The relation between the physico-chemical properties and lymphotropicity of microsphere-in-oil (S/O) emulsion was investigated in comparison with water-in-oil (W/O) emulsion and sesame oil without any pharmaceutical modification. S/O emulsion gave the best lymph node accumulation of tripalmitin (¹⁴C), used as an oil tracer, after intramuscular injection. W/O emulsion was next most effective. Increase of injection volume and massage of the injection site accelerated the lymphatic transport of S/O emulsion, suggesting that hydrostatic tissue pressure plays a role in lymphatic delivery. Sesame oil solution of tripalmitin (¹⁴C) injected without any pharmaceutical modification was not effectively transferred to the lymph nodes, but addition of surfactants to the oil resulted in an increased accumulation of tripalmitin (¹⁴C) in the lymph nodes. Measurements of surface and interfacial tension of various formulations revealed that addition of surfactants caused a marked decrease of interfacial tension against water, which would promote dispersion of the formulations into smaller oil droplets. Microscopic observations of the thigh muscle after intramuscular injection of various oil formulations revealed a correlation between their extent of dispersion and their lymphotropicity. These results suggst that S/O emulsions have physico-chemical properties suitable for effective lymphotropic transport.

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Various TRH analogs, γ-butyrolactone-γ-carbonyl-His-Pro-NHR, γ-butyrolactone-γ-carbonyl-N^3-im-methyl-His-Pro-NHR, 2-ketopiperidine-6-carbonyl-His-Pro-NHR and 3-oxoperhydro-1, 4-thiazine-5-carbonyl-His-Pro-NHR (R=H, methyl, ethyl, n-propyl, n-butyl, n-amyl, n-hexyl, β-phenethyl) were synthesized and pharmacologically tested in the hope of separating the central nervous action from endocrine activity. Among them, γ-butyrolactone-γ-carbonyl-His-Pro-NH₂ was found to have potent central nervous actions in antagonizing pentobarbital sleep and in potentiating apomorphine-induced circling in mice, and only nominal TSH-releasing activity in rats.

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New 1 : 1 adenine-ZnCl₂ and 2 : 1 adenine-CaCl₂ complexes were obtained from water, ethanol, or 90% ethanol solution. The infrared and proton magnetic resonance spectra of the complexes were characterized to investigate the binding site of the metal to adenine by comparison with those of other adenine-metal complexes. On the basis of these data, it is suggested that Zn or Ca is coordinated with the N (1) site of neutral adenine base.

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We synthesized several 2- and 8-substituted derivatives of adenosine 3', 5'-cyclic monophosphate (c-AMP), and examined the effects of these compounds on the isolated atrial muscle preparation of the guinea pig. We found that several 8-substituted c-AMPs could produce definite positive inotropic and chronotropic effects, even when applied from outside the cell, while all the 2-substituted c-AMPs produced negative inotropic and chronotropic effects. Pretreatment of the preparation with aminophylline resulted in a potentiation of the positive inotropic and chronotropic effects of 8-substituted c-AMPs, while the negative inotropic and chronotropic effects of 2-substituted c-AMPs were abolished. The mechanisms by which these effects were produced are discussed.

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An enzymatic method was developed for the simple and rapid assay of lactobacilli. A glucose-hexokinase system was used to remove extracellular ATP, and intracellular ATP was assayed with the luciferin-luciferase system using luciferase commercially prepared from fireflies. Freeze-dried powders of Streptococcus faecalis (SF), Lactobacillus bifidus (LB) and erythromycin-resistant Lactobacillus acidophilus (LA-E) were used as the samples. Prior to assay the freeze-dried powder of lactobacilli was incubated at 37°in a glucose-peptone-yeast extract medium to increase the amount of intracellular ATP. Using the proposed system, the intracellular ATP levels of SF, LA-E and LB were about 100, 80 and 800 times those obtained with the conventional system, respectively. The detection limit was 1.0×10⁵ cells/0.1 ml. Reproducibility was 6.0-9.9% (coefficient of variation). This method was found to be extremely sensitive compared with the conventional luciferin-luciferase system and is convenient because it does not require sterilization and can be completed within a day. This method seems suitable for routine application in survival tests of lactobacilli.

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A novel fluorescent labelling agent, 5-dimethylamino-2-oxidoisoquinolin-1-yl diazomethane (3) was designed and synthesized for the fluorescent labelling of the phosphate moiety of nucleotides and nucleic acids. Starting from 1-cyano-5-nitroisoquinoline (4), 1-carboxy-5-nitroisoquinoline (5) was obtained after hydrolysis with hydrochloric acid. Esterification of 5 with methanol in the presence of sulfuric acid afforded 1-methoxycarbonyl-5-nitroisoquinoline (6). Catalytic hydrogenation of 6 followed by treatment with formic acid-acetic anhydride gave the 5-formamido derivative (8). Methylation of 8 with methyl iodide in the presence of sodium hydride afforded the 5-N-methylformamido derivative (9). Reduction of both the ester group and formyl group with aluminum hydride followed by treatment with chloranil and acetic anhydride provided 1-acetoxymethyl-5-dimethylaminoisoquinoline (11). N-Oxidation of 11 with m-chloroperbenzoic acid followed by selective removal of the oxido group at the 5-position by reaction with carbon disulfide afforded 1-acetoxymethyl-5-dimethylaminoisoquinoline-2-oxide (13). After deacylation of 13, selenium dioxide oxidation of the hydroxymethyl group followed by reaction with p-toluenesulfonyl hydrazide gave 5-dimethylamino-1-formylisoquinoline-2-oxide p-toluenesulfonyl hydrazone (16). On treatment of 16 with sodium ethoxide, the desired compound (3) was obtained. Reaction of 3 with p-nitrobenzoic acid gave the crystalline p-nitrobenzoyl ester. Treatment of uridine 5'-phosphate with 3 gave uridine 5'-(5-dimethylamino-2-oxidoisoquinolin-1-yl) methylphosphate (17). This labelled nucleotide was highly fluorescent, with an excitation maximum of 353 nm and an emission maximum at 523 nm. The fluorescence characteristics of 17 were compared with those of the model compound (13) and uridine 5'-(5-dimethylaminoisoquinolin-1-yl)methylphosphate (18).

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Species differences in the metabolism of paeonol (2-hydroxy-4-methoxyacetophenone : I) in mice, rats, guinea pigs and rabbits were studied by the RI tracer technique. After the administration of a single dose of 20 mg (6-70μCi)/kg paeonol [carbonyl-¹⁴C] (I-¹⁴C), radioactive metabolites excreted in the urine were analyzed. The excretion of I-¹⁴C was rapid in all species ; that is, about 83-98% of administered radioactivity was excreted in the urine and feces during two days after the administration. The ¹⁴C-activity excreted in feces was 10.6% in mice, 4.7% in rats, 2.9% in guinea pigs, and 0.8% in rabbits. It is clear that the urine is the major excretion route in all species. 2, 5-Dihydroxy-4-methoxyacetophenone (II), resacetophenone (III) and unchanged I were detected in all species as urinary metabolites. These metabolites were found as the free form, β-glucuronide, sulfate and enzyme-resistant conjugate. The major metabolite in each species was as follows ; mice, II 43.1% ; rats, II 62.0% ; guinea pigs, III 58.1% ; rabbits, III 42.7% and II 42.2%. All the metabolites were conjugated in rats, while the total amount of unconjugated metabolites was 50-60% of total metabolites in other species. Although β-glucuronide and sulfate in rats accounted for 19.9% and 8.2%, respectively, only small amounts of these conjugated forms were excreted in other species. Enzyme-resistant conjugate accounted for 60.2% in rats, but only 35-43% in other species.

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A series of 3-(aminomethylene)-3H-indoles (1a-e and 6a-8a) was prepared by the condensation of 3-indolecarbaldehydes (2-5) with secondary amines. Among them, 3-(1-pyrrolidinylmethylene)-3H-indoles (1a and 6a-8a) were obtained as stable colorless prisms, while 3-(piperidinomethylene)-3H-indole (1b) and 3-(morpholinomethylene)-3H-indole (1c) readily polymerized in refluxing benzene. Reaction of 1 with electrophiles, such as alkyl and acyl halides, gave 1-substituted 3-aminomethylene-3H-indolium salts, which were converted to the corresponding 1-substituted 3-indolecarbaldehydes by hydrolysis. Reaction of 1 with active methylene compounds proceeded under mild reaction conditions to afford the corresponding condensation products in good yields. Successive reaction of 1 with electrophiles and nucleophiles provided a convenient route to 1, 3-disubstituted indole derivatives.

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The fluorescent reagents, N-(4-dimethylamino-1-naphthyl) bromoacetamide (2a) and N-(4-anilino-1-naphthyl) bromoacetamide (2b), were covalently introduced into 4- and 2-thiouridines at the sulfur site as a model fluorescent probe. The fluorescence properties of the products are discussed.

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The absolute configuration of boschnaloside was established by chemical correlation of asperuloside tetraacetate with boschnaloside tetraacetate. During this work it was also found that catalytic reduction of asperuloside tetraacetate gave either of the two C-8 epimers (4) and (5), depending upon the kind of catalyst used, PtO₂ or Pd-C. The absolute structure of boschnaloside was also confirmed by its degradation to 3(R)-cis, cis-boschnialinic acid. Finall, conversion of genipin into boschnaloside was also achieved.

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Intraperitoneal administration of the extract from Rhei Rhizoma decreased the urea-nitrogen (BUN) concentration in rat serum. An attempt was made to extract and partially purify the active substance(s), monitoring the BUN-decreasing activity. The activity was detected in the aqueous extract. After partition between butanol and water, biological activity was detected in both fractions. Further purification was carried out by dialysis and Sephadex LH-20 column chromatography, by which small molecular compounds, including anthraquinone glycosides, were eliminated. The BUN-decreasing activity was detected in fractions 3-2-1B, 3-2-2B, and 4-2-3-S1.

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Proton migration in a model of the proton cryptate was studied from a quantum chemical point of view. The barrier to proton transfer in the model is 13 kcal/mol. This result is consistent with experimental findings which suggest that the proton is jumping back and forwards in the proton cryptate. Since the barrier height for proton transfer is 41 kcal/mol in the absence of the three ether groups, the three oxygen lone pairs play a significant role in lowering the barrier to proton transfer. A migration map of the proton transfer is presented, and the paths of migration are shown. Moreover, the migration paths are analyzed by the energy decomposition method.

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A method was developed to study the correspondence of color change with structural change of the polyenyl cation in the color reaction of cholesterol in strong acid. Five unsaturated steroids having conjugated systems consisting of 3, 5-diene (I), 3, 5, 7-triene (II), 4, 6, 8(14)-triene (III), 3α, 5-cyclo-6, 8 (14)-diene (IV) and 2, 4, 6-triene (V), and four bisteroids having 3, 3'-linkages and conjugated systems consisting of 3, 3', 5-triene (VI), 3, 3', 5, 5'-tetraene (VII), 3, 3', 5, 5', 7-pentaene (VIII) and 3, 3', 5, 5', 7, 7'-hexaene (IX) were prepared and their electronic spectra were measured in trifluoroacetic acid (TFA), in a CHCl₃ solution of TFA, in H₂SO₄ and in 80% H₂SO₄. The absorption maxima observed immediately after the preparation of the solution and those observed after standing for various times at room temperature were correlated with the chemical species presumed to be produced in strong acids. The following chemical conversions were found to take place : ready conversion of I to VI and VII, and of VI to VII ; very rapid conversions of both II and IV to III ; ready conversion of V to a dimeric molecule ; conversion of III to a dimeric molecule. The color change of cholesterol in strong acid was found to be mainly due to the formation of VI and VII and to their chemical conversions.

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The interaction between Triton X-100 and phosphatidylcholine liposomes was studied by the spin labeling technique and turbidity measurement. The order parameter (S) of spin labeled stearic acid and the turbidity both decreased upon addition of Triton X-100. The effect of Triton X-100 depended on the species of phosphatidylcholine. In egg phosphatidylcholine liposomes, the effect of Triton X-100 was biphasic. In the range of low Triton/phospholipid molar ratios (below 0.4) there was a gradual decrease in the S value without any appreciable decrease in the turbidity. This suggests that Triton X-100 fluidizes the membrane without forming small fragments. Above a molar ratio of 0.4 there was a drastic decline in the turbidity, indicating that the membrane was solubilized. In dimyristoylphosphatidylcholine liposomes, biphasic behavior was not observed. The S value and the turbidity decreased noticeably upon addition of a little Triton X-100. The sensitivity of dimyristoylphosphatidylcholine liposomes to Triton X-100 was much greater than that of egg phosphatidylcholine liposomes. Phase separation between phosphatidylcholine-rich regions and Triton-rich regions may occur in view of the S values reached upon the addition of Triton X-100 to both phosphatidylcholine liposomes. This was also indicated by the spectral change of spin labeled phosphatidylcholine liposomes caused by Triton X-100.

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The interaction of unionized and ionized species of prostaglandins (PGE₁, PGE₂ and PGF_2a) with lauromacrogols was studied by equilibrium dialysis and potentiometric titration. The magnitude of interaction was analyzed in terms of the partition coefficient (P), moles of drug bound per mole of surfactant (K) and moles of drug bound per micelle (K'). The order of the interaction constants with lauromacrogol containing 23 mol of ethylene oxide was PGE₁>PGE₂>PGF_2a, which corresponds to the order of their partition coefficients between water and cyclohexane. The effect of oxyethylene chain length on the interaction was examined with PGE₁ and PGF_2a. Regardless of species, the K value increased with increasing oxyethylene chain length. The P and K' values decreased with increasingly hydrophilic micellar phase. This is due to an increase of the partial molar volume and a reduced aggregation number, leading to an increase of micellar concentration with increasing oxyethylene chain length. The measures of the degree of interaction are desired to be closely related to the changes of physicochemical characteristics of micelles, i.e., to the amounts of drugs in the hydrophobic and hydrophilic regions available for drug binding.

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The insulin-like activity-possessing protease (ILAPP), which was partially purified from Pronase, showed a high but short-lived increasing effect on the vascular permeability of mice, whereas only a low hemorrhagic activity was detected at the same dose. Soybean trypsin inhibitor and leupeptin both inhibited this increasing effect, whereas they exerted no influence on the hydrolytic activity of ILAPP towards casein or succinyl-L-alanyl-L-alanyl-L-alanine p-nitroanilide under the same conditions. The vasodilative activity assayed in a dog was very low, and no kinin-forming activity was detected. This enzyme showed a kinin-inactivating activity when preincubated with bradykinin. Its hydrolytic activity was higher towards acetyl-L-alanyl-L-alanyl-L-alanine methyl ester than towards derivatives of basic amino acid methyl esters. These results suggest that ILAPP differs from trypsin and trypsin-like enzymes in the specificity. The possibility that the insulin-like activity of ILAPP is attributable to kinins produced directly by ILAPP may therefore be excluded.

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Two prenylcoumarins, XII and XIII, were isolated from the root of Angelica pubescens MAXIM. (Umbelliferae) growing in Japan, together with eleven known coumarins, osthol (I), bergapten (II), glabra-lactone (III), angelol (IV), psoralen (V), xanthotoxin (VI), isopimpinellin (VII), byak-angelicin (VIII), coumurrayin (IX), 7-methoxy-8-senecioylcoumarin (X) and scopoletin (XI). The structures of XII and XIII were elucidated as 8-(3-hydroxyisovaleroyl)-5, 7-dimethoxycoumarin and 5-isopentenyloxy-7-methoxy-8-senecioylcoumarin, respectively, by chemical and spectral studies. XIII is a new natural prenylcoumarin, and was named angelin.

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Saikosaponins a and d inhibited adrenaline-induced lipolysis in fat cells isolated from epididymal adipose tissue of rats. Saikosaponins a, b₁, b₂, b₄, c and d inhibited ACTH-induced lipolysis in the fat cells. Insulin-stimulated lipogenesis in adipose tissue was significantly reduced by saikosaponin d. Propranolol (a β-blocker) inhibited not only lipolytic actions induced by adrenaline and ACTH but also lipogenesis stimulated by insulin. In contrast to propranolol, saikosaponins b₁, b₂, b₄ and c selectively inhibited ACTH-induced lipolysis without affecting the lipogenetic effect of insulin. The structure-activity relationship of saikosaponins is discussed.

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Four metabolites of the antitumor agent 1-(tetrahydro-2-furanyl)-5-fluorouracil, formed in vitro by rat liver microsomes, were isolated by thin-layer chromatography or high-performance liquid chromatography. On the basis of mass spectrometry, ¹H-NMR spectral analysis, and comparison with authentic samples, these metabolites were identified as 1-(trans-4-hydroxytetrahydro-2-furanyl)-5-fluorouracil, 1-(cis-4-hydroxytetrahydro-2-furanyl)-5-fluorouracil, 1-(trans-3-hydroxytetrahydro-2-furanyl)-5-fluorouracil and 1-(4, 5-dehydrotetrahydro-2-furanyl)-5-fluorouracil. These metabolites were also found in the plasma and urine of rats after administration of 1-(tetrahydro-2-furanyl)-5-fluorouracil.

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The protected tetrasaccharide (7) was synthesized in 79% yield by condensation of 1, 6-anhydro-2, 2', 3, 4', 6'-penta-O-benzyl-β-lactose (4) with the acetylated oxazoline derivative of lacto-N-biose I (6). The protecting groups of 7 were removed by the following series of reactions to provide lacto-N-tetraose (13) : debenzylation, acetylation, acetolysis, and de-O-acetylation. The synthetic product (13) was crystallized from aqueous ethanol as hygroscopic fine needles, mp 225-228°, [α]²¹_D+27° (4 min)→+21.3°(3 hr) (c=0.45, H₂O). The homogeneity and the mobility of 13 were confirmed by gel permeation chromatography using a Bio-Gel P-4 column. The specific rotation and IR spectrum of 13 were in good agreement with those of the natural product reported by Kuhn, Gauhe, and Baer [Chem. Ber., 86, 827 (1953)]. ¹³C-NMR spectral data for 1, 6-anhydro-β-lacto-N-tetraose (9) are also presented.

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A 13, 15-diazasteroid system was synthesized from 2-(6-methoxynaphthyl) ethyl tosylate (steroidal segment of the A, B rings) and ethylene urea (steroidal segment of the D ring) in 24.3% overall yield. The tosylate formed a 1 : 1 adduct with ethylene urea in 44% yield, using 2 molar equivalents of ethylene urea in the presence of sodium hydride in benzene. The adduct was cyclized to 13, 15-diazasteroid hydrochloride in 69.7% yield by prolonged heating in the presence of phosphorus pentoxide in phosphorus oxychloride. The hydrochloride was neutralized to provide the 13, 15-diazasteroid in 79.1% yield by treatment with potassium hydroxide solution. The biological activity of the hydrochloride of the 13, 15-diazasteroid is now being examined.

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(±)-PGH₁ analogs (9, 11-epoxymethano PGH₁, 11, 9-epoxymethano PGH₁, 9, 11-epoxycarbonyl PGH₁, and 11, 9-epoxycarbonyl PGH₁) were synthesized from 9- or 11-substituted prostaglandin derivatives or their synthetic intermediates. These analogs showed interesting biological activities, different from those of PGH₂ analogs.

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The reaction of sulpyrine with nitrite under mild conditions similar to those of the human stomach gave three N-nitroso compounds. The main products were 4-(N-methyl-N-nitroso) aminoantipyrine (I) and 1-diketobutyryl-1-phenyl-2-methyl-2-nitrosohydrazide hydrate (III), and 1-acetyl-1-methyl-2-nitroso-2-phenylhydrazine (II) was formed as a minor product. The possible pathways of formation of I and III are discussed.

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Clivia miniata REGEL. (Amaryllidaceae) was found to contain a novel alkaloid, clivacetine (2), as well as lycorine (3), clivatine (4), clivimine (5), and clivonine (6). Clivacetine was identified as O-acetoacetylclivonine (2) and has an interesting structure from a biogenetic viewpoint, since it is a possible biosynthetic precursor of clivimine (5).

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5-Amino-3-phenylisoxazole-4-carbaldehyde (III) was synthesized by the Vilsmeier reaction of 5-amino-3-phenylisoxazole (I) via N, N-dimethyl-N'-(4-formyl-3-phenylisoxazol-5-yl) formamidine (II). Reaction of III with formamidine and ethyl imidates afforded 3-phenylisoxazolo [5, 4-d] pyrimidine (IVa) and its 6-substituted derivatives (IVb-e), respectively. Condensation of the oxime (V) of III with ethyl orthoformate gave 3-phenylisoxazolo [5, 4-d] pyrimidine 5-oxide, which was readily deoxygenated with PCl₃ to give IVa. Treatment of IVa with Grignard reagents gave 4-substituted 4, 5-dihydro derivatives (VIIa, b) which were aromatized with K₃Fe (CN)₆ to yield 4-substituted isoxazolo [5, 4-d] pyrimidines (VIIIa, b).

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Epoxidation and allylic oxidation of cholesteryl acetate were studied using hydroperoxide (ROOH : R=H, ethyl, iso-propyl, tert-butyl, or cumyl) and a metal complex [Fe (acac)₃ or Mo (CO)₈] in benzene or acetonitrile. In the presence of the iron catalyst, formation of the epimeric C (5, 6)-epoxides predominated in the reactions with H₂O₂, while allylic oxidation giving C (7)-ketone, epimeric C (7)-alcohols and C (7)-alkyl peroxides was favored in the reactions with alkyl hydroperoxide, regardless of the polarity of the solvent. The stereoselective β-epoxidation was independent of the structure of ROOH, but the ratio of allylic oxidation to epoxidation was dependent on it. When the iron catalyst was replaced by the molybdenum catalyst, β-epoxidation decreased and solvent effects on the reaction were sufficiently marked to exclude allylic oxidation in the non-polar medium.

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Treatment of 1H-1, 2-benzodiazepines having an electron-withdrawing group such as methoxycarbonyl or cyano in the 3-position with lead tetraacetate resulted in the formation of the corresponding 5-acetoxy-5H-1, 2-benzodiazepines, which are the first reported examples of 5H-1, 2-benzodiazepines. The 5H-diazepines, on treatment with base, underwent tautomerization to the 1H-isomers, and on photolysis gave 3-acetoxyindole via the tricyclic valence isomers. Furthermore, treatment of a 5H-diazepine with methanol or acetic acid gave the corresponding 4-methoxy- or 4-acetoxy-4, 5-dihydro-1H-1, 2-benzodiazepine, respectively, by 1, 4-addition of the reagent.

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A new coumarin glycoside (1) was isolated from the root and rhizoma of Glehnia littoralis FR. SCHMIDT et MIQ. (Umbelliferae), together with fourteen known coumarins (4-17). The structure of 1 was elucidated as osthenol-7-O-β-gentiobioside by chemical and spectral studies.

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Two crystalline color reaction products were isolated from the reaction mixture of butylurea with 1-phenyl-1, 2-propanedione in dioxane containing HCl. One of them (A) was found to be identical with 1, 3'-dibutyl-5, 5'-diphenyl-2, 2'-dioxo-4, 4'-diimidazolylmethane (I) which was isolated previously. The other one (C) was found to be isomeric with compound (B) which was also isolated previously. It was found that the only difference between B and C is the positions of their butyl side chains. In order to decide the positions of the side chains of B and C, the differences between the physico-chemical properties expected to be shown by 3, 3'-dibutyl-(II) and by 1, 1'-dibutyl-5, 5'-diphenyl-2, 2'-dioxo-4, 4'-diimidazolylmethane (III) were estimated theoretically. From the results of this estimation the structures II and III could be assigned to B and C, respectively.

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From the aerial parts of Lindsaea chienii CHING a new diterpenoid glycoside (lindokaurenoside C) has been isolated together with lindsaea acid, creticoside A, trans-cinnamic acid and p-hydroxy-trans-cinnamic acid. From spectroscopic evidences and chemical reactions the structure of the new glucoside has been established as ent-2α, 13-dihydroxykaura-16-ene 2-O-β-D-glucoside.

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A sensitive enzyme immunoassay for the determination of neocarzinostatin (NCS), a proteinaceous antitumor antibiotic, has been developed. Horseradish peroxidase was used as the labeling enzyme and was conjugated with NCS according to Nakane's method. Separation of bound and free fractions was achieved by a double antibody solid phase method using Sepharose 4B gel coupled with purified IgG of goat anti-rabbit IgG serum. Thyramine and hydrogen peroxide were used as substrates for the fluorophotometric assay of peroxidase activity. A satisfactory standard curve for NCS was obtained in the range of 10 to 1000 pg/assay tube, corresponding to the range of 0.5 to 50 ng/ml of sample solution. This enzyme immunoassay could be applied to the determination of NCS in serum after its administration to rabbits. NCS in various tissue homogenates could also be assayed by this method.

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From the aerial parts of Dennstaedtia wilfordii (MOORE) CHRIST. three new pterosins (pterolacton A, B and dennstopterosin) and two new pterosides (pterolacton A 3-O-β-D-glucoside and pterolacton A 3-O-β-D-(4'-p-coumaroyl)-glucoside) have been isolated together with pterosin A, D and L. Their structures have been established from spectroscopic evidences and chemical reactions.

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Two alternative deprotecting procedures were employed for the synthesis of porcine vasoactive intestinal polypeptide (VIP). Besides HF, TFA-thioanisole was found to cleave all the protecting groups employed, Z, Bzl and Mts, suppressing a newly found side reaction, i.e., acid-catalyzed succinimide formation from Asp residues with the free carboxyl group. A small amount of the N-terminal His residue linked to Ser-Asp was found to be released on standing at pH 6.

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From the fronds of Bolbitis subcordata (COPEL.) CHING a new C-methylleucoanthocyanidin was isolated and shown to be (2R, 3S, 4S)-5, 4'-dimethoxy-6, 8-dimethyl-3, 4, 7-trihydroxy-flavan.

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Anodic oxidation of 2-hydroxy-3-methoxy-5-methylbenzaldehyde (II) in acetonitrileacetic acid (3 : 1) containing sodium acetate gave an acetoxylated product in which the acetoxyl group is attached to the side-chain methyl group. On the other hand, oxidation of the Schiff bases (I) derived from II under the same conditions gave cyclohexenetriones. The latter products were shown to be formed by further oxidation of the initially formed acetoxylated Schiff bases, in which the acetoxyl group is introduced into the ring meta to the hydroxyl group.

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A facile preparation of 5-(3-indolylmethylene) hydantoins, which are useful intermediates for syntheses of tryptophan and its derivatives, was achieved by condensation of hydantoin with 3-(aminomethylene)-3H-indoles formed in situ by the reaction of indoles with Vilsmeier-Haack reagent followed by neutralization with anhydrous bases.

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The new sterols 24(ξ)-methyl-25(ξ)-cholest-5-ene-3β, 26-diol and 24(ξ)-ethyl-25(ξ)-cholest-5-ene-3β, 26-diol were isolated, together with p-coumaric acid, apigenin and amentoflavone, from pollen grains of Podocarpus macrophylla D. DON.

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Methods for the preparation of p-amidinophenyl esters from amino acid derivatives are described. Blocking of the amidino function with the benzyloxycarbonyl group and its deblocking by catalytic hydrogenation after coupling were satisfactory. p-Amidinophenyl esters are of special interest because of their susceptibility to the hydrolytic enzyme, trypsin, and some parameters of the hydrolysis of these compounds by trypsin are presented.

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Various 9-substituted 8-phenyltheophyllines (III) were prepared in two steps starting from 5, 7-dimethyl-2-phenyloxazolo [5, 4-d] pyrimidine-4, 6 (5H, 7H)-dione (I). Thus, treatment of I with amines afforded 5-(N-substituted benzamidino)-1, 3-dimethylbarbituric acids (II). The reaction of II with thionyl chloride or phosphorus oxychloride gave III.

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Nitration of 4, 7-dibromobenzo-2, 1, 3-thiadiazole (I) with fuming nitric acid afforded 4, 7-dibromo-5, 6-dinitrobenzo-2, 1, 3-thiadiazole (II) and, unexpectedly, the tribromo derivative : C₆Br₃N₃O₂S (III). Compound II was readily converted by treatment with sodium azide into bisfuroxanobenzo-2, 1, 3-thiadiazole (IV), which was reduced with triethylphosphite to give bisfurazanobenzo-2, 1, 3-thiadiazole (VII). Compound VII was also synthesized starting from 4-bromo-6, 7-(2', 1', 3'-oxadiazole) benzo-2, 1, 3-thiadiazole (VIII). Reduction of VII with sodium hydrosulfite gave 4, 5-diamino-6, 7-(2', 1', 3'-oxadiazole)-benzo-2, 1, 3-thiadiazole (XI), which was cyclized to 4, 5-(2', 1', 3'-oxadiazole) benzo [1, 2-c : 3, 4-c'] bis [1, 2, 5] thiadiazole (XII) by treatment with N-sulfinylaniline. The transformation of XII to benzo [1, 2-c : 3, 4-c' : 5, 6-c"] tris [1, 2, 5] thiadiazole (XIV), via 4, 5-diaminobenzo-[1, 2-c : 3, 4-c'] bis [1, 2, 5] thiadiazole (XIII), is also described.

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A sensitive phosphorimetric method is described for the assay of urinary free and total 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG), a major metabolite of norepinephrine. Free MHPG, after separation from urine by Amberlite CG-50 column chromatography, is oxidized with periodate to vanillin, which is then measured phosphorimetrically in a mixture of ether and ethanolic potassium hydroxide. Total MHPG is measured after β-glucuronidase and arylsulfatase mediated hydrolysis of conjugated MHPG. The assay requires less than 2 ml of urine to provide test, blank and standard samples, and offers lower limits of determination of 5 and 30 ng/ml for free and total MHPG, respectively.

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An aqueous solution of sodium ε-amino-n-caproate can be used for efficient and simple separation of aldehydes, overcoming the difficulties associated with the NaHSO₃ method.

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Alkylations of N, N-dimethyl-(Ia) and N, N-diethyladenine (Ib) with methyl iodide, ethyl iodide, and benzyl bromide in N, N-dimethylacetamide in the presence of potassium carbonate gave the corresponding N, N, 9-trialkyladenines (II) in 54-74% yields, as well as minor amounts of N, N, 3-trialkyladenines (III). The alkylation of I without base gave the latter compounds (III) in 76-90% yields.

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A sensitive method for the detection and determination of phenylthiohydantoin amino acids using hypochlorite-thiamine reagent is described. Phenylthiohydantoin amino acids were developed on a Kieselgel plate and N-chlorinated on the plate by spraying sodium hypochlorite. Excess hypochlorite was removed by aeration and the plate was sprayed with the alkaline thiamine reagent. The chlorinated thiohydantoin gave and intensely fluorescent spot. An application of this method to Edman degradation is also described.

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Two 1, 3-bridged 1, 2, 3, 4, 5, 6-hexahydro-2, 6-methano-3-benzazocine derivatives (1 and 2) have been synthesized starting from 3, 6-dimethyl-8-methoxy-1, 2, 3, 4, 5, 6-hexahydro-2, 6-methano-3-benzazocine 3 as shown in Chart 1. Compounds 1 and 2 showed no significant analgesic activity.

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Copper (I) halide-catalyzed decomposition of phenolic α-diazoketones gave spiro [4.5]-deca-6, 9-diene-2, 8-dione in high yield.

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A dipeptide named kyotorphin, H-Tyr-Arg-OH, and its retro isomer were synthesized using N^G-mesitylene-2-sulfonylarginine. Two corresponding stereoisomers containing D-Arg were also synthesized. Among these, H-Tyr-D-Arg-OH was found to possess an analgesic effect 6 times higher than that of the corresponding L-isomer.

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11-Hydroxy-Δ⁸-tetrahydrocannabinol, which is an active metabolite of Δ⁸-tetrahydrocannabinol, produced a spectral change on interacting with liver microsomes of rabbits and rats (λ_max 395 nm ; λ_min 428 nm). The apparent dissociation constants (Ks) were found to be 402.1 and 76.6 μM for microsomes of untreated rabbits and rats, respectively. Pretreatment with phenobarbital enhanced the spectral change caused by 11-hydroxy-Δ⁸-tetrahydrocannabinol with microsomes of both species, as well as increasing the cytochrome P-450 content. Furthermore, 11-hydroxy-Δ⁸-tetrahydrocannabinol interfered with the spectral change caused by Δ⁸-tetrahydrocannabinol or hexobarbital, but not with that caused by aniline. These results are consistent with the view that 11-hydroxy-Δ⁸-tetrahydrocannabinol is further metabolized by the microsomal monooxygenase system involving cytochrome P-450.

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The sulfoxide of Boc-Cys (S-acetamidomethyl)-OH was prepared by oxidation with sodium perborate. Mercuric acetate and iodine failed to cleave the S-protecting group from the sulfoxide. Hydrogen fluoride and methanesulfonic acid partially converted the sulfoxide to S-p-methoxyphenylcysteine in the presence of anisole. A reducing reagent, thiophenol, converted the sulfoxide to acetamidomethyl phenyl sulfide and N^α-Boc-S-(phenylthio) cysteine.

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The relationship between antifungal activity and the energy of the lowest unoccupied molecular orbital (LUMO) of several phenolic compounds was examined using MINDO/3 method. Among the phenols examined, all the compounds which are highly active in inhibiting the growth of fungi possess a low-lying LUMO as compared with the compounds poor in antifungal activity. From these results, it is conceivable that the LUMO energy of a molecule should be one of the useful indices for screening antifungal drugs.

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