An attempt was made to prepare Cu, Ni, Co (II), Mn (II), Zn, and Cd complexes of m-amino- and p-amino-benzoic acids, as well as these of ο-aminobenzoic acid for comparison, and elemental analysis, including metal elements by atomic absorption spectroscopy, infrared spectral analysis, DTA and TGA were carried out. These complexes have 2 : 1 ratio of meta1 to ligand and their -NH2 and -COOH groups are assumed to participate in the complex formation.
Weak acid and neutral fractions separated from the extracts of male and female human urine showed an inhibitory action against contraction of smooth muscle preparations. Their inhibitory action was tested on contraction of isolated guinea-pig ileum induced by histamine and isolated rat uterus by oxytocin. It was suggested that one of the principles of inhibitory activity in the urine might be the metabolites of steroid hormones.
To obtain the most suitable conditions for thin-layer chromatographic screening procedure for detection of doping drugs, 8 solvents and 9 reagents were compared, using 28 neutral, basic, and acidic drugs. Disturbance of separation by tartaric acid added to urine extracts to prevent loss of drugs by evaporating is proved to be eliminated by over-spotting of concentrated NH4OH on the original line before running. The method elaborated in the present investigation is as follows : The methanol solution of the residue from chloroform extract, corresponding to 8 ml of urine, containing 2 mg of tartaric acid, is spotted on silica get plate containing a fluorecent substance. After over-spotting of concentrated NH4OH, the plate is developed with chloroform -methanol (9 : 1). The spots of drugs are visualized by the following means : (1) Ultraviolet irradiation, (2) 1% HgNO3, (3) Mandelin's reagent, (4) Dragendorff's reagent. Urine samples must be frozen when analysis is not done immediately. A plate containing mixed fluorecent substances is suitable for this method. The sensitivity was not inferior to any other method by ultraviolet spectra, color test, and microcrystal test using 200 ml of urine.
Methyl 1-substituted 5-oxo-3-pyrazolin-3-yl acetates (IV) were synthesized by the reaction of methyl 3-oxoglutarate (I) with 1-substituted hydrazine (III). An attempt was made to introduce a methyl group into 2-position of methyl 1-phenyl-5-oxo-3-pyrazolin-3-yl acetate (IVa) but it was found that a pyrazole-type compound (XI) was also produced besides the pyrazolone type (VIIIa) through this process. On the basis of this result, 1, 2-disubstituted 5-oxo-3-pyrazolin-3-yl acetic acids (VIII) were synthesized by introduction of the corresponding substituent into 2-position of IV. 1-Substituted 5-oxo-3-pyrazolin-3-yl acetic acid (V) and VIII were easily pyrolyzed and changed into 3-methylpyrazolone type compounds (the former into VI and the latter into IX). The activity of VIII as an antipyretic, analgesic, and antispasmodic was tested but they proved to be no better than the well-known medicines.
Pharmacological action of 1-phenyl-5-methoxypyrazol-3-ylacetic acid (IVa) was investigated and it proved to have a high analgesic and anti-inflammatory activity. There-fore, methylation of 5-position in methyl 1-phehyl-5-oxo-3-pyrazoline-3-yl acetate (Ia) was attempted. On the basis of its result, 1, 5-disubstituted pyrazol-3-yl acetic acids (TV) were synthesized by introducing the corresponding substituent into the 5-position of I. During this reaction, V was also produced and some methods to separate V from TV were attempted. It was then found that TV showed a considerably different hardness of decarboxylation according to various substituents in 1-position.
The pharmacological properties of 3, 4-dihydroxy-5-methoxy-benzoylaminoguanidine (AG-2H) on the cardiovascular systems were examined. In the rat and cat, AG-2H (1-5 mg/kg) caused a temporary pressor response which was inhibited by adrenergic α-blocking agents (phentolamine and phenoxybenzamine), but not by hexamethonium and reserpine, while the pressor response was potentiated by cocaine. AG-2H showed a vasoconstriction on the isolated rabbit ear, which was inhibited by phentolamine. The positive inotropic action of the isolated rat and guineapig heart induced by AG-2H (1-3 mg/heart) was slightly inhibited by propranolol. On the other hand, the positive inotropic action of norepinephrine was suppressed by a large dose of AG-2H (5 mg/heart). On the heart of the open chest cat, AG-2H showed an increment of contractile force synchronized with the pressor response. This action was not accompanied by a positive chronotropic action. In the normal and Ca2+-free Krebs-Henseleit solution, AG-2H as well as ouabain slightly prolonged the plateau phase of action potential in the guinea-pig papillary muscle. In the Ca2+-free medium, epinephrine which was effective as other drugs in the normal one, prolonged the plateau phase markedly. From these results, it is suggested that AG-2H posseses mainly a direct sympathomimetic activity on the blood pressure and a musculotropic activity on the heart muscle.
Fluorometric determination of tripelennamine hydrochloride (I) in mixed anti-cold pharmaceutical preparations was studied, and a selective assay method with high sensitivity was established, The fluorescent substance of this reaction was extracted and isolated as an iodide. This substance, mp 150-151°, has the formula C14H15N2I, and its chemical structure was found to be 1-benzyl-2, 3-dihydroimidazo1o[1, 2-α]pyridinium bromide, and the fluorescence is produced by ring formation to a quaternary ammonium compound, accompanied with separation of dimethylamino group, from the result of a moderate oxidation of I by BrCN. This assay method is as follows : Tripelennamine is extracted with petroleum ether from the sample solution containing about 4 mg of I and the extract is evaporated with small amount of citric acid. The residue is dissolved in EtOH to make 1000 ml. To 4 ml of this ethanolic solution, 10 ml of pH 8 buffer solution and 4 ml of 2.5% BrCN solution are added and the mixture is heated at 50° for 30 min. The fluorescence of cold solution is determined at 415 mμ, exciting at 340 mμ.
After diazotization of aminoketone derivative (V) in a usual way, the diazotized salt was decomposed without a catalyst at 70° for 1 hr. The chromatography of the resulting products on silica gel did not afford a phenolic ketone (VII), but gave a seven-membered product (VI), acetyl derivative (VIII), and dienone (IX). On the other hand photo-Pschorr reaction gave only the phenolic ketone (VII).
For the quantitative determination of prostaglandin, prostaglandins dehydrogenase dependent on nicotinamide adenine dinucleotide (NAD+) was purified from swinelung. This enzyme catalyzed the oxidation of secondary alcohol group at C-15 in prostaglandin to a ketone, and the coenzyme NAD+ was reduced to NADH. For higher sensitivity, this assav method was coupled with the reduction of resazurin, as catalyzed by diaphorase in the presence of a catalytic amount of NADH, and resorufin formed during this enzymic reaction was measured by a very sensitive fluorometry. This method made it possible to measure 4×10-11 mole of prostaglandin.
The amine-cleavage of 3-phenoxy-1, 2-propylene sulfides (II) synthesized from P.G.E. (I) with KSCN gave only sec-thiol derivatives. A mixture of sec-thiol and prim-thiol derivatives was obtained by the reaction of II with ethylmagesium bromide. The reaction of II with Grignard reagents containing more than three carbons afforded sec-thiol derivatives. The reaction product of ο-nitro-substituted II with Grignard reagents was identified as prim-thiol derivatives.
A new phloroglucinol derivative, filixic acid ABA (III), C32H36O12, mp 163-166°, was isolated from the rhizomes of Dryopteris dickinsii (FR. et SVA.) C. CHR. (Aspidiaceae). The structure of this compound was deduced from a detailed study of infrared, ultraviolet, and nuc1ear magnetic resonance spectra, and alkaline cleavage, and confirmed by its synthesis. Another phloroglucinol derivative was identified as filixic acid BBB (I).
The reaction of ethyl 3-methylindole-2-carboxy1ate (IV) with SO2Cl2 afforded ethyl 3-methyloxindole-3-carboxylate (VI) and not the compound (V). which had been reported by Elks, et al.2) Furthermore, the reaction of IV with Pb(OAc)4 gave an intermediate indolenine derivative (XIV), which underwent rearrangement by aqueous AcOH into the corresponding oxindole (VI).
The metabolic fate of 2-amino-3-ethoxycarbonyl-4, 5, 6, 7-tetrahydrothieno[2, 3-c]pyridine (Nor-Y-3642) and benzoic acid was investigated to determine the metabolic pathway of 2-amino-3-ethoxycarbonyl-6-benzyl-4, 5, 6, 7-tetrahydrothieno[2, 3-c]pyridine (Y-3642) reported in the previous paper.1) When 35S-Nor-Y-3642·HCl was administered intraperitoneally to mice and rats, approximately 75% of the given radioactivity was excreted as 35S-Nor-Y-3642-N-glucuronide and 35S-2-amino-4, 5, 6, 7-tetrahydrothieno[2, 3-c]pyridine-3-carboxylic acid (35S-Nor-Y-3642-COOH) besides the unchanged compound in the urine. Furthermore, 35S-2-amino-3-ethoxycarbonyl-6-acetyl-4, 5, 6, 7-tetrahydrothieno[2, 3-c]pyridine (35S-N6-Ac-Nor-Y-3642) was found as one of the metabolites of 35S-Nor-Y-3642 in the bile of rats. On the other hand, when 14C-benzoic acid was intraperitoneally administered to mice and rats, almost all of the given ridioactivity was excreted as 14C-hippuric acid and 14C-benzoyl glucuronide together with a small amount of the unchanged form in the urine. These results suggest that Nor-Y-3642 and benzoic acid are the metabolic intermediates of Y-3642 in mice and rats.
Gas chromatographic method for determination of urinary metabolites derived from an officinal dose of 3-deoxyestrone (I) and 16α-chloroestrone methyl ether (II) hat been devised. Separation of each of three principal metabolites was satisfactorily attained when they were transformed into the trifluoroacetyl derivatives and then subjected to gas chromatography under the appropriate column conditions (Table I). The structures of the metabolites were confirmed by gas chromatography-mass spectrometry (Fig. 2). Preliminary purification by thin-layer chromatography proved to be effective for removal of interference due to undesirable substances, less and more polar than the metabolites. The calibration curve for quantitation of each metabolite was constructed. employing androst-5-ene-3β, 16β, 17β-triol tris (trifluoroacetate) as an internal standard. The amount of the metabolites excreted in urine after oral administration of I and II to man was determined by the procedure thus established.
The base obtained by the Mannich reaction of 6-azauracil was proved to be an N-substituted compound, differing from that obtained from uracil. When piperidine is used as the base, the reaction progresses to a 1, 3-di substituted compound, while the use of morpholine gives only 1-substituted compound. The Mannich reaction of 1-methyl-and 3-methyl-6-azauracil was also found to give. N-substituted compounds. None of the compounds obtained (I to VI) showed any marked antibacterial activity, prophageinducing activity, or anticancer action against a transplanted tumor.
Complexes of 5'-cytidylic acid (5'-CMP) with zinc, lead, cobalt, cadmium, and manganese were isolated as crysta11ine precipitate, and mercury and silver complexes were isolated as amorphous ones from the so1ution of pH 3.2-6.5. Ratio of metal. to ligand was found to be 1 : 1 except in Ag-5'-CMP and Hg-5'-CMP, whose compositions were 2 : 1 and 3 : 2, respectively. The infrared spectra of these complexes were compared with those of sodium, potassium, calcium, ahd barium salts of 5'-CMP. Metal coordination to the phosphate group was suggested from the infrared absorption band near 980 cm-1. Coordination of mercury, cobalt, zinc, and cadmium with the cytosine ring moiety was also assumed from the spectral changes in the region of 1700-1600 cm-1, compared with those of salts of 5-'CMP and cytosine-metal complexes isolated.
The Diels-A1der type adduct (II or VI) was formed on the reaction of 2, 3-diphenyl-5, 6-dihydropyrazine (Ia) with diethyl fumarate or with N-methylmaleimide. The stereostructures (IIa and VIa) were proposed from the nuclear magnetic resonance measurements.
The inconsistency of asebotin in Pieris japonica was checked by thin-layer chromatography. Asebotin was proved to be contained in Pieris japonica collected in warm regions while phlorizin is found in those collected in cold regions.