DL-Aspartic acid [3-14C] is distributed in approximately 10-2% (24-hour value) in the 80% ethanol extracted fraction of various organs of the rat, the distribution concentrated mostly in the spleen. Incorporation of labeled aspartic acid into caronsine and anserine, and the dilution value of specific activities are about the same as in the case of β-alanine, and incorporation of radioactivity is greater into carnosine than into anserine. Under the same conditions, incorporation of L-leucine [U-14C] into carnosine and anserine not taking place, incorporation of aspartic acid and β-alanine into these peptises are specific. β-Aspartylhistidine is decarboxylated by the carnosine-synthesizing enzyme system but there is no formation of carnosine or anserine, and β-aspartylhistidine is found to be hydrolyzed and further decomposed. β-Aspartylhistidine decreases the amplitude of systolic heart beat of excised toad heart but does not affect peripheral blood, differing from carnosine, and antagonizes oxytocin action in uterine muscle. From these experimental evidences, it is concluded that aspartic acid is a biological precursor of carnosine and anserine, but there is little possibility than β-aspartylhistidine serves as a direct precursor of carnosine.
5-Monoiodohistidine was synthesized and its conjugation with β-alanine. L-aspartic acid, L-glutamic acid, L-leucine, and DL-β-aminoisobutyric acid was followed in vitro with radiolabeled 5-monoiodohistidine-131I as the substrate. It was found that only β-alanine underwent conjugation, as in the case of using 14C-labeled β-alanine and histidine, and biosynthesis of 131I-carnosine was observed. This reaction is markedly proceeded by the enzyme system from the liver and kidney, and weakly by that from skeletalmuscle and small intestine, and this enzyme preparation is very labile in any case. This experiment seems to suggest that the assay of the ability for biosynthesis of carnosine by the use of 131I-labeled monoiodohistidine would be facile and it may be considered that the final step in the biosynthesis of carnosine in vivo is the β-peptide bonding between β-alanine and histidine.
Anti-inflammatory action of vitamin B1 and its derivatives was examined by three methods : (1) Rat hind paw edema, (2) intraperitoneal dye leakage of the mouse, and (3) vascular permeability of the guinea pig skin. Rat hind paw edema induced by a-chymotrypsin was inhibited by vitamin B1, T.T.F.D. and T.A.T.D. inhibited rat paw edema induced by acetic acid and a-chymotrypsin. B.T.M.P. and S.T.M.P. were effective in the rat paw edema induced by carrageenin. The leakage of dye into the peritoneal cavity in the mouse was inhibited by vitamin B1 and its derivatives except S.T.M.P. when acetic acid was an irritant. They showed significant inhibition of the increase in vascular permeability of the guinea pig skin induced by acetic acid. From these results, it is concluded that when inflammation is induced by acetic acid, vitamin B1 and its derivatives are effective with all the three methods used here. In addition, it was found that the anti-inflammatory action of vitamin B1 derivatives can be divided into two proto-types : (a) T.T.F.D. and T.A.T.D. of disulfide type, (b) B.T.M.P. and S.T.M.P. of S-acyl type.
The reaction of phenyl glycidyl ether with ethanol at about 80° in the presence of sodium hydroxide, sodium ethoxide, or triethylamine afforded mainly 1-ethoxy -3-phenoxy-2-propanol in 50-65% and 2-ethoxy-3-phenoxy-propanol in only 1.8%. By the identification of oxidative products of these compounds with 1-ethoxy-3-phenoxy-2-propanone or 2-ethoxy-3-phenoxypropionic acid, which was obtained by another independent synthetic method, the structures of these compounds was proved to be the primary or the secondary alcohol mentioned above. The reaction of phenly glycidyl ether with ethanol in the presnece of sulfuric acid resulted in the increase of primary alcohol. Without the catalyst, the reaction velocity was very low and at elevated temperature, a large amount of 3-phenoxy-1, 2-diethoxypropane was formed. Thioethanol reacted well with phenyl glycidyl either even at lower temperature than ethanol did and afforded 1-ethylthio-3-phenoxy-2-propanol. Higher thiols, however, showed poorer yields. The reaction of phenyl glycidyl ether with dialkylaminoethanol or dialkylaminothioethanol also gave corresponding basic etheralcohols. When phenylthioglycidyl ether was reacted with ethanol or thioethanol, 1-ethoxy-3-phenylthio-2-propanol or 1-thioethyl-3-phenylthio-2-propanol was obtained in a good yield. These compounds are expected to be effective as a muscle relaxant.
A new spectrophotometric method is proposed for the determination of saccharin sodium. The present method is based on solvent extraction into nitrobenzene of the ionpair formed between tris (1, 10-phenanthroline)-iron (II) and the anion of saccharin. The maximum absorbance of the extract in the organic phase is at 516 mμ. To obtain optimum conditions for the determination of saccharin sodium, various factors were investigated, A maximum extraction is obtained at pH 4-8, when an excess of at least 20-fold (molar) of the phenanthroline-iron (II) chelate to saccharin is present. The color intensity of the extracted species remains constant at room temperature for several hours after separation of the organic layer. A linear relationship is obtained over the tested range of 10-6 to 3.2 × 10-5M of saccharin in aqueous solution. The effect of several other food additives on this method was investigated. Saccharose, glucose, dulcin, citric acid, and sodium chloride do not interfere with the determination in a moderate amount. Sodium cyclamate gives a slight positive error.
The retention indices of various compounds were measured on Apiezon-L, Squalane, and SE-30 as the stationary phase, and the following fact was found : Retention indices of normal paraffins were expressed approximately by the quadratic function of boiling points, i.e., I = 0.602×10-2·K2-0.730 K+131. The relation between boiling point and retention index in various compounds corresponds similarly to that between boiling point and retention index of normal paraffins, except that alcohols do not follow the above equation on Apiezon-L and Squalane, and amines on SE-30. The validity of applying this formula for the determination of boiling points of various compounds was investigated.
Chemical method for the determination of ecdysterone (I) and inokosterone (II) was examined. Both I and II show ultraviolet absorption with a maximum at 250 mμ and the use of this wave length for their determination was attempted. Since this absorption maximum disappeared on the application of sodium borohydride, I and II can be determined from the decrease of this absorption. Application of sulfuric acid to I and II produces fluorescence and the fluorescence intensity of II increases markedly when made ammonia alkaline. Utilization of this fact made it possible to determine II in a mixture of I and II. Linearity was established between the mixture of I and II, and a difference in the fluorescence intensity in sulfuric acidity and ammoniac alkalinity, and this fact was utilized to determine the mixing ratio of I and II in their mixture.
Relationship between chemical constitution and antitumor activity of isoquinoline derivatives was examined, and the following results were obtained. These compounds were classified into five types according to the substituent at 1-position of the isoquinoline ring, and their in vitro activity against HeLa cells and in vivo activity against Ehrlich ascites carcinoma were tested. A parallelism was found between the in vitro and in vivo activities and the alkyl type compounds, especially the ones possessing tertiaryalkyl or cycloalkyl group in 1-position showed a marked inhibitory effect, while the other four types were only slightly active. These data suggested that the antitumor activity of isoquinoline derivatives was mainly determined by the chemical structure of the substituent at 1-position and in the alkyl type compounds the activity increased as the number of carbon atoms in the alkyl increased.
Based on the results previously reported, several derivatives of 1 -alkyl substituted isoquinoline were tested for antitumor activity using Ehrlich carcinoma (ascites and solid), sarcoma-180 (ascites and solid), SN-36 leukemia (ascites), and NF-sarcoma (solid) in mice. These compounds showed marked inhibitory effect on all the above tumors, although their activities were stronger in the case of ascites than in solid type. Especially marked inhibitory effects were observed with 1-(1, 1-dimethylpropyl)-3-methyl-6, 7-methylenedioxyisoquinoline·HCl (B-19), 1-(1, 1-dimethylpentyl)-6, 7-methylenedioxyisoquinoline·HCl (B-21), 1 -[(1-methylcyclopentyl)-methyl]-3-methyl-6, 7-methylenedioxyisoquinoline·HCI (B-36) and 1-[(1-methylcyc1ohexyl)methyl]-3-methyl-6, 7-methylenedioxyisoquinoline·HCl (B-36). By the effect of these compounds, number of Ehrlich ascites carcinoma cells in the ascites fluid decreased 1 hour after their administration and recovered after 24-48 hours, while the increase of degenerating cells was observed at 1-9 hours. It seemed that the antitumor activity of 1-alkyl substituted isoquinoline derivatives depended on the chemical structure of 1-alkyl residues and the actitity was greatest with tertiary or cyclo-alkyl groups.
In order to prepare 5-nitro-2-furylvinylene derivatives in a better yield and in a simpler manner, the Wittig reaction was carried out to prepare novel nitrofurans. Of the derivatives thereby obtained, 2-bromo-5-[5-nitro-2-furylvinylene] furan, methyl 5-[5-nitro-2-furylvinylene] furan-2-carboxylate, and 4-[5-nitro-2-furylvinylene] benzonitrile were obtained in stereoisomers which were successfully separated and purified.
Baeyer-Villiger rearrangement of 1-amino-2-hydroxypropiophenones (VI-XI) derived from 1-phenyl-2-amino-1, 3-propanediols (IV and V) was examined. Treatment of phenyl ketone (VI) and p-acetamidophenyl ketone (XI) with peracetic acid in the presence of sulfuric acid afforded serine. Since optically active VI derived from the natural type of IV gave D-serine, asymmetric carbon in VI was found to have the same configuration as L-phenylalanine. This conclusion agreed with Honjo's report.10) Other ketones (VII-X) gave benzoic or p-nitrobenzoic acid.
Reddish coloration took place upon mixing riboflavin and sulfanilamide in high concentrations, and the nature of interactions between riboflavin and compounds related to sulfanilamide including p-aminobenzoic acid was investigated. New absorption bands were observed in the vicinity of 370 and 500mμ in the difference spectra of mixed systems of riboflavin and sulfanilamide related compounds. Analysis of the data of these two difference bands through |the Benesi-Hildebrand equiation revealed that 1 : 1 complexes were formed. Furthermore, it was also revealed from the values of K, ΔH, and ΔS, as well as solvent effects that sulfanilamide and p-aminobenzoic acid behave quite similarly in interacting with riboflavin. A reciprocally correspondent relationship was found to exist between the two difference bands ; whereas the band at about 500 mμ was more distinct in aqueous medium, the one at about 370 mμ was more distinct in non-aqueous media. It was presumed that charge-transfer interaction which is dominant in non-aqueous media (occurrence of the band around 370 mμ) proceeds to the extent of formation of riboflavin semiquinone (occurrence of the band around 500 mμ). It was found from the effects of pH on absorbances at the longer wavelength maxima that undissociated species of both riboflavin and sulfanilamide compounds are more favorable for the complex formation reactions. These results for complex formation in aqueous medium has been confirmed also through the examination of changes in solubility of riboflavin which took place in the presence of sulfanilamides compunds. Possible correlations of sulfanilamides-riboflavin complexing to the metabolic disturbance of vitamin B2 which is incidental to administration of sulfonamide drugs have also been referred to.
Reaction of 1-(methylsulfonyl)4-phenylphthalazine (I) and active methylene compounds was carried out in benzene, in the presence of sodium amide. As the so-called active methylene compounds, ethyl cyanoacetate (IIIa), ethyl acetoacetate (IIIb), ethyl malonate (IIIc), malononitrile (IIId), and phenylacetonitrile (IIIe) were used. In addition, reaction was also examined with ethynylbenzene (IIIf), acetonitrile (IIIg), 2-picoline (IIIh), and ethyl acetate (IIIi). As would be expected, reaction with IIIa to IIIf respectively afforded ethyl a-cyano-4-phenyl-1-phthalazineacetate (IVa), ethyl α-acetyl-4-phenyl-1-phthalazineacetate (IVb), ethyl 4 -phenyl-1-phthalazinemalonate (IVc), 4-phenyl-1-phthalazinemalononitrile (lVd), α, 4-diphenyl-1-phthalazineacetonitrile (IVe), and 1-phenyl-4-(phenylethynyl)phthalazine (IVf) (Chart 1 and Table I). From the reaction witln IIIg, a compound considered to have been formed by the reaction of the product corresponding to the above type, 4-phenyl-1-phthalazineacetonitrile (IVg), with I was obtained in the forms of bis(4-phenyl-1-phthalazinyl)acetonitrile (IVg') (Chart 2 and Table I). In the case of IIIh, this compound did not take part in the reaction, and the reaction of I and sodium amide took place to form 1, 1'-imino-bis[4-phenylphthalazine] (VI). Reaction of I with IIIi afforded 1-ethoxy-4-phenylphthalazine (VII) (Chart 2). Hydrolysis of IVg' gave 1, 1'-methy1enebis[4-phenylphthalazine](VIII) (Chart 3). In a previous paper, 1) the by-product formed by the reaction of I and methyl ketone in the presence of sodium amide was assumed as 4, 4'-diphenyl-1, 1'-biphthalazine but this assumption was found to be wrong, and the product is now corrected to VIII. The route of its formation is presumed as shown in Chart 4.
In the metabolic study of butyl aryl ethers in vivo, it has been shown in this laboratory that dealkylation is only minor and penultimate hydroxylation is the major one. In connection with this study, metabolism of 4-butylaminoantipyrine was investigated to learn whether it also undergoes the penultimate hydroxylation rather than the dealkylation of butyl chain. Contrary to this expectation, the major pathway of this drug was found to be dealkylation in all of the animals tested (rabbits, guinea pigs, mice, and rats), excreting 4-aminoantipyrine and its acetylated metabolite, 4-acetamidoantipyrine, as the main metabolites. However, it has also been recognized that there is a small amount of other urinary metabolite. The structure of this minor metabolite has been identified as 4-(2-butenylamino)antipyrine by the comparison with the authentic sample chemically synthesized. Since it seems very likely that this metabolite may be formed from the penultimate hydroxylated metabolite by dehydration, the presnet study would support the hypothesis that, as in C- and O-alkyl compounds, the penultimate hydroxylation takes place also in N-alkyl compounds if they have a considerably long alkyl chain such as butyl group.
4-(Methylsufonyl)cinnoline (I) gradually undergoes decomposition when left in air and changes into 4-cinnolinol (IV). I easily undergoes decomposition into IV when heated with water and the decomposition is further facilitated when heated with a dilute acid. When I is heated in aqueous solution of sodium hydroxide or in methanolic solution of sodium methoxide, IV or 4-methoxycinnoline (V) is formed. Application of aniline, hydrazine, or hydroxylamine to I respectively gives 4-anilino-(VI), 4-hydrazino-(VII), or 4-hydroxyamino-cinnoline (VIII) (Chart 2). The methylsulfonyl group in I is substituted also with active methylene (methine) carbanion. For example, reaction of I, in the presence of sodium amide, with ethyl acetoacetate (IXa), ethyl malonate (IXb), ethyl cyanoacetate(IXc), malononitrile(IXd), phenylacetonitrile(IXe), or ethynylbenzene (IXf) respectively affords ethyl 4-cinnolineacetate (Xa'), ethyl 4-cinnolinemalonate (Xb), ethyl α-cyano-4-cinnolineacetate (Xc), 4-cinnolinemalononitrile (Xd), α-phenyl-4-cinnolineacetonitrile (Xe), or 4-(phenylethynyl)cinnoline (Xf) (Chart 3 and Table I).
Irradiation (medium-pressure mercury lamp, Pyrex filter) of cholest-4-ene-3, 6-dione (I) in cyclohexane resulted in the efficient hydrogen abstraction by I from the solvent to give cholestane-3, 6-dione (II) and 4ξ-cyclohexyl-5ξ-cholestane-3, 6-dione (III). On irradiation in benzene, I also afforded II.
Changes in heat resistance, fungicidal resistance, respiration quotient, and variation of intracellular phosphorus with the germination of conidia were measured in Cochliobolus miyabeanus. Incubation of the conidia in distilled water at 30° results in the formation of a germ tube in 30-90 minutes, and the heat and fungicidal resistances decrease at this period, and the respiration quotient shows the minimum value. From these facts, the stage of germination of this conidia can be divided into three stages of the first stage of germination (0-30 min), germ-tube formation (30-90 min), and germ-tube elongation (after 90 min). There was no great change in the distribution of phosphorus compounds in the cells during these three stages.
Treatment of 3-acetyl-4-hydroxy-6-methyl-1H-2-pyridone (V) with hydroxylamine gives its oxime (VI) whose treatment with polyphosphoric acid at room temperature results in the Beckmann rearrangement to afford 3-acetamido-4-hydroxy-6-methyl-1H-2-pyridone (VIII). Further heating of VIII with polyphosphoric acid results in its transition to 2, 6-dimethy1-5H-oxazolo[4, 5-c]pyridin-4-one (IX), which is also obtained on heating VI with polyphosphoric acid directly. IX also transits to VIII when left with hydrochloric acid at room temperature, while hydrolysis of IX with heating gives 3-amino-4-hydroxy-6-methyl-1H-2-pyridone (X). Treatment of dehydroacetic acid oxime (XIV) with polyphosphoric acid gives 3-acetamido-4-hydroxy-6-methyl-2H-pyran-2-one (XVI) and 2, 6-dimethyl-4H-pyrano[3, 4-d]oxazol-4-one (XVII), and the latter transits to IX on treatment with ammonia. Marcus and others5) reported that the reaction of XIV and diketene gave its cyclized product, 3, 6-dimethyl-4H-pyrano[3, 4-d]isoxazol-4-one (XV) but re-examination of this reaction showed that the product is a cyclized oxazolo compound (XVII) via the Beckmann rearrangement.
The Wittig reaction was carried out on cyanomethylenetriphenylphosphorane (I) and α-bromocyanomethylenetriphenylphosphorane (IV) as the phosphoranes and 5-nitrofurfural (IIa) as the aldehyde, and also the reaction of I with furfural (IIb), 5-methylfurfural (IIc), and 5-bromofurfural (IId). Examinations were also made on the solvent effect and the salt effect of Lewis bases on the formation ratio of cis and trans compounds. Identification of the components was made through elemental analyses and the infrared and NMR spectra, and the formation ratio of cis-and trans-olefins was determined by gas chromatography. It was thereby found that the cis compound was formed in larger amounts in benzene solvent.
It is known that, in general, chloromethylation of phenol derivatives gives only the polychloromethylated product or polymer. However, by protection of the hydroxyl group with a benzoyl group, monochloromethylated products (III and VII) were obtained under normal condition in a high yield.