Snake post-synaptic neurotoxins can be classified into the following types : (i) shortchain neurotoxins ; the neuromuscular block is easily reversible. (ii) long-chain neurotoxins ; the neuromuscular block is almost irreversible. Judging from these results, shortchain neurotoxin is more useful for the isolation of acetylcholine receptor from the electric organ of fishes, muscles and thymus. Long-chain neurotoxin seem to be a useful tool to detect anti-acetylcholine receptor antibodies in the sera of patients with myasthenia gravis. Snake neurotoxins serve to clarify the function of neurotransmitters at the cholinergic receptor site in the level of molecular biology, and these results may be helpful in the elucidation of the pathogenesis of myasthenia gravis.
The coordination sites of tetracyclines to Cu2+ in a solution at pH 5 were examined by the methods of circular dichroism (CD) spectra, absorption spectra and optical rotation. The results obtained were as follows : 1) The CD spectrum of tetracycline (TC)-Cu2+ complex showed a positive peak at 400 nm, whereas that of tetracyclinenitrile (TN)-Cu2+ complex didn't show any positive peaks in the range over 350 nm. 2) Bathochromic shifts in the absorption spectra of TC and TN induced by the addition of Cu2+ at pH 5 were observed. 3) With an increase in the molar ratio of Cu2+ to TC or oxytetracycline (OT), the optical rotation changed from negative to positive. On the other hand, the optical rotation of TN-Cu2+ complex was independent of the molar ratio of Cu2+ to TN. 4) The CD spectrum of TC-Cu (ethylenediamine)2+ or OT-Cu (ethylenediamine)2+ complex showed a positive peak at 400 nm or 390 nm. 5) The curves obtained by the plots of the optical rotation against the molar of Cu (ethylenediamine)2+ to TC or OT were similar to that obtained by the plots against the molar ratio of Cu2+ to OT. Therefore, at pH 5, tetracyclines may bind to Cu2+ at both C11-C12 β-diketone site and C2 carboxamide site through bending of A ring over B and C rings.
Condensation products of 3-acetyltetramanilide (3-(1'-anilinoethylidene) pyrroline-2, 4-dione) with 13 kinds of substituted benzaldehydes were prepared. The compounds synthesized were tested for the antitumor activities against Yoshida sarcoma cells in vitro and Ehrlich ascites carcinoma in vivo. 3-Acetyl-5-benzylidenetetramanilide was shown to be effective both in vivo and in vitro. The relative reactivities of condensation reactions of 3-acetyltetramanilide, 3-acetyltetronanilide and 3-acetylthiotetronanilide with benzaldehyde under a variety of catalysts were examined.
Four anti-tumor antibiotics were isolated from a new strain of Pseudomonas. Two of which were identified as bactobolin and N-acetylbactobolin respectively. The other two constituents, bactobolin B and bactobolin C, were shown to be new compounds and these structures were elucidated from the spectral data and the results of chemical degradation. Antimicrobial activities of these compounds were also discussed.
Some N-acylbactobolins (VI) were synthesized, and N-L-alanylbactobolin was identified as bactobolin B. The product by Edman's degradation reaction on bactobolin was thought to be a key intermediate for synthetic studies, named bactoboamine which was converted to N-acyl derivatives (IX). Antimicrobial activities of VI and IX were also discussed.
Assay of lysozyme chloride have been conducted by bacteriolytic method, by using Micrococus lysodeikticus as substrate. But the reproducibility of this method is invariably poor because of enzymatic processes in a heterogeneous system. We began to research the determination of lysozyme chloride by reversed phase high performance liquid chromatography. And we succeeded in retaining lysozyme chloride to reversed phase stationary, which is a basic condition for the determination by HPLC. We present here a HPLC system capable of retaining lysozyme chloride to reversed phase stationary by adding alkylsulfonic acid or perchloric acid as anion type counter ion to mobile phase. The effects of HPLC conditions on retention of lysozyme chloride were also examined. The good reproducibility could be suggested, when it was applied to the determination of lysozyme chloride.
An arginine ester hydrolase, ME-2 was isolated from the venom of Trimeresurus mucrosquamatus by gel filtration on Sephadex G-100 and ion exchange chromatographies on CM-Sephadex C-50, DEAE-Sephadex A-50, DEAE-Sephacel. By these procedures, 6.7 mg of purified preparation was obtained from lg of crude venom. This enzyme hydrolyzed arginine esters, such as N-tosyl-L-arginine methyl ester (TAME) or N-benzoyl-L-arginine ethyl ester (BAEE), but did not hydrolyze casein, hemoglobin, N-tosyl-L-lysine methyl ester (TLME), N-acetyl-L-tyrosine ethyl ester (ATEE), N-tosyl-L-argininamide (TAA) or N-benzoyl-L-argininamide (BAA). The esterolytic activity was inhibited by benzamidine but not by diisopropyl fluorophosphate (DFP) or trasylol. The purified preparation was homogeneous as judged by disc electrophoresis on polyacrylamide gel and isoelectric focusing. The molecular weight of ME-2 was determined to be approximately 29800, and the isoelectric point was found to be pH 5.62 by isoelectric focusing with carrier ampholyte. The esterolytic activity of the final preparation was 779.8 units/mg. This enzyme had capillary permeability-increasing and kinin-releasing activities. This protein was stable to heat treatment, and between pH 5 and 9. Its Michaelis constant (Km) for TAME and inhibition constant (K1) for benzamidine were found to be 4.27×10-3 M and 0.194×10-3 M, respectively. This protein contained a small amount of carbohydrates.
We measured the blood oxygen tension (Po2) in the rat aorta in connection with cardiac lesions caused by the subcutaneous administration of sympathomimetic amines. The degree of cardiac lesions caused by l-isoproterenol, which acts predominantly on the β-receptor, or dl-norepinephrine, which acts predominantly on the α-receptor, was dose dependent, and these drugs decreased the Po2 in the same dose range. The decrease in Po2 due to dl-norfenefrine, which causes very little cardiac lesion and acts on the α-receptor, was not significant. Moreover, it is ascertained that cardiac lesions and a decrease in Po2 caused by l-isoproterenol could be prevented by pretreatment with propranolol and that cardiac lesions and a decrease in Po2 caused by dl-norepinephrine could be prevented by pretreatment with phentolamine.
The inhibitory effects of ginseng saponin on adrenal atrophy, thymus atrophy and the decrease of serum K+ concentration induced by cortisone acetate in unilateral adrenalectomized rats were studied. Ginseng saponin was isolated from the roots of Panax ginseng C. A. MEYER by using of Amberlite XAD-2 resin. Ginseng saponin (200 mg/kg, ×7 days, p. o.) was found to be antagonistic to the suppressive action of cortisone acetate (10 mg/kg, ×7 days, i. m.) on compensatory adrenal hypertrophy in rats after unilateral adrenalectomy and this finding was supported by histochemical studies. Ginseng saponin was also found to be antagonistic to the decrease of thymus weight and serum K+ concentration induced by cortisone acetate in these rats. On the other hand, the same dose of ginseng saponin alone had no effect on compensatory adrenal hypertrophy, thymus weight and serum K+ concentration in these rats. In histochemical studies, ginseng saponin showed no effect on the improvement of fatty metamorphosis induced by cortisone acetate in the liver of these rats. These actions of ginseng saponin, except for the effect on serum K+ concentration, were approximately the same as those of glycyrrhizin but different from those of saiko saponin isolated from the roots of Bupleurum falcatum LINNE.
Pharmacological properties of 3-(di-2-thienylmethylene)-5-methyl-trans-quinolizidinium bromide (HSR-902), 2-(di-2-thienylmethylene)-5-methyl-trans-quinolizidinium bromide (HSR-911), 3-diphenylmethylene-5-methyl-trans-quinolizidinium bromide (HSR-405) and 2-diphenylmethylene-5-methyl-trans-quinolizidinium bromide (HSR-402), new antispasmodic agents, were compared with those of atropine and butylscopolamine bromide, and the following results were obtained. These antispasmodic agents shifted the dose-response curve of acetylcholine chloride (ACh) in parallel to the right without decreasing the maximal response in isolated guinea pig ileum, and the plots of Schild were found to be linear. The order of potency to antagonize the spontaneous motility of guinea pig ileum was as follows : HSR-911≒atropine>HSR-402>HSR-902»HSR-405> butylscopolamine bromide. The order of potency to antagonize the contraction of rat stomach induced by vagus nerve stimulation was as follows : HSR-911>HSR-402>HSR-902»HSR-405>atropine»butylscopolamine bromide. The order of potency to mydriatic activity of rat or mouse was as follows : atropine>HSR-911≒HSR-402»HSR-405>HSR-902>butylscopolamine bromide. The order of potency to inhibit pilocarpine induced salivation of guinea pig or mouse was as follows : atropine»HSR-911>HSR-402>HSR-902>HSR-405>butylscopolamine bromide. These results indicated that antagonisms of these antispasmodic agents against ACh were competitive, and the selectivity to antispasmodic activity of HSR-902 might be the highest among these antispasmodic agents including atropine and butylscopolamine bromide.
Reaction of acetoacetamide with urea and benzaldehyde gave 5-carbamoyl-6-methyl-4-phenyl-1, 2, 3, 4-tetrahydro-2-pyrimidone. Similarly, acetoacetanilide or 3-aminocrotonamide reacted with urea and aldehydes to give the corresponding tetrahydropyrimidone derivatives.
The working conditions for rapid analysis of 10-hydroxy-Δ2-decenoic acid (I) in royal jelly by means of capillary tube isotachophoresis were studied. The leading electrolyte composed of 0.01 M hydrochloric acid in 30% methanolic solution was adjusted to pH 5 with L-histidine, while the terminal electrolyte of 0.01 M 2 (N-morpholino) ethanesulfonic acid monohydrate in 30% methanolic solution was adjusted to pH 7 with tris (hydroxymethyl) aminomethane. I was determined within 10 minutes without any interference. Both sebacic acid and 10-hydroxydecanoic acid co-existing as side fractions were also identified. This method is simple and fast as well as sensitive and quantitative, eliminating the need of pretreatment of the sample.
A high pressure liquid chromatography (HPLC) by use of ultraviolet absorption detector was examined for the determination of pindolol in the plasma and urine. The pindolol extracted from biological fluid and the disopyramide used as an internal standard, were chromatographed as ion pairs with heptanesulphonic acid by use of Nucleosil 7C18 reverse phase column. The coefficients of variation for the peak height ratio were 2.7% (50 ng/ml) to 4.3% (10 ng/ml) in the plasma and 2.3% (20 μg/ml) to 3.8% (5 μg/ml) in the urine. The detection limits of pindolol were about 2 ng/ml in the plasma and 0.5 μg/ml in the urine. The plasma levels and the urinary excretion rates of pindolol after oral administration of 10 mg pindolol to a healthy male volunteer could be measured by this method. This method is more sensitive and reproducible than the fluorometric method and may be used for a clinical pharmacokinetic study and a bioavailability study of pindolol.
The 1, 3-cycloaddition of diazomethane to α-methylenephenylacetate, apoatropine, aposcopolamine and atropic acid methyl ester, was studied and 3-phenyl-3-alkoxycarbonyl-1-pyrazolines obtained. Alkamine esters of these compounds were rather stable, however, methyl ester of 1-pyrazoline derivative was changed to 1H-2-pyrazoline derivative and 3-phenylpyrazole under basic conditions. And also, methyl ester of 1-pyrazoline derivative was rearranged to 1H-2-pyrazoline derivative in acidic solutions. When chloroform solution of methyl ester of 1-pyrazoline derivative was kept standing at room temperature of heated at 50°, 1-phenyl-1-cyclopropanecarboxylic acid methyl ester was obtained.
The species differences in the metabolism and excretion of tulobuterol (I) were investigated in rat, dog, guinea pig and rabbit after oral administration of 14C-tulobuterol. The urinary metabolites were analyzed by thin-layer chromatography. The urinary excretion of radioactivity within 72 hr. was estimated and about 50% of the administered dose were excreted in the urine of rat, dog and guinea pig, whereas relatively a small amount (24.6% of dose) in the urine of rabbit. The excretion of 14CO2 into expired air within 48 hr. was 19.6% and 35.8% of dose in guinea pig and rabbit, respectively, and these values were larger than 0.8% in rat. Major metabolites excreted in the urine were 3-hydroxy-(III) and 4-hydroxy-5-methoxytulobuterol (IV) in rat, IV and o-chloromandelic acid (VI) in dog, an unchanged drug, 1-(o-chlorophenyl)-1, 2-ethandiol (V) and IV in guinea pig, and VI in rabbit. On the basis of these results it was clarified that there were marked species differences in the metabolism and excretion of I.