaClO2 solution to acid solution (below pH 4.7) of thiamine, followed by NaOH solution and butanol, and vigorous shaking results in the formation of thiochrome which transits to the butanol layer. When pure thiamine solution is used, the amount of thiochrome formation is the same as that by the ferricyanide method and can be utilized for the determination of thiamine. In this reaction, it is necessary to add NaClO2 in acid solution of thiamine. If the acidity of this mixture is weak, formation of thiochrome is poor. If a mixture of NaClO2 and NaOH is added to thiamine, there will be no formation of thiochrome. Acidification of NaClO2 results in the formation of ClO2 and the use of aqueous solution of ClO2and NaOH also gives rise to thiochrome but not quantitatively as in the case of NaClO2. ClO2 is a strong oxidizing agent but thiamine is fairly stable to ClO2 and a part of it is decomposed to 2-methyl-4-amino-5-aminomethylpyrimidine but the majority remains unchanged, as was proved by paper chromatography. Formation of thiochrome is effected by ClO2+NaOH when the alkali is added and the formation of thiochrome in acid medium was not detected by paper chromatography.
Passage of ClO2 gas in thiothiamine (3-[2′-methyl-4′-aminopyrimidyl-(5′)]-methyl-4-methyl-5-β-hydroxyethylthiazole-2-thione) or its solution results in the formation of thiochrome and the formation of thiamine is hardly detected. Addition of NaClO2 to the acid solution of thiothiamine results in their reaction with generation of ClO2 to form thiamine and thiochrome. Thiochrome is quantitatively formed by the addition of an aqueous solution of ClO2 in a diluted solution of thiothiamine and this reaction also occurs in acidity. Addition of chlorine water to the diluted solution of thiothiamine results in the formation of thiamine and thiochrome, while that of HClO3 gives only thiamine. Formation of thiochrome was confirmed by extraction with butanol after alkalization while that of thiamine was determined by the ferricyanide-sodium hydroxide method. Thiothiamine is effective as thiamine in animal body but is more easily changed to thiochrome in acid or neutral medium than thiamine that its biological effect must also be weaker.
In order to test the presence of a curare-like action, Decamethonium and its derivatives were prepared. In the preparation of the bis (quaternary salts), the reaction of decamethylene dihalide and the corresponding tertiary amines may be influenced by the steric effect rather than their basicity. In the present case, decamethylene bis (tertiary amines) were prepared first from decamethylene dichloride and the corresponding secondary amines and the subsequent treatment of the purified bis (tertiary amines) with alkyl halide yielded the required bis (quaternary salts).
In order to examine the influence of the nitrogen and a substituent in the nucleus on the curare-like effect, decamethylene-bis [1-(3, 4-dimethoxyphenyl)-6, 7-dimethoxy-1, 2, 3, 4-tetrahydroisoquinoline] dimethiodide and bis [1-(3, 4-methylenedioxyphenyl)-3-methyl-6, 7-methylenedioxy-1, 2, 3, 4-tetrahydroisoquinoline] dimethiodide were prepared. In the reaction of decamethylene diiodide and tetrahydroisoquinoline derivatives, which were prepared by the well-known method, long heating was necessary because of the steric effect. The compounds obtained may be a stereoisomeric mixture since they possess two or four asymmetric carbons.
In order to examine the influence of the substitutent in the 1-position and the benzene nucleus on the curare-like effect, α, θ-bis(2-methyl-1, 2, 3, 4-tetrahydroisoquinolyl-1)octane derivatives were prepared. Sebacic di-β-amides were prepared by the dehydration between sebacic acid and 3, 4-substituted β-phenylethylamines. Bis(3, 4-dihydroisoquinolines) were prepared by the Bischler-Napieralski method and the catalytic reduction of their dimethiodide after chlorination with silver chloride yielded bis(2-methyltetrahydroisoquinoline), which were derived to their dimethiodide with methyl iodide. By the recrystallization of the dimethiodides, 6, 7-methylenedioxy and 6-methoxy compounds were isolated into two stereoisomers.
Resolution of the stereoisomers of α, θ-bis (2-methyl-1, 2, 3, 4-tetrahydroisoquinolyl-1) octane derivatives was examined in order to find out the influence of stereoisomers on the curarelike effect. α, θ-Bis (6, 7-methylenedioxy-2-methyl-1, 2, 3, 4-tetrahydroisoquinolyl-1) octane was separated into racemic and meso-forms by oxalic acid and derived to the respective dimethiodide. The dimethiodide of the racemic compound was resolved into optically active compounds by silver d-camphorsulfonate. 6-Methoxy compound was also solated into two isomers by oxalic acid but 6, 7-dimethoxy compound failed to yield the isomers by the similar method, giving only one form.
6, 7-Substituted hydroquinone bis (2-methyltetrahydroisoquinolyl-(1)-methyl) ether dimethiodides were prepared in order to examine the influence of the substitutents in the 1-position and the benzene nucleus of the d-tubocurarine molecule on the curare-like effect. The preparation of the compounds followed the method similar to that described in the previous paper. The compounds prepared may be a mixture of stereoisomers since they possess two asymmetric carbons.
Resolution of the stereoisomers of 6, 7-substituted hydroquinone bis (2-methyl-1, 2, 3, 4-tetrahydroisoquinolyl-(1)-methyl) ethers was examined in order to find the influence of stereoisomers on the curare-like effect. 6, 7-Methylenedioxy compound was isolated into the racemic and meso-forms by oxalic acid and derived to the corresponding dimethiodides. The racemic amine and dimethiodide were resolved into the respective optically active compounds with d-tartaric acid and d-camphor silver sulfonate. 6, 7-Dimethoxy compound was also resolved into two isomers with oxalic acid.
7, 7′-Ethylenedioxy-bis (1-veratryl-6-methoxy-2-methyl-1, 2, 3, 4-tetrahydroisoquinoline) dimethiodide was prepared as a curarising agent. In reducing 1-veratryl-6-methoxy-7-benzyloxy-3, 4-dihydroisoquinoline methiodide or methochloride, use of platinic oxide as a catalyst effected reduction but not debenzylation while the objective was attained with the use of palladium-carbon and platinic oxide.
Examinations were made on the synthesis of diphenylene dioxide by the thermal condensation of the potassium salt of o-bromo-, o-chloro-, and o-iodophenol with copper catalyst. It was thereby clarified that, as in the case of the Ullmann reaction, the reaction proceeded most smoothly when the halogen in halophenols was bromine, unless there was an atomic group, such as the nitro radical, that activated the nuclear chlorine atom, and that the yield is the best when metallic potassium was used, rather than potassium hydroxide, in the preparation of the potassium salt, apart from the economic question. The corresponding iodo compound was found to be impracticable since the liberation of iodine occurred at below the reaction temperature.
Formation of a molecular compound between several kinds of chlorophenol derivatives and β-dimethylaminoethyl benzhydryl ether (I) was examined by plotting a fusion curve by thermal analysis. It was thereby found that molecular compounds were formed from 2, 2′-thio-bis (4-chlorophenol) (II), 2, 2′-thio-bis (4, 6-dichlorophenol) (III), and 5-chlorosalicylic acid (IV) with the respective melting points of 92-95°, 120°, and 87°. It was also confirmed that the molecular compounds (X) and (XI) were formed respectively from (II) and (I), and (III) and (I) in 1:1 ratio only and not in 1:2 ratio. Further, the adduct (IX) of p-chloro-o-benzylphenol (VIII) and (I) was not obtained in a crystalline form but its formation was confirmed by its infrared by its infrared absorption spectral measurement.
The sample solution was prepared from the 1:100 aqueous digest solution of the leaves of Geranium nepalense Sweet, which was diluted to four volumes. To 5cc. of this solution, 5cc. of the extemporaneously prepared 0.02g./cc. methylene blue solution containing molar equivalent of sodium hydroxide was added and the whole volume was divided into two parts. At the same time, two samples of 5cc. of the 0.01g./cc. methylene blue solution were taken as the standard. All these sample solutions were submitted to capillary analysis at the same time, in a neighboring place, and under the same conditions. The elevation ratio of methylene blue in such capillary images, indicated by Rb, gave the value of Rb≤0.4 for the leaves of the Geranium JP. As a result of examinations, it was found that, as long as the filter paper used was the same, the value of Rb was proportional to the content of tannic acid in Geranium. It was also found that the relationship between this Rb value and percentage of tannic acid content could be indicated by the following formula: y=39.6-43x, where y indicates the tannic acid content, and x, the Rb value. The constants become definite with the filter paper used when the temperature is 11-35° and the relative humidity about 71-90%. Therefore, the content of tannic acid in the Geranium can be assumed from the Rb values measured by using this equation.
Application of one mole of thiocyanic acid to isonicotinic acid hydrazide (II) gives the adduct (III) of (II), while the heating of this mixture yields isonicotinic acid thiosemicarbazide (I) and 3-γ-pyridyl-Δ2-1, 2, 4-triazoline-5-thione (IX), formed by the dehydrationcyclization of (I). Application of two moles of thiocyanic acid to (II) results in the formation of the adduct (VII) of (II) with two moles of thiocyanic acid and the adduct (VIII) of (I) with thiocyanic acid. Application of heat in this case yielded (VII) and (IX).
By heating 1-isonicotinylthiosemicarbazide (I) in tetralin or alkoxide, 3-γ-pyridyl-Δ2-1, 2, 4-thiazoline-5-thione (II) was obtained, while the treatment of (I) with conc. sulfuric acid yielded 2-amino-5-γ-pyridyl-Δ2-1, 3, 4-thiadiazole (III). 5-Methylthio-3-γ-pyridyl-4 H-1, 2, 4-triazole (V) was obtained by heating or treatment with alkali phosphate of isonicotinyl-S-methylisothiosemicarbazide (IV) and oxidation of (V) with potassium permanganate yielded the 5-methylsulfonyl compound (VI).
By heating isonicotinylhydrazine (I) with carbon disulfide in pyridine, 5-γ-pyridyl-Δ4-1, 3, 4-oxadiazoline-2-thione (III) and 4-isonicotinylamino-3-γ-pyridyl-Δ2-1, 2, 4-triazoline-5-thione (V) were obtained. (III) was also obtained on heating (II) (R=K or R=CH3) in decalin. Treatment of (II) (R=CH3) with conc. sulfuric acid gave 2-methyl-thio-5-γ-pyridyl-1, 3, 4-thiadiazole (VI) which was derived to the methylsulfonyl compound (VII) by oxidation. Application of hydrazine hydrate to (II) (R=CH3) gave 4-amino-3-γ-pyridyl-Δ2-1, 2, 4-triazoline-5-thione (IX) whose desulfuration yielded 4-amino-3-γ-pyridyl-1 H-1, 2, 4-triazole. Methylation of (III) to (XIII) and its oxidation chiefly gave 5-γ-pyridyl-Δ4-1, 3, 4-oxadiazoline-2-one (XII).
The amides, dialkylamides, and dialkylaminoethanol esters of ar-α- and ar-β-tetralylacetic acid and ar-β-tetrahydronaphthoic acid were synthesised. Some of the derivatives thereby obtained were found to possess curarimimetic, analgesic, local anesthetic, and temperature depressing actions, although such actions were not so marked.
Microdetermination of nitrous acid was successfully established by introducing into the azotometry, the nitrogen gas generation by the reaction of nitrous acid and ammonium sulfaminate. The value of purity determination of potassium nitrite by this method agreed well with the result of potassium permanganate titration. Optimal determination range: 20-200 γ of nitrous acid.
Microdetermination of reducing sugars was successfully established by the application of the determination method of reducing sugars using potassium ferricyanide to azotometry Amixture of potassium ferricyanide and reducing sugars is warmed for 30 minutes on a water bath at 100°, and the amount of potassium ferricyanide consumed is determined by the hydrazine-azotometry. The ratio of the value of potassium ferricyanide consumed converted into the nitrogen volume, V, and the amount of sample, M, i.e. K (V/M), is constant for aldohexose, ketohexose, and aldopentose. By the utilization of respective K values, reducing sugars can be determined. Optimal determination range: 10-120γ
Microdetermination of sulfanilamides was established by measuring by azotometric method the nitrogen gas generated from the reaction of the diazonium salt of sulfanilamide with sodium azide in an acid medium. The results of determination of several sulfanilamides agreed well with the results of diazo titration. Optimal determination range: 4×10-6-5×10-7M.
S-Alkylthiocysteines were obtained by the reaction of ethyl and allyl benzenethiosulfonates with cysteine in hydrated alcoholic solution. By a similar method, several thiamine alkyl disulfides, 2-[2′-methyl-4′-aminopyrimidyl-(5)]-methyl-5-hydroxy-Δ2-pentenyl-(3) allyl disulfides, were also prepared.
On heating 2-amino-4-methylthiazole and ethyl acetoacetate at 120-130°, an acetoacetylated compound (Ia) is formed which yields a pyrimido [2, 1-b] thiazole compound (A) on being heated with conc. sulfuric acid at 50-60°. Since (A) shows absorption curve practically similar to that of 3-methylpyrimido [2, 1-b] thiazol-5-one, the structure of (A) was assumed to be 3, 7-dimethylpyrimido [2, 1-b] thiazol-5-one, which indicates that a transition had occurred during cyclization by conc. sulfuric acid. This is exactly the same as the reaction that occurs on the cyclization of 2-acetoacetylaminopyridine with conc. sulfuric acid reported by Adams, et al.. As for the structure of (Ia), it may be assumed as a compound with the side-chain amino group acetoacetylated, from its absorption spectrum. Treatment of 2-acetamino- and 2-benzoylamino-4-methylthiazole with conc. sulfuric acid does not yield any rearrangement products, and 2-acetoacetylaminothiazole and its 4-phenyl derivative do not undergo cyclization with conc. sulfuric acid.
Examinations using paper chromatography and microbiological assay were carried out on the growth factor contained in cotton wool, with properties similar to panthothenic acid, nicotinic acid, biotin, and vitamin B12. Rf values of the pantothenic acid- and nicotinic acid-activities agreed with those of pantothenic and nicotinic acids but the B12-activity did not agree with Rf value of the crystalline B12. The crystalline biotin, when developed with water-butanol mixture, was distributed into two factors with Rf 0.1-0.3 and 0.6-0.8, active to Lactobacillus arabniosus, and a larger amount collected in the spot with a larger Rf value. The biotin-activity of the cotton-wool factor agreed in Rf values with the two tactors but a larger amount collected in the spot with a smaller Rf value.
After testing the availability of several azo compounds, which have nitro and amino groups in ortho- or para-position, p-nitro-p′-aminoazobenzene was selected as the most excellent indicator for the titration of phenols in ethylenediamine and numerous samples of phenols were titrated satisfactorily with this indicator. For phenols with negative substituents such as nitrophenol, it is preferable to use pyridine as a solvent and azo violet as the indicator.
Hydrogen peroxide solution was applied to isosafrole in formic acid and the oil thereby obtained, assumed to be isosafrole glycol ester, was submitted to hydrolysis and dehydration with sulfuric acid from which the objective 3, 4-methylenedioxybenzyl methyl ketone was obtained in approx. 60% yield.
3, 4-Methylenedioxybenzyl methyl ketone was derived to its oxime and reduced or was submitted to the Leuckart reaction to form β-3, 4-methylenedioxyphenylisopropylamine (I), reacted with 3-benzyloxybenzoic chloride to the acid amide, and cyclized by the usual method to 1-(3′-benzyloxyphenyl)-3-methyl-3, 4-dihydro-6, 7-methylenedioxyisoquinoline (III). Dehydrogenation of (III) gave, through 1-(3′-benzyloxyphenyl)-3-methyl-6, 7-methylenedioxyisoquinoline (VI), the objective 1-[3′-(β-diethylaminoethoxy)-phenyl]-3-methyl-6, 7-methylenedioxyisoquinoline (VII). Hydrolysis of (III) gave (IV) which was reacted with diethylaminoethyl chloride to another of the objective, 1-[3′-(β-diethylaminoethoxy)-phenyl]-3-methyl-3, 4-dihydro-6, 7-methylenedioxyisoquinoline (V).
Water-insoluble steroidal saponins were isolated from the methanolic extracts of the powdered rhizome of Dioscorea nipponica Makino, D. gracillima Miq., and D. tenuipes Franch. et Sav. Purification of these saponins by liquid chromatography through alumina yielded the same chief component from the portion eluted with a chloroform-methanol (5:1) mixture. The substance and its acetate showed the same properties, such as the melting point, specific roatation, and others, with pure dioscin obtained by the similar purification of a sample of dioscin, a water-insoluble saponin isolated from D. tokoro Makino and its acetate. All these four samples were found to form diosgenin, glucose, and rhamnose by hydrolysis. It has thereby been clarified that the chief component of the water-insoluble saponins contained in the foregoing three domestic Dioscoreaceae plants is dioscin.