Anthelmintic activities of various compounds were tested by the oral administration of 1g. of each to human adults. Alkylresorcinol derivatives, such as 2, 4-dihydroxy-5-alkylbenzaldehyde and 2, 4-dihydroxy-3-chloro-5-alkylbenzaldehyde, were found to be ineffective. 2-Chloro-3-methyl-4-thiocyano-6-isopropylphenol was slightly effective, but 4-chloro-3-methyl- and 3-methyl-4-thiocyano-6-isopropylphenol were practically ineffective as an anthelmintic. Of diphenylmethane derivatives, 2, 4-dihydroxy-5-chloro-phenylmethane showed a worm expulsion rate of 83% in the first test, but the rate dropped to 33% in the second trial. The exceptionally high rate shown in the first, test is assumed to have been due to the presence of a minute amount of impurity in the compound. 2-Chloro-4-cyclohexylphenol showed the expulsion rate of 66.6% although the substance possessed a slight unpleasant odor and taste. Diallyl sulfide, on which much expectation was placed, failed to show any anthelmintic activity. A good expulsion rate of 33.3% was shown by phenyldihydroquinazoline, used as its citrate, and it might give a good result with repeated use. Assuming a certain relationship to exist between the decrease of the melting point and the anthelmintic action, fusion compounds of several anthelminties, and a substance obtained by the further addition of digenea extract, expecting an additive action, were prepared. A fusion product of ethyl phenyl thiocarbamide and butyl phenyl thiocarbamide, cyclohexylresorcinol, chlorocyclohexylresorcinol or hexylresorcinol, or with the addition of digenea extract were tried but only the substance prepared br the addition of digenea extract, hexylresorcinol, and cinnamon powder to the fusion product of the two esters showed a worm expulsion rate of 60, 70 and 100%, the others being totally ineffective.
A simple method for the microestimation of proteins by the use of tetrabromophenolphthalein ethyl ester (T. B. P.) is described. A mixture of 5.0cc. of the test solution and 0.1cc. of 0.05% T. B. P. solution is adjusted to pH 3.8 with 0.1 N acetic acid, and its optical density is measured by a photometer. From the protein concentration-absorption coefficient curve (Fig. 5), the quantity of protein (casein value) is calculated. The limit of estimation is approximately 1 γg/cc.
Nitration of benzo-(f)-quinoline with sulfuric and nitric acids at a low temperature results in the formation of 7-, 9- and 10-nitro derivatives in approximately 40:8:13 ratio. Nitration of henzo-(f)-quinoline N-oxide with conc. sulfuric acid and potassium nitrate at 60° chiefly yields 7-nitro compound with a trace of 10-nitro compound as a byproduct. In either case, the substitution occurs on the outermost benzene nucleus. Furthermore, no specific polar effect of N-oxide group can be witnessd in this case. These two points are similar to the case of benzo-(h)-quinoline and its N-oxide. In connection with the present series of studies, 6-bromobenzo-(f)-quinoline was synthesized.
The genuine glycoside, rhodexin C, was hydrolyzed with crude enzyme extracted from fresh leaves of Rhodea japonica and the formation of rhodexin B and glucose in the product was confirmed by paper partition chromatography. Further, glucose content was determined by the Bertrand method and the decomposition rate was calculated.
An Attempt was made to utilize the decomposition of thiaminase by sulfurous acid for the synthesis of allied compounds but it could not be made successfully. For example, reaction of N-benzylpyridinium chloride and aniline, in the presence of sulfurous acid, failed to yield the desired N-benzylaniline. Reaction of N-[2′-amino-4′-hydroxypteridyl-(6′)]-methyl-4-methyl-5-β-hydroxyethylthiazolium iodide and N-(p-aminobenzoyl)-glutamic acid, in the presence of sulfurous acid, also failed to give any indication of the formation of folic acid. Therefore, the relationship between this decomposition reaction and chemical structure was examined, and as a result, it was found that compounds in which the 2-position of the pyrimidine nucleus is substituted with ethyl radical (III) or the 4-position with methylamino radical (II) are decomposed almost similar to the case of thiamine (I), but that hydroxythiamines are not decomposed at all. In this case, the amino radical at 4-position has an important significance. Compounds in which the thiazole nucleus is substituted with methyl (IX), ethyl (X) or carbethoxyl (XI) in the 5 position, are also decomposed, the decomposition being promoted by the addition of aniline. However, the compounds in which the pyrimidine nucleus has been substituted with nitrobenzyl (VI), (VII), (VIII), or in which the thiazole nucleus has been substituted with benzothiazole (XII) are entirely indifferent to the decomposition. The effect of the addition of thiamine analogs during this reaction to the decomposition of thiamine was examined from which it was learned that nitrobenzyl homologs (VI), (VII), (XIV) and 2′-methyl-4′-aminopyrimidyl homologs (XI) and (XVI) tended to inhibit decomposition, while o-aminobenzyl homologs (XIII) and (XV), and benzothiazole homologs (XII) promoted decomposition. Hydroxythiamine (IV) seemed to give no effect.
Body weight, normal body temperature, temperature decrease and variability in body temperature, with and without parenteral administration of glucose-calcium chloride injection, were examined with rabbits fixing it on its back, and measuring these points either consecutively or in an interval of 3 and 7 days. It was then found that by using a rabbit well accustomed to this back posture before the test, by using the animals in 3-day intervals, and by the injection of the solution after the temperature has reached the lowest level, the decrease of body temperature due to the glucose-calcium chloride injection can be suppressed.
Influence of external temperature on the body temperature of a rabbit fixed on its back was examined. The decrease of atmospheric temperature to 0°C failed to effect any marked decrease of body temperature in unfixed rabbits. The rabbits were fixed on its back at this temperature, and when the body temperature has decreased considerably, pyrogen and a substance which normally effects lowering of body temperature in rabbits were administered. As a result, it was found that the body temperature failed to rise by the administration of pyrogen in minimum pyrogenous amount, but the lowering of body temperature was also found to be small.
Culture filtrate of Pseudomonas aeruginosa No. 151 and Escherichia coli were administered to rabbit fixed on its back, consecutively and in 4- and 7-day intervals, in a minimum pyrogenous quantity. The temperature rise was found to decrease gradually by repeated administration, daily or in 4-day intervals, but almost constant in 7-day intervals.
1) The seasonal variation in santonin content and the yield of crop were examined with the second year growth of Artemisia kurramensis Qaz., sown at Kasukabe in April 1950, during July 21, 1951, to November 2, 1951 (Table I). The results showed that the yield of crop harvested on August 21 was the largest with 246.7kg per 10 ares on air dry basis, with the highest santonin content of 2.341%, which is enough to rival that of Flos Cinae (flowers of A. Cina Willk.). 2) In the same way, the seasonal variation in santonin content and the yield of crop were examined with the first year growth of the same plant, sown in March 1951, during September 20, 1951, to March 18, 1952 (Table II). 3) In general, both the santonin content and the yield of crop increase extremely in the second year growth, when sown in spring. 4) The cultivation of A. kurramensis in Japan is largely dependent on the amount and number of days of rain during summer, especially in July and August. 5) Some knowledges were obtained on the breeding of seedlings of the autumn-sown plant.
Synthesis of alkynol from aldehyde and acetylene has been made possible by W. Reppe. 3, 4-Methylenedioxyphenylethynylcarbinol (I) was synthesized by the ethynylation of piperonal and acetylene under high pressure and high temperature. Hydration of (I) in the presence of mercury sulfate, yielded 3, 4-methylenedioxyphenyl-acetyl carbinol (II) which, by catalytic reduction in methanolic solution of methylamine, finally gave 3, 4-methylenedioxyephedrine. Its pharmacological action was compared with that of ephedrine by a simple animal test.
Eleven kinds of N, N-dimethyl-N′-(2-thiazolyl)-ethylenediamine derivatives were prepared by the respective condensation of 2-arylaminothiazoles, 2-arylmethylaminothiazoles and 2-arylamino-4-methylthiazoles with dimethylaminoethyl chloride, in the presence of sodium amide. N, N-Dimethyl-N′-benzyl-N′-(2-thiazolyl)-ethylenediamine was obtained by the condensation of N, N-dimethyl-N′-(2-thiazolyl)-ethylenediamine and benzyl chloride. Of the compounds tested as to their pharmacological activity, N, N-dimethyl-N′-benzyl-and N, N-dimethyl-N′-(p-methoxybenzyl)-N′-(2-thiazolyi)-ethylenediamine were found to possess remarkable antihistaminic activity.
N, N-Dimethyl-N′-(p-halobenzyl)-N′-(2-thiazolyl)-ethylenediamines were prepared by the condensation of 2-(p-halobenzylamino)-thiazoles and dimethylaminoethyl chloride. Of the compounds obtained, those with fluorine showed the strongest antihistaminic activity.
Since the sulfathiazole exhibited an antibacterial action against avian type tubercle bacilli among the various sulf as tested, it was assumed that the thiazole nucleus might possess a peculiar affinity towards acid-fast bacilli, above the fact that the thiazole nucleus possessed comparatively weak toxicity. For these reasons, some thiazole derivatives possessing simple substituents were prepared and their antibacterial activities were tested (Cf. Table I). Compounds of the general formula X-C6H4-X1-R in which factors that were assumed to be responsible for antibacterial activities were taken up and the electronic conditions of X and X1 were discussed according to the report of Ishidate and Okano. The relationship between the structure and the antibacterial activities was compared with a few benzene and pyridine derivatives.
In accordance with the method of Ochiai and Nagasawa, 5-alkyl ketone derivatives of 2-hydroxy-4-methylthiazole were prepared by the Friedel-Crafts reaction, and the relationship between the number of carbon atoms in the alkyl chain and the antibacterial activities was examined. As shown in Table I, the antibacterial activities increase with the increasing number of carbons between 1 to 5 carbons against Mycobacterium tuberculosis H 37.
Since 2-mercaptob-5-nitropyridine showed a strong antibacterial activity against avian type tubercle bacilli among the thiol compounds tested, its activities against various bacteria were tested (Table III). It was learned that the pyridine derivatives possessing hydroxyl or amino group in the p-position of the nitro group hardly showed any antibacterial activity (Cf. Table I), while pyridines possessing thiol group alone showed activities against both gram-positive and -negative bacteria. None of these compounds were found to be antagonized by p-aminobenzoic acid, methionine, cysteine or cystine (Table II), and were not precipitated by other than Cd... Studies are being continued as to the antibacterial activities, toxicity and the mechanism of such activities of allied. compounds.
Near-ultraviolet absorption of quinoline N-oxide and its derivatives in various solvents was determined and following results were obtained: 1) In solvents of weak polarity not possessing any active hydrogen, bathochromic effect is very great compared to the corresponding quinolines but, with the increase in the polarity of solvents, hypsochromic effect increases that in acid solution or in conc. sulfuric acid, the absorption becomes almost equal to that of corresponding quinolines. From these results, the contribution of the electronic state of the N-oxide group in the molecule is discussed. It was concluded that the chief reason is the solvation effect was comparatively examined with the solvation effect of pyridine N-oxides. 2) The near-ultraviolet absorption appearing in naphthalene and quinoline compounds was explained by the directional properties of electronic vibration.
By the adrenal ascorbic acid depletion method of Sayers, et al., the ACTH activity was examined with the pituitaries of bonito (Katsuwonus vagans), tuna (Thynuus orientalis) and sei whale (Balaenoptera borealis). The crude prolactin fractions were found to be toxic but the toxicity disappeared upon acid-heat treatment and only the ACTH activity could be observed. Since the number of rats used under the present experimental conditions were limited, the results could not be discussed quantitatiyely but it can be qualitatively defined that fish pituitaries also possess ACTH activities. The extract from the bonito pituitary by the method of Li, et al. possessed 12.5 γ/100g. efficacy.
Used against soy sauce, propyl p-hydroxybenzoate, as a control, failed to prevent growth of the mold after 5 days in approx. 0.005% concentration, or 6 days at approx. 0.007% concentration, while among the 16 kinds of compounds of the 1st group tested, the growth of the mold was inhibited during the 60-day test period by vitamin K3 in approx. 0.005% concentration, and by 2-cyclohexylresorcinol, 6-chloro-2-cyclohexylresorcinol, 2, 4-dihydroxy-3, 5-dichlorovalerophenone and 6-tert-amyl-2-naphthol in approx. 0.001% concentration. Of the 12 kinds of compounds of the 2nd group, mold growth was prevented for the 60-day period by 2, 4-dihydroxyvalerophenone in approx. 0.007% concentration and by the sodium salt of 4-tert-amylresorcinol in approx. 0.005 % concentration, while propyl p-hydroxybenzoate used as the control allowed growth of the mold on the 5.5th day in approx. 0.01% concentration. The above five compounds. of the 1st group possessed a strong mold-preventing action against soy sauce while the two compounds of the 2 nd group also possessed a fair amount of moldprevention.
Several N-substituted acid amides were prepared by the following sequence of reactions: R⋅C(:NH)⋅OC2H5+R′⋅NH2→R⋅C(:NH)⋅NHR′+C2H5OH (1) R⋅C(:NH)⋅NHR′+H2O→R⋅CO⋅NHR′+NH3 (2) As can be seen from the results given in the Table, both reactions proceed with difficulty when R is an aromatic group as compared with those where R is an aralkyl. Thus “Eupaverin” base was produced in a much better yield than the “Neupaverin” base.
Effect of allyl- and propenyl-phenols against Trichophyton was examined and o-allyl-and o, o′-diallylphenols, diallylresorcinol and o-eugenol were found to have stronger activity than eugenol and were found to be effective at 1:16, 000 dilutions or over. The introduction of the allyl radical resulted in the increase of the efficacy with the increasing number of the allyl groups, the isomerized propenyl compounds being still more effective.
Efficacies of 37 kinds of salicylic acid derivatives against Trichophyton and Achorion were examined and it was found that 3-nitro-5-chlorosalicylic acid and 2′, 4′-dichlorophenyl 3, 5-dichlorosalicylate were effective up to 1:32, 000 dilution, while salicylanilide, 3, 5-dichlorosalicylanilide and 3, 5-dichlorosalicylmethylamide were effective up to 1:128, 000 dilutions. It was also found that the fungi did not easily acquire resistance to these chemicals and also that these compounds were not irritating to the skin as a 2% ointment.
Alkyl esters of coumarin- and 6-nitrocoumarin-3-carboxylic acids were prepared from coumarin-3-carboxylic acid (I) and 6-nitrocoumarin-3-carboxylic acid (II), respectively, and alcohols with hydrogen chloride gas as the condensing agent. Ethyl p-(coumarin-3-carboxy)- and ethyl p-(6-nitrocoumarin-3-carboxy)-aminobenzoates were prepared by the application of ethyl p-aminobenzoate on the respective chloride of (I) and (II).
As one of the steps in preparing glucuronic acid from glucose, 1, 2-cyclohexylidene-3, 5-benzylidene-D-glucuronic acid was obtained in a good yield by the electrolytic oxidation of 1, 2-cyclohexylidene-3, 5-benzylidene-D-glucose in 10% aqueous solution of potassium hydroxide with potassium permanganate as the carrier.
Of the 74 kinds of compounds tested, following 10 were able to inhibit the growth of tubercle bacilli (human type) in vitro in concentrations above 160, 000 dilutions, whereas the same degree of growth inhibitory action was shown in 160, 000-320, 000 dilutions of sodium p-aminosalicylate used as a control: Phenanthrenequinone, thymoquinone, thymoquinone mono (aminoguanyl) hydrazone nitrate, p-benzoquinone, p-benzoquinone mono-(aminoguanyl) hydrazone, 1, 4-naphthoquinone, 1, 4-naphthoquinone mono (aminoguanyl) hydrazone nitrate, 2-methyl-1, 4-naphthoquinone, 2-methylanthraquinone, and p-sulfonamidobenzaldehyde thiosemicarbazone. Phenyl p-aminosalicylate seemed to be hopeful as it was able to inhibit the growth of tubercle bacilli in 1, 600, 000 to 6, 400, 000 dilutions. Of the lichen components tested, none showed remarkable tuberculostatic actionn except that some inhibftion of the growth of tubercle bacilli was witnessed with 80, 000 dilution of methyl evernate, and in 40, 000 dilutions of methyl and propyl esters of atranorin and lecanoric acid, and isobutyl and amyl esters of evernic acid.
1) Reaction of 4-nitropyridine N-oxide with phosphorus trichloride in chloroform results in the formation of N-(4′-pyridyl)-4-pyridone by the secondary reaction of 4-nitropyridine and phosphoryl chloride formed by the first reaction. 2) 4-Nitropyridine undergoes similar decomposition when allowed to stand with thionyl, sulfuryl or acetyl chloride, or warmed with these reagents in chloroform. 3) 4-Nitropyridine undergoes similar decomposition to N-(4′-pyridyl)-4-pyridone by a long standing by itself. 4) Heating this N-(4′-pyridyl)-4-pyridone with phosphoryl chloride in a sealed tube to 120-130° results in the formation of approximately two moles of 4-chloropyridine.
A method of isolating a minute amount of proteins by the electrophoresis, using a filter paper dipped in buffer solution, was examined. The placing of the filter paper between two sheets of glass plate was found to be advantageous in keeping the buffer solution constant. Good isolation was effected with whole serum proteins, serum globulin, and ACTH.
It was found that, as a colorimetric determination of isonicotinic acid hydrazide, the colorimetry of Molybdane Blue formed by the addition of phosphomolybdic acid, utilizing the reducibility of the hydrazide, gave the simplified and fairly accurate mode of determination, and that this method could be used for the determination of the hydrazide content in blood and in preparations. However, the colorimetry is interfered by the presence of reducing substances such as vitamin C and urea which must be eliminated by preliminary treatment.