Since the analgesic substance, 1-ethyl-2-dimethylaminomethylcyclohexyl benzoate (I), and diphenhydramine (II) show synergetic action, synthesis of aminocyclohexane derivatives that have diphenhydramine-like activity was attempted. For this purpose, N, N-dimethyl-3-cyclohexylpropylamines (X), cyclohexyl dialkylamino-acetates (XIII, XIV, XV), N-cyclohexyl-2-dimethylaminoacetamides (XIX), and N-cyclohexyl-(XX), N-benzoyl-N-cyclohexyl-(XXI), and N-benzyl-N-cyclohexyl-N′, N′-dimethylenediamines (XXII) were synthesized.
For the purpose of elucidating the relationship between effective partial structure of morphine skeleton and analgesic action, several compounds possessing the A-C rings in the morphine skeleton as the basic structure were synthesized. The compounds prepared were 1-dimethylaminoalkyl-1-phenylcyclohexanes (XIII, XVIII, XXV, and XXXII) and 2-dimethylaminoalkyl-2-phenylcyclohexanols (XIV, XXI, XXIX, and XXXV).
Ethyl cyclohexylidenecyanoacetates (VI), obtained by the Cope condensation of cyclohexanone derivatives (V), was reacted with the Grignard reagent of phenyl bromide to form ethyl α-cyano-1-phenylcyclohexaneacetates (VII) and their decarboxylation afforded the 1-phenylcyclohexaneacetonitriles (VIII). Hydrolysis of (VIII) gave 1-phenylcyclohexaneacetic acids (XI), while the reduction of (VIII) followed by dimethylation, or the condensation of (XI) and dimethylamine followed by reduction, afforded N, N-dimethyl-1-phenylcyclohexaneethylamines (X and III). 1-(1-phenylcyclohexyl) trimethylamine (IV) was prepared by the Curtius degradation of (XI) and subsequent dimethylation.
In order to make further studies on the relationship between analgesic action and effective partial structure of morphine, N, N-dimethyl-2-alkyl-2-phenylcyclo-hexylamines (XII and XIII), 1-(2-ethyl-2-phenylcyclohexyl) trimethylamine (XVII), and N, N-dimethyl-2-ethyl-2-phenylcyclohexaneethylamine (XXV) were prepared.
2-Alkyl (or aralkyl)-2-phenyl-6-dimethylaminocyclohexanones (VIII and IX) and dimethylaminomethylcyclohexanone derivatives (XXII to XXIV) were prepared as compounds allied to N, N-dimethyl-2-ethyl-2-phenylcyclohexylamine (XIX). Starting with these aminoketones, secondary and tertiary dimethylaminocyclohexanol derivatives (X and XI) and dimethylaminomethylcyclohexanol derivatives (XXV to XXX) were prepared, as well as their esters (XV, XVI, XXXI to XXXVIII, and XLII).
The present series of work was undertaken in order to make a systematic examination of old and new poisons based on the difference in chemical structure of each poison during separation and detection by paper chromatography. The separation of hypnotics by paper chromatography is affected by the reaction of the developing solvent and the Rf values differ in each case. The chromatographic separation of barbituric acid, thiobarbituric acid, and diphenylhydantoin derivatives is affected by the concentration of ammonia when a mixture of ammonia and organic solvent is used as the developing solution. This is due to the ease or difficulty of enolization of these chemicals and the enolization is governed by their chemical structure. Therefore, the poisons can be separated by changing the reaction of the developing solvent. According to experimental result, increase of ammonium concentration to over 10% will effect separation of barbiturate, thiobarbiturate, diphenylhydantoin, and bromoacylurea.
The hypnotics can be discriminated by spot tests on a filter paper by the silver nitrate, copper complex salt, or copper-pyridine complex salt methods by the utilization of difference in their chemical structure. The silver nitrate method is capable of discrimination between barbiturate, thiobarbiturate, and bromoacylurea, while the coloration with copper complex method is common to those having structural units of -N-CO-N-, -N-CS-N-, and -N-C=N-, and is discrimiated from bromoacylurea. Copper-pyridine complex method gives coloration of compounds possessing easily enolizable =CO and =CS, separating them from other hypnotics not possessing such units. The limit of detection by the foregoing three methods is around 5-15γ.
Although it is generally accepted that the combustion gas emerging from the platinum-carbon and reduced copper layers in oxygen determination is washed by alkali to remove the acidic gases before the generated carbon monoxide is oxidized to carbon dioxide, an attempt was proposed to simplify the whole apparatus by packing the oxidant closely after the copper layer as shown in Fig. 1. However this was not usable on nitrogen-containing samples because of the generation of considerable amount of hydrocyanic acid which was found to be oxidized to carbon dioxide more easily than carbon monoxide. Therefore, the alkali tube must be absolutely necessary before the oxidation of carbon monoxide, to remove the hydrocyanic acid with other acidic gases. A further investigation is described on a suitable temperature of reduced copper packed in order to fix the sulfur, concluding that it keeps a complete performance even at about 600°.
The cationic active agents (benzalkonium chloride, benzethonium chloride) and nonionic surface active agents (Tween 20, 40, 60, and 80, Span 20, 40, 60, and 80, Emalgen 106 and 408) were found to affect suppressively the crystal growth of 4, 4′-diaminodiphenyl sulfone, and a minimum point in granular size appeared on the curve between the concentration of the surface active agents and granular size of the sulfone. The concentration of the active agents to effect minimum particle size is approximately constant for a given type of the agent but differs somewhat according to the structure of the active agents. The surface tension value of the aqueous solution of surface active agents and that of the agent saturated with 4, 4′-diaminodiphenyl sulfone was invariably lower in the latter. The critical micelle concentration of aqueous solution of surface active agents was the same as that of the agent saturated with the sulfone, but was significantly different in the case of sodium lauryl sulfate and benzalkonium chloride. Since the concentration of the surface active agents effecting minimum particle size was at a point far higher than the critical micelle concentration, the micelle formation of the agent is not an essential factor of crystal growth suppression and the priority of the agent in competition with the sulfone molecule in the field of adsorption to crystal nucleus must be the primary cause. The static bond between the ionic active agent and polar group of diaminodiphenyl sulfone must be advantageous for exchange adsorption phenomenon.
In order to synthesize pyrimidine-2-stibonic acid, the diazo reaction, which occupies one of the most important step in the route to its formation, was examined in detail and a new modified procedure was devised. By the use of isoamyl nitrite as the diazotization agent and dehydrated ethanol as the solvent, with a shortened reaction time, the intermediate, pyrimidine-2-stibonyl tetrachloride, was obtained in a good yield. On warming this intermediate with dilute alkali, pyrimidine-2-stibonic acid was obtained in almost quantitative yield. This modified procedure was applied to the synthesis of eight kinds of stibonic acid reported in the past literature and the stibonic acids were obtained in better yield than that reported through the stibonic tetrachloride intermediate.
Examination, of alkaloids and steroids in Sinomenium acutum REHD. FT WILS. afforded a tertiary phenolic base, isosinomenine (I), and a substance containing nitrogen, forming orange yellow needles, m.p. 201-202°, as the new bases, and β-sitosterol and stigmasterol as the steroids.
1) Using the second year growth of Artemisia kurramensis QAZ., sown in the spring, comparative examinations were made between transplantation (transplanted plot) in the spring and without such transplantation (perennial plot) (Table I). Both the growth and yield of crop were better in the perennial plot than in the transplanted plot, but there was no significant difference between the two in santonin content. 2) Comparative cultivation experiments were made with the second year growth of this plants, sown in the autumn, among the plots of different planting distances, i.e. 4125, 6204, and 7050 plants in 10 ares (Table II). The growth and yield of crops were both the best in the least number of plants i.e. 4125 plants in 10 ares (planting distance, 80×30cm.). The maximum santonin content was found during the middle to latter part of July, which is the same period as that of the second year growth sown in the spring. 3) Both tle first and second year growths showed a marked influence of acclimatization. The growth, yield of crop, and santonin content tended to be better as the generation of acclimatization was higher (Tables III and IV). 4) The highest santonin content in the first year growth, sown and transplanted in the spring, was in the early part of August, being about two weeks later than that of the second year growth. This is considered to be the effect of transplantation in the spring, but there seem to be no essential difference in the period at which the santonin content becomes the highest between the first and second year growths.
1) The seed setting of Artemisia kurramensis QAZ. is considered to require adequate dryness and temperature rather than the short day effect. It was found that although the plant did not bear seeds in the open air field in this area (Kasukabe, Saitama Pref.), seeds could be obtained by culture in a green house (Table I). 2) Cutting was found to be the best in December and woody twigs were found to be better than herbaceous shoots for the cuttings. It was found that immersion of the cuttings for 24 hours in 5, 000-50, 000 diluted solution of potassium α-naphthaleneacetate or β-indoleacetate was quite effective in acceleration of rooting (Table II). 3) Santonin content of Artemisia kurramensis, sown in March, 1954, was followed in 10-day intervals from August 2, 1954, to October 12, 1955 (Table III and Fig. 1). The maximum content during the first year growth was in the latter part of August (1.41%), the content decreased gradually, and became nil during the latter part of February to the beginning of April. The santonin content increased suddenly from the middle of April to reach the maximum in the second year growth during the middle of July (2.33%), and decreased thereafter. The annual variation of santonin content seems to go parallel with the vegetative growth of the plant. 4) The santonin content of third, fourth, fifth, and sixth year growths was examined and they were all similar to that of the second year growth (Table IV). 5) Individual santonin content in the third year growths was determined on the same individuals whose santonin content was determined in the previous year (Table V). There was a significant positive correlation of santonin content between the second and third year growths and it was considered that the amount of santonin produced by the plant is hereditary. From such results, plant breeding of Artemisia kurramensis by family selection was started.
1) It was found that the cultivation of Ammi visnaga LAM. in Japan should start with autumn sowing during the middle of October, at the latest, from the point of growth, yield of fruit, and content of khellin (Table I). 2) The optimal soil moisture in the said cultivation seems to be in the range of 60-70% of the water capacity from the point of growth, yield of fruit, and khellin content (Table II). 3) This plant grows favorably in the soil with pH ranging from 6.5 to 7.5, with neutral reaction as the median point. 4) It was confirmed through experimental cultivation extending from 1951 to 1955 that the cultivation of this plant should be made in a place with small amount of rainfall during the summer and a well-drained soil rich in clay (Tables II and III). 5) Comparative cultivation was carried out with five foreign strains with different origin (khellin content of the original seed, 0.61-0.94%) and the khellin content of the fruit from these cultivated plants was 0.32-0.51%, indicating a fair decrease of the content. However, this was thought to be due to the profuse generation of diseases during August, before flowering, and unhealthy fruit had been mixed (Table IV). 6) In one individual plant, the fruits that ripened earlier seemed to have higher content of khellin (Table V).
Coloration reaction of cardiotonic glycosides like digitoxin, strophanthin, and ouabain with 3, 5-dinitrobenzenesulfonic acid in alkaline solution was studied and optimal conditions for colorimetric determination was established as follows: A mixture of 4cc. of methanolic solution of the glycoside (below 10mg%) and 0.5cc. each of 2% aqueous solution of potassium 3, 5-dinitrobenzenesulfonate and 5% aqueous solution of potassium hydroxide is allowed to stand at room temperature for 10minutes and the optical density of the resultant solution is read by photoelectric colorimeter, using a 530-mμ filter.
The decomposition of thiamine (I) into 2-methyl-4-amino-5-pyrimidylmethanesulfonic acid (II) by sulfite is a specific reaction in thiamine and probably originates in the fact that the nitrogen in its thiazole ring is quaternary. In order to prove this fact, the following experiment was carried out. Thiamine is present in the thiol form (IV) in a strong alkaline solution and the nitrogen in it is tertiary, not submitting to the specific cleavage by sulfite. The nitrogen in the thiazole ring of thiothiamine (III) is also tertiary and sulfite cleavage does not take place in a concentrated solution of thiothiamine. However, addition of sulfite to a dilute (0.1mg%) solution of thiothiamine causes its conversion into thiamine and this conversion is almost complete by the use of sulfurous acid. The formation of thiamine from thiothiamine is greater, the larger the amount of sulfurous acid, but is suppressed with excess of this acid. The formation of thiamine by sulfurous acid decreases with increasing concentration of thiothiamine and this is a specific reaction that takes place only in a dilute solution of thiothiamine. On the other hand, sulfite cleavage of thiamine hardly occurs in a dilute solution of thiamine and that after the use of a great excess of sulfite.
The formation of thiamine from thiothiamine by sulfurous acid is possible only in a dilute solution and cannot be concluded as the reductive action of sulfurous acid. Since the amount of thiamine formed does not increase with increasing concentration, it seems more likely to be oxidation by dissolved oxygen. Therefore, the amount of thiamine formed from thiothiamine was measured by the addition of a definite amount of SO2 to nitrogen-saturated solution (dissolved oxygen, 1.5p.p.m.), oxygen-saturated solution (dissoloved oxygen, 34.5p.p.m.), and distilled water (dissolved oxygen, 9.5p.p.m.). As was anticipated, the amount of thiamine formed was the largest in oxygen-saturated solution and the smallest in nitrogen-saturated solution. Aeration does increase the formation of thiamine but the amount is about the same as when oxygen-saturated solution is used. Compared to autoxidation of sulfurous acid in a medium not containing thiothiamine, the oxidation of sulfurous acid is somewhat accelerated in the presence of thiothiamine but the amount of thaimine formed is not correctly in proportion to the oxidation of sulfurous acid. On the other hand, the effect of dissolved oxygen in sulfite cleavage of thiamine is to stabilize thiamine in a dilute solution while thiamine is more easily decomposed in nitrogen-saturated solution.
It was found that, among 4-(2-hydroxyethylmercaptomercuri) benzoic acid (I), 4-ethylmercaptomercuribenzoic acid (II), 4-(4-carboxyphenylmercuri) thiosalicylic acid (III), 3-chloro-4-(4-carboxyphenylmercuri) thiosalicylic acid (IV), and 2-chloro-4-(p-tolymercuri) thiosalicylic acid (V), all except (II) were able to inhibit succinic acid oxidation of the intact cells and cell-free extracts of Escherichia coli in about the same degree, while (II) inhibited that oxidation of intact-cells better. Of these compounds, the activity of (V) was the weakest. As for their action on succinic acid oxidatin of Staphylococci, (I) and (II) strongly inhibited the action while the inhibition by the others was weak. From the comparison of the straight line in the inhibition-concentration relationship by probit measure, it was considered that the action of organic mercurials was something apart from their bonding with the SH group in enzyme proteins.
In order to investigate the effect of abnormality in dietary calcium and phosphorus content on the excretion of radioactive strontium, albino rats, about 2 months old, were maintained on diets low in phosphorus (P, 0.187%), low in calcium (Ca, 0.011%), and low in phosphorus and calcium (P, 0.189%, Ca, 0.015%), and on a normal control diet (P, 0.582%, Ca, 0.601%). The rats were injected subcutaneously with radioactive strontium and then treated with dihydrotachysterol given orally. In addition to the metabolism of radiostrontium, calcium and phosphorus metabolism were examined. In the groups of low calcium diet and of low phosphorus and calcium diet, rate of radiostrontium excretion was not markedly changed, though calcium balance had become negative and the amount of calcium in carcasses was reduced. On the other hand, greatly enhanced excretion of radiostrontium, mainly into urine, was observed in the group of low phosphorus diet, which was merely slightly low in the amount of phosphorus as compared with the severely deficient diet reported effective to activate radiostrontium excretion by Copp, et al. In the low-phosphorus group no decrease of carcass calcium was observed, though slight decrease in calcium retention was found. Therefore, it is evident that normal bone structure was not markedly injured in this group. Active excretion of radiostrontium in the group of low phosphorus diet seems to be related to increased activity of urinary excretion of calcium and to active absorption of calcium in the intestines. Skeletal disturbance in low calcium and in low calcium and phosphorus diets fail to decrease radiostrontium accumulation into the skeleton. Dihydrotachysterol did not have any significant effect on radioactive strontium excretion.
The existing procedure for the determination of alkoxyl group, both gravimetric and volumetric, was too complicated and required a long period of time. A new micro-determination with a simple procedure and good precision, which might be termed a combustion method, was devised. By this procedure, the sample is heated with hydriodic acid in air stream, the alkyl iodide formed by decomposition is introduced into a combustion tube connected to the decomposition apparatus, and burned with platinum contact as a catalyst. The iodine formed by combustion is collected by a silver gauze heated in the same combustion tube and weight-increase of the silver gauze is determined. This procedure of analysis takes only 30 minutes and accuracy of analysis is within ±0.2%.
Antipyrine reacts with nitrous acid to form nitrosoantipyrine whose reduction with zinc dust and hydrochloric acid results in the formation of aminoantipyrine. By the utilization of this reaction, antipyrine can be determined by Iwasaki's azotometry. Two following procedures are possible for this determination: 1) Reacting antipyrine with a definite excess of nitrous acid and determining the excess nitrous acid by Iwasaki's azotometry. 2) Reacting antipyrine with nitrous acid to form nitrosoantipyrine, reducing this with zinc dust and hydrochloric acid to form aminoantipyrine, and determining this aminoantipyrine by Iwasaki's azotometry using the diazo method. Antipyrine was determined by these procedures with an accuracy of ca. ±1%.
Bis (4, 5-dimethyl-2-thiazolyl) sulfide (XVI) was obtained by the reaction of 2-mercapto-4, 5-dimethylthiazole (XIV) and 2-chloro-4, 5-dimethylthiazole (XV). The reaction of 2-(2-mercapto-4-methyl-5-thiazolyl) ethanol (I) and (XV) afforded 4, 5-dimethyl-2-thiazolyl 4-methyl-5-(2-hydroxyethyl)-2-thiazolyl sulfide (XVII), as well as bis [4-methyl-5-(2-hydroxyethyl)-2-thiazolyl] sulfide (V) and (XVI). The reaction of 2-(2-chloro-4-methyl-5-thiazolyl) ethanol (IV) and (XIV) gave (XVII) as well as (XVI) and (V). The reaction of ethyl 2-chloro-4-methyl-5-thiazolecarboxylate (IX) and (XIV) afforded only 4, 5-dimethyl-2-thiazolyl 4-methyl-5-ethoxycarbonyl-2-thiazolyl sulfide (XX), while the reaction of ethyl 2-mercapto-4-methyl-5-thiazolecarboxylate (VIII) and (XV) gave bis (4-methyl-5-ethoxycarbonyl-2-thiazolyl) sulfide (X) and (XVI).
It was found that the two optical isomers, oxyacanthine and repandine (both having structural formula (I)), belonging to the bisbenzylisoquinoline-type base, could be separated from each other from their mixture by multi-buffered paper chromatography. A mixture of ether and benzene (1:1 in volume) was used as the developing solvent and taking a larger unbuffered space than usual (cf. Fig. 1, No. 7), the papergram was colored by the use of platinum chloride-potassium iodide reagent. It was thereby shown that the two bases gave spots of different color, oxyacanthine at pH 5.6 of reddish violet color and repandine at pH 5.8 of grayish violet color.
Piperidine and diethylamine were found to be effective basic catalyst in the condensation of vanillin and aliphatic aldehydes and new compounds, α-ethyl-, α-pentyl-, and α-hexyl-4-hydroxy-3-methoxycinnamaldehydes, were prepared. From this result, mistaken observations appearing in the reports of Weizmann, Verkade, and Uyeno were pointed out.
In the past determination of alkoxyl group, decomposition of the sample with hydriodic acid and distillation of alkyl iodide were carried out in carbon dioxide gas or nitrogen stream. As a new procedure of determining alkoxyl group, replacement of these gases with air was experimented and it was found that the gas could be replaced sufficiently with air.