A new reaction was found by which 5-cholroorotic acid (III) was obtained via the intermediate (II) by the treatment of ethyl orotate (I) with pyridine and thionyl chloride, a nucleophilic reagent. Also, both 5-chlorouracil (V) and 5-chloromethylthiourail (VIII) were formed respectively by a similar reaction of uracil (IV) and methylthiouracil (VII). It is considered that this reaction is an electrophilic reaction with the complex of thionylpyridinium chloride, and that an electron releasing group at the 4-position tends to lower the yield of 5-chloro compound, while an electron-withdrawing group gives a higher yield of 5-chloro compound. The reaction of II or III with amine bases gave their substituted compounds, in which substituents at the 4- and 5-positions did not lie coplanar to the pyrimidine ring. Furthermore, it was found that the treatment of guanidine derivative (XIV) with cone. sulfuric acid afforded the amino form (XX) of pyrimido[5, 4-d]pyrimidine derivative.

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A parotin-like substance, fraction 100-1, was separatedin a highly purified form from mixed human saliva by mild procedures which comprised acetone fractionation, chromatography on ECTEOLA- and CM-cellulose, and Sephadex G-100 gel filtration. This substance produced hypocalcemia and leucocytosis preceded by transient leucopenia in rabbits with a single intravenous injection of 0.1 mg/kg. It also elicited depression of the serum acid phosphatase activity in a dose of 0.15 mg/kg. It has a molecular weight of 350000 and contains 21.6% galactose and 19.6% glucosamine. The relation of this substance to other patotin-like substances from human saliva is discussed.

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Saliva-parotin-A was submitted to chemical cleavage with N-bromosuccinimide, performic acid, thioglycollic acid, or cyanogen bromide. The cleavage products were fractionated by Sephadex G-75 gel filtration, and the resultant fractions were assayed for hypocalcemic and leukocytosis-promoting activities. In most cases, the first peak possessed the biological activity. An active fraction (CNBr-I) from the cyanogen bromide cleavage product was tested for stability against heat, acid, and alkali, and for susceptability to chemical modifications with a variety of reagents. It was inferred that a histidine residue, or residues, might participate in the biological activities of CNBr-I. This fragment was a glycoprotein which contained 61.75% amino acid residues, 30.60% hexose, and 2.16% glucosamine. Its molecular weight was calculated to be 20000 by the Archibald method.

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The relationship between the serum citrate-lowering and circulating leukocytes-increasing actions of Saliva-parotin-A (SPA) in rabbits was investigated concerning the effective dose and duration of the action. Under a control feeding, the serum citrate concentration decreased significantly at 24 hr after intravenous injection of SPA and recovered progressively to the control level after 48 to 72 hr. Under the same condition, the circulating leukocyte count increased significantly at 24 and 48 hr, and decreased to the control level at 72 hr. From disagreement between the two actions in the effective dose and the duration of the action, it may be concluded that the citrate-lowering and leukocytes-increasing activities of SPA differ from each other in their mechanisms of action, and that there is no direct relationship between them.

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Piperidinium iodide and some alkyl-substituted piperidinium iodides were treated with lithium-diphenyl adduct in anhydrous tetrahydrofuran in order to study the possible reductive cleavage reaction of the piperidine ring. 1, 1-Dimethyl-2-nonylpiperidinium iodide (I₄) was found to undego reductive ring cleavage and yield 5-(N, N-dimethylamino)-tetradecane (2%) and N, N-dimethyltetradecylamine (36%), together with N-methyl-2-nonylpiperidine (21%) and 1-dimethylamino-5-tetradecene (36%). In the reaction of I₄ with lithium aromatic hydrocarbons, the ease of reductive ring cleavage increased with decrease in the electron affinity of the hydrocarbons.

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The effects of calcitonin-like factor and calcitonin were compared in rats treated under various conditions. The hypocalcemic effect of this factor was stronger in rats fed with low-calcium diet than those fed with normal diet, and its effect did not alter by prior parathyroidectomy, though it was abolished by thyroparathyroidectomy. Calcitonin-like factor had a weak hypophosphatemic effect since it did not decrease serum phosphate under various conditions, especially this factor did not diminish hyperphosphatemia induced by parathyroidectomy, and the addition of phosphate did not increase the hypocalcemia effect of this factor. It may be suggested that this factor is different from calcitonin in hypophosphatemic action and this factor did not act by releasing calcitonin from the thyroid gland. This factor inhibited the amount of total hydroxyproline released from metaphysis into the medium, so that calcitonin-like factor seems to give the hypocalcemic effect primarily through the inhibition of bone resorption.

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In continuation of the preceding work, content of free amino acids in 28 kinds of domestic lichens was determined by the amino acid autoanalyzer (Hitachi Model KLA-2) and total nitrogen by the semimicro-Kjeldahl method. The lichens examined in the present series contained from 11 to 17 kinds of amino acid. In a total of 103 kinds of lichens examined in the previous (75) and present (28) series, content of alanine and glutamic acid was generally larger than that of other amino acids. The lichens that contained more than 100 μmoles of amino acid per 100 g of the lichem were ca.40% of the previous 75 kinds and ca. 61% in the present 28 kinds in the case of alanine, and ca.17.3% of the previous 75 kinds and co.25% of the present 28 kinds in the case of glutamic acid. As in the previous work, taurine was found in Cetraria endocrocea (ASAHINA) SATO, Cladonia bellidiiflora (ACH.) SCHAER, Cladonia granulans VAIN., and Ramalina crassa (DEL. ex NYL.) MOT. The total nitrogen of 1.88-2.70% in 3 kinds of Lobaria in Stictaceae and of 2.64% in one kind of Pannaria in Pannariaceae was 1argeer than that in other lichens. A large amount of total nitrogen was also found in 4 kinds of Lobaria and 1 kind of Pannaria in the previous work. In most of the 28 kinds of lichens examined in the present work, some Ninhydrin-positive substances other than amino acids constituting the protein were detected but no detailed examination was made on them.

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In an effort to increase the sensitivity and specificity in the assay of salivary galnd hormones, a radioimmunoassay of parotin was attempted. The radioimmunoassy was carried out by incubation of a mixture of anti-parotin serum, ¹³¹I-parotin, and parotin sample, followed by separation of antibody-bound ¹³¹I-parotin (B) from free ¹³¹I-parotin (F) by paper electrophoresis with hydrodynamic flow. The radioactivity of B and F was counted, and the amount of parotin was determined by the ratio of B/F. The sensitivity was 2 mμg. Although S-parotin showed cross reaction with parotin to some extent, negative cross reactions were found for bovine serum γ-globulin and for other substances, such as the hypocalcemic substance from B. subtilis K and the MSH like substance from human which are active in decreasing calcium. This radioimmunoassay method can be used for determination of parotin present in the parotid gland, submaxillary gland, serum and urine of guinea pigs, and in urine and saliva of human, for which the previous methods of bioassay cannot be used directly. The present method confirmed that the content of the hormone in serum and urine of guinea pigs decreases suddenly after sialoadenectomy. The partial agreement between the results of the immunoassay of parotin and the bioassay is understandable from the fact that the anti-parotin serum neutralizes the hypocalcemic activity but not the leucocyte activity.

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For the purpose of examining the color reaction of alcohol with vanadium (V) complex of oxine analog, 5- or 7-methylthio-8-hydroxyquinoline derivatives (XIX to XXII) were synthesized by the Skraup method from methylthioacetylaminoaryl acetates (XIV, XVI-XVIII). The acetates (XIV, XVI-XVIII) were easily prepared in two steps (as shown in Chart 1) from nitrohydroxyaryldimethylsulfonium perchlorate (V-VIII) synthesized by nitration of hydroxyaryldimethylsulfonium perchlorate (I-IV) in perchloric acid. The addition of a slight excess of ethanol changed the toluene solutions of vanadium complexes of the novel compounds (XIX-XXII) from greenish black to brownish red. The absorption spectrum of the brownish red solution derived from XXI had a maximum (shoulder) at 485 nm, which was close to that obtained by the addition of ethanol to vanadium (V) complex of 5, 7-diiodo-8-hydroxyquinoline (Fig.2). In this color reaction, the hyperchromic effect of methylthio group in the quinoline ring of oxine is comparable to that of iodine group.

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The effect of saliva-parotin-A (SPA) on the serum and prostatic phosphatase activities was investigated in intact and parotidectomized rabbits. The enzyme activities were measured with p-nitrophenylphosphate as substrate for serum phosphatases and with β-glycerophosphate for prostatic acid phosphatase. A single injection of 100 μg/kg of SPA decreased significantly the serum acid phosphatase activity after 5.5 hr, but had no effect on the serum alkaline phosphatase activity. The prostatic acid phosphatase activity towards β-glycerophosphate decreased slightly, but the decrease was not statistically significant against the control due to deviation of the measured values. No difference was found in the serum acid and alkaline phosphatase activities between intact and parotidectomized rabbits. The serum acid phosphatase activity in immature rabbits was higher than that in adults and became lower as the body weight increased. A marked decrease in the serum acid phosphatase activity was observed when SPA was given to parotidectomized immature rabbits. These results suggest that the depression of the serum acid phosphatase activity by SPA may be attributed to the decrease in the release of the enzyme from bone cells, and/or that SPA may affect indirectly the acid phosphatase activity level in serum, into which the enzyme is released from leukocyte cells.

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As modification of β-(3-hydroxy-4-methoxyphenyl) ethylbenzene (I) having the essential part of sweet phyllodulcin molecule, β-(3-methoxy-4-hydroxyphenyl) ethylbenzene (II), 3-hydroxy-4-methoxyphenylmethylbenzene (III), 3-(3-hydroxy-4-methoxyphenyl) phthalide (IV), γ-(3-hydroxy-4-methoxyphenyl) propylbenzene (V), δ-(3-hydroxy-4-methoxyphenyl) butylbenzene (VI), and β-(3-hydroxy-4-methoxyphenyl) cyclohexylethane (VII) were synthesized. II, III, IV, and VI were tasteless, while V and VII revealed a moderate sweet taste. Consequently, it seemed that not only the position of the hydroxyl and methoxyl groups but the distance between the two phenyl groups in these molecules may be important factors for the appearance of sweet taste, and π-electrns of the phenyl group in I may not be necessary for interaction with the receptor site.

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Column chromatographic separation of the ether extract of the bulbs of Scilla japonica afforded four fractions, consisting of paraffins (Fr.I), esters (Fr.II), aldehydes (Fr.III), and alcohols (Fr.IV). The composition of these fractions was determined, qualitatively and quantitatively, by gas-liquid chromatography and mass spectrometry. The results are summarized in Table I and II.

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Chemical correlation of carboxyacetylquercinic acid (1a) and its congeners with lanosterol was examined and, taking into consideration of the circular dichroism (CD) of the first two the stereochemistry of these compounds was established.

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In order to obtain 2, 6-dicyano-7-ethoxycarbonyl-1, 5-dioxo-1, 2, 3, 5-tetrahydroindolizine as an intermediate for the synthesis of camptothecin, Michael condensation of methyl 3-cyano-4-ethoxycarbonyl-1, 2-dihydro-2-oxopyridine-6-carboxylate (V) with acrylonitrile was carried out but failed to afford the objective substance. A similar condensation of V with t-butyl acrylate also resulted in failure. Ethyl 5-cyano-4-ethoxycarbonyl-5-oxo1, 6-dihydro-2-pyridine glyoxylate (VIII) was synthesized by the reaction of 3-cyano-4-ethoxycarbonyl-1, 2-dihydro-2-oxopyridine-6-carboxylic acid chloride with ethyl t-butylmalonate. Although cyclization of VIII to ethyl 6-cyano-1, 2, 3, 5-tetrahydro-1, 3, 5-trioxo-7-indolizine carboxylate (IX) was unsuccessful, Friedlander reaction of VIII with 2-aminobenzaldehyde gave the expected 8-cyano-7-ethoxycarbonyl-9, 11-dihydro-9, 11-dioxoindolizino[1, 2-b]quinoline (X) in one step.

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Effects of Saliva-parotin-A (SPA) and various hormones on the calcium level and acid phosphatase activity in serum were investigated in intact rabbits. The serum calcium level decreased significantly by a single injection of SPA (100 μg/kg), ACTH (3.2 U/kg), or vasopressin (10 U/kg) but not one of thyroca1citonin (0.3 U/kg), insulin (0.4 IU/kg), or cortisone (30 mg/kg). The decrease in the serum acid phosphatase activity was statistically significant for SPA, but not for vasopressin and cortisone. Thyrocalcitonin, insulin, and ACTH had no effect on the serum acid phosphatase activity, The possibility that SPA affects indirectly the calcium level and acid phosphatase activity in serum is not excluded.

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From the volatile oil of the subterranean part of Asarum caulescens (Aristolochiaceae), linalool was isolated and identified. Two kinds of new sesquiterpene alcohols, germacra-1(10), 4, 7 (11)-trien-9 α-ol and 2-methyl-2-vinyl-3-isopropenyl-5-isopropylidenecyclohexanol, both C₁₅H₂₄O, were also isolated and their structures were determined from chemical and spectroscopic evidences. The volatile oil of A. europaeum ocuring in Europe is known to contain the aromatic compounds largely, and that of Asiasarum and Heterotropa occuring in the eastern Asia also contains the aromatic compunds mainly, but the volatile oil of A. caulescens does not contain them at all.

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Studies were made on the rapid dietary induction of hypercholestereolemia in rats, with a view to employing experimental conditions thereof in our first screening of orally active hypocholesterolemic agents. The experimental results confirmed that the sex differences had no influence, but the strain, age, body weight of the animals, and diet had a great effect on elevation of serum cholesterol levels. Marked hypercholesterolemia was induced when such strains of rats as showing a high sensitivity to exogenous cholesterol were fed a high fat diet containing 2% cholesterol and 1% cholic acid for a period of 3 days. The hypocholesterolemic activity of natural and synthetic substances was determined in the 3-day oral assay. The most active compound was diethylstilbestrol, followed by L-thyroxine, eritadenine, and β-estradiol. Nicotinic acid, clofibrate, and fradiomycin proved active when given in large doses.

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Reaction of diketene with aliphatic imido esters (Ia-c) affords 2-alkyl-2-ethoxy-6-methyl-3, 4-dihydro-2H-1, 3-oxazin-4-one (IIa-c), which is converted into 5-acetyl-2, 6-dialkyl-4(3H)-pyrimidone (IIIa-c), N-acylacetoacetamide (IVa-c), and 2-alkyl-6-methyl-4(3H)-pyrimidone (Va-c). On the other hand, aliphatic imido ester, which has an electron-attracting moiety such as ethyl cyanoacetimidate (XII) and ethyl 1-ethoxyformimidoyl acetate (VI), does not produce oxazin derivatives but gives 6-ethoxy-4-methyl-2-pyridone-3-carbonitri1e(XIII) and ethyl 2-ethoxy-4-hydroxy-6-methylpyridine-3-carboxylate (VII), which is considered to have an enamine structure. In the presence of a catalytic amount of acetic acid, VI yields 2-ethoxycarbonylmethylene-6-methyl-3, 4-dihydro-2H-1, 3-oxazin-4-one (XVII : R=CO₂Et). XVII is trasnformed to ethyl N-acetoacetylmalonamate (XVIII) and ethyl 2, 6-dihydoxy-4-methylpyridine-3-carboxylate (XIX).

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Oleanolic acid was incubated with Colletotrichum phomoides. Chromatography of the fermentation product led to the isolation of three metabolites which have been established as II, III and IV.

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The metabolites and metabolic pathway of a new anticonvulsant drug, sodium dipropylacetate (DPA), in rat were investigated using ¹⁴C-labelled DPA. Most of the metabolites in urine and bile was glucronide and free DPA was as little as one-seventh of the total metabolites. In feces, only free DPA was detected, and the possibility of enterohepatic circulation of DPA was presumed. A part of dosed DPA was excreted in expired air in the form of CO₂. This degradative reaction took place in liver mitochondria and required CoA and oxygen. It was stimulated by ATP and EDTA, and inhibited by various enzymic reaction inhibitors such as malonate, Antimycin-A, chloropromazine, PCMB, and 2, 4-dinitrophenol. Therefore, this degradation is not a one-step reaction, decarboxylation, but must be β-oxidation of a fatty acid.

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Some experiments were carried out on the method for evaluation of mixing properties of some medicinal granule preparations, using lactose, crystalline lactose, corn starch, and synthetic aluminium silicate. These medicinal powders (A) were labeled with radionuclide, indium-113 m. Medicinal granule preparations (B) were lactose granules of 100 -200, 65-100, and 42-65 mesh, and commercial pharmaceutical granular preparations of 42-50 (I), 48-100 (II), 20-35 (III), and 28-36 (IV) mesh. Mixing of A and B was carried out with a YMQ-type dispensing mixer, three tines for 5 sec each. Mixing ratio was 1 : 4, 1 : 2, 1 : 1, 2 : 1, and 4 : 1 for A to B. Permitted limit of good mixing properties was decided to be 6.18% of the coefficient of variation of A and B in 10 samples. Experimental results indicated that lactose granules of 100-200 and 65-100 meshes, and I showed good mixing with A in various mixing ratios. Other medicinal granules showed poor mixing with crystalline lactose and synthetic aluminium silicate among A, and showed different mixing ability according to physical properties of B. This method of evaluation is effective for predicting the mixing ability of medicainal granules and general medicinal powders in dispensing.

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The structural characteristic effects of apoludin (I), isoapoludin (II), burrodin (III), and their related pseudoguaiane sesquiterpenelactones were studied by means of mass spectra. Structure of this type of unknown sesquiterpenelactones could be inferred from these mass spectral data.

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The sesquiterpene hydrocarbons of the essential oil of Agastache rugosa O. KUNTZE were examined. The sesquiterpene hydrocarbons, comprising 0.48% of the oil, were found to contain mainly l-caryophyllene, together with small amounts of α-ylangene, β-elemene, β-humulene, β-farnesene, γ-cadinene, calamenene, and two other unidentified compounds.

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Eight alkaloids have been isolated from the trunk and leaves of Lindera oldhamii HEMSL., and three of these have been identified with d-dicentrine (I), N-methylnandigerine (N-methylhernangerine) (V), and N-methylovigerine (N-methylhernovine) (VII). A new alkaloid, O-methylbulbocapnine, C₂₀H₂₁O₄N, mp 129-130°C, [α]³¹_D+248°C (CHCl₃), has been proved to have the structure VI ; and the occurrence of an oxoaporphine base, dicentrinone (II), has been evidenced.

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