Five triterpene glycosides, named α-, β1-, β2-, β3-, and β4-fatsin, were obtained from the leaves of Fatsia japonica DECNE et PLANCH. α-Fatsin (mp 251-253°(decomp.), [α]25D+32.2°), β1-fatsin (mp 210-215°(decomp.), [α]25D+17.6°), and β2-Fatsin (mp 239-241°(decomp.), [α]25D+16.6°) were characterized as hederagenin 3-O-β-D-glucopyranosyl (1-4)-α-L-arabinopyranoside (I), 3-O-β-D-glucopyranosyloleanolic acid (V), and hederagenin 3-O-α-L-arabinopyranoside (IX), respectively. Hydrolysis of β3-Fatsin afforded oleanolic acid and glucose, and the sapogenin of β4-Fatsin was proved to be 3β, 16α-dihydroxyolean-12-en-28-oic acid (echinocystic acid).
A molar equivalent of pyridine N-oxide was added to 6-methyl-4(3H)-pyrimidinone (I) and 4(3H)-quinazolinone (V), and heated with addition of platinum-added palladium carbon, catalyst(Pd/Pt-C) by which products were compounds (III, IV, VI, and VII) formed by reaction at respective 2-position. III, IV, and VII are new compounds hitherto unreported in literature. These were derived to IIIa, IVb, and VIIb to determine their structure. The same reaction of I and V with 0.5 mole of pyridine results in the formation of III and VI in approximately double the yield of the former. It seems possible to synthesize various nonsymmetric compounds by the use of a different N-oxide.
1-Cyano-4-imino-2-methylthio-4H-pyrido [1, 2-c] pyrimidine (III) was obtained from 2-pyridineacetonitrile and dimethyl cyanamidedithiocarboxylate, and the reactivity of III was examined. The reaction of III with an amine, acetic anhydride, and methyl iodide gave the corresponding products (X, IV, and V), while III or 3-cyano-4-imino-4H-pyrido [1, 2-α] pyrimidine (IX) and dimethyl acetylenedicarboxylate afforded 1, 9-diazacycl-[3, 3, 3] azine derivative (XII) and 1, 4-diazacycl [3, 3, 3] azine derivative (XIII). The reaction of XII or XIII with an amine resulted in the formation of corresponding products (XIV, XV).
N-Alkyl-1-(3, 4-dimethoxyphenyl)-1-(2-, 3-, or 4-substituted phenyl) methylamines (XI-XLVI) were synthesized from 3, 4-dimethoxybenzaldehyde and 2-, 3-, or 4-substituted bromobenzene using the Grignard reaction and halogenation, followed by amination with various alkylamines. Of the compounds synthesized and tested, XVIII was found to possess a weak antitussive activity, and XVI and XIX showed a marked local anesthesic activity.
N-Alkyl-1-(2, 3-dimethoxyphenyl)-1-(2-, 3-, or 4-substituted phenyl) methylamines (XI-XLI) were synthesized from 2, 3-dimethoxybenzaldehyde (II) using the Grignard reaction, followed by halogenation and amination with various alkylamines. Of the compounds synthesized and tested, XXI, XXII, and XXVII were found to possess a weak antitussive activity, and XI, to XV, XVII, XXI, XXII, and XXVII showed a marked local anesthesic activity.
Some derivatives of 2-chlorophenyl-2-dimethoxyphenylglycine were synthesized. N-Acetyl-2-(2-chlorophenyl)-2-(3, 4-dimethoxyphenyl) glycine (VIII) was cyclized to 4, 4-diphenyloxazolone (IX) on heating with Ac2O. Cleavage of IX with alkylamines gave the corresponding amines (XI and XII). IX was converted into imidazolones (XIII) by heating with alkylamines in benzene. Ammonolysis of IX with NH3 gave N-acetylglycinamide (XIV), which cyclized to imidazolone (XV) in the presence of NaOEt.
(S)-(+)-8-methyl-5, 6, 7, 8-tetrahydroindane-1, 5-dione (VI) and (S)-(+)-8a-methyl-1, 2, -3, 5, 6, 7, 8, 8a-octahydro-3, 8-naphthalenedione (VII) were asymmetrically synthesized from 2-methylcyclopentane-1, 3-dione (IV) and 2-methylcyclohexane-1, 3-dione (V) using L-proline derivatives. Asymmetric synthesis of VI or VII from 2-methyl-2- (3-oxobutyl) cyclorentane-1, 3-dione (X) or 2-methyl-2- (3-oxobutyl) cyclohexane-1, 3-dione (XI) was also carried out under various reaction conditions. The reaction path of this asymmetric synthesis is discussed.
Optically active (-)-bisdehydroestrone methyl ether (V) was synthesized by two methods ; one is synthesis of (-)-V from optically pure S-(-)-II, and the other is the asymmetric cyclization of triketone (X) with L-proline pyrrolidide enamine method.
A new type agglutinin was separated from eggs of Rana nigromaculata nigromaculata HALLOWELL. This agglutinin was named "Egg agglutinin". This agglutinin has a property of basic protein and ability to agglutinate AH-109A cells, but this agglutination can be inhibited specifically by Gangioside and Globoside-1 isolated from human red blood cells of Type A. This agglutination reaction was not interfered after the digestion of AH-109A cells with neuraminidase, but was completely inhibited after trypsin treatment.
Carbon-13 nuclear magnetic resonance spectra of six barbituric acid derivatives were measured. In addition, extended Huckel calculations were carried out on 18 barbituric acid derivatives, and the following examinations were made. (1) Shielding constants, based on Pople's theory, were calculated. (2) Additive equations on atomic populations of sp3-hybridized carbons were calculated. (3) Monoionized state of 5-ethylbarbituric acid was estimated from the standpoint of total energy.
It was found that a seties of α-hydroxyaldehydes produce a fluorescence by reaction with adenine in acetic acid solution, and its intensity decreases with increasing number of carbon atoms of the aldehydes. This reaction is specific for α-hydroxyaldehydes and not for simple aldehydes. It is also specific for adenine derivatives in which the primary amino group in position 6 is not substituted, and other nucleic acid bases do not give fluorescence by this method. This fluorogenic reaction is due to the formation of a double bond of Schiff bases, and it was assumed that oligomers of glyoxal exhibit the fluorescence by reaction with adenine after fission of its ring structure, resulting in an aldehyde group.
The membranes used were the gelatin membrane and the mixed membrane composed of gelatin and chondroitin sulfate-C (chs), and these were cross-linked by formaldehyde in order to make them insoluble. The weight ratio of chs to gelatin in the mixed membrane was 0.342. pH dependence of the swelling was measured in the range of pH 2.8-8.1 using McIlvaine buffer solution. The result obtained from the gelatin membrane was different from that of the mixed membrane. In the former, the degree of swelling increased with decreasing pH, while in the latter, it became maximum at the vicinity of pH 4.9 which is the isoionic point pH of gelatin. These results satisfied the following equation of the swelling rate, e.g., d(WW/WM)/dt=k(WW/WM), where (WW/WM) is the weight of water per gram of membrane constituent, and k is the constant of the swelling rate. The value of k, in various pH solutions was thus obtained. The swelling of these membranes was anisotropic, e.g., the thickness increased but the radius hardly increased. In the case of the mixed membrane, the anisotropy of swelling was so pronounced that the radius decreased in the acidic region. From these results, orientation of the molecules in the membrane was discussed.
The inhibitory effect of various substances on an immobilized derivative of carboxypeptidase CN (AEC-CPase CN), which was prepared by conjugating the enzyme to AE-cellulose through glutaraldehyde, was examined. Like the native enzyme, AEC-CPase CN was inactivated by methanol and this inactivation became more pronounced as the concentration of methanol increased. AEC-CPase CN was slightly more stable to the attack of Pronase than the native enzyme. Enzymic activity of AEC-CPase CN was completely loss by the addition of low concentrations of sodium dodecyl sulfate and cetyltrimethylammonium bromide, whereas Triton X-100 and Tween 80 caused only a partial loss of the enzymic activity even at high concentrations. Of six phenylalanine analogs tested, β-phenylpropionic acid, a competitive inhibitor, showed the most inhibitory effect. Pretreatment of AEC-CPase CN with 25 mM β-phenylpropionic acid almost completely protected the enzyme from inactivation by diisopropylfluorophosphate, while the attack of p-bromophenacyl bromide and HgCl2. was only partially blocked by the same pretreatment. The denaturating effect of methanol, heat, and pH was not prevented by the pretreatment with β-phenylpropionic acid.
Isoquinoline 2-oxide (I) reacts with cyanogen bromide in ethanol to give ethyl N-(1- and 4-isoquinolyl) carbamates (II and III) accompanied with some by-products (IV, V, or V' and VI). The amount of II is always larger than that of III and the best yield (42.6%) of II is obtained from the reaction at room temperature for 5 days. 1-Chloroisoquinoline 2-oxide (VII) gives no carbamate and 4-bromoisoquinoline 2-oxides (VIII) afforded 2-isoquinolylcarbamate (IX) in a small yield. While 1-hydroxyisoquinoline 2-oxide (XI) gives only the 4-bromo derivative (XII) by the usual procedure, 4-ethoxycarbonylamino-isocarbostyril (XIII) is produced in 22% yield when sodium acetate is added to the reactants. Although pyridine 1-oxides generally resist this reaction, only 4-hydroxypyridine 1-oxide is highly reactive and ethyl N-(4-hydroxy-3-pyridyl) carbamate hydrobromide (XVIII·HBr) is obtained in 57 % yield. Reactions of 1-aminoisoquinoline 2-oxide (XIV) and 2-aminopyridine 1-oxide (XX) are quite similar to that of 2-aminoquinoline 1-oxide, and the corresponding oxadiazolo derivatives (XV and XXI) are produced without the participation of ethanol.
The constituents of the fresh aerial parts of Dioscorea tenuipes FRANCH. et SAVAT. (Dioscoreaceae) collected at the Ozeki pass, Shiga, in September and October, were extracted and separated with special care not to cause secondary alteration. Seven new substances, tentatively designated G1, G2, G3, G4, S1, S4, and SE1, were isolated together with five known compounds ; phytosterols, taraxerol, neotokorogenin, diotigenin, and tenuipegenin. G2, mp 214-215°, [α]D -48.6°, and G3, mp 211-212°, [α]D -41.2°, were characterized as diotigenin 2-acetate (I) and its 2, 4-diacetate (X), respectively, which represent the first spirostanol (steroidal sapogenin) acetates found in nature.
The structures of two steroidal substances, G1 and SE1, isolated from the fresh aerial parts of Dioscorea tenuipes FRANCH. et SAVAT. (Dioscoreaceae), were investigated. G1 (mp 221-222°, [α]D -10.0°, C29H46O6) and SE1 (mp 172-174°(decomp.), [α]D +134.4°, C36H60O12·2H2O, Ehrlich reaction +) were characterized as diotigenin 4-acetate (I) and the corresponding furostanol 26-O-glucoside, 4β-acetoxy-22-methoxy-5β-furostane-2β, 3α, 26-triol 26-O-β-D-glucopyranoside (VIII), respectively. VIII is the first reported furostanol glucoside corresponding to spirostanol acetate and is most likely to have been formed secondarily from the 22-hydroxy analog (IX), mp 165-166°(decomp.). IX is regarded as the prototype compound of I. The mass spectrum of VIII and compositions of the major fragment ions are presented.
From the ether-soluble fraction of the water extracts of the roots of Scutellaria baicalensis GEORGI (Japanese name "Wogon"), two new flavone derivertives, named skullcapflavone-I and -II were isolated along with β-sitosterol, campesterol, stigmasterol, wogonin, baicalein, and oroxylin-A (5, 7-dihydroxy-6-methoxyflavone). The chemical structures of skullcapflavone-I (V) and -II (VI) were established as 2', 5-dihydroxy-6, 8-dimethoxyflavone and 2', 5-dihydroxy-6, 6', 7, 8-tetramethoxyflavone, respectively, by chemical and physical analyses.
Gas chromatographic method for the determination of bile acids in bear gall was developed. Neutral lipid-free methanolic extract from bear gall was treated with 2.5 N sodium hydroxide in an autoclave. Bile acids thus obtained were esterified by diazomethane and then their hydroxyl groups converted to trimethylsilyl ethers in anhydrous pyridine. Gas chromatography using 0.5% cyclohexanedimethanol succinate column resulted in the quantitative separation of cholanic acid derivatives. The calibration curve was constructed with the use of lithocholic acid as an internal standard.
An attempt was made to determine quantitatively the naphthaldehyde content in naphthoic acid by the use of oscillographic polarography. Use of a NH4OH -NH4Cl buffer solution as the supporting electrolyte enables quantitative determination of naphthaldehyde to within 0.01% of accuracy. However, when 1-naphthaldehyde and 2-naphthaldehyde are present, together the very proximity of their peak potentials, being -1.29 V. vs. SCE and -1.32 V. vs. SCE, respectively, makes it difficult to determine both quantitatively at the same time. Their qualitative determination, on the other hand, is quite feasible.
3, 4, 5, 6-Tetrachloro-3', 4', 5', 6'-tetrahydroxyfluoran methyl ester (I) formed a blue complex with silver (I) in the presence of excess 1, 10-phenpthroline (II) and polyvinylpyrolidone (PVP) in a neutral solution. The color was applied for spectrophotometric determination of Ag [I], CN-, I-, and S2O32-. The absorbance of I-II-Ag [I] solution was measured at 630 on or 650 nm against I-II solution for the determination of Ag [I]. The difference in the absorbance of I-II-Ag [I] solution and of I-, CN-, or S2O32- solution was measured at 630 nm against water for the determination of CN-, I-, and S2O32-. Linear calibration curves were obtained for 0 to 2.8 μg of Ag [I], 0 to 3.0 μg of CN-, 1.0 to 7.5 μg of I-, and 0 to 6.5 μg of S2O32- using I and II.
In order to find optimal cultivation conditions for Stevia rebaudiana Bertoni, which contains a diterpene-type sweet substance, stevioside, several experiments have been undertaken since 1971. The seasonal variation in the content of stevioside and its content in the samples cultivated at 5 different places in Japan were examined. In addition, 28 samples divided by their morphological characteristics were examined for their stevioside productivity.