A partial modification was made on the colorimetric determination of homosulfanilamide, using 2, 4-dinitrofluorobenzene as the reagent, reported earlier. This method was used for the measurement of its blood level and a good result was obtained. In this method, the sample blood is completely hemolyzed, deproteinized with trichloroacetic acid, and the supernatant is separated. This is neutralized, the reagent adjusted to pH 8 is added, and heated to effect coloration. The pigment thereby formed is extracted with benzene and optical density of the benzene layer is measured. The value obtained by this method is that obtained after removal of metabolites of homosulfanilamide. Blood level of homosulfanilamide after its administration to men and rabbits was measured. In the case of human beings, administration of 5g. in a single dose resulted in the maximum blood level of ca. 4.5mg% after 1hour while the administration of 1g. to a rabbit resulted in the maximum blood level of ca. 3.5mg% after 1hour. Combined use of pharmaceutics with low toxicity, having inhibitory action of monoamine oxidase, such as 20mg. of β-phenylisopropylhydrazine, resulted in a blood level of ca. 10mg% after 2hours and 10.2mg% after 3hours. The same marked elevation in the blood level was observed after concurrent administration of isonicotinic acid hydrazide. Consequently, pharmaceutics which can be used as substrate for monoamine oxidase, like homosulfanilamide, can markedly increase the biological activity of monoamine oxidase inhibitor with comparatively low toxicity.
The biological metabolites of homosulfanilamide were found to be p-carboxy-benzenesulfanilamide and acetamidomethylbenzenesulf anilamide. Examinations were made on the determination in urine to measure unchanged homosulfanilamide alone and this was found to be possible by a slight modification of the method reported earlier for the measurement of its blood level. The separatory determination of the metabolites was effected by the use of paper chromatography. Homosulfanilamide was administered to men and the value of the metabolites excreted was determined. p-Carboxybenzenesulfanilamide was found to be excreted in the largest amount and a part of it was excreted as p-acetamidomethylbenzenesulfanilamide, together with the unchanged homosulfanilamide. Experiments with rabbits showed that combined use of β-phenylisopropylhydrazine of comparatively small toxicity or isonicotinic acid hydrazide resulted in decreased excretion of p-carboxybenzenesulfanilamide compared to the single administration of homosulfanilamide and the amount of unchanged homosulfanilamide was found to be 2-6 times higher than that in single administration.
Homosulfanilamide was found to be decomposed in the body and, in order to examine the location of this decomposition, mode of its decomposition in the digestive tract and various organs was examined. Decomposition in the digestive tract was examined by the use of simulated gastric juice, intestinal juice, and enteric bacteria (Escherichia coli communior, Streptococcus faecalis R, and Lactobacillus acidophilus) but there was no evidence of decomposition and a marked decomposition was found to be effected by intestinal mucosa enzyme. Further examination using the homogenate of rabbit liver and by the perfusion test with excised rabbit liver showed a marked decomposition to take place in both cases. Decomposition of homosulfanilamide on addition of β-phenylisopropylhydrazine or isonicotinic acid hydrazide was examined and it was found that the decomposition of homosulfanilamide was only 30-40% on addition while that of homosulfanilamide was 70% after about 5hours. However, in the test with intestinal mucosa enzyme, effect of these inhibitors was not observed.
(1) o- or p-Chloroaniline and acetoacetic ethyl ester were condensed at room temperature in the presence of one drop of hydrochloric acid to afford ethyl 3-(o- or p-chloroanilino) crotonate, which was cyclized by heating in paraffine into 2-methyl-8-chloroquinolinol (I) and 2-methyl-6-chloro-quinolinol (II). (I) and (II) were reacted with phosphorus oxychloride to dichloro derivatives, (III) and (IV) respectively, which were derived to the compound (V) and (VI) by reacting with thiourea. (V) and (VI) were also obtained from the reaction of (I) and (II) with phosphorus pentasulfide respectively. When o-chloro-, o-bromo- and p-chloro-aniline were condensed with acetoacetic ethyl ester in xylol to give the anilide derivatives (VII), (VIII) and (IX), followed by the cyclization with sulfuric acid, (IX) afforded (X) but the other compounds gave the starting materials unchanged. (X) was reacted with phosphorus oxychloride to (XI), which was prepared with thiourea and 2-chloro-6-nitrolepidine into (XII) and (XIII). (2) 4-Chloroquinoline 1-oxide was reacted with aniline, p-chloroaniline, p-phenetidine, o- and p-toluidine and p-anisidine to afford (XV-XX). On the other hand, quinoline 1-oxide, which was nitrated with potassium nitrate in sulfuric acid into 4, 8-dinitroquinoline 1-oxide, which was reacted with hydrochloric acid into (XXIII), followed by the reaction with p-chloroaniline into (XXV). The anti-candida test was attempted for (V), (VI), (XII), (XIII) and (XV), and 4-nitroquinoline 1-oxide was found to show strong anti-candida activity.
For the purpose of obtaining synthetic analgesics, 1-(2-dialkylamino- or -morpholino-acyl)-1-indanamine, its N-methyl and 2-phenyl derivatives (XIX to XLII), and 1-(2-dialkylamino- or -morpholino-acyl)-3-(1-indanyl)ureas (XLVII to LV) were synthesized, having 1-indanamine and 2-phenyl-1-indanamine as the parent ring and by the introduction of dialkylaminoacyl and alkylaminoacylurea groups as the substituent in the nitrogen atom. Pharmacological action of these compounds was examined and they were all found to have better analgesic effect than aminopyrine, with weaker toxicity than that in about half of the synthesized compounds.
1) The 32 kinds of carboxylic acid derivatives of pyrrole and pyrrolidine tested with the exception of L-proline were found to excite the motion of hog ascaris (Ascaris suilla) and Allolobophora foetida SAVIGNY with later paralysis. Of these compounds tested, kainic acid had the strongest action, followed by 5-isobutyl-4-methyl-3-carboxy-2-pyrroleacetic acid and 5-isobutyl-4-methyl-2, 3-pyrroledicarboxylic acid, and that of acetylkainic acid and dihydrokainic acid was weak. Appearance of motor paralysis by the 2, 3-pyrroledicarboxylic acid derivative is comparatively early and the action is similar to that of the aqueous extract of Digenea simplex Ag. 2) Effect of kainic acid and 5-isobutyl-4-methyl-2, 3-pyrroledicarboxylic acid, which had strong motor excitation action, on the oxodoreductase of hog ascaris was examined and it was found that both these compounds inhibited dehydrogenase action but not peroxidase. 3) Kainic acid and 5-isobutyl-4-methyl-2, 3-pyrroledicarboxylic acid caused denaturation of iron-containing substance in the epithelial cells of digestive organs in hog ascaris, vacuolization of mitochondria, and other retrograde metamorphosis, such as the metamorphosis and break-down of nucleus, out no marked change in dihydro-kainic acid. Comparative examination was made with dithiazanine.
Hog ascaris was found to contain a fair amount of cytochrome c and this suggests the possibility that the electron-transfer route is in a system passing through cytochrome c. Kainic acid and many of its allied substance were examined to see whether they inhibited electron transfer of hog ascaris and all were found to have such action, especially strong in kainic acid, L-proline, and many of pyrroledicarboxylic acids. Especially strong activity was found in pyrroledicarboxylic acids with isobutyl or phenyl group in 5-position of the pyrrole ring. 5-Isobutyl-4-methylpyrrol-2, 3-dicarboxylic acid (B. C. P.) and Dithiazanine were found to have stronger inhibitive action than kainic acid, vanillic acid was about equal to and N-acetylkainic acid was weaker than kainic acid. Kainic acid lactone and dihydrokainic acid had only a very weak inhibitive action. Kainic acid was found to inhibit oxidation of reduced diphosphopyridine nucleotide but not succinic acid dehydrogenase of hog ascaris.
Paper chromatography of artificial sweet flavours have been studied. Saccharin sodium was identified as a violet spot when colored with α-naphthylamine·cupric acetate solution over the developed paper. As this spot was observed to be a black spot on a pale blue fluorescent paper under ultraviolet light, the limit of identification of saccharin sodium has been increased more sensitively than the usual method. On the other hand, when the hydrolyzed solution of cyclamate sodium and dulcin with hydrogen peroxide in hydrochloric acid acidic media (the hydrolyzed compound were identified to be cyclohexylamine and p-phenetidine respectively) were developed and colored with quinhydrone solution, both showed violet-red spot, thus it was able to identify both cyclamate sodium and dulcin simultaneously. The developing solvent mixtures employed were AcOEt-HAc-H2O (4:1:2), AcOEt-HCOOH-H2O (4:1:2) and AcOEt-EtOH-AcOH-H2O (4:2:1:2) and the limit of identification was 15γ of cyclamate sodium, 20γ of dulcin and 3γ of saccharin sodium. The result of applying this method to the food-stuff was found to be favourable.
The tosylated o-nitroaniline (I) was condensed with 2-diethylaminoethyl chloride and N-(2-diethylaminoethyl)-2-nitroaniline (IV) was obtained, after the detosylation with conc. sulfuric acid. The nitro group of (IV) was reduced with stannous chloride and hydrochloric acid to N-(2-diethylaminoethyl)-o-phenylenediamine (V), which was condensed with carbon disulfide into 1-(2-diethylaminoethyl)-2-benzimidazolethiol (VI). When the potassium-salt of (VI) was reacted with the corresponding halides in either ethanol, benzene or xylene, 11 kinds of new substances were obtained. The analgetic effect of these derivatives was studied by the D'Amour-Smith method, and the substitution of hydrogen in the thiol group with methylene was found to give a decrease of the action generally, though (VI) showed a slight action.
Growth inhibitory action of 3-acetyl-4-hydroxycoumarin (AHC) and dehydroacetic acid (DHA) against lactobacillus and yeast was examined AHC inhibited the growth of these microörganisms both in the logarithmic and stationary phases of their multiplication. The antimicrobial activities of AHC and DHA became stronger at lower pH range. Taking account of pK′ values (AHC, 4.29; DHA, 5.12), it seems that the nondissociated form of AHC or DHA is more active than the dissociated form. The competitive effect of vitamins, amino acids, and metal ions to the growth inhibitory activity of AHC was also examined, but no restoration of the inhibition was observed by these substances except in the case of pyridoxamine which showed a slight recovery of the activity.
The inhibitory effect of 3-actyl-4-hydroxycoumarin (AHC) and dehydroacetic acid (DHA) on the biosyntheses of nucleic acids and proteins, as well as on the fermentation or the respiration of lactobacillus and yeast was examined. At a concentration of AHC or DHA where the syntheses of nucleic acids and proteins were markedly inhibited, the fermentation of lactic acid by L. arabinosus was rather promoted (about 7%). The inhibition of respiration of Sacch. cerevisiae was found much less than that of the synthesis of nucleic acids and proteins. These effects of AHC or DHA were similar to those reported for 2, 4-dinitrophenol as a growth inhibitor. Accordingly, it is probable that the inhibitory activity of AHC or DHA against these microörganisms depends on an uncoupling of the reactions concerned with anabolic and catabolic metabolism, e.g. oxidative phosphorylation.
The roots of Panax ginseng (P. shinseng), P. quinquefolium, and P. japonicus, which are used as crude drugs, contain saponins on which some studies have been reported since earlier times. None of the constituents of the first two roots have fully been elucidated, while oleanolic acid was found as a sapogenin of P. japonicus. In the present study, a prosapogenin and a sapogenin (panaxadiol) C30H52O3, m.p. 250°, were isolated from the roots of P. ginseng. A comparative study suggested that panaquilon (Garriques), panaxin (Kotake), and ginsenin (Yonekawa), which were isolated earlier workers, would be similar saponins while panacon (Garriques), α-panaxin (Kotake, et al.), and a compound, m.p. >270° (Asahina, et al.) would be identical with the present prosapogenin, m.p. 330°. Panaxadiol seems to be identical with panax sapogenol obtained by Kondo, et al., though it has not been established. From the sapogenin fraction of P. japonicus, a small amount of panaxadiol has also been isolated.
Some reactions have been studied to elucidate the chemical structure of panaxadiol, C30H52O3, a sapogenin of Ginseng roots. Panaxadiol contains an active hydroxyl, a hindered hydroxyl, and an ether oxygen, but no double bond. On oxidation of panaxadiol monoacetate with chromium trioxide, a ketonic compound, panaxanolone acetate, was obtained, whose ketone group was reduced by the Wolff-Kishner method into methylene to give panaxanol. By the action of hydrochloric acid in glacial acetic acid the ether oxygen bridge in panaxanol was cleaved to afford anhydropanaxanol acetate from which anhydropanaxanol, C30H50O, was formed on hydrolysis.
As the derivative of pregnanediol, a therapeutic for menstrual and climactic disturbances, 6α-methylpregnanediol was synthesized. Pregnanediol diacetate was derived to 6α-methylprogesterone via dihydroxyallopregnan-6-one diacetate. Reduction of 6α-methylprogesterone over palladium-strontium carbonate resulted in the formation of a mixture of 5α- and 5β-compounds, in a ratio of 1:3 in ether and in 2:3 in ethanol. Reduction of 6α-methylpregnanedione gave objective substance. Some considerations were made on the mechanism of partial reduction and the effect of 6α-methyl group.
In order to test therapeutic effect on menstrual and climactic disturbances, allopregnane-3α, 20α-diols substituted with hydroxyl, bromine, methyl, or fluorine in 2β-position were prepared from their 2α, 3α-epoxide.
It is reported that 2, 6-lutidine 1-oxide (I) afforded 6-methyl-2-pyridinemethanol (II) and 2, 6-dimethyl-3-pyridinol (III) in the reaction with acetic anhydride and that the obtained (II) was reacted again with acetic anhydride to afforded 2, 6-pyridine-dimethanol (VI). An investigation of the reaction confirmed that (I) produced 2, 2′-ethylenebis (6-methylpyridine) (IV), besides (II) and (III), and that (V) afforded 6-methyl-picolinaldehyde (VII) and 3-hydroxy-6-methyl-2-pyridinemethanol (VIII). Further attempt of reacting 2, 6-dimethyl-3-pyridinol 1-oxide (IX) with acetic anhydride produced (VIII) and 5-hydroxy-2-methyl-2-pyridinemethanol (X).
The reaction of ketene with N-oxides of eight sorts of pyridine and quinoline homologs, such as 4-picoline 1-oxide (I), 3-picoline 1-oxide (II), 2, 6-lutidine 1-oxide (III), 4-nitropyridine 1-oxide (IV), 4-nitro-2-picoline 1-oxide (V), 4-nitro-2, 6-lutidine 1-oxide (VI), quinaldine 1-oxide (VII) and, lepidine 1-oxide (VIII) has been investigated. Though (II), (IV) and, (V) were recovered unreacted, (I), (III) and, (VI) afforded the corresponding alcoholic and phenolic products, respectively. This reaction was considered to proceed similary to that with acetic anhydride and deoxygenated products have been obtained in each reaction. None of the phenolic derivatives were produced from the reaction of (VII), which afforded only alcoholic derivatives such as 2-quinolinemethanol and α, α′-bis (2-quinolinemethanol) (IX), however, (VIII) gave only phenolic products of 3-hydroxylepidine.
A production of tertiary base has been investigated by the reaction of excess LiAlH4 in dehydrated tetrahydrofuran with non-phenolic quaternary base, dl-laudanosine (III) methiodide, O, O-dimethylcorytuberine (IV) methiodide and dl-canadine (V) methiodide, and with biscoclaurine-type non-phenolic quaternary base, O-methyldauricine (VI) dimethiodide, isotetrandrine (VII) dimethiodide and insularine (VIII) dimethiodide, applying the Cope method. Methiodide of (III), (IV) and (V), having comparatively simpler structure, gave the corresponding tertiary base in better yields, however, only a few amount of desired tertiary base was produced from biscoclaurine-type base of being rather complicated in its structure. On the other hand, the reaction of aporphine type, phenolic quaternary base, corytuberine (X) methiodide and isocorydine (XI) methiodide, ended in almost complete recovery of the starting material.
The Hofmann degradation of isotetrandrine (I) gave two kinds of new methine crystals of having m.p. 149.5-151° and 107°, isolated by a buffer extraction at pH 4.0, besides the known α-isotetrandrine methine (II). The contact reduction of the respective derivatives gave tetrahydromethine (IV) and (IX), which were subjected to the ring opening reaction with metal-sodium in liquid ammonia. An investigation of the phenolic and non-phenolic base have approved that the structure of the methine of m.p. 149.5-151° was (III) and that of m.p. 107° (VIII).
Substances excreted into human urine after oral administration of sulfisoxazole was examined and a new type glucuronide, besides N4-glucuronide, was isolated. Its various properties were examined and its structure was assumed. This substance decomposes into sulfanilamide by the action of alkali hydroxide and into sulfisoxazole and glucuronic acid on being heated with dilute hydrochloric acid, the bonding ratio of the base to acid being 1:1. This substance was assumed to be the glucuronide of sulfisoxazole and the position of the bonding of glucuronic acid could be either in nitrogen of the isoxazole ring, or N1 or N4 of the sulfonamide group. Since the N4-glucuronide has already been found, this was assumed to be the N1-glucuronide. Comparison of ultraviolet spectra and paper chromatographic behavior of the synthesized N1-glucuronide and this substance showed disagreement. Application of crude β-glucuronidase to this substance resulted in partial decomposition to sulfisoxazole and glucuronic acid, and since this behavior is similar to those of sulfisoxazole N1-glucuronide and sulfaphenazole glucuronide, there is a possibility that this substance might be an O-glucuronide. It was therefore assumed that this substance is a glucuronide formed by bonding of glucuronic acid to the nitrogen in the isoxazole ring of sulfisoxazole.
A gas chromatographic method for some barbituric acid derivatives, antipyretics, and phenothiazine derivatives is described employing an apparatus with hydrogen flame ionization detector and a column containing 1% SE-30 Silicone polymer as the liquid phase.
It has been certified in buffer solution that glucuronic acid ethyl ester was converted firstly, in the water solution, into glucuronolactone, which later shifted to a equilibrium with glucuronic acid.
Examinations were made on Kitagawa's method for preparation of canavanin from Canavalia ensiformis and a method was devised for the use of ion-exchange resin to prevent occlusion of deaminocanavanin, which is responsible for lowering the purity and biological activity of canavanin, and to simplify the procedure. This new method was found to give the free-form canavanin of high purity, easily and in a good yield, directly from its flavianate.
As an alkaloid component from Aristolochia debilis SIEB. et ZUCC. (Japanese name “Umanosuzukusa”), a very small amount of the quaternary base, magnoflorine (I), had been reported to be contained in it. Further detailed examination certified that another quaternary base, cyclanoline (II) was contained in this plant.
Methyl glucopyranuronate (I) has been synthesized in liquid state from glucuronolactone (II) and methanol by the catalyst of sodium methylate. Ethyl glucopyranuronate (III) has been prepared similary from (II) and ethanol in the presence of sodium ethylate but for the isolation of pure crystalline state it was required to treat them from intensely colored reaction solution. Authors have found that in the case of ester syntheses, anionic exchange resine possessed an excellent catalytic activity and succeeded in isolating (I) in crystalline state. In the synthesis of (III), the reaction solution had almost colorless, therefore the unreacted (I) could be recovered and the crystals of (III) was obtained easily as crystals, simply by concentrating the reaction solution.
Mannan of Candida albicans was isolated by modified Vogel's method and purified by preparation of its copper complex. This material showed [α]D+53°, molecular weight of 4500 (Akiya-Barger method and from reducing end-group assay). Its infrared spectrum showed no β-configuration of mannopyranose. Periodate oxidation products of this mannan were identified with glyoxal (as p-nitrophenylosazone, m.p. 310°, and by infrared spectrum) and glyceraldehyde (as p-nitrophenylhydrazone, paper chromatography and electrophoresis). From these results, it was concluded that the mannan has been 1-2, 1-6, or 1-2-6 α-linkages, but not 1-3 or 1-4 linkages.
Twenty three kinds of 2-thiophenecarboxaldehyde derivatives were synthesized and their antibacterial action was examined with human-type tubercle bacilli H37Rv, sensitive strain, and a strain resistant to 100 γ of INH, 250 γ of PAS, and 250 γ of streptomycin. 2-Thiophenecarboxaldehyde salicyloylhydrazone showed antibacterial action against both sensitive and resistant strains at 6.25 γ and 4-alkylthiosemicarbazones at 12.5 γ, while 4-arylthiosemicarbazones were all ineffective. 2-(2-Thienylmethyleneamino) phenol showed a good antibacterial action at 1.25 γ against both sensitive and resistant strains.
Colorimetric quantitative determination of dihydrocodeine by ceric ammonium nitrate, 2, 4-dinitrophenylhydrazine has been studied. Though such an easily oxidizable substance as aminopyrine, sulpyrine and phenacetine interfere the determination, a presence of amynophylline, methoxyphenamine hydrochloride of five times as much of dihydrocodeine and nearly half volume of chloroprophenpyridamine maleate do not interfere the determination. This method follows the Beer's law within the concentration of 30-300γ/cc. with an accuracy of σ=0.95% (n=6). Method of Determination: It will be carried out similarly to that of codeine. The concentration of ceric ammonium nitrate is 0.8% and the reaction period of 2, 4-dinitrophenylhydrazine is 25 min.