In an attempt to find some effects, if any, of a few organic acids on metabolism, various changes appearing in the urine of the writer himself after ingestion of various organic acids were followed. With the constant diet of noodles, ingestion of citric or acetic acid was found to elevate the pH value of the urine, which attained the maximum 2 hours after taking such acid. At the same time, the amount of lactic acid in the urine was found to be markedly decreased. The same tendencies were not so marked in the case of the ingestion of tartaric and lactic acids. Considering from such results, it is thought that the ingestion of citric or acetic acid somehow affected the progress of the Krebs cycle in the body, probably effecting better oxidation of pyruvic acid, with attendant decrease of the formation of lactic acid from pyruvic acid, and the elevation of urine pH by the decrease of acid substances in the urine.
Effect of the ingestion of citric or acetic acid on the creatine and creatinine, total nitrogen, ammonia nitrogen, amino nitrogen, urea, and uric acid in the urine was examined by human tests of the writer himself. It was thereby found that the ingestion of citric or acetic acid did not give any marked changes on the amount of creative, while the decrease was noted in the amount of creatinine, ammonia nitrogen, and uric acid, and a tendency for the increase in the amount of total nitrogen, amino nitrogen, and urea. From these results, the decrease in the amount of creatinine and increase in the amount of urea were found to be significant and the ingestion of citric or acetic acid was thought to influence metabolism in the human body.
Effect of the ingestion of citric or acetic acid on the amount of calcium in the urine was examined by the writer himself. As a result, these acids were found to effect increase of calcium in the urine while the calcium decreased on the concurrent ingestion of dlsodium phosphate than in the case of citric or acetic acid. The changes in the amount of calcium in the urine was assumed to be due to the effect of citric or lactic acid ingestion on metabolism in the human body. The decrease in the amount of calcium in the urine with concurrent ingestion of a phosphate seemd to be of special significance.
Water of pH 9, giving positive tetrabromophenolphthalein (TBP) reaction, was passed through a column (1×10cm.) filled with 10g. of ion exchangers, Cl-type Amberlite IRA-400, H-type Amberlite IR-120, or Na-type Amberlite IR-120, at the rate of 2cc./minute. The effluent water was found to give negative pyrogen test and weak TBP reaction. The same adsorption treatment of pyrogenpositive serum through a column of Na-type Amberlite IR-120 also resulted in the removal of pyrogen from the serum with only a small loss of antibody value.
Fructose methylphenylhydrazone-2, considered to be the isomer of glucosone methylphenylhydrazone to date, consumes over 5 moles of periodic acid in oxidation and forms by catalytic reduction, not the corresponding glucosaminol and mannosaminol, but fructose itself. On the contrary, glucosone (fructosone) methylphenylhydrazone consumes 4 mols of oxygen by periodic acid oxidation to form glyoxalic acid methylphenylhydrazone and absorbs 2 moles of hydrogen by catalytic reduction with palladium-carbon to form isoglucosamine. The fact indicates that the hydrazine residue in such hydrazones is bonded at C1.
Some attempts were made to elucidate the partial structure of limonin by the summarization of known facts and reëxaminations were made on few points. Starting with etiolimonic acid, the ozonolytic product of limonin, desoxyetio-limonic acid and dihydroetiolimonic acid were newly prepared in crystalline state. Examination of the infrared absorption spectra of limonin derivatives reconfirmed the presence of a lactone, carbonyl, and an oxime, and the presence of a hydroxyl group in dihydroetiolimonic acid and dihydrolimonin.
Analgesic, sedative, and anticonvulsant activities and toxicity of ethyl 1-methyl-4- (3′, 4′-dimethylphenyl) piperidine-4-carboxylate (Dimethylpethidine) and 1-methyl-4-(3′, 4′, -dimethylphenyl) piperidyl-4-methyl (or ethyl) sulfone were compared with those of Pethidine. Analgesic action of Dimethylpethidine was slightly weaker than that of Pethidine but the anticonvulsant action was about equal, analgesic action was stronger, and toxicity was weaker, LD50 being 0.29mg./g. mouse by subcutaneous injection. The compound obtained by the substitution of ethoxycarbonyl group in Dimethylpethidine with methyl (or ethyl) sulfone possessed weaker analgesic and anticonvulsant action than those of Pethidine, with equal or stronger analgesic action, and weaker toxicity.
Spectroscopic studies were made on the complex formation of Eriochrome Black T with Cu2+, Co2+, Ni2+, Zn2+, Cd2+, Mg2+, Ba2+, Sr2+, and Cr3+. The complex formation of these metallic ions is affected to a great extent by the pH and the measurements were carried out at pH 3.4, 5.9, 8.2, 10.0, and 13.5. Cu, Co, and Ni ions were found to form complex salts at any pH but Zn, Cd, and Mg ions formed complex salts only in the alkaline range and not in the acid range. Ba and Sr ions failed to form complex salts in the concentration of 3×10-4 mole used in the present experiments. Cr ion did not form the complex in the alkaline range and formed a substance on the acid range but whether this is a complex salt or a simple sulfonate remains unclarified, though the absorption curve of the product was different from that of Eriochrome Black T. The composition of such complex salts were found to be in 1:4 molar ratio of Cu and Co ion with Eriochrome Black T, 1:2 molar ratio of Ni, Zn, and Cd ions with T, and 1:1 molar ratio of Mg with T, in a solution of pH 10.
In order to study the relationship between the fluidity of granules and weight deviation of tablets during tabletting, granules were coated with vaseline to obtain granules of different fluidity. Actual tests with such granules indicated that the fluidity of granules which affects weight deviation is chiefly static friction (Fig. 1), since greater its coefficient, the greater becomes the weight deviation (Fig. 3). The coefficient of static friction increases with the increasing amount of powder (incluing lubricant powder) in the granules (Fig. 4). From these results, it seems more advantageous to have the granules as small as possible and its limit seems to be 100 mesh (Fig. 5). The lubricant powder effects decrease of the coefficient of static friction of sticky granules but such ability differs with different grades (Fig. 6). It seems, from these results, that it would be significant to measure the static friction coefficient of granules in checking the granular quality of a lot in the plant.
The expectorant effect of crude saponins obtained from the seeds of tea (Thea sinensis L.), camellia (Camellia japonica L. var hortensis Makino), and Camellia Sasanqua Thunb. was examined by measuring the amount of increased secretion of tracheal mucus in puppies. It was thereby found that the crude saponin from tea and Sasanqua was stronger than that of Panax japonicum, with higher fish index. The expectorant effect of camellia saponin was about equal to that of the Panax saponin but with far higher fish index. It seems that the hemolytic index and expectorant effect are not parallel between different saponins.
2-o-Hydroxybenzylidenehydrazono-4-thiazolidone and its 5-substituted derivative possess strong antibacterial action against the tubercle bacilli but their allied compounds in which no hydroxyl group is present in the ortho-position of the benzene ring are weak in such action. Allied compounds possessing allyloxyl, bromo, or carboxyl group in the ortho-position were prepared and their antibacterial action was tested but they all had only a weak activity, especially those with the carboxyl group. The compounds (II) to (V) in Table I were obtained by the usual method of condensing aldehyde thiosemicarbazones with ethyl chloroacetate or ethyl α-bromopropionate. 2-o-Carboxybenzylidenehydrazono-4-thiazolidone (VI) was obtained from 2-α-methylbenzylidenehydrazono-4-thiazolidone by decomposition with hydrochloric acid to 2-hydrazono-4-thiazolidone hydrochloride, followed by application of phthalaldehydic acid.
It has already been shown that o-hydroxybenzylidenehydrazono-4-thiazolidone (I) is a powerful growth inhibitor of tubercle bacilli. 2-Salicyloylhydrazono-4-thiazolidone (II), with a very similar structure with a hydroxyl group in the ortho-position of the benzene ring bonded to -CONH-N=group, lacks the antibacterial functional group, -CH=N-N-, present in (I) so that its antibacterial action is very weak. Ten kinds of 2-acylhydrazono-4-thiazolidones possessing -CO-NH-N=structure as in (II) were prepared and were submitted to antibacterial tests. The actions were found to be very weak with the exception of 2-isonicotinoylhydrazono-4-thiazolidone (VIII), whose comparatively high antibacterial action was assumed to be due to the isonicotinoyl residue. The syntheses of these compounds followed two methods. The one started with the heating of carboxylic acid hydrazide hydrochlorides with ammonium thiocyanate in chlorobenzene as a solvent, the thiocyanate was submitted to rearrangement to form the acylthiosemicarbazide, which was condensed with ethyl chloroacetate to obtain the desired 2-acylhydrazono-4-thiazolidones. The other was a new method devised by the writers in which 2-thio-4-thiazolidone and carboxylic acid hydrazide were reacted in methanol solution, in weakly alkaline state, with hydrogen peroxide which effected concurrent desulfurization and the desired reaction to the objective substance. This method is recommended because of a good yield of the products.
Condensation of the active methylene group at 5-position of 4-thiazolidone derivatives with aromatic aldehydes results in the formation of 5-arylidene-4-thiazolidones. These substances contain α, β-unsaturated ketonic bonding, =C=C-C=O, in their molecule so that they should react with the SH-system enzymes requisite for bacterial metabolism and may inhibit the growth of bacteria. Under such expectations, 2-thlo-, -oxo-, or -imino-5-arylidene-4-thiazolidones were prepared and were submitted to in vitro screening for tubercle bacilli. Of these compounds tested, 2-oxo derivatives showed the most powerful action, followed by the 2-thio derivatives, while 2-imino derivatives showed a much weaker action than the foregoing two. The effect of hydroxyl and nitro groups as a substituent in the benzene ring and the position thereof on the antibacterial action did not follow any definite order. The compounds already appearing in the existing literature were prepared as described and the unknown substances were prepared in a good yield by the condensation of 4-thiazolidones with aromatic aldehydes in acetic acid with the use of sodium acetate or in dilute sodium hydroxide solution with the addition of ammonium chloride.
Examinations were made on the fact that the yeast ribonucleic acid causes non-ionization phenomenon of the silver ion and on such abiltiy in nucleic acids. The results obtained showed that there is a quantitative constancy between nucleic acid and silver (RNA-Na:Ag=1:0.16mg.). The amount of silver in the sample of RNA-Ag product and nondialyzability of silver bonded to nucleic acid were determined. It was thought that the polynucleotide construction of nucleic acid molecule was necessary for the appearance of non-ionizability of nucleic acid to silver ion. It was also found that the electropherograms of nucleic acid and RNA-Ag product are identical while the shape of the ultraviolet absorption curve of RNA-Ag product is the same as that of nucleic acid but was shifted to a longer wave length region. It is therefore concluded that the non-ionization of silver salt by nucleic acid is due to complex formation.
In connection with the synthesis of terracinoic acid, intramolecular cyclization of β-phenylpropionic acid and its esters, indicated by the general formula C6H5-CRR′-CH2-COOR″ (R=R′=R″=H or CH3), with polyphosphoric acid was examined. The yield was found to be no different from that by the Friedel-Crafts method. The methoxyl group was found not to be affected by polyphosphoric acid but did affect yield of indanone, especially in the case of ortho-substituted, with which the formation of indanone was not found (Table I). Polyphosphoric acid was found to be a suitable condensation agent since it did not affect the methoxyl group and the ester can be used per se for the cyclization reaction. However, the separation of the ester and the indanone was difficult when the boiling points of the two were similar, as in the present case.
As a possible intermediate for the preparation of antipyrine derivatives of therapeutic value, 4-f ormylantipyrine was prepared by treating antipyrine either with dirnethylformamide or formyl methylanilide in the presence of phosphoryl chloride. The yields were 88.1% and 53.2%, respectively. As an aldehyde, this compound condensed with malonic acid, nitromethane, hippuric acid, and diketopiperazine with separation of water. However, benzoin and Cannizzaro type of reactions failed to give the expected compounds, (VI) and (VII) being the chief products, respectively. Several other derivatives of 4-formylantipyrine were also prepared, none of which, however, showed properties of any interest from physiological view point.
In an attempt to synthesize N-methyl-3-hydroxy-(4b, 8a-ethanoiminomethano)-4b, 5, 6, 7, 8, 8a, 9, 10-octahydrophenanthrene (B), its intermediate 9-phenethyl-N-methyl-1, 2, 3, 4, 6, 7, 8, 9-octahydroisoquinoline was prepared by the method shown graphically in the text, proving the feasibility of the synthesis of (B).
A possible use of ion exchange resins for isolation of Aureomycin in urine was investigated. It was found that Aureomycin and components of urine were retained on strong cation exchangers, Amberlite IR-112, but it seemed possible to remove urine materials by the following procedures. Aureomycin can be eluated with 1N HCl-EtOH solution (8%) while the use of 1.5N AcOH-EtOH or 0.2N HCl-EtOH eluates the components of urine alone, not Aureomycin. A colorimetric determination of Aureomycin present in the eluate involves reaction of this antibiotic with boric and sulfuric acid mixtures, as previously described.
The Aureomycin-borate complex in concentrated sulfuric acid might be composed of two components, according to the increasing concentration of sulfuric acid used, if borates are present in excess. The one somewhat stable chelate compound in 78% H2SO4 showed the maximum absorbance at 555mμ and the other stable one in higher concentration of sulfuric acid than 91%, showed the maximum at 530mμ. The structure and nature of these two chelate compounds were presented and the possible mechanism was discussed. The molar ratio of Aureomycin to boron was 2:1 in the former and 1:1 in the latter.
Ten kinds of dialkylethynylcarbinol derivatives and two of their hydrogenated derivatives were prepared and their head-drop dose, HD50, and toxicity, LD50, were measured. It was thereby found that all the compounds tested possessed hypnotic action, without causing any side-effects such as convulsion. Approximate comparison of the effect of each carbinol derivative with 1/HD50 of 3-methylpentyn-1-ol-3 as 1, for convenience, showed that the effect of 3-ethylpentyn-4-ol-3 and 3-methylpentanol-3 was greater than that of 3-methylpentyn-1-ol-3 while all the others were weaker. In general, the hypnotic action was comparatively strong when the sum of the carbon atoms in the two alkyl groups in the foregoing dlalkylethynylcarbinol derivatives was 3-6, and weak when such was 2 or above 7, the strongest being that in which both alkyls were ethyl. When one of such alkyl groups were fixed to methyl, ethyl, propyl, or butyl, and the other alkyl changed from methyl to butyl, the strongest hypnotic action appeared when the latter was an ethyl group. The presence of the ethynyl group was considered to be a requisite to a certain extent in the appearance of the hypnotic action.
Nine kinds of alkylcoumarin derivatives were prepared and their hypnotic effect and toxicity were compared with those of 4-methylcoumarin as the standard. It was thereby found that the alkyl group (methyl to butyl) in the 3-position of the coumarin ring effected hypnotic action when the number of carbon atoms in the alkyl was odd and toxic action when such number was even, with the peak toxicity with the ethyl group. The effect of the methyl group into the coumarin ring was found to be in the decreasing order of 8-, 3-, and 4-methyl, while that of the toxicity was in the increasing order of 4-, 8-, 3-, 5-, 7-, and 6-methyl compounds. Side effects, such as convulsion, was not observed in 3-propyl-, 4-methyl-, and 8-methylcoumarins, while sedative action was observed in 3-methyl, 4-methyl-, 5-methyl-, 8-methyl-, 3-propyl-, and 3-butylcoumarins. In general, the difference between HD50 and LD50 seemed to be slight in alkyl coumarins, with the exception of 4-methyl- and 8-methylcoumarins.
The aqueous extract of Digenea simplex Ag. yielded, besides the amino acid already reported, two substances which give yellow ninhydrin reaction. One of them was isolated as colorless, long needles, m.p. 275° (decomp.), C10H15O4N⋅H2O, which was confirmed, by paper chromatography, infrared spectral analysis, and mixed fusion, to be α-kainic lactone, obtained by the acid treatment of kainic acid. By saponification, the lactone was derived to hydroxykainic acid, C10H17O5N, as colorless needles, m.p. 260° (decomp.).
Skyrin, iridoskyrin, islandicin, a pigment of m.p. 287° (decomp), and a pigment of m.p. 273° (decomp) were isolated from the ether extract of the mycelium of Penicillium islandicum Sopp, D strain. It is assumed that the pigment of m.p. 287° (decomp) is closely related in its chemical structure to the pigment of m.p. 273° (decomp), which has a toxicity LD50 of 5mg./15g. mice (per os).
Nineteen kinds of styryl-type dyes were prepared by the condensation of tertiary salts of 2-methyl-6′-chloro (methoxy, ethoxy) pyrido [2′, 3′:5, 4] thiazoles and γ-picoline with aromatic aldehydes. A kind of styryl-type dye was obtained by the application of salicylaldehyde on bis [4-methylthiazolyl-(2)] methane monomethiodide. Two kinds of aminovinyl compounds were obtained by the fusion of the quaternary salt of 2-methyl-6′-chloro (ethoxy) pyrido [2′, 3′:5, 4] thiazoles with diphenylformamidine.
The principle which effects decrease of serum calcium level in rabbits, contained in equine submaxillary gland, was found to be collected in a fraction of 12.5% sodium sulfate concentration from the pH 5.4-precipitate. This fraction, similar to parotin and S-parotin, were found to effect the circulating leucocyte counts in rabbits, the effect being the strongest among the various extracts of the equine submaxillary gland. Electrophoretic test of this fraction showed its purity to be 74.07% and the fraction was positive to general protein reaction. The maximum absorption in the ultraviolet spectrum appeared at 276mμ. This fraction was similar in many points to bovine S-parotin that it was designated as equine S-parotin.
Sixteen kinds of amino acids were detected as the componental amino acids in S-parotin of 96.1% purity by electrophoretic test. The terminal amino acids in S-parotin were found, by the DNP method, to be alanine, histidine, and arginine. DNP-S-parotin did not have the potency of decreasing serum calcium level in rabbits while the elementary analysis of S-parotin gave the values of 49.53% carbon, 7.35% hydrogen, and 14.05% nitrogen. The fractionation of the principle which decreases the serum phosphorus level was obtained with 71.9% purity, which contained 9.49% nitrogen, and its isoelectric point was pH 4.0. The effect of S-parotin on the amount of pyruvic acid in the blood was similar to that of parotin but the principle which decreases serum phosphorus level effected marked decrease of pyruvic acid in the blood 30 minutes after its injection, the amount of the acid returning to the normal after 6 hours.
There is a parotin-like substance in the hog subtaxillary gland, similar to those in the bovine and equine submaxillary glands. This was designated as the hog S-parotin. The fraction of 15% ammonium sulfate (pH 5.4-precipitate) showed the strongest activity of decreasing the serum calcium level in rabbits and also showed effect on circulating leucocyte counts in rabbits and ultraviolet absorption spectrum similar to those of the bovine and equine S-parotin. This fraction was found to have over 90% purity by electrophoretic analysis.
The hog S-parotin gave elementary analytical values of 45.85% carbon, 7.40% hydrogen, and 14.33% nitrogen, and was positive to almost all of the general protein reaction. Its componental amino acids were 17 kinds, same as in parotin, and contained valine which was not present in the bovine and equine S-parotin. Polarography of the hog S-parotin showed a half-wave potential by the second wave of bivalent cobalt of -1.35 V. This S-parotin did not affect the inorganic phosphorus level in rabbit serum.
Requisite conditions for the preparation of furylacrylic acid and 5-nitrofurylacrylic acid have been examined. The results indicated that furylacrylic acid can be obtained in 71.2-75.80% yield by reacting a mixture of 1 mole furfural, 1.2 moles acetic anhydride, and 1.4 moles anhydrous potassium acetate at 145-150° for 1.5-2.5 hours, and that 5-nitrofurylacrylic acid can be obtained in 54.4-55.3% yield by the addition of 1 mole of furylacrylic acid to a mixture of 4 moles of 80% nitric acid and 6.3 moles acetic anhydride maintained at -2° to -5°.
Standard curve showing relationship between the unit of impure insulin from whales and the value obtained by the colorimetric method gave a straight line. Comparison of determination of whale insulin by the present method and the bioassay specified in Japanese Pharmacopoeia showed that they gave approximately agreeing results, with an average error of ±0.7 unit.
It was found by admixture that 5-butylpicolinic acid synthesized by the present writer was identical with fusaric acid, extracted from the culture filtrate of Gibberella fijikuroi of rice plant. 5-Propyl- and 5-pentylpicolinic acids were prepared from 2-methyl-5-ethylpyridine by its derivation to the 5-cyano compound, to 5-propionyl and 5-valeroyl derivatives by the Grignard reaction, to 5-propyl and 5-pentyl derivatives by the Wolff-Kishner reduction, reacted with benzaldehyde to change the 2-methyl to 2-styryl, and finally oxidized with potassium permanganate in acetone. It was found by the examination of the reaction of the cyano group in 3-cyano- and 2-methyl-5-cyanopyridine with the Grignard reagents that there was no great difference in the yield of the ketone formed when the alkyl group of the reagent was methyl, ethyl, propyl, or butyl.
In the ultraviolet spectra of hydroxyanthraquinone derivatives, the absorption appearing in shorter wave lengths region than 250mμ is very important, though it has been neglected to date. When anthraquinones have one, two, three, and four hydroxyl groups, λmax appears at 222.5, 225, 230±2.5, and 236mμ, respectively. These maxima are independent of the nature of the hydroxyl groups, as at α or β, but the intensity value depends chiefly on the number or α-hydroxyls. The intensity of maximum absorption at 270mμ (ca.) depends on the β-hydroxyl group and the log ε value above 4.1 shows the presence of a β-hydroxyl group and the value below 4.1 denies it. Therefore, it would be possible to predict the number of hydroxyls and α:β ratio from the ultraviolet absorption curve. Further, hydroxyanthraquinone derivatives with 1, 4-dihydroxyl in the molecule exhibit a shift in the visible region and the absorption maximum appears at above 480mμ.
N1-Acylation of 4-acetaminonaphthalene-1-sulfonamide using condensation agents other than pyridine, such as trimethylamine, quinoline, or dimethylaniline, with acetone and sodium hydrogen carbonate, was compared with that of the pyridine method and a few new observations were made.
From the acid ether-soluble portion of the extracts of the stems of Berbevis Thunbergii DC. var. Maximowizii Franch and of Berberis amurensis Rupr. var. japonica (Regel) Rehd. forma Bretschneideri (Rehd.) Ohwi, which contain berberine, ferulac acid, which exhibits blue fluorescence under ultraviolet light, and vanillinic acid, which does not show fluorescence, were detected by paper chromatography. There was a compound in the neutral portion of the latter plant which exhibited blue fluorescence.