High pressure, catalytic reduction of p-nitrosalicylic aryl and aralkyl esters in methanol, ethanol, or benzene, with Raney nickel as the catalyst, gave the esters of p-aminosalicylic acid in a good yield. The similar reduction of the sodium, calcium, or barium salt in water or dilute ethanol also gave the PAS salts in a good yield. The esters of PAS obtained were phenyl ester (I), guaiacol ester (II), m-cresol ester (III), thymol ester (IV), m-methyl-p-isopropylphenyl ester (V), and benzyl ester (VI). PAS hydrazide was also obtained. Antibacterial action in vitro against H37Rv strain of tubercle bacilli was five times that of PAS-sodium 2 H2O in (I) and (III), and 3.3 times in (II). With H37Rv R-SM strain, (I) and (III) were 15 times stronger, and (II), 7.5 times stronger. The action was about the same against H37Rv R-PAS strain. However, (V), the isomer of (IV), was found to be 250 times that of (IV) aginst H37Rv strain and 200 times against H37Rv R-SM strain. The antibacterial action of PAS hydrazide was about 1/15 that of isonicotinic acid hydrazide against H37Rv strain, and 1/13 aginst H37Rv R-SM strain.
In the glycoside synthesis by the Helferich and Shishido method, the reaction of β-pentaacetyl-D-glucose and phenols by fusion, with zinc chloride as a catalyst, results in the formation of a large amount of tetraacetyl-β-compound as a by-product besides the objective α-compound. The amount of the β-compound formed as a by-product was found to vary with the kind of phenols used. In order to better the yield of the α-compound, examinations were made on the use of zinc bromide, instead of zinc chloride, as a catalyst, variation of the reaction temperature, and the use of α- as well as β-pentaacetyl-D-glucose as the starting material. It was thereby found that, irrespective of the use of α- or β-pentaacetyl compound, the extension of the reaction time to 60-90 minutes and the use of phenol were found to give the maximum yield of the tetraacetyl-α-phenyl-D-glucoside, the yield being 35% with zinc chloride catalyst and 45% with zinc bromide catalyst.
By catalytic hydrogenation with PdO as a catalyst in alcoholic solution at oridinary temperature and pressure, dictamnine (I) and skimmianine (II) absorb one mole of hydrogen to yield respectively dihydrodictamnine, C12H11O2N, long pillars, m.p. 96-97° (picrate: yellow prisms, m.p. 183°), and dihydroskimmianine, C14H15O4N, prisms, m.p. 163° (picrate: yellow needles, m.p. 183°). Dihydroskimmianine does not absorb bromine in glacial acetic acid solution while (II) is brominated. Dihydrodictamnine and dihydroskimmianine are therefore 4-methoxy-(V) and 4, 7, 8-trimethoxy-2, 3-dihydrofuro [2, 3-b] quinoline (IV), respectively. (II) is cleaved to 3-ethylcarbostyril compound (VII) by catalytic hydrogenation with PtO2-catalyst, but (IV) cannot be hydrogenated with the same catalyst. This suggests that hydrogenolysis of the furo [2, 3-b] quinoline bases with PtO2-catalyst results in the formation of (VII) through 3-vinylcarbostyril (VI), the fission of the furan ring occurring in the 1:2-position. On hydrogenation over Raney nickel, (II) yields a mixture of (IV) and (VII). From the above facts, dihydroacronycidine by Lahey, Lamberton and Price obtained from acronycidine (III) by catalytic reduction over Raney nickel must be 4, 5, 7, 8-tetramethoxy-2, 3-dihydrofuro [2, 3-b] quinoline (VIII). Tetrahydro and hexahydro derivatives of acronidine by Lamberton and Price are considered as (Xa-Xb) and (XIa-XIb), respectively.
Since a group of compounds that show analgesic action possess common functional group-CH2-CH (A)-CH3, a series of butanols possessing a functional group-CH (A)-CH-(A)-CH3, with a phenyl or thienyl in α-position, were prepared. Acylation of the hydroxyl in these compounds was not successful.
Since there are several compounds in the series of 2-thienyl-or 5-bromothienyl ketones possessing N-methylpiperidyl or -pipecolyl group that show powerful analgesia, synthetic methods for such ketones were examined.
The synthetic methods were examined for 3, 5-bis [di (2′-thienyl) methylene]-1, 2, 6-trimethylpiperidine, a compound in which the 2-position of the powerfully analgesic 3-dimethylamino-1, 1-di (2′-thienyl)-3-methylpropylene-(1) is bonded with a methylene group and one of the N-methyl is conjugated. It was found that this compound is easily obtained by the dehydration of the dihydroxy compound prepared by the application of thienyl lithium on 3, 5-diethoxycarbonyl-1, 2, 6-trimethylpiperidine.
In order to establish the chemical isolation method for the glycosides of Digitalis purpurea, perfect isolation of each glycoside by paper chromatography was examined as the fundamental study. A mixture of the glycosides was separated into two groups, one of true glycosides and the other of digitoxin class and genin group. Each of these was again chromatographed under different conditions and each glycoside was successfully separated with sufficient difference of Rf values.
Starting with o-Xylene of approximately 95% purity, it was derived through the chloromethyl compound (II) to the cyanide (III) and condensed with methyl-bis (β-chloro-ethyl) amine, in the presence of sodium amide, to obtain 1-methyl-4-(3′, 4′-dimethylphenyl)-4-cyanopiperidine (IV). Subsequent saponification and esterification yielded ethyl 1-methyl-4-(3′, 4′-dimethylphenyl)-piperidine-(4)-carboxylate (V). On the other hand, (II) was derived through the sulfide (VII and VIII) and the sulfone (IX and X), with subsequent condensation with methyl-bis (β-chloroethyl) amine to 1-methyl-4-(3′, 4′-dimethylphenyl)-piperidine-(4) methyl sulfone (XI) and ethyl sulfone (XII).
The rhizome of Veratrum stamineum Maxim. was extracted by the ammonia-ether-chloroform method and the alkaloids were isolated by the utilization of the difference in the solubility of the hydrochloride and the sulfate. Jervine, veratramine, and the so-called amorphous base were obtained in the respective yield of approximately 0.1%, 0.025%, and 0.28%.
In order to find an antagoinist for the toxicity of procaine, β-diallylaminoethyl p-aminobenzoate was prepared. The free base has b.p9 218-220° and gives two kinds of hydrochlorides, monohydrochloride of m.p. 158-160° and dihydrochloride of m.p. 200-20° (decomp.). Lethal dose, LD50, of this compound is 21.8mg./10g. by subcutaneous injection in mouse (Behrens-Kärber method), being far weaker in toxicity than procaine or cocaine. The toxic symptoms are the same as those of procaine. Though no antagonism against procaine was detected, its local anesthetic action was strong, being 2 times that of procaine and about 1/2 that of cocaine in surface anesthesia, and twice that of procaine and 1/5 to 1/2 of cocaine in filtration anesthesia. The toxicity of this compound against respiration, blood pressure, and heart action in rabbits is 1/3 to 1/2 that of procaine. Its concentrated solution effected contraction of the blood vessels while a diluted solution effected dilation, and the addition of epinephrine always effected contraction of the blood vessels. The aqueous solution of this compound was unchanged by high-temperature sterilization.
The leaves and trunk bark of Xylosma apactis Koidz. possess comparatively strong bitterness and their components were examined. A substance assumed to be a new glycoside, m.p. 206°, was obtained and it was found to possess the same structure as hydroxypopuloside, i. e. salireposide. The properties of this substance were entirely different that the two are not identical. Moreover, where salireposide is decomposed by emulsin, the new glycoside is not decomposed by it though it is decomposed by Takadiastase. From these facts, the new glycoside was assumed to be the anomer of salireposide, being in a relation of α- and β-isomers. This new glycoside was named xylosmoside.
The sesquiterpenoid lactone obtained from the rhizome of Inula helenium was reduced with sodium amalgam and dihydroisoalantolactone was isolated as the product. Allyl oxidation of the latter with selenenium dioxide yielded 3-hydroxydihydroisoalantolactone, which was derived to 3-oxotetrahydroalantolactone and compared with tetrahydrosantonin.
A series of new compounds, α-alkoxymethylene-β-alkoxypropionitrile, were obtained by the formylation of β-alkoxypropionitrile in benzene with ethyl formate and sodium alkoxide followed by the alkylation of its product, α-sodioformyl-β-alkoxypropionitrile, with alkylation agent such as alkyl sulfate. In this case, if the reaction is carried out in an anhydrous solvent, cis-form compound is first obtained which transits to the trans-form by treatment with alkali. The determination of these ci-and trans-compounds could be made through their physical constants but the structure of α-methoxymethylene-β-ethoxypropionitriles was confirmed by the measurement of the dipole moment of the two isomers. By the detailed examination of the formylation of β-alkoxypropionitrile the yield of the product from this reaction was increased. α-Sodioformyl-β-ethoxypropionitrile was also obtained by the addition of ethyl formate to acrylonitrile in the presence of metallic sodium but the yield by this method was poor.
A series of 2-methyl-4-amino-5-alkoxymethylpyrimidine were obtained in a good yield by the condensation of cis-α-alkoxymethylene-β-alkoxypropionitrile and acetamidine in alcohol. The best yield was obtained in those possessing ethoxyl group in the β-position, the yield being approximately 80%, and those with methoxyl group in the β-position followed with 56% yield. Further, if this reaction is carried out in methanol or in the presence of an excess of sodium alkoxide, the compounds of this series are not obtained and a large amount of the substance of m.p. 203-204°, identical with the condensation product of α-alkoxymethylene-β-alkoxypropionitrile and acetamidine, is obtained.
Condensation of trans-α-alkoxymethylene-β-alkoxypropionitrile and acetamidine gave a substance of m.p. 203-204° which was found to be 2-methyl-4-amino-5-acetaminomethylpyrimidine. Some colorless needle crystals of m.p. 173° (dehydrate) were obtained as an intermediate during this reaction. The substance was found to be identical with the produtc of a reaction between 2-methyl-4-amino-5-aminomethylpyrimidine and acetoimidomethyl ether and was confirmed as 2, 7-dimethyl-5, 6-dihydropyrimido [4, 5-d] pyrimidine. It was thereby clarified that in this reaction, the alkoxyl groups in the α-and β-positions in α-alkoxymethylene-β-alkoxypropionitrile are consecutively substituted with acetamidine, followed by cyclization to the afore-mentioned pyrimidopyrimidine and the latter finally undergoes hydrolysis to 2-methyl-4-amino-5-acetaminomethylpyrimidine. By the utilization of this reaction, the important intermediate of vitamin B1, 2-methyl-4-amino-5-aminomethylpyrimidine, can be obtained in an industrially advantageous manner.
Formylation of alkyl β-alkoxypropionate with ethyl formate and sodium alkoxide, in dehydrated benzene, gave alkyl α-sodioformyl-β-alkoxypropionate. which was alkylated with alkyl sulfate to alkyl cis-α-alkoxymethylene-β-alkoxypropionate. Treatment of these cis-compounds with alkali effected their transition to the trans-type. It became clear therefore that the same reaction occurs in this case as in the similar reaction of α-alkoxymethylene-β-alkoxypropionitrile reported before. The structures of the cis-and trans-compounds hereby obtained were easily assumed from the fact that their respective physical constants were in the same relationship as in the corresponding propionitriles. The structures were further confirmed by the measurement of the dipole moments of the two isomeric ethyl α-methoxymethylene-β-ethoxypropionates. In order to comparatively examine the effect of the alkoxyl group in the β-position of these compounds on dipole moment, ethyl and butyl α-sodioformylacetates were methylated with dimethyl sulfate and ethyl trans-α-methoxymethyleneacetate and butyl cis-α-methoxymethyleneacetate were prepared.
Reaction of alkyl α-alkoxymethylene-β-alkoxypropionate and acetamidine yields 2-methyl-4-hydroxy-5-alkoxymethylpyrimidine in a good yield from either cis- or trans-compound. Reaction of alkyl α-methoxymethyleneacetate and acetamidine also gives 2-methyl-4-hydroxypyrimidine in a good yield from either cis- or trans-compound.
Reaction of β-alkoxypropionitrile or alkyl β-alkoxypropionate with alcohols, with a small amount of alkali hydroxide or sodium alkoxide as a catalyst, results in the transetherification between the alkoxyl group in β-position and the alcohol. The reaction of the propionate was also carried out with mineral acid as a catalyst and transesterification also occurred in this case. It was also clarified that transetherification occurred between the alkoxyl in the α- and β-positions with alcohol by the same reaction in α-alkoxymethylene-β-alkoxypropionitrile and alkyl α-alkoxymethylene-β-alkoxypropionate. In this case, however, the product was found to be the trans-type alone from the transetherification, irrespective of the cis- or trans-type used as the starting material. Transesterification also occurred in propionic acid esters and the ease of such substitution was found to be in the order of the ester, α-alkoxymethylene, and β-alkoxy group. The reaction also occured in propionic acid esters by the use of mineral acid as a catalyst.
γ-Amino- and γ-alkylamino-propanols were obtained by the direct addition of ammonia, liquid ammonia, or primary and secondary amines to allyl alcohol, with sodium alkoxide or ammonium acetate as a catalyst, by heating in a pressurized vessel for 7-10 hours at 100-150°. Aminopropanols were obtained in 20-60% yield in the reaction with ammonia and primary amines, but dipropanolamine was also formed as a by-product. Moreover, some non-basic oily substance, assumed to be the polymer of allyl alcohol, was also obtained. By the addition of an amine to allyl p-nitrobenzoate, dialkylaminopropyl p-nitrobenzoate was obtained in one step.
Capsaicin was isolated in a pure state from the acetone extract of 1-5g. of powdered Capsicum, by paper partition chromatography using benzine saturated with methanol as the developing agent. The blue coloration was formed by 3% phosphomolybdic acid and 0.1N sodium hydroxide and its extinction coefficient (E) was measured after 1 hour by the Beckman Model DU spectrophotometer at 730mμ (in 1cm. cell). Quantitative determination of capsaicin was attempted by calculating the content from the standard concentration-extinction coefficient regression equation, γ=1241 E-40.02 (cf. Table III), for capsaicin. The content of capsaicin was determined in the two commercial kinds of Japanese Capsicum of the highest pure line, “Yatsuhusa” (Capsicum annaum L. var. fasciculatum Irish.) and “Takanotsume” (C. annuum L. var. parvoacuminatum Makino) and a few kinds of Capsicum tinctures of market products (cf. Tables VI and VII). Vanillin, used by North as the standard for the determination of Capsicum, was also submitted to the same quantitative determination and it was found usable in place of capsaicin. The relationship between capsaicin and vanillin is as follows: Capsaicin content (γ)=Vanillin content (γ)/23.00
Distribution and generation of capsaicin were examined and it was found that it was not present in any plant organs other than the fruit. It was found that the epidermal cells of dissepiment in fruit undergoes segmentation and proliferation in the direction perpendicular to the surface and capsaicin forms first in oily state between its outer cell wall and cuticle. Capsaicin crystals precipitate out as colorless tetragonal to hexagonal plates in the secretion organs of dried fruit or fresh fruit sealed with glycerol water. From microchemical studies, the secretion chiefly consits of capsaicin and fatty oil and it is assumed that it is formed as a lipid-capsaicin complex in nature.
1) 1-Phenyl-2-methylaminopropane (I), 1-phenyl-2-aminopropane (II), and ephedrine (III) color red to scarlet with sodium hypobromite and pyridine. 2) (I) colors orange to red with potassium ferricyanide, sodium nitroprusside, and sodium carbonate, and then hydrochloric acid and phenylhydrazine. 3) (II) colors reddish violet with 1 drop each of 5% sodium nitroprusside and acetone, and 20-30mg. of sodium hydrogen carbonate, followed by 30 minutes' warming. Limit of detection being 50 γ of (II) hydrochloride. This is a characteristic coloration for aliphatic primary amines (free and salts) and amino acids. 4) (III) colors violet on heating with 2 drops of 1% alcoholic solution of ninhydrin and 1 drop of pyridine.
In electrochromatography, five kinds of different filter paper holders were used and the velocity of migration, area of colored band, and migration distance of each position on the filter paper were measured under identical conditions so as to examine the separatory ability and equality of rate of each apparatus. It was thereby found that the best separation is effected by placing the paper in a convex form, with the middle higher, while for uniform mobility, the filter paper should be cooled directly by closely adhering the lower surface on a cooling vat.
The bacterial component of Streptococcus faecalis was investigated through paper chromatography. As free amino acids, aspartic and glutamic acids, and a comparatively large amount of alanine were detected. From the hydrolysates of the bacteria with hydrochloric acid and with baryta, the following were determined as the amino acid constituting the the bacteria: Aspartic acid, glutamic acid, serine, glycine, threonine, alanine, valine, leucine, phenylalanine, histidine, lysine, arginine, tyrosine, and tryptophan. Polysaccharides were extracted from the bacteria and its hydrolysate was submitted to paper electrophoresis from which glucose was detected. It was confirmed through paper chromatography that by washing the bacteria seven to eight times with physiological saline solution, the bacteria are freed of the substances from the cultural medium.
The rhizome of Veratrum grandiflorum Loes. fil., growing in Nagano Prefecture, was digested with cold alcohol and the effective principle was isolated and purified by the usual method and the so-called amorphous base was obtained as a pale yellow base in a high yield of 0.9% (0.36% from V. viride Ait.). Unexpectedly, however, jervine and veratramine were not obtained. The extraction and purification of the principle from the rhizome of Veratrum japonicum Loes. fil., growing in Sadogashima were made in a similar manner but the yield of the principle was only 0.02% and no other alkaloids were obtained.
Tetraploid plant (n=18) of insect flower (Chrysanthemum cinerariaefolium Bocc.) were successfully brought up by the treatment with colchicine in 1947, and these were cultivated during 1947 to 1953 in comparison with the diploid with following results: 1) The dimensions of the stomata was larger in the tetraploid than the diploid and there was a stochastically significant difference. The number of stomata per field of vision was smaller in the tetraploid than the diploid, there being also a significant difference. 2) The size of the flowers was larger and the number of ray flowers was more numerous in the tetraploid than the diploid, there also being a significant difference. 3) The content of pyrethrin in the air-dried flowers was also larger in the tetraploid than the diploid, there also being a significant difference. This fact shows the possibility of increasing the content of effective principles in medicinal plants by the artificial chromosome doubling and thereby the breeding superior varieties. 4) No significant difference was observed in the content of pyrethrin I and II between the tetraploid and diploid plants. 5) It was stochastically confirmed that the pyrethrin content of dried flower decreased by storage.
Based on the paper chromatographic analysis of protamine zinc insuline carried out by Robinson, determination of purity of crude insuline powder was attempted. It was found that the insuline band (Rf 0.47) and the crude protein band (Rf 0) on such a chromatogram were separated, as in the Robinson method, but a substance appeared at Rf 0.40. This was assumed to be an insuline-like substance but not possessing potency equal to that of insuline. Determination of insuline through the combination of this chromatogram elution and photoelectrocolorimetry was found to give results with mean error of within 1 UV/mg. with that of bioassay.
Reaction of 2, 6-lutidine 1-oxide and acetic anhydride gave 2-acetoxymethyl-6m-ethylpyridine and 3-acetoxy-2, 6-lutidine both of which were respectively hydrolyzed by hydrochloric acid to 2-hydroxymethyl-6-methylpyridine and 3-hydroxy-2, 6-lutidine.
Reaction of quinaldine 1-oxide and acetic anhydride gave quinaldyl alcohol in 30% yield. The same reaction with lepidine 1-oxide gave three products, i.e. 2-hydroxylepidine, 3-hydroxylepidine, and lepidyl alcohol, the yield of the last one being too small to permit the use of this method for its preparation.