1-Sulfanilamidophthalazine, 1-sulfanilamido-4-methyl-phthalazine, and 1-sulfanil-amido-3-methyl-4-(3H) phthalazinone were obtained by (a) fusion of 1-halogeno derivative and potassium salt of sulfanilamide, or (b) condensation of 1-amino derivative with p-acetamidobenzenesulfonyl chloride in pyridine and hydrolysis of the acetyl compound thereby obtained. 1-Amino- and 1-amino-4-methylphthalazines were prepared from their 1-chloro derivatives by heating with phenol in the presence of potassium carbonate to 1-phenoxy derivative and its fusion with ammonium acetate. 1-Amino-3-methyl-4-(3H) phthalazinone was prepared by the chlorination of 2-methyl-1, 4-phthalazinedione with phosphoryl chloride to 1-chloro-3-methyl-4-(3H) phthalazinone and its amination in the presence of bronze powder as a catalyst, or condensation of phthalonic acid with methylhydrazine to 3-methyl-4-oxo-3, 4-dihydro-1-phthalazinecarboxylic acid and its Curtius degradation.
1-Phenyl-2-methyl-3, 4-bisbromomethyl-3-pyrazolin-5-one, m. p. 187° (decomp.), was prepared by the application of 2.2 moles of N-bromosuccinimide to 1-phenyl-2, 3, 4-trimethyl-3-pyrazolin-5-one or 1.15 moles of the imide to 1-phenyl-2, 4-dimethyl-3-bromomethyl-3-pyrazolin-5-one. Although the melting point of the product differs from that (175°) given in literature, its analytical values agreed well and its structure was confirmed by the following reactions. The 3, 4-bisbromomethyl compound was reacted with several kinds of aliphatic and aromatic primary amines and new pyrroline derivatives, which were negative to secondary amine reactions, were obtained.
Measurement of viscosity was carried out in a small and narrow range of the rate of shear on aqueous solution of sodium carboxymethylcellulose added with granules of procaine penicillin G, chloramphenicol, chloramphenicol palmitate, and talc, whose particle size distribution is known. They all showed a typical structural viscosity and the three, with the exception of talc, showed dependence on shear rate. There was a linear relationship between the logarithms of viscosity, ηap and rate of shear. Viscosity increases with increasing amount of the granules added but the absolute value varies with the kind of the granule, though independent of the rate of shear. With the increasing amount of the granules, dependence of consistency on shear rate increases and structural viscosity becomes marked. This is assumed to be due to the formation of a floccule with the exception of talc.
Measurement of viscosity was carried out in a large and wide range of rate of shear on a suspension of five kinds of granules, including the four mentioned in the preceding paper and barium sulfate. All were found to be a Bingham body and showed a simple plastic flow as long as the amount of the granules added was small but exhibited hysteresis loop in the viscosity curve as the amount of granules increased, and the thixotropic coefficient, M, increased with the increase of yield value and plastic viscosity. In the present series of experiments, barium sulfate alone showed exceptional characteristics and both the yield value and plastic viscosity tended to decrease when the amount added was small.
Measurements of viscosity were carried out on the same samples as those described in the preceding paper. In suspension of substances like talc, which does not have a strong thixotropic character, ηap increases with the increasing amount of the granules added and spinnability, which is a lower-limit type, also increases. A transition point, at which the spinnability changes to the upper-limit type, is observed when ηap becomes too large. When the amount of the granules added increases further, ηap also continues to increase but spinnability decreases while still in the upper-limit type and finally disappears altogether. In a suspension with strong thixotropic character, ηap increases with increasing addition of the granules at first and spinnability, which is a lower-limit type, also increases. With further increase in the amount added, the length of spinnability becomes definite, irrespective of the velocity of drawing and in spite of increasing ηap. This is probably due to the marked appearance of thixotropic character with the increased addition of granules. Further increase in the amount results in the decrease of spinnability, irrespective of the velocity of drawing.
Condensation of 1, 3, 4, 6-tetraacetyl-β-D-glucosamine (I) and phthalic anhydride affords 1, 3, 4, 6-tetraacetyl- N-phthaloyl-β-D-glucosamine (II) whose reaction with hydrogen bromide in glacial acetic acid and acetic anhydride mixture gives 3, 4, 6-triacetyl-1-bromo compound (III). Reaction of (III) with methanol in the presence of silver carbonate results in the formation of methyl 3, 4, 6-triacetyl-N-phthaloyl-β-D-glucosaminide (IV), which is also obtained on boiling (II) with 5% methanolic hydrochloric acid until the optical rotation becomes definite and subsequent acetylation. Condensation of (II) and phenol results in the formation of phenyl 3, 4, 6-triacetyl-N-phthaloyl-β-D-glucosaminide (IX), whether the catalyst used is p-toluene-sulfonic acid or zinc chloride. As shown by the foregoing, substitution reaction of C-1 position in (II) chiefly results in the formation of a β-bonding and the formation of α-bonding is markedly obstructed. This is thought to be due to the bulky phthalimide group in C-2 position that causes a great resistance to the formation of a half-chair form (XIII), assumed to be the intermediate in the formation of α-bonding from C-1 conformation (XII).
By the resolution of DL-1-phenyl-2-dimethylaminoethanol, (+)- and (-)-1-phenyl-2-dimethylaminoethanethiol were synthesized. Using these as a catalyst, asymmetric syntheses of methyl mandelate and 1-naphthylglyoxalate were carried out and it was found that the product obtained with (+) catalyst is a levorotatory compound and that with (-) catalyst is a dextrorotatory compound.
Using synthesized (+)- and (-)-2-[N-methyl-N-(α-methylphenethyl)amino]ethanethiol as a catalyst, methyl mandelate and methyl 1-naphthylglyoxalate were respectively prepared from phenylglyoxal and 1-naphthylglyoxal, but it was not an asymmetric synthesis. It was thereby assumed that the presence of a mercapto group in proximity of asymmetric carbon atom is necessary for this kind of asymmetric synthesis.
Spasmodic action of 10 homologs of 2-methyl-4-amino-5-hydroxymethylpyrimidine (OMP) was compared with that of OMP and it was found that four homologs, 2-methyl-4-amino-5-ethoxymethylpyrimidine (IV), 2-methyl-4-amino-5-pyrimidinecar-boxaldehyde (V), 2, 5-dimethyl-4-aminopyrimidine (III), and 2, 6-dimethyl-4-aminopyrimidine (IX), possessed specific spasmodic action. The activity of (IV) and (V) was similar to that of OMP, and (IV) showed equal activity in about equimolar amount of OMP while (V) caused spasmodic action only in pyridoxine-deficient mice. The activity of other two homologs is suppressed by phenobarbital and ethylurethan but is not suppressed by pyridoxine, diphenylhydantoin, and mephenesin, while the action does not increase by pyridoxine-deficient diet, These effects are different from the spasmodic action of OMP. It was assumed from the series of present work that the specificity of the chemical structure responsible for the spasmodic action of OMP is the presence of a methyl in 2-position, amino in 4-position, and radical easily converted to hydroxymethyl group in 5-position.
Examination on spasmodic action of thiosemicarbazide on mice revealed that the action is suppressed by Phenobarbital, ethylurethan, and diphenylhydantoin, and also by pyridoxine, pyridoxamine, and pyridoxal. This spasmodic action was found to be increased by breeding the mice for one week on a pyridoxine-deficient diet or by oral administration of 1mg./10g. body weight of INAH, which would supplement the experiments of Parks and others.
The chorionic gonadotropin (HCG) extracted from human placenta was obtained in nearly homogeneous state by column chromatography through Hyflo Super-Cel. However, this method of isolation was found to give a poor yield and it was difficult to obtain the product in a large amount, which made it impossible to carry out detailed studies on chemical and physicochemical properties. Further studies on its method of purification were carried out and it was found that the concurrent use of fractional precipitation by ethanol and salting-out gave human chorionic gonadotropin, thought to be homogeneous, in comparatively high yield.
Application of 10%, 20%, or 48% hydrobromic acid to 5-bromo-6-hydroxyauinoline (I) or 7-hydroxy-8-bromoquinoline (II) resulted in recovery of the starting material in either case. The same reaction carried in the presence of phenol was found to cause quantitative debromination in the case of (I) and 48% hydrobromic acid and a debrominated compound was obtained in a high yield from (II) and 20% hydrobromic acid. The same was true in the case of 5-bromo-6-aminoquinoline and 20% hydrobromic acid with phenol. Debromination did not occur in the case of 6-hydroxy-7-bromoquinoline (V) and hydrobromic acid, even in the presence of phenol.
Debromination of bromine in the 5-position of 5, 7-dibromo-6-hydroxyquinoline (III) with hydrobromic acid and phenol was examined and 6-hydroxy-7-bromoquinoline (II) was found to be obtained in a high yield when (III) is heated for 2 hours with 48% hydrobromic acid and phenol, with agitation. (II) is also obtained in a high yield when (III) is heated with phosphoric and sulfuric acids and glycerol.
The principle extracted and purified from human placental chorion, responsible for the Mainini reaction and considered to be almost homogeneous by electrophoresis and ultracentrifugation, was examined for its amino acid composition. By the two-dimensional paper partition chromatography, 13 kinds of amino acid were detected as the componental amino acid. The degree of acid hydrolysis of the protein fraction (MG) was examined by varying the period of hydrolysis and it was found that the splitting of valine and isoleucine was insufficient in 24 hours and that aspartic acid, threonine, and serine were decomposed to a fair extent in 72 hours (Table I and Fig. 4). Determination of the amino acid was made by column chromatography with Dowex 50 and Amberlite XE-64, and by determination of 17 kinds of amino acid, the composition of amino acids in MG was decided (Table II). The isoelectric point of MG is around pH 4.1 but the number of basic amino acid residue is larger than that of acidic amino acid residue (Table III). This was assumed to be due to the fact that the sugar in MG is attached to basic amino acid residue to reduce its basicity.
2-Methyl-4-amino-5-hydroxymethylpyrimidine (OMP) effects characteristic movement (spasm) in mice and this is suppressed by pyridoxine. Relationship between chemical structure and growth inhibiting action was examined with 11 kinds of compounds related to OMP, using Saccharomyces carlsbergensis and it was found that while the methyl at 2- and amino at 4-position of the pyrimidine ring are requisite conditions for growth inhibiting action, the substitution of 5-position with bromine, methyl, or aldehyde also causes such action. Growth inhibition of these compounds were found to be recovered by the addition of pyridoxine. Further, compounds bonded to nicotinic acid or nicotinamide in 5-position were found to loose its antipyridoxine action, with weakening of nicotinic acid or nicotinamide action.
Hydnocarpyl acetate (III), chaulmoogryl acetate (III′), isohydnocarpyl acetate (VIII), and isochaulmoogryl acetate (VIII′) derived from the componental fatty acids of hydnocarpus oil, hydnocarpic acid (I) and chaulmoogric acid (I′), were each oxidized with potassium permanganate in acetic acid and 4 kinds of ω-hydroxyketonic acids were obtained; 4-oxo-15-hydroxypentadecanoic acid (V), 4-oxo-17-hydroxyheptadecanoid acid (V′), 5-oxo-16-hydroxyhexadecanoic acid (X), and 5-oxo-18-hydroxyoctadecanoic acid (X′). The melting point of these acids and their derivatives are listed in Table I.
Colorimetric determination of glucuronic acid was examined. Each ten cc. of 40% sulfuric acid and a solution obtained by 10g. of urea dissolved in 40% sulfuric acid and made up to 100cc. were each added to 1cc. of a solution containing ca. 0.1% as glucuronic acid, heated in a boiling water bath for 10 minutes, and cooled for 1 minute in an ice bath. After 10-20 minutes, optical density was measured at 430 mμ, with the solution with sulfuric acid alone as the blank. The content of glucuronic acid in the sample is calculated from the standard curve prepared by the same procedure using pure glucuronolactone. This method of determination is hardly affected by other sugars, accuracy is good, and reagents are very stable that it is thought to be suitable for the determination of glucuronic acid preparations. Moreover, the use of a sample added with sulfuric acid as a blank enables the prevention of error to a minimum. If the specific absorbancy, A330/A440, comes to within 2.0, then the presence of glucuronic acid may be considered in the case of a single substance.
Application of α-haloketone to dithiocarbazate (II) should give variety of cyclization products but the actual product obtained was found to be a six-membered 1, 3, 4-(4H)-thiadiazine derivative (IV), which undergoes rearrangement to a five-membered thiazoline derivative on treatment with an acid. This five-membered compound was found to return to the six-membered compound on benzoylation.
Solubility of diphenylhydantoin sodium was measured as well as the viscosity of its saturated solution and solubility curve and viscosity-temperature curve were prepared. It was found that the temperature at the point of break in the two curves corresponded to the transition point of diphenylhydantoin sodium hydrate.
A crude muco-complex containing polysaccharides was obtained by ethanol fractionation from acetone-dried cell body of Micrococcus lysodeikticus. This mucocomplex was separated from polysaccharides by precipitation with acrinol. The polysaccharide is composed of D-rannose alone and the muco-complex from amino acid and sugar portions.
In order to examine the change of 2-methyl-4-amino-5-hydroxymethylpyrimidine (OMP) in vivo, a method of determining the amount of OMP in the urine by the combination of paper ionophoresis and measurement of optical density in the ultraviolet region was established. Excretion of OMP in the urine of rabbits administered with OMP was examined and it was found that 76% of OMP injected was excreted as 2-methyl-4-amino-5-pyrimidinecarboxylic acid during 7 hours after intravenous injection and only 10% of OMP was detected in the urine. The peak of excretion of these two compounds was found 1 hour after administration and 75% of the amount administered was excreted within 3 hours. Discussions were made on the significance of these evidences.
The equilibrium constant, K, and degree of shift of ultraviolet absorption at the time of formation of hydrogen bond between pyridine 1-oxide (I), 3-picoline 1-oxide (II), 2, 6-lutidine 1-oxide (III), and isoquinoline 2-oxide (IV) and ethanol in heptane were examined and following results were obtained. 1) In any of these N-oxides, formation of a hydrogen bond is evident and π-π* and π-π** absorptions show a marked blue shift. This degree of shift is indicated in Table II. Such general properties were considered in connection with the dipole moment →μg and →μe at the ground and excited states, as well as from the electron migration effect. 2) The value of K upon formation of hydrogen bond with ethanol was 14.9, 16.4, 6.06, and 14.0, respectively, for (I), (II), (III), and (IV). Considerations were made on the relationship between the structure and ability to form hydrogen bonds from the magnitude of these values.
Reduction of 4, 8-dimethylnonanoic acid with lithium aluminum hydride afforded (III) and its reaction with red phosphorus and bromine afforded a bromide (IV), which was derived to (V) by heating with ethyl 1, 1, 2-ethanetricarboxylate, in the presence of sodium ethoxide. The free acid (VI), obtained by the saponification of (V), was heated at 190-200° to form an acid anhydride (VII), which was then derived to a dicarboxylic acid (VIII) by heating with alkali. The s-benzylthiuronium salt and p-phenylphenacyl ester of (VIII) were found identical by mixed fusion with the respective derivatives of nupharanedioic acid, derived from deoxynupharidine. This has proved that the structure (A) proposed by the writers for anhydronupharanediol is correct.
A new alkaloid, nupharamine, was obtained from nuphar, a dried rhizome of Nuphar japonicum DC. The alkaloid comes as a colorless, viscous liquid of b.p1 130-134°, [α]D8: -35.4°, C15H25O2N; forms a picrolonate of m.p. 167.5-168°, diacetyl compound of b.p0⋅25 166-169°, and N-methylnupharamine methiodide, m.p. 203.5°, on warming with methyl iodide. From its infrared spectrum, the presence of a furan ring was presumed. This base is a secondary amine and possesses one hydroxyl and one bridged oxygen that forms the furan ring. Presence of two C-methyl groups was predicted in nupharamine.
Nitration of N-methyl-15-aza-des-N-morphinan with fuming nitric acid in glacial acetic acid afforded two kinds of nitration products. The main product was found, through reduction and diazotization to the hydroxy compound, to be 3-hydroxy compound, proving that the nitro group had entered the 3-position.
Comparative examinations were made in relation to chemical structure on the fact that cyanoacetic acid hydrazide (CAH) shows almost the same antitubercular action as that of isonicotinic acid hydrazide (INAH). Based on the facts that CAH does not effect this action by secondary intramolecular cyclization to 3-amino-2-pyrazolin-5-one, similarity of the size of the CAH molecule and its basicity to INAH, antibacterial activity lies in the acid hydrazide group in both, and that CAH is ineffective to INAH-resistant strains, it was considered that the action mechanism of CAH is approximately the same as that of INAH and that CAH is a simplified form of INAH.
ω-Monohydrazide of acidic amino acids, obtained by the substitution of the ω-acid amide group of asparagine and glutamine with an acid hydrazide group, were prepared and their antitubercular action was examined. These compounds showed antitubercular action in 30 γ/cc. concentration against H37RV strain. On the other hand, 5-oxo-2-pyrazolin-3-carboxylic acid, an intramolecular cyclization product of α-keto-ω-acid hydrazide, derived from aspartic acid hydrazide, showed activity similar to that of the original aspartic acid hydrazide. From such a fact, considerations were made on the possible presence of an amino acid oxidase system or direct transamination of enzyme system in the human-type tubercle bacilli.
Twelve kinds of 2-pyrazolin-5-one derivatives, substituted in the 1, 3, or 4 position were synthesized and relationship between their structure and antitubercular action was examined. With the exception of 3-methyl derivatives, compounds substituted in the 1, 3, or 4 position were found to have generally weaker antibacterial action. The action of the 3-methyl derivative against human-type tubercle bacilli, H37RV strain, was equal to that of 2-pyrazolin-5-one and was effective up to a concentration of 3 γ/cc.
5-Oxo-2-pyrazoline derivatives substituted with acetamide and hydrazide groups at 3 or 4 position were prepared and their action against human-type tubercle bacilli H37Rv was tested, but no inhibitory action was observed, even in 100γ/cc. concentration, in Kirchner medium added with bovine serum.
The phenolic base of Coptis japonica had been said to be columbamine but detailed examination revealed it to be jatrorrhizine. A minute amount of another phenolic base (reduction product, m. p. 272°) was also obtained.
A multi-buffered paper chromatography was applied to the tertiary base in Stephania japonica MIERS and the base was fixed in a buffered zone of special pH value (Fig. 1, Nos. 1-7). The non-phenolic bases indicated 5.4 for insularine (No. 1), 4.4 for epistephanine (No. 2), 3.2-3.0 for stephanine (No. 3), and 2.8 for protostephanine (No. 4), hasubanonine (No. 5) passed through the buffered zone of pH 2.0, and phenolic bases indicated 5.2 for hypoepistephanine (No. 6) and 3.0 for homostephanoline (No. 7). This process was utilized for the separation of a large amount of bases: the resinous residue, obtained after through separation of crystalline bases, was dissolved in chloroform and the solution was shaken with MacIlvaine buffer (double strength), corresponding to the fixed zone on buffered paper (Fig. 1, Nos. 8-11) to transit the bases into the buffer solution, The buffer solution was treated as usual and free bases were obtained. The residue from non-phenolic bases afforded epistephanine, protostephanine, and stephanine, the latter failing to be isolated by methods other than the present one. The residue from phenolic bases yielded by the same treatment, hypoepistephanine and homostephanoline.
Alkaloids in the root and stems of domestic Xanthoxylum piperitum (LINN.) DC. (Japanese name “Sansho”) and X. piperitum (LINN.) DC. var. inerme MAKINO (Japanese name “Asakura-zansho”), Rutaceae family, were examined. From the root and stem of X. piperitum and from the root of var. inerme, quaternary aporphinetype magnoflorine (I) was detected and it was also found that a small amount of a base forming a styphnate of m. p. 260° was contained in the root and stems of X. piperitum and in the stems of var. inerme, while the latter did not contain magnoflorine. The amount of tertiary bases contained in either of the plants was found to be extremely small. It is an interesting fact that while Australian Xanthoxylum and Fagara species contain menisperine (II), the domestic Xanthoxylum spp. contain magnoflorine (I).
Components of Aristolochia Kaempferi WILLD. (Japanese name “Ohba-umanosuzu-kusa”) and A. debilis SIEB. et ZUCC. (Japanese name “Umanosuzukusa”) were examined. As a basic component, an aporphine-type quaternary base, magnoflorine (II), and aristolochic acid (3, 4-methylenedioxy-8-methoxy-10-nitrophenanthrene-1-carboxylic acid) as an acid component, were isolated from both plants.
The presence of sedoheptulose was examined by paper chromatography in 9 kinds of Sedum genus, 3 kinds of Bryophyllum genus, and 1 Cotyledon genus of the Crassulaceae family and the results obtained are given in Table I.
It was found that several kinds of alkaloid were contained in the trunk bark of Neolitsea sericea (BLUME) KOIDZ., of domestic Lauraceae family, From its phenolic bases, an aporphine-type tertiary base, boldine, was obtained and this was derived to d-glaucine by the application of diazomethane.
Effect of hydrogen bond on the near-ultraviolet spectrum of antipyrine was quantitatively examine in a three-componental system with ethanol in heptane as a solvent. It was thereby learned that a distinct hydrogen-bond formation existed between antipyrine and ethanol at 1:1 ratio, the value of equilibrium constant, K, being 5.42. The absorption band at 280.5mμ undergoes a blue shift of 713cm-1 by the formation of a hydrogen bond but the band at 239.5mμ undergoes red shift of 1020cm-1. Such experimental results were considered from structural point of antipyrine as a mesoionic compound. The band at 239.5mμ was thought to be chiefly due to the benzene ring in the antipyrine molecule.
Reaction of 1 mole of ethyl DL-S6- and -S8-monoacetyl-dihydro-α-lipoate with 1 mole of aniline and/or benzylamine was found to effect facile migration of the acetyl group to the amine in S8-acetyl compound but the migration was difficult in the case of S6-acetyl compound and aniline, showing clearly the difference between S6- and S8-acetyl/derivatives. This fact agrees well with that found with ethyl DL-diacetyl-dihydro-α-lipoate.