In order to study the mechanism of electrolytic reduction of α-nitroölefins, polarographic examination of 1-nitro-1-pentene and controlled-potential electrolyses at pH 2 and 6, and at various potentials were carried out. Representative polarograms are shown in Figs. 1 and 2, and their data are listed in Table I. The polarograms obtained were different with 10-2 and 10-3-10-4M solutions. In controlled-potential electrolysis, the potential corresponding to the first wave in 10-2M solution of pH 2 afforded valeraldoxime as the 4-electron reduction product, and at the potential corresponding to the second wave, formation of a substance giving positive reaction to N-alkylhydroxylamine was observed. It was considered that a 6-electron reduction product, N-pentylhydroxylamine, had been formed in this case. The result was almost the same at pH 6 as at pH 2, but in controlled-potential electrolysis in lower potential corresponding to plateau-A of the first wave, at -0.70 V υs. S. C. E., colorless labile oil of b. p3 68°, with molecular weight of ca. 260 (by Rast method), was obtained besides valeraldoxime. This substance was considered to have been formed by the condensation of an intermediate and the starting material.
From the fact that 5-bromo-, 3, 5-dibromo-, 5-chloro-, and 3, 5-dichlorosalicylaldehyde, obtained by bromination or chlorination of salicylaldehyde, are stable and reactive, the possibility of their use as a reagent for primary amines was examined. It was thereby found that these four kinds of halogenosalicylaldehyde would be utilizable as an analytical agent for primary amines since they easily form the Schiff bases which can be separated by simple means. Conditions for these separation and identification were examined using 16 kinds of aromatic amines and two kinds of aliphatic amines. Melting point, crystal form, and elemental analytical values of their Schiff bases were listed in Tables I-IV).
Selenium dehydrogenation of monogynin, C15H20O3, (m.p. 138°, [α]13.5D-164°(CHCl3), oxime, m.p. 201-202° (decomp.)) affords 1-methyl-7-ethylnaphthalene and 1-methyl-7-ethyl-2-naphthol, and hydrogenation of monogynin gives α-tetrahydrosantonin by absorption of 1 mole of hydrogen. From these results, the empirical formula (A) is given to monogynin but the position of the double bond is still unknown.
Dehydration of ketomibulactone, C15H20O4, afforded dehydroketomibulactone, m.p. 69-72°, C15H18O3, whose hydrogenation gave α-tetrahydrosantonin as one of the reaction products. Oxidation of the ozonide of acetylmibulactone with chromium trioxide afforded an acid lactone, m.p. 214°, C17H24O8, and its alkaline hydrolysis followed by acidification effected decarboxylation and formation of a lactone, m.p. 216°, C14H22O5. This fact indicates that a carbonyl in the acid lactone is present in the position β to the lactone ring. The aldehyde, C15H20O4, formed by the oxidation of mibulactone with lead tetraacetate, forms bromoform by potassium hypobromite and this shows that it has a CH3CO group. Consequently, the structural formula (I) is proposed for mibulactone.
Comparative examinations were made on analgesic activity of phenothiazine, 1, 2, 3, 4-tetrahydrophenothiazine, 3-phenylbenzo-1, 4-thiazine, and 2, 3-dihydrobenzo-1, 4-thiazine derivatives by electrical stimulator (rectangular-wave). Analgesic activity of this kind of compounds is very weak that the threshold amount of morphine was used at the same time, in accordance with the modified Haffner method, thereby raising the threshold value, and comparative tests were effectively carried out. The reaction of mice to electrical stimulation takes the logarithmic normal-type, so that logarithmic transformation of the threshold value and correction of the value after administration with that before administration enabled a more uniform analyses. Phenothiazine derivatives were generally more effective than 1, 4-thiazine and tetrahydrophenothiazine derivatives. Of the latter case, 3-phenyl-4-(3-dimethylaminopropyl)-6-chlorobenzo-1, 4-thiazine was the most effective.
Examinations were made on the concurrent determination of sulfur and halogen, and the use of manganese dioxide as an agent in removing nitrogen oxides outside the combustion tube. With further improvement, these results were applied to the analyses of carbon and hydrogen, and it became possible to determine carbon, hydrogen, sulfur, and halogen at the same time. In this method, the sample is burned in oxygen atmosphere with platinum catalyst, the sulfur oxide and halogen formed are absorbed in absorption funnel, filled with somewhat coarse hairy silver formed during electrolysis of silver. The increase in its weight is weighed, silver sulfate absorbed in the funnel is washed off with water, and analytical value of sulfur is calculated from the decrease in weight. The analytical value of halogen is calculated from the amount of insoluble silver halide remaining in the funnel. Nitrogen oxide is removed by manganese dioxide provided between carbon dioxide-absorption tube and water absorption tube. The air supplied through the branch of combustion tube is preliminarily passed through a layer of sodium asbestos-anhydron and this removes the error in carbon and hydrogen determination. By using two sets of absorption tubes and absorption funnels alternately, the total analysis time can be limited to around 60 minutes, and good analytical values can be obtained for carbon, hydrogen, sulfur, and halogen.
N-Methyl (V) and N-benzyl (VIII) derivatives of ethyl 2-(ethoxycarbonylmethylaminomethyl) isovalerate compound were obtained from ethyl 2-aminomethylisovalerate (II). (V) was obtained by N-methylation of (IV) with formaldehyde and (VIII) was obtained by condensation of (II) and benzaldehyde, reduction of its product, and further condensation with ethyl iodoacetate. These were submitted to the Dieckmann reaction using sodium hydride and the reaction afforded ethyl 1-methyl-4-isopropyl-3-pyrrolidone-2-carboxylate (IXa) and ethyl 1-benzyl-4-isopropyl-3-pyrrolidone-2-carboxylate (IXb). Hydrolysis of (IX) with dil. hydrochloric acid easily effected ketonic fission and 1-methyl-4-isopropyl-3-pyrrolidone (Xa) and 1-benzyl-4-isopropyl-3-pyrrolidone (Xb).
Fifty-five kinds of quinoxaline derivatives (31 of which were new compounds) were prepared and their antibacterial activity in vitro was tested with Mycobacterium tuberculosis H37RV. These compounds tested were carboxylic acid, aldehyde, chloro, hydrazino, hydroxy, and mercapto derivatives. Strong antibacterial action was found in mercapto derivatives, rather than in quinoxaline system compounds corresponding to INAH and Tibione.
dl-N, 4′-Dimethylisococlaurine (VI) is the phenolic by-product obtained on cleavage of isotetrandrine (I) with metallic lithium in liquid ammonia, with dioxane as a solvent. The racemic compound of (VI) was synthesized by the route of (IX)→(X)→(XI)→(XII)→(VI), obtained as a free base of m.p. 45° and a picrate of m.p. 138°. Infrared absorption spectra of the two showed them to have identical structure and the structure of (VI) was thus synthetically confirmed. Attempted reduction of 3, 4-dihydroisoquinoline compound (X) to (XII) in one step with palladium-carbon catalyst afforded a benzoyl compound (XIII) as a by-product and its structure was identified.
Crebanine (II), contained in Stephania capitata SPRENG. and Stephania sasakii HATAYA, together with stephanine (I), was synthesized by the route shown in Chart 1. The synthetic studies on crebanine, independently being carrie dout by the present writers and Govndachari of India, reached the same conclusion and the aporphine-type structural formula (II), proposed by Tomita and his collaborators, for crebanine was synthetically proved to be correct.
Separatory determination of β-lysine and roseonine, formed by acid hydrolysis of racemomycin-B, an antibiotic produced by a mutant strain of Streptomyces racemochromogenus, was attempted and separation of carbon dioxide, β-lysine (I), and roseonine (II) was effected. (I) and (II) were well separated by exchanger resin column using Amberlite IRC-50 (Na form). It was found by this that the molar ratio of the formation of carbon dioxide, β-lysine, and roseonine is 2:3:2 from 1 mole of racemomycin-B. Since this ratio is different from that of roseothrlcin or geomycin, the antibiotics of the streptothricin series, racemomycin-B was assumed to be a new antibiotic different from these substances. It was considered appropriate to give the formula C60H128O32N20 to racemomycin-B from these experimental results.
Coloration of hesperidin and eriodictin by the indophenol reaction gives solutions whose absorption curves are almost without difference. Therefore, colorimetric determination of these substances must be preceded by their separation. Hydrolysis of citrin and paper chromatography of the hydrolyzate with distilled water saturated with benzene as the developing solvent, by the one-dimensional ascending method, gives the spots of the hydrolyzate of hesperidin and eriodictin respectively at Rf 0.44 and 0.11 (20-21°). By cutting out the papergram at respective Rf values, extraction with 20% aqueous solution (v/v) of pyridine, and addition of dimethyl-p-phenylenediamine hydrochloride solution and sodium hypochlorite solution to the extract to effect oxidative condensation in Kolthoff-Vleeschhauwer buffer of pH 8.2 gives a blue solution. Extraction of this blue pigment with isobutanol and measurement of its optical density at 620mμ successfully effected separatory determination of hesperidin and eriodictin in citrin.
The acetone-dried powder of soil bacteria KT 84 (Pseudomonas sp.) effected asymmetric hydrolysis of N-benzoyl-, N-dichloroacetyl-, and N-chloroacetyl-DL-phenylalanine respectively affording N-benzoyl- (V), N-dichloroacetyl- (VI), and N-chloroacetyl-D-phenylalanine (VII) in a good yield, besides L-phenylalanine. The acetone-dried powder of KT 83 easily hydrolyzed (V), (VI), and (VII) to afford acylated D-phenylalanine in a good yield. This soil bacteria also hydrolyzed N-bezoyl-, N-dichloroacetyl-, and N-chloroacetyl-L-phenylalanine. The acetone-dried powder and bacterial suspension of KT 84 did not hydrolyze N-benzoyl-DL-phenylglycine but effected asymmetric hydrolysis of N-dichloroacetyl-and N-chloroacetyl-DL-phenylglycine to form the corresponding D-phenylglycines (XIII and XIV) and L-phenylglycine. The acetone-dried powder and bacterial suspension of KT 83 hydrolyzed (XIII) and (XIV) to form D-phenylglycine, and also hydrolysed N-dichloroacetyl-, and N-chloroacetyl-L-phenylglycine. N-Dichloroacetyl derivative of phenylglycine is more easily hydrolyzed than the N-chloroacetyl derivative. The acetonedried powder of KT 84 asymmetrically hydrolyzed DL-phenylglycine amide to form L-phenylglycine and D-phenylglycine amide (XVII) in a good yield. D-Phenylglycine was prepared by the hydrolysis of (XVII) with 10% HBr. The acetone-dried powder of KT 84 effected asymmetric hydrolysis of N-benzoyl-p-nitro-DL-phenylalanine (XVIII) and afforded p-nitro-L-phenylalanine and N-benzoyl-p-nitro-D-phenylalanine. The N-benzoyl-p-amino-D- and -L-phenylalanine were synthesized. While KT 84 hydrolyzed only the L-form benzoyl derivatives of p-nitro- and p-amino-phenylalanines, KT 83 hydrolyzed both L- and D-forms.
The acetone-dried powder and bacterial suspension of the soil bacteria KT 84 (Pseudomonas sp.) did not hydrolyze the N-benzoyl derivative of threo-β-phenyl-DL-serine but did effect asymmetric hydrolysis of its N-dichloroacetyl derivatives to produce threo-β-phenyl-L-serine (II) and N-dichloroacetyl-threo-β-phenyl-D-serine (III). The N-dichloroacetyl and N-benzoyl derivatives of erythro-β-phenyl-DL-serine were asymmetrically hydrolyzed by KT 84 and respectively formed N-dichloroacetyl- (VI) and N-benzoyl-erythro-β-phenyl-D-serine, besides erythro-β-phenyl-L-serine (V). The acetone-dried powder and bacterial suspension of KT 83 hydrolyzed (III) and (VI), respectively forming threo- and erythro-β-phenyl-D-serines, and also hydrolyzed N-dichloroacetyl-threo- and -erythro-β-phenyl-L-serines. Both KT 84 and 83 hydrolyzed the erythro-type of N-dichloroacetyl-β-phenylserine more easily than its threo type. The dichloroacetyl derivatives of β-phenylserine were prepared by the chloral hydrate method. The acetone-dried powder and bacterial suspension of KT 84 effected asymmetric hydrolysis of N-dichloroacetyl-threo- (XIII) and -erythro-β-p-nitrophenyl-DL-serine (XVI) to form, respectively, threo-β-p-nitrophenyl-L-serine (XIV) and N-dichloroacetyl-threo-β-p-nitrophenyl-D-serine (XV), and erythro-β-p-nitrophenyl-L-serine (XVII) and N-dichloroacetyl-erythro-β-p-nitrophenyl-D-serine (XVIII). The acetone-dried powder of KT 83 hydrolyzed (XV) and (XVIII) to respectively form threo- (XIX) and erythro-β-p-nitrophenyl-D-serine (XX), and also hydrolyzed N-dichloroacetyl-threo- and -erythro-β-p-nitrophenyl-L-serines.
The acylase of soil bacteria KT 84 was extracted in a cell-free state and its cell-free acetone powder was prepared. The bacterial mass, acetone-dried powder, cell-free extract, and cell-free acetone powder of KT 84 all effected asymmetric hydrolysis of α, ε-di-N-benzoyl- and α-N-acetyl-ε-N-benzoyl-DL-lysine affording ε-N-benzoyl-L-lysine and α, ε-di-N-benzoyl-D-lysine, and ε-N-benzoyl-L-lysine and α-N-acetyl-ε-N-bezoyl-D-lysine, respectively. The benzoyl derivative of ε-N-benzoyl-L-lysine was more easily hydrolysed than its acetyl derivative in this case. Eleven kinds of α-N-acyl derivative of ε-N-benzoyl-DL-lysine were synthesized and hydrolytic action of cell-free extract of KT 84 was examined by ninhydrin colorimetry. The results showed that (1) m- and p-nitrobenzoyl, dichloroacetyl, and benzoyl derivatives were more easily hydrolyzed than other acyl derivatives; (2) m- and p-nitrobenzoyl derivatives were hydrolyzable but o-nitrobenzoyl was hardly hydrolyzed, while p-aminobenzoyl was far more resistant to hydrolysis than p-nitrobenzoyl derivative; (3) cinnamoyl and phenylacetyl derivatives are less hydrolyzable than benzoyl derivative; and that (4) chloroacetyl, acetyl, and formyl derivatives are much less hydrolyzable than dichloroacetyl derivative.
The acetone-dried powder of soil bacteria KT 84 effects asymmetric hydrolysis of N-benzoyl-DL-leucine and -DL-valine to form respectively L-leucine and N-benzoyl-D-leucine (IV), and L-valine and N-benzoyl-D-valine, besides benzoic acid. L-Leucine and L-valine so obtained were led to their N-benzoyl derivatives. N-Benzoyl-L-leucine is more easily hydrolyzed by KT 84 than N-benzoyl-L-valine. The acetone-dried powder of KT 83 hydrolyzed (IV) to form D-leucine and benzoic acid.
The acetone-dried powder of soil bacteria KT 84 effected asymmetric hydrolysis of N-benzoyl-DL-glutamic acid and -DL-aspartic acid to respectively form L-glutamic acid and N-benzoyl-D-glutamic acid (IV), and L-aspartic acid and N-benzoyl-D-aspartic acid, besides benzoic acid. N-Benzoyl derivatives were prepared from these L-glutamic and L-aspartic acids. The acetone-dried powder of KT 83 hydrolyzed (IV) to form D-glutamic and benzoic acids.
The acetone-dried powder of soil bacteria KT 84 effected asymmetric hydrolysis of N-benzoyl-DL-methionine, N-acetyl-DL-methionine, and di-N-benzoyl-DL-cystine to respectively form L-methionine and N-benzoyl-D-methionine (V), L-methonine and N-acetyl-D-methionine, and L-cystine and di-N-benzoyl-D-cystine in a good yield. N-Benzoyl-L-methionine is more easily hydrolyzed by KT 84 than its N-acetyl derivative. The acetone-dried powder of KT 83 hydrolyzed (V) to form D-methionine and benzoic acid.
The acetone-dried powder, bacterial mass, and cell-free acetone powder of soil bacteria KT 84 effected asymmetric hydrolysis of N-benzoyl derivatives of serine, threonine, allothreonine, 2, 4-diaminobutyric acid, and ornithine to respectively form, besides benzoic acid, L-serine and N-benzoyl-D-serine, L-threonine and N-benzoyl-D-threonine, L-allothreonine and N-benzoyl-D-allothreonine, L-2-amino-4-benzamido- and D-2, 4-dibenzamido-butyric acids, and δ-N-benzoyl-L-ornithine and α, δ-di-N-benzoyl-D-ornithine. The yield of L-threonine was very poor and formation of glycine was detected in this case.
Sodium carboxymethylcellulose (Na-CMC) is being used as a vehicle for pharmaceutical suspension and dynamic behavior of Na-CMC gel, related to the appearance of preparations, was examined. Measurement of static viscoelasticity showed that its behavior can be explained mechanically as that of 3-element model of a combination of the Maxwell element and Voigt element, and values of each componental element were calculated. Thixotropy of Na-CMC gel was measured and its mechanical model examined, and these two elastic elements were found to show different behaviors. Further, as a reference, viscosity of dilute solutions was determined and abnormality of viscosity was recognized. Application of a few theories of dilute solutions of high-molecular substances indicated full of inconsistent results.
Sodium carboxymethylcellulose (Na-CMC) possesses viscoelastic properties and should naturally show the Weissenberg effect. Relationship between this effect and thixotropy was examined with thixotropic substances, such as Na-CMC gel. Measurement of the Weissenberg effect while continuously changing the number of revolution indicated the presence of a hysteresis loop with up and down curves, as was also recognized in flow curve. The Weissenberg effect was found to change in accordance with destruction and thixotropy of the gel and its qualitative explanation was well explained by exactly the same method of explanation as in thixotropy.
Viscosity of dilute aqueous solution of methylcellulose was measured to help examine the phenomena of turbidity and gelation of methylcellulose solution with rise in temperature. Temperature dependence of [η], Huggins' k′, and root-mean-square end-to-end distance, √r2, was examined and it was thought that solvation decreases with rise in temperature in dilute solution and curving of the methylcellulose molecule. Compounds with larger degree of polymerization had larger k′ value but scarcely any difference was recognized in substances with low degree of polymerization.
Rate of free and collective sedimentation of powdered pharmaceutics in aqueous solution of methylcellulose and sodium methylcellulose was examined. Sedimentation velocity of granules was calculated from the Stoke's formula for free sedimentation and the value was compared with specific sedimentation volume. It was thereby learned that the aqueous soution of sodium carboxymethylcellulose and methylcellulose had great efficiency to disperse granules. Sedimentation velocity of a suspension that undergoes collective sedimentation was found to be expressed by a definite curve, as indicated in Eq. 2, by the use of a dimensionless number, irrespective of the kind of granules present or the degree of flocculation.
Reaction of ethyl 2-oxocyclohexanecarboxylate or N, N-bis(2-ethoxycarbonylethyl)-methylamine with hydrazobenzene respectively afforded 1, 2-diphenyl-4, 5, 6, 7-tetrahydro-3-indazolinone or 1, 2-diphenyl-5-methyl-4, 5, 6, 7-tetrahydro-1H-pyrazolo [4, 3-c] pyridin-3(2H)-one. Reaction of antipyrine with 2-haloacyl halide or anhydride, in the presence of anhydrous aluminum chloride in carbon disulfide afforded 4-(2-haloacyl) antipyrines, which were further reacted with dialkylamine, piperidine, or morpholine to give 4-(2-dialkylaminoacyl)-, 4-(2-piperidinoacyl)-, and 4-(2-morpholinoacyl) antipyrines.
Phosphodiesterase, phosphomonoesterase, 5′-nucleotidase, and lecithinase-A are markedly stable in cobra venom and can be preserved over a long period. Two kinds of phosphodiesterase are present in fresh cobra venom immediately after collection. Two kinds of strongly basic protein are also present in the cobra venom and the two kinds of substance related to nucleic acid, having absorption maxima at around 252 and 257mμ, present in the venom can be separated from each other by column chromatography using carboxymethylcellulose.
Glycyrrhizin (glycyrrhizinic acid) in liquorice or liquorice preparations was separated by paper chromatogram and extracted with 50% by volume of ethanol or 0.05N ammonia water. By measuring optical density of the solution so prepared at 252mμ in the former and at 258mμ in the latter, determination of glycyrrhizin was established.
Attempts were made to synthesize (2, 3-dimethoxy-6-nitrophenyl) acetic acid (XV) by various routes shown in Chart 1 and it was found that the route starting with 2, 3-dimethoxy-6-nitrobenzoic acid (I) and going through (III), (XIII and XIV), to (XV) by the Arndt-Eistert reaction is the most reliable.
Sequoyitol and hinokiflavone were detected as the components of fallen leaves of Metasequoia glyptostroboides. Paper chromatographic examination revealed the presence of glucose, fructose, galactose, and sucrose. This is the first instance of a detection of sequoyitol from plant leaves.
Utilizing anodic synthesis, a kind of Kolbe reaction, 10 kinds of straight-chain fatty acids, besides those already reported, were synthesized. Electrolytic oxidation of phenylacetic acid and levulic acid respectively gave dibenzyl and 2, 7-octanedione. As an example of electrolytic oxidation, 1-naphthol was prepared from naphthalene. It was found that methanol used as the reaction solvent was also oxidized in this case to formaldehyde.
It is known that nitration of acetophenone is accompanied by the formation of a substance of m.p. 146-148° as a by-product in 5-10% yield when cooling efficiency is poor. Syntheses of furoxans proved that this by-product is bis (m-nitrobenzoyl) furoxan. Application of dilute nitric acid to p-bromoacetophenone in acetic acid afforded bis (p-bromobenzoyl) furoxan.
For the qualitative estimation of carbonyl compounds, aliphatic, aromatic, and terpenic carbonyl compounds listed in Table I were derived to 2, 4-dinitrophenylhydrazones and submitted to paper partition chromatography. As shown in Table II, paper chromatography using liquid paraffine as the stationary phase and methanol as the mobile phase effected good qualitative estimation of aliphatic and terpenic carbonyl compounds. Further examinations on solvent system are necessary for the estimation of aromatic carbonyl compounds by this method.