Cleavage reaction of 1, 6-dimethyldibenzo-p-dioxin (I) with metallic sodium in liquid ammonia gives two kinds of phenolic compound, one of which was found to be 2-hydroxy-3-methylphenyl o-tolyl ether (II) and the other, 2-hydroxy-6-methyl-phenyl m-tolyl ether (III). Cleavage reaction of 1, 6-dimethoxy-3, 8-dimethyldibenzo-p-dioxin (VIII) with sodium in liquid ammonia, in the presence of sodium hydride, was found to inhibit the formation of the abnormal product, biphenyl derivative. Summarizing the results obtained to date, some considerations were made on the reaction conditions and mechanism of cleavage reaction of dibenzo-p-dioxin derivatives with sodium in liquid ammonia.
The quantitative method of vitamin D by the coloration with SbCl3, which is followed by the separation from vitamin A coexisted through Superfiltrol absorption chromatography, is easily influenced by the copresence of vitamin A and impurities in vitamin A. For the separation from the vitamin A impurities, Al2O3 has been recognized to be superior and so double layer column chromatography, i.e. over the Seperfiltrol (moisture: 10%), 0.7×7.0cm., Al2O3 (moisture: 4%), 0.7×3.0cm. was packed, was used and eluated by the hexane: ether (7:1) mixture. The quantitative measurement of vitamin D has become available within 10% error in the case of V.A/V.D (I.U.)≤10, even when vitamin A parmitate oil, contained in natural vitamin coexisted. In order to sensitize the separation, the triple layer column chromatography, i.e. Al2O3 (moisture: 5%), 1.0×2.5cm. was placed below the Superfiltrol (moisture: 15%), 1.0×1.0cm., and above the Superfiltrol (moisture: 10%), 1.0×3.5cm., has been applied to determine vitamin D in as much as 20 times of vitamin A coexistence. This method will be applicable to determine vitamin D in as much as 200 times of vitamin A coexistence, when moisture of the adsorbent, as well as the activity will be regulated properly.
On the paper-electrophoresis of human serum by normal method, the fraction containing serum cholinesterase activity was observed to be localized mostly between α2- and β-globulin peaks. On the zone-electrophoresis through starch colomn by the modified method of Smithies, the fraction invariably travelled with haptoglobin, which was localized between the fractions of slow α2- and β-globulin.
Physicochemical properties of gougerotin, a new antibiotic, were examined and it was assumed that gougerotin is a basic substance with molecular formula C16H25O8N7 and has a peptide-like bond. Since it forms a dipicrate, it is assumed to have two amino groups, one of which was found to be a primary amino group, determined by the Van Slyke method. It also has an amide-form nitrogen. The ultraviolet spectrum of gougerotin is very similar to that of cytidine and it was assumed that gougerotin has a moiety similar to pyrimidine nucleoside in its structure.
Hydrolysis of gougerotin with 6N hydrochloric acid was found to give three kinds of constituent components, while hydrolysis with conc. hydrochloric acid at room temperature results in partial hydrolysis. Each of these constituent components was isolated by the utilization of difference in their solubility and they were found to be D-serine, sarcosine, and C substance, an unknown pyrimidine nucleoside. Besides these, formation of one mole of ammonia was observed. Partial hydrolysate was separated by ion-exchanger chromatography and it was found to be the C substance bonded with D-serine.
Physicochemical properties of C substance (C10H14O6N4), a constituent component of gougerotin, were examined. Its ultraviolet spectrum was very similar to that of cytidine and from its pK′ value, this substance was assumed to be a pyrimidine nucleoside closely related to cytidine. The substance was also found to have a primary amino group determined by the Van Slyke method and a carboxyl group, as the functional groups, besides the amino group in the cytosine ring.
Catalytic reduction of the C substance over Adams platinum oxide afforded CR substance, C10H17O6N3, and one mole of ammonia. The C substance is resistant to acid hydrolysis but its reduced product, CR substance, is very labile to acids and easily undergoes hydrolysis to form tetrahydro-2 (1H)-pyrimidinone. This fact suggested the presence of a cytosine ring and the same catalytic reduction was carried out on cytosine and cytidine, from which ammonia was detected and the cytosine ring was reduced to tetrahydro-2 (1H)-pyrimidinone ring. This reaction is a new method for the reduction of cytosine derivatives and will prove to be one of useful means for determining the position of substituents in cytosine derivatives. Since cytosine was isolated by hydrolysis of C substance by boiling with nitric acid, C substance was found to be a compound having a cytosine ring and a side chain with a carboxylic acid and amino group
Based on the assumption that C substance has a cytosine ring bonded with a sugar, various derivatives were prepared to determine the position of sugar bonding. Deami-nation of C substance with nitrous acid and hydrolysis of its product afforded uracil. Methylation of C substance with dimethyl sulfate and hydrolysis of its product gave N, 3-dimethylcytosine, while methylation with diazomethane, followed by hydrolysis afforded 3-methylcytosine. These experimental facts indicated that neither the nitrogen in 3-position of cytosine nor the primary amino group in 4-position of the cytosine ring in the C substance had any substituent. Since the ultraviolet spectrum of the C substance was similar to that of cytidine, it was assumed that the C substance had a sugar bonded to the 1-position of its cytosine ring.
In order to examine the structure of the sugar moiety of the C substance, acetyl-ated (II and V) and formylated (III) derivatives of the C substance (I) and CR substance (IV) were prepared and their properties were examined. Functional groups in the sugar portion that are acylated are two hydroxyls and one amino group. The sugar portion becomes very unstable when (I) and (IV) are hydrolysed and difficult to isolate. Therefore, (I) and (IV) were deaminated to uronic acid derivative (VI) and hydrolysed under mild conditions by the use of cation-exchange resin, from which glucuronolactone (VIII) was isolated in crystalline form and identified with an authetic specimen. Reduction of this uronic acid derivative, followed by hydrolysis, and examination of its product by paper chromatography revealed a spot corres-ponding to glucose (X). The experimental evidences indicate the sugar moiety of the C substance is a uronic acid of hexosamine and takes the glucose configuration by deamination.
In order to elucidate structure of the sugar portion in the C substance, CR substance (I) was converted to its methyl ester (II), acetylated, and reduced by sodium borohydride, followed by hydrolysis, resulting in the isolation of hexosamine (V). Comparison of its behavior to various coloration reagents with glucosamine and 3-amino-3-deoxy-D-glucose showed that (V) is different from either of these. (V) failed to crystallize and, therefore, it was acetylated to N-acetylhexosamine (VI). Methanolysis of (V) and acetylation of its product afforded crystalline methyl hexosaminide tetraacetate (VIII). These experimental results proved that the sugar portion in the C substance is a uronic acid of hexosamine.
In order to determine the position of the amino group in the sugar portion of the C substance, Ninhydrin oxidation of hexosamine and CR substance was carried out but pentose was not detected in either of these reaction solutions, showing that there are no amino group in 2- or 5-position of the hexose. Periodate oxidation of the C substance (VII), CR substance (VIII), seryl-C substance (IX), and N-acetylated derivatives (XI and X), and examination of their products showed that the sugar moiety in the C substance takes a pyranose structure and that the amino group is in its 3-position. Consequently, the planar structure of the C substance was assumed to be 1-(1-cytosinyl)-3-amino-1, 3-dideoxyhexopyranuronic acid (VII).
The sugar portion in the C substance is known to take the glucose configuration on deamination with nitrous acid and various properties of N-acetylhexosamine and methyl hexosaminide tetraacetate, the crystalline derivatives of hexosamine, were compared with the corresponding derivatives of 3-amino-3-deoxy-D-glucose, but they were found to be different. This shows that the sugar portion of the C substance takes the allose configuration. The N-glycoside linkage between cytosine and the sugar moiety in the C substance was proved to be β-oriented by oxidation of the C substance with periodic acid to diglycol aldehyde derivative (IX) and its comparison with the corresponding derivative of cytidine. It is concluded from these evidences that the structure of the C substance, is 1-(1-cytosinyl)-3-amino-1, 3-dideoxy-β-D-allopyranuronic acid, as indicated in Fig. 2.
Seryl-C substance, formed by bonding of D-serine and the C-substance, obtained by partial hydrolysis of gougerotin, was examined to elucidate its structure. Hydrazine decomposition and DNP method showed that the carboxyl group in D-serine takes part in this bonding and that the amino group in the cytosine ring of the C substance is in a free state, since catalytic reduction of the seryl-C substance results in the formation of ammonia and deamination of this substance with nitrous acid, followed by hydrolysis, affords uracil. It was thereby assumed that seryl-C substance is 1-(1-cytosinyl)-3-D-serylamino-1, 3-dideoxy-β-D-allopyranuronic acid (I), in which the carboxyl group of D-serine is bonded in acid-amide structure to the amino group in the sugar portion of the C substance.
In order to examine the free amino group in gougerotin, the DPN method was applied. Although the amino group in serine was found to be in a free state, DNP-sarcosine was labile to acid hydrolysis and it was impossible to estimate whether sarcosine is an N-terminal group or not. Consequently, gougerotin was methylated with dimethyl sulfate and hydrolyzed, by which both serine and sarcosine disappeared, showing that the amino group in sarcosine does not take part in the bonding. Consequently, the carboxyl group in sarcosine would be bonded to the amino group in cytosine and it was assumed that the carboxyl group in the uronic acid is forming an acid amide. This assumption was found to be correct by the deamination of gougerotin with nitrous acid, followed by hydrolysis of its product, and catalytic reduction or hydrazine decomposition of gougerotin. Therefore, gougerotin structure was assumed to be 1-(N-sarcosyl-1-cytosinyl)-3-D-serylamino-1, 3-dideoxy-β-D-allopyranuronamide (I).
Photodecomposition of thioproperazine (I) was carried out by irradiation of its dimethanesulfonate in 0.72% aqueous solution with fluorescent light (twelve 20-W coolwhite light tubes, ca. 5000 luces). It was found by paper partition chromatography, using a solvent system of butanol-acetic acid-water (4:1:5) that (I) (yellow fluorescence spot, Rf 0.80) is completely decomposed in 24 hours. At the same time, a marked change took place in the ultraviolet absorption spectrum of the aqueous solution. Ampules filled with 5% aqueous solution of the dimethanesulfonate of (I) were continuously irradiated for 72 hours and 10-[3-(4-methyl-1-piperazinyl)propyl]-2-phenothiazinesulfonic acid (II) was isolated and identified as the chief decomposition product, the yield of (II) being about 80% of the product. Photodecomposition of the dimethanesulfonate of (II) afforded 10-[3-(4-methyl-1-piperazinyl)propyl] phenothiazine.
Stability of thioproperazine to light in aqueous solution was quantitatively evaluated and its stabilization was attempted. The control solution containing 0.72% thiopro perazine dimethanesulfonate, 0.05% sodium sulfide, 0.1% ascorbic acid, and 0.6% sodium chloride, and the solutions containing various additives in 0.02 mole concentration were each adjusted to pH 3.5, filled in 1cc. ampules, and sealed in nitrogen atmosphere. These ampules were irradiated with fluorescent light (ca. 5000 luces) in a cabinet provided with twelve 20-W fluorescent tubes of cool white light. Thioproperazine in the irradiated solution was determined colorimetrically, using cobalt thiocyanate reagent. Lower, unsaturated fatty acids, especially maleic acid, showed marked stabilizing effect and unsaturated acids not containing a conjugated double bonds or saturated acids did not show such effect. Pyridinecarboxylic acids and their amides and esters also showed a good effect, especially those having a substituent in the 4-position. In hydroxy-, amino-, and alkyl-pyridines, those substituted in the 3-position were effective. Saturated bases like piperidine and piperazine were ineffective. Photodecomposition of 10-[3-(4-methyl-1-piperazinyl)propyl]-2-phenothiazinesulfonic acid was also prevented by the addition of similar compounds.
3-Piperidino-1, 1-diphenyl-1-butanol and 3-piperidino-1, 1-diphenyl-1-butene were obtained as a decomposition product by heating ethyl 3-piperidino-1, 1-diphenylbutyl sulfone in dil. HCl. When water solution of tartaric acid salt of l-form was heated, it afforded l-3-piperidino-1, 1-diphenyl-1-butanol and d-3-piperidino-1, 1-diphenyl-1-butene. The production velocity of ethyl 3-piperidino-1, 1-diphenyl butyl sulfone and 3-piperidino-1, 1-diphenyl-1-butene will be accelerated by an increase of the hydrogen ion concentration, the production velocity of the former being greater. This finding showed that ethyl 3-piperidino-1, 1-diphenyl butyl sulfone may rarely produce 3-piperidino-1, 1-diphenyl-1-butene directly in the case of the decomposition without through a production of 3-piperidino-1, 1-diphenyl-1-butanol.
Reduction of 2, 3-methylenedioxy-9, 10-dimethoxydibenzo [a] acridizinium chloride (I) with lithium aluminium hydride in dioxane or tetrahydrofuran afforded dihydroberberine (II) in a good yield. Application of methyl iodide to (II) in dichloromethane or chloroform solution gave 13-methyl-2, 3-methylenedioxy-9, 10-dimethoxydibenzo [a]-5, 6-dihydroacridizinium iodide (VIII), and 13-methyl-2, 3-methylenedioxy-9, 10-dimethoxydibenzo [a, g] quinolizidine (VII), and that of ethyl iodide gave 13-ethyl-2, 3-methylenedioxy-9, 10-dimethoxydibenzo [a, g] quinolizidine (IX). The substance obtained on application of methyl iodide to acetoneberberine (V) had been believed as 8-acetonyl-13-methyl-dihydroberberine hydroiodide (VI) but it was proved to be 13-methyl-2, 3-methylenedioxy-9, 10-dimethoxydibenzo [a]-5, 6-dihydroacridizinium iodide (VIII) from its analytical values and from infrared and ultraviolet spectra. The same examinations were made on the reaction products obtained from application of ethyl iodide and propyl iodide to (V). Reduction of (VIII) with sodium borohydride gives (VII) but the same reduction of 13-ethyl-2, 3-methylenedioxy-9, 10-dimethoxydibenzo[a]5, 6-dihydroacridizinium iodide (X) afforded (IX) or the dihydro compound (XII) according to reaction conditions. Reduction of (VIII) and (X) with lithium aluminium hydride in tetrahydrofuran afforded the corresponding dihydro compounds (XI and XII).
The substance (II) of m.p. 223-224° (decomp.), obtained by oxidation of acetone berberine (I) with potassium permanganate was assumed, from its infrared and ultraviolet spectral characteristics, to be acetonyl-13-hydroxy-2, 3-methylenedioxy-9, 10-dimethoxydibezo [a, g] quinolizidine. Boiling of (II) with mineral acid afforded 13-hydroxy-2, 3-methylenedioxy-9, 10-dimethoxydibenzo [a]-5, 6-dihydroacridizinium salt (VI) and its treatment with alkali hydroxide gave berberine phenolbetain (VII). Reduction of (VII) with sodium borohydride or lithium aluminium hydride gave 13-hydroxy-2, 3-methylenedioxy-9, 10-dimethoxydibenzo [a, g] quinolizidine (dl-ophiocarpine) (VIII). Reduction of (II) with sodium borohydride failed to form the expected (VIII) and gave 8-(2-hydroxypropyl)-13-hydroxy-2, 3-methylenedioxy-9, 10-dimethoxy dibenzo [a, g] quinolizidine (IV).
Thioctamide (IV) has been synthesized from thioctic acid by a comparatively complicated method. In the present series of work, thioctamide was obtained easily by the application of sodium thiosulfate and sodium disulfide to 6, 8-dichloroöctanamide (III), prepared from 6, 8-dichloroöctanoic acid (I). It was found, however, that the thioctamide (IV) so obtained is impure judged from its ultraviolet absorption spectrum and its reason and method of purification were examined. It was found that the molar ratio of sodium disulfide and sulfur to (III) affected the purity of (IV) and thioctamide was finally obtained.
A new method of synthesis for thioctamide was devised by application of acid halogenation agent to thioctic acid (I) and reaction of thioctoyl chloride so obtained with ammonia and amines. Thioctamide and derivatives thereby formed were found to be pure judged by their ultraviolet absorption spectra.
In order to develop a more potent new antipyretic-analgesics reported in the preceding paper, (amino acid-type derivatives of salicylic acid system, especially N-dimethylcarbamoylmethyl-o-methoxybenzamide (III)), glycine dimethylamide-type compounds of salicylic acid, substituted with nitro, chloro, or acetamido group in its benzene ring, were synthesized by several routes, considering the relation of PAS and acetophenetidine to salicylic acid.
Considering that the irreversible inhibition of α-methylphenethylhydrazine (JB-516) to monoamine oxidase is weakened in the presence of substrate tyramine, inhibitory effect of JB-516 on monoamine oxidase under various conditions, in the presence of tyramine and amphetamine, was measured manometrically. It was thereby clarified that JB-516, amphetamine, and tyramine act on the same active center and that the time of addition of the substrate has a great effect on the result of measurement of inhibitory effect of irreversible competitive inhibitor.
Dipole moment and ultraviolet spectrum of quinoxalines and their N-oxide compounds (I to X) were measured to examine whether the N-oxide group is in 1- or 4-position. The dipole moment of quinoxaline itself has a value of 0.82D and this value was used to analyze the observed moments of quinoxaline N-oxides. It was thereby found that 2-methylquinoxaline N-oxide (IV) was 1-oxide and that 2-methoxy-(VI), 2-ethoxy- (VIII), and 2-ethoxy-3-methyl-quinoxaline N-oxide (X) were all 4-oxid. es These facts were in good agreement with ultraviolet spectral data. In 2-alkoxyquinoxalines and their N-oxides, the dipole moment values agreed with the structure with the alkyl group oriented towards nitrogen in 1-position and this may be due to the static attraction between the lone-pair electrons of nitrogen in 1-position and hydrogen of the alkyl group. This may be interpreted as the nature of steric interference experienced in N-oxidation.
Various properties of essential oil of plants were examined with essential oil of eight kinds of Lauraceae plants and standard samples of various essential oils. Separation of each component by gaschromatography, identification, and quantitative determination were carried out, as well as the examination of various pharmacological effects on circulation system, such as the action of samples on the excised heart of a toad, rabbit heart, rabbit respiration and blood pressure, and on blood vessels of toad hind legs. Essential oil was separated well into each component by gas chromatography, using polyethylene glycol 6000 and silicone DC 200 as the stationary phase and determination was made in comparatively high accuracy. The components obtained agreed well with those given in the past literature but there were few discrepancies. In pharmacological test, these essential oils were generally inhibitive and depressive against heart and blood pressure, but the action was weak in camphor and the essential oil from the camphor tree, some of which showed acceleration of the heart action and elevation of blood pressure. In the perfusion test of toad hind leg, non-water-soluble factor of the essential oil seemed to have a grave effect on the test and physiological action of the essential oils could not be confirmed.
The degree of coloration, specificity and stability of pentose with various compositions in different heating period of the various polyphenol reagents in the mixed solvent of hydrochloric acid and acetic acid have been investigated. The activity of resorcinol and orcinol in diphenol group has been observed as a coloration reagent and the isomers of triphenol were found to be effective for coloration reagent. Because of the instability of hydroxyhydroquinone, its triacetate was used for the test. All of the compounds examined possess two hydroxyl groups commonly at meta position, which were considered to be related closely to the coloration reaction. An optimal condition of polyphenols for the colorations is: resorcinol reagent showed a low degree of coloration; phloroglucinol and hydroxyhydroquinone reagent showed unstable coloration; orcinol reagent was superior to the specificity, though the degree of coloration was low; and pyrogallol reagent had a high degree of coloration, being inferior to the specificy. It was concluded that the phenolic compounds mentioned above were available to the quantitative colorimetry of pentose.
In the course of studies on thiamine disulfide, a new compound, O-benzoylthiamine disulfide was synthesized by the oxidation of O-benzoylthiamine in alkaline solution or by the benzoylation of thiamine disulfide in pyridine. O-Benzoylthiamine disulfide occurs odorless, colorless prisms, m.p. 146-147°. Its infrared and ultraviolet spectra are shown in Chart I. On hydrolysis of O-benzoylthiamine disulfide with 10% hydrochloric acid, 4-amino-5-aminomethyl-4-methylpyrimidine, bis (1-acetyl-3-hydroxypropyl) disulfide, benzoic acid, and formic acid were formed.
A small amount of the tertiary phenolic base, m.p. 128°, isolated from Nandina domestica f. shinananten HORT. (Japanese name “Shina-nanten”), which was reported previously, has been identified as isoboldine (I).
The lotus seeds discovered from the prehistoric dwellings at Kemikawa in Chiba Prefecture in 1951, purported to be about 2000 years old, were sown and germinated. This ancient lotus has since been cultivated and was named “Ohga-hasu” to honor its finder and cultivator, Dr. I. Ohga. Scientific name given to this ancient lotus is Nelumbo nucifera GAERTN., the same as ordinary lotus. Examination of alkaloids in the leaves and leaf petioles of this lotus yielded the aporphine-type bases, roemerine (II), and nornuciferine (III), and benzyl-tetrahydro-isoquinoline-type base, armepavine (IV), from the leaves, and roemerine (II) and nornuciferine (III) from the leaf petiole. Presence of nuciferine (I) in the leaves and petiole was presumed from the result of paper chromatography but was not isolated due to the small amount of the fresh leaves available. These results indicated that the alkaloids contained in this ancient lotus are the same as those in ordinary domestic lotus.