Dehydration rate of water, contained in minute amount of below 1/1000, in oils and fats and isotopic exchange between hydrogen and tritium were examined by the measurement of radioactivity of tritiated water (radioactivity, 2mc./ml.) with a liquid scintillation spectrometer. Dehydrated sesame oil, J.P., was mixed with tritiated water, this was suctioned through an aspirator at a definite temperature, and a sample was taken out at definite intervals to measure the radioactivity, using Packard Tricarb liquid scintillation spectrometer, Model 314. Correlation between I (c. p. m.) and t (hr.) was found to be expressed by the formula I=K/tn (where K is a constant). At 130°, n was 0.59 and at 220°, n was 0.85. Isotopic exchange reaction between hydrogen and tritium hardly took place in around 60 hours.
The function of oxygen in photosantoninic acid, C30H42O9, one of the ultraviolet irradiation products of santonin, was disclosed by the reaction shown in Chart 1 and by the examinations of infrared and NMR spectra. This photo-product has one mole of water of crystallization difficult to remove, and is a heptacyclic compound possessing two ketones, one ether likage, and a γ- monohydroxy-dicarboxylic acid grouping, as is expressed in the partial structure (XVII).
It was proved that photosantoninic acid has two five-membered ring ketones both of which conjugated with different cyclopropane and one of which has an α-methylene. The secondary hydroxy at a γ-position relative to one of the carboxyl groups was shown to be adjacent to a cyclopropane conjugated with a ketone. These conclusion are summarized by the partial structure (XXX) or (XXXI).
Photo-reaction of santonin to photosantonihic acid proceeds through the intermediate formation of lumisantonin (I), and the change of I to photosantoninic acid is considered to be a base-catalyzed dimerization process because the latter has the twofold composition (C30H40O8) to that of lumisantoninic acid (II), C15H20O4. Based on the conclusion from the preceding paper, a mechanism of dimerization of I shown in Chart I was proposed and the resulting structural formula (III) (conformational formula (IV)) was assigned to photosantoninic acid. This mechanism requires 6β (H) configuration of I, and 6-epimer of I should not give a dimerization product. Therefore, three isomers of I, i.e, lumi-11α(H)-santonin (VI), lumi-6-epi-santonin (VII), and lumi-6-epi-11α(H)-santonin (VIII), were prepared and their dimerization was attempted. In accord with the above assumption, VI formed a dimer of m. p. 233° named metaphotosantoninic acid, while the others remained unchanged. The reaction sequence shown in Chart 2, carried out to prove the relative positions of the two ketone groups, also supported the formula (III).
Examinations were made on tertiai y bases contained in the tuber of Corydalis ambigua CHAM, et SCHLECHT, var. amurensis MAXIM. (Japanese name “Ezo-engosaku”), growing wild in the Hokkaido and northeastern parts of Japan, and one hind of phenolic base and four kinds of nonphenolic bases were isolated. The phenolic base was assumed to be d-corybulbine. Two of the four nonphenolic bases were identified as d-corydaline and protopine. One of the remaining two was found to be a tetrahydroberberine-type base of m. p. 144-146° and the other was a base of m. p. 194-195°. There is a fairly great difference in the bases contained in this plant and Corydalis ambigua CHAM. et SCHLECHT growing in China.
In order to introduce formylmethylene group into carbonyl compounds, formylmethylenetriphenylphosphorane (Wittig reagent) was newly synthesized from triphenylphosphine. Alternative and stepwise condensation of the newly synthesized formylmethylenetriphenylphosphine with nitro-2-thiophenecarboxaladehyde, 5-nitro-2-furaldehyde, and p-nitrobenzaldehyde afforded five kinds of corresponding polyene aldehydes. Attempt was made to synthesize 5-nitro-2-thiophenepropylaldehyde from α-bromo-5-nitro-2-thiopheneacrolein by dehydrobromination but the objective propiolaldehyde was not obtained and only α-methoxy-5-nitro-2-thiopheneacrolein was formed.
State of binding between five kinds of sulf anilamides used widely at present, sulfisomidine, sulfathiazole, sulfaphenazole, sulfadimethoxine, and sulfisoxazole, with crystalline bovine serum albumin was examined by the equilibrium dialysis method. In a phosphate buffer solution of pH 4.9-8.0, the number of binding sites on the bovine serum albumin is constant irrespective of the kind of sulfanilamide or pH value. Strength of this bond is fairly different with the kind of sulfanilamides, which can be divided into two groups by the strength of binding; one group consisting of sulfaphenazole, sulfadimethoxine, and sulfisoxazole which have a maximum value in this pH range, and another group of sulfisomidine and sulfathiazole which do not have the maximum value and the value increases with increasing pH. It was, however, clarified that the strength of binding decreased even in sulfathiazole and sulfisomidine with further increase in pH value.
A solution of calcium pyruvate isoniazone dissolved in water shows increase of electric conductivity with passage of time and the value reaches an equilibrium after a long time. This phenomenon was found to be due to hydrolysis of this isoniazone into isonicotinic acid hydrazide and calcium pyruvate, and confirned by the measurement of electric conductivity and absorption spectrum. This hydrolysis is a first-order reaction and the velocity constant at 25° becomes approximately equal by these two methods of measurement (1.450×10-3). Equivalent conductivity of calcium pyruvate isoniazone was calculated from specific conductivity of its aqueous solution. It was also found that there is no effect of neutral salts in this hydrolysis and that the activation energy of this hydrolysis is ca. 11 kcal./mole.
In order to find the effect of pH on the hydrolytic reaction of INAH derivatives (isoniazones of calcium pyruvate, sodium pyruvate, sodium glucuronate, glucose, and sodium o-phthalaldehydate), hydrolysis rate of these derivatives at 25° and 36° was measured in buffer solutions of pH 2.08 to 7.02. It was found that a relation of k=k0⋅[H3O+] exists between the rate constant k and [H3O+], and the values of k, k0, half-life t1/2, and equilibrium constant K were determined. A close correlation was found to exist between this hydrolytic reaction and pharmaceutical effect.
Seeds of Australian solanaceae plant, Duboisia myoporoides R. BR., containinig a large amount of tropine-type alkaloids, were imported and examinations were made on the growth in Japan, determination of components such as hyoscyamine and scopolamine, antispasmodic effect, and toxicity. Fresh seeds imported from Australia in 1961 showed germination rate of 26.5% in an average, at room temperature. Open-air growth of seedlings was generally good in southern Izu peninsula and the growth seemed to be especially good in shaded place, protected from wind and not too dry. These results showed that the plant will be able to bear the winter weather in Japan. Amount of total alkaloid was 1.13% in winter and 1.53% in summer, while that of hyoscyamine was 11% and 15%, and that of scopolamine was 25% and 22%, respectively in winter and summer. Antispasmodic effect of the extract was due mainly to the alkaloids contained in it and the effect of the alkaloid was represented almost completely by the above two alkaloids. Toxicity was very strong and the acute toxicity (LD50) of the extract was of the same order of atropine while that of total crude alkaloid was about 1/10 of it. These experimental results showed that the cultivation of Duboisia myoporoides is possible and it would be the best of cultivate an individual with high content of total alkaloid and of hyoscyamine.
Securinine, the alkaloid from Securinega suffruticosa (PALL.) REHD., and virosecurinine, the alkaloid from Securinega virosa PAX. et HOFFM. were found to be the optical antipodes by comparison of their ORD curves as well as other physical properties of the free bases and their derivatives.
The crude drug Chinese Byaku-shi, the dried root of Angelica dahurica BENTH. et HOOK. var. pai-chi KIMURA, HATA et YEN (Umbelliferae), afforded five kinds of furocoumarins, oxypeucedanin (I), isoimperatorin (II), imperatorin (III), phellopterin (IV), and byak-angelicin (V), and Jananese Byaku-shi, the dried root of A. dahurica BENTH. et HOOK. var. dahurica, afforded I, III, and IV in addition to V and byak-angelicol (VI), which had been found first by Noguchi from the same drug, by the process shown in Chart 1. In the present experiments, it was found that through column chromatography over silicic acid (Mallinckrodt) VI was converted into V by the opening of the epoxide ring in the side chain. By thin-layer chromatography, a spot identified as VI was detected from the ether extract of Chinese Byaku-shi but V was not detected, contrary to the result mentioned above. Therefore, it is considered that almost all of V isolated from Chinese Byaku-shi had originally been contained in the drug in the form of VI. The present result seems to support the proposal of Kimura and others assigning a local variety of A. dahurica BENTH. et HOOK. for the original plant of Chinese Byaku-shi.
The coumarin derivative constituents of Angelica formosana BOISS. and A. anomala LALL. (Umbelliferae), which have been assigned as the original plants of some kind of Byaku-shi (Chinese crude drug), were examined in order to compare with the coumarin-constituents of Japanese and Chinese Byaku-shi, dried roots of A. dahurica BENTH. et HOOK. var. dahurica and var. pai-chi KIMURA, HATA et YEN, respectively, reported in the preceding paper. Dried root of A. formosana afforded six kinds of furocoumarins, isoimperatorin (I), imperatorin (II), bergapten (III), phellopterin (IV), oxypeucedanin (V), and oxypeucedanin hydrate (VI), and two kinds of white crystalline substances, m. p. 159-160°, C11H10O5 (VII), and m. p. 187-188°, C14H14O4 (VIII), which were presumed from IR spectra as coumarin derivatives having hydroxyl group in their molecules. On the other hand, dried root of A. anomala afforded only two kinds of coumarins, III and umbelliferone (IX), in a small amount and pale-yellow viscid oil which did not crystallize up to the present. The present result seems to suggest that A. formosana is suitable to be regarded as an allied plant of Byaku-shi, but A. anomala should not be assigned for the drug.
Dried leaves of Cirsium martimum MAKINO was extracted with methanol, methanol was evaporated from the extract, and a glycoside was obtained as almost colorless needles, m.p. 243° (melting once at 173-179°) in 0.2% yield. Its molecular formula agrees with C23H24O11⋅2H2O; [α]D23 -70.8°. Hydrolysis with conc. hydrochloric acid afforded 1 mole each of a genin as slightly yellow needles, m.p. 257°, C15H8O4(OCH3)2, and glucose. The glycoside was named cirsimarin, and the genin, cirsimaritin. Demethylation of cirsimaritin gives scutellarein and its permethylation of gives scutellarein tetranethyl ether, so that cirsimaritin is scutellarein dimethyl ether. Decomposition of cirsimaritin with 3% hydrogen peroxide gives p-hydroxybenzoic acid, and that with 20% potassium hydroxide gives p-hydroxybenzoic acid and 4, 5-dimethoxyresorcinol. Methylation of cirsimaritin with diazomethane affords a trimethyl ether, m.p. 1.83°, which agreed with scutellarein-4′, 6, 7-trimethyl ether, m.p. 183°, UV λmaxEtOHmμ (log): 279 (4.24), 336 (4.42). Hydrolysis of permathylated cirsimarin with 10% sulfuric acid affords colorless needles, m.p. 228° C15H7O3(OCH3)3. Decomposition of this aglycone, m.p. 228°, with 3% hydrogen peroxide gives p-hydroxy-benzoic acid. It follows, therefore, that cirsimaritin is scutellarein 6, 7-dimethyl ether. The structure of cirsimarin is cirsimaritin 4′-glycoside.
Long white needles, m. p. 215° (decomp.), were obtained in 0.07% yield from the fresh leaves of Salix bakko KIMURA. This substance corresponds to the molecular formula of C27H30O15⋅2H2O, and colors greenish brown to ferric chloride, faint pink to orange with magnesium and hydrochloric acid, and faint pink with zinc and hydrochloric acid. Its hydrolysis with 35% hydrochloric acid produced one mole each of chrysoeriol, glucose, and arabinose. The position of sugar bonding was proved to be 7, since complete methylation of the glycoside with diazomethane and subsequent hydrolysis afforded luteolin 3′, 4′, 5-trimethyl ether, m. p. 281-282°. This glycoside is therefore chrysoeriol 7-glucoparabinoside and was named bakkoside.
The B-homo-ketone formed by the Tiffeneau reaction of cholestan-6-one and -7-one is the same and the fact proved that the structure of this B-homo-ketone is 5α, 8β-7-one. In order to examine the physiological disturbances and therapeutic effect on climactic disturbances based on this method, B-homo-5α-pregnane-3α, 20α-diol was synthesized.
Examinations were made on the constituents of crude drugs used frequently as aromatic stomachics, and seven kinds of essential oils including Japanese peppermint oil and fennel oil, for their yield and various properties, qualitative and quantitative determination of their componental constituents by gas chromatography, direct action on excised mouse intestine, antispasmodic action, their action on the rate of transfer of intestinal content. some considerations were then made on the acceleration of gastrointestinal functions and gas elimination, which are considered to be the main pharmacological effect of these crude drugs. Polyethylene glycol 6000 was used as the stationary phase in gas chromatography and a fairly good result was obtained in the analysis of essential oil components. These results were highly similar to the data published to date in literature. These essential oils and their components generally showed antispasmodic action. They did not show any action of accelerating the transfer of intestinal content but rather suppressed such a movement in some cases. The effect of essential oils on human body is more likely to be thought secondary effect via the olfactory and taste senses.
Action on blood pressure, pharmacological actions in general, acute toxicity were comparatively examined in guanethidine (I) and the compound formed by changing its eight-membered ring to a seven-membered ring, [2-(hexahydro-1-azepinyl)ethyl]guanidine sulfate (II). A lasting hypotensive action on rabbits and albino rats was seen in I but it raised blood pressure of cats. II showed lasting depression of blood pressure in rabbits and cats and temporary depression in rats but raised the blood pressure of dogs. Hypotensive action of II in rabbits was not inhibited by hexamethonium or atropine but was inhibited by reserpine. Hypertensive action of adrenaline and noradrenaline in rabbits, cats, and dogs is augmented by II. II showed increased amplitude of excised toad heart in a low concentration but showed no marked action on excised vascular muscle, peripheral blood vessel, skeletal muscle, and excised intestine. LD50 of II in mice was 97.4mg./ kg. by intravenous injection, 887 (776-1088)mg./kg. by subcutaneous injection, and 2000-3000mg./kg. by oral administration, the toxicity being not so strong.
Extract of the fruit of Catalpa ovata G. DON was fractionated in order to find the active principle in the fruit which is used as a diuretic. From a fraction found active by pharmacological test, catalposide and another glucoside, m. p. 199-202°, which was found to be des-p-hydroxybenzylcatalposide were isolated by chromatography. Presence of these constituents in another active fraction was also detected by paper partition chromatography. According to the pharmacological test, these glucosides, are regarded to play important roles as the active principles. β-Sitosterol was isolated from the unsaponifiable matter of seed oil. Other constituents, including seed oil, wax, were also investigated and a flavonoid, p-hydroxybenzoic acid, and citric acid were obtained.
Govindachari assumed the formula (II) for the quaternary iodide, C22H24O4N2I2, a product obtained by oxidation with manganese dioxide-sulfuric acid of tiliacorine, the alkaloid of Tiliacora racemosa COLEBR., an Indian plant of Menispermaceae family. In order to examine the structure of tiliacorine, for which Govindachari presumed formula (I), the quaternary iodide (II) was synthesized from 2, 7-bis(2-aminoethyl)-4, 9-dimethoxy-dibenzo-p-dioxin (III) by the route shown in Chart 1 and 2, 9-dimethyl-6, 13-dimethoxy-1, 2, 3, 4, 8, 9, 10, 11-octahydro-p-dioxino[2, 3-h:5, 6-h′]diisoquinoline, having a structure formed by bimolecular condensation of 2-methyl-1, 2, 3, 4-tetrahydroisoquinoline skeleton, was synthesized.
In order to examine the electronic effect of sulfur atom on the neighboring group, reactions of some ω-hydroxyalkyl phenyl sulfides (general formula p-R-C6H4-S(CH2)nOH, R=CH3, H, Br, NO2; n=2, 3, 4, 5) with p-tosyl chloride were carried out. As shown in Table I, ω-chlorination occurred in the case of n=2; R=CH3, H, Br and n=4; R=CH3, H, while only ω-tosylation took place in the Other cases. Considering the electronic effect of para-substituents, this result implies that the sequential order of reactivity of the hydroxyl group would be β>δ>γ≅ε in respect to sulfur atom and it can be explained from the following interpretation. In the case of n=2, the sulfur atom would exert an inductive effect along the carbon chain to produce an intermediate cation with ease and p-tosyl chloride would attack as a nucleophilic reagent. On the other hand, in the case of n=4, the inductive effect would not exert any appreciable influence through the four carbon atoms, but ω-carbon would be favorably located to accept the released lone-pair electrons of sulfur across the space, which would facilitate the formation of an intermediate cation. It was also found that vacuum distillation of some ω-(p-tosyloxy)alkyl phenyl sulfides gave α, ω-bis (phenylthio) alkane as the distillate and α, ω-bis (p-tosyloxy) alkane as the residue. This result indicates the formation of sulfonium salt and its subsequent thermal decomposition, as illustrated in Chart 2.
The inhibitory effect of liver catalase by 3-amino-s-triazole (3-AT) in vivo was observed commonly in mouse, rat, guinea pig and rabbit. The normal value of liver catalase activity is shown in the following order; guinea pig>rat≅mouse>rabbit and the inhibitory effect by 3-AT was found to be rabbit>mouse>rat>guinea pig, which was considered to be approximately in a reverse order. Then, the related compounds to 3-AT in the chemical structure, such as 3-ureido-s-triazole (3-UT), 3, 5-diamino-s-triazole (3, 5-DAT), 4-amino-s-triazole hydrochloride (4-AT), Nitron, 5-amino-1, 2, 3, 4-tetrazole (5-ATT) and 2-amino-1, 3, 4-thiadiazole (2-ATD) were examined in its inhibitory effects. The strongest effect was found in the original compound, 3-AT, after which 3-UT and Nitron followed. Any inhibitory effect has not been observed in the rest of four compounds.
A simple evaporator for flasks under reduced pressure was designed which needed no rotating joint. It moves in a rotary oscillation at an angle of about 110°, and an outlet is connected with a condenser by a rubber tube. The angle of rotation can be increased by combining the oscillator with a pair of gears, but the simplest one may be useful for usual purpose of evaporation with minimum foaming or bumping in the flasks when it is oscillated about 150 times per minute.
When a water solution of N-acetylglucosamine (GlcNAc), N-acetylgalactosamine (GalNAc) and N-acetylmannosamine (ManNAc) is passed through the resin columns of Amberlite IR-120 (H+ form) and IR-4B (OH- form), followed by the condensation of each eluate under reduced pressure, the epimerization was observed in a part of the eluates. GlcNAc produced two kinds of N-acetylaminosugars unidentical with ManNAc and GalNAc, GalNAc produced a kind of unidentical N-acetylaminosugar and also ManNAc gave two kinds of N-acetylaminosugars unidentical with GlcNAc and GalNAc. However, the epimelized N-acetylhexosamines and N-acetylaminosugars were found to be so small in amount that any disturbance was not observed on the paper partition chromatogram of the original N-acetylhexosamines for the identification.
The reaction products formed by the application of a mixture of conc. sulfuric acid and nitric acid (sp. gr. 1.42) on several kinds of dibenzo-p-dioxin derivatives were examined. 1, 6-Dibromodibenzo-p-dioxin (V) remained almost inert to the action of sulfuric and nitric acid mixture for a long time but dibenzo-p-dioxin (I) and 2, 7-dimethyldibenzo-p-dioxin (III) easily formed the corresponding dinitro compounds, 2, 7-dinitrodibenzo-p-dioxin (II) and 2, 7-dimethyl-3, 8-dinitrodibenzo-p-dioxin (IV).