The role of electroorganic chemistry (EOC) as a tool for the synthesis of bioactive compounds was described. The generation of active species in the EOC, and some chemical characters of the species were explained in the first place. In the next place, the use of these species in the synthesis of some bioactive compounds was described.
Two new sterol glycosides, mp 270.5-275.5°C (dec.), [α]D-42.4°, C39H64O14 (I) and mp 214-217°C (dec.), [α]D-58.3°, C45H74O18 (II), were obtained from aerial parts of Fritillaria thunbergii MIQ. On the basis of the results of spectral and chemical investigations, I and II were characterized as 3-O-β-D-glucopyranosyl-6, 22-dioxo-5α-cholestan-3β, 26-diol 26-O-β-L-glucopyranoside and 3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl-6, 22-dioxo-5α-cholestan-3β, 26-diol 26-O-β-D-glucopyranoside, respectively.
A mixture of stigmasterol, campesterol, β-sitosterol (1) and its 3-O-β-D-glucoside (2), succinic acid (3), thymidine [obtained in the form of diacetate (4)] were isolated from the fresh bulbs of Fritillaria thunbergii MIQ. (Liliaceae). The crude drug "Bai-mo", which was prepared from the above bulbs by treatment with lime followed by bleaching in the sun, provided 5α, 6β-dihydroxy-cholestan derivatives corresponding to 1 and 2, together with 1, 2, 3, and 4.
Amino compounds of diphenyl ether (or sulfide) and naphthyl ether were condensed with 2, 4-(or 3, 4-) dichlorophenyl isocyanate in benzene to form their carbanilides (1-25). Amino compounds of naphthyl ether were thiocyanated with ammonium thiocyanate and bromine in acetic acid to form their thiocyanato compounds (26-29). Amino compounds of naphthyl ether and diphenylamine (or sulfide) were condensed with salicylic acid or 5-chlorosalicylic acid using phosphorus trichloride in xylene to form their salicylanilide (30-37). Antimicrobial tests of the synthesized compounds showed that carbanilides (10, 12, 13, 20, 21) had a strong antimicrobial activity against Staphylococcus aureus.
The structures of 22-hydroxy furostanol bisglycosides, T-e (2), T-j (3) and T-h (4), which are recognized as real substances of "nolonin"and correspond to proto-type of pennogenin glycosides, have been substantiated by 13C nuclear magnetic resonance spectra. Moreover, two new sapogenols, G-I and G-II, have been isolated from the hydrolysate by using H2SO4 in 50% EtOH of the MeOH extractives of the underground parts of Trillium kamtschaticum PALL. and their structures could be deduced as the formulae 8 and 9, respectively.
An improved column fractionation procedure was developed for the analysis of biological samples. According to the simulation of the solute distribution in two phases for various extraction models, a multi-step liquid-liquid partition technique employing packed columns was found more effective than the usual funnel separation-batch method and the droplet current distribution procedure. As an alternative to the open-bed extraction column used for the pre-treatment of the biological fluids, a closed-bed column system was introduced through the incorporation of a ultraviolet monitor. Drugs and their metabolites in urine were extracted quantitatively using diethyl ether as a carrier solvent. The aqueous liquid-liquid distribution column having a high theoretical plate number was prepared using diatomaceous earth consisting of small size particles and applying a slurry packing procedure.
A method for high sensitivity analysis of D-penicillamine in the blood was developed by use of a high-performance liquid chromatography with twin-electrode voltammetry detector. This method enabled us to determine low concentration of D-penicillamine in the human blood (detection limit 20 ng/ml) after oral administration (100 mg/ml).
The thiobarbituric acid (TBA) method of lipid hydroperoxide requires Fe catalyst to decompose the hydroperoxide to TBA-reactive secondary products during the reaction. In order to characterize the role of Fe catalyst, FeCl3, ferritin, hemin, defatted tissue homogenate and whole tissue homogenate, were employed for the reaction of methyl linoleate hydroperoxide (MLHPO) with TBA. The results are as follows : 1) Non-heme iron (FeCl3, ferritin) could play as an effective catalyst at a wide range of concentration, while hemin in high concentration suppressed the color intensity of optimum condition. The maximum efficiency of Fe catalyst existing in tissue homogenate lies between non-heme iron and hemin. 2) The TBA reaction catalyzed by FeCl3 or ferritin was inhibited by EDTA, but not by BHT. The reverse effect of BHT and EDTA was observed in hemin or tissue homogenate catalysts. 3) The TBA reaction catalyzed with fresh FeCl3 proceeded regardless the absence of oxygen, while the TBA value obtained with aged FeCl3, ferritin, hemin or tissue homogenate was significantly depressed under anaerobic conditions. 4) The catalytic activity in rat liver tissue was distributed similarly among all subcellular fractions, but 105000×g pellet showed the highest effect per unit of protein.
Administrations of loop diuretic agents, furosemide and piretanide from the femoral vein and portal vein infusion of several doses to rats were carried out, and then, plasma concentrations and cumulative amounts excreted in the urine and bile of the drug were determined. The relationship between doses and the area under the plasma concentration-time curves (AUC) after intravenous administrations was nonlinear over the dose range from 10 to 30 mg/kg. Active renal excretion process was also supported, and then, it was observed that AUC's after intraportal infusion of each drug varied in accordance with alteration of infusion time in a dose of 10 mg/kg.
The pharmacokinetics of deslanoside was studied in four healthy volunteers and five patients with cardiac failure. It was found that the homogeneous enzyme immunoassay (EMIT[○!R]) for digoxin was able to apply for the determination of deslanoside serum levels. The curve of the average serum levels of deslanoside in four healthy volunteers after intravenous administration was typically adapted to two compartment model, and a half life time (t1/2) and an apparent volume of distribution (Vd) were calculated to be 41 hours and 10 l/kg, while t1/2 and Vd in the cases of five patients were 61 hours and 6.4 l/kg on average, respectively. An equation of β=0.69×10-4Cer+0.0089 (γ=0.69) was obtained in an investigation to know a relationship between elimination rate constant (β) and creatinine clearance (Cer), indicating good correlation between renal function and urinary excretion. Metabolism of deslanoside to digoxin was examined in analysis of urine collected during 24 hours after administration of three healthy volunteers by means of a combination of high performance liquid chromatography and homogeneous enzyme immunoassay, and convertion ratio of deslanoside to digoxin was found to be 4-7%. Toxic serum level of deslanoside was able to be defined as 2.0-2.5 ng/ml by 22 measurements of deslanoside serum levels in 12 patients with cardiac failure.
The simplified method for the determination of phenytoin (DPH), phenobarbital (PHB), carbamazepine (CBZ), primidone (PRM), valproic acid (VPA), and ethosuximide (ES) was established. In this method, the serum containing these drugs was acidified and extracted with chloroform containing hexobarbital and cyclohexanecarboxylic acid as internal standards. Then, some aliqout of the extract was evaporated to dryness in vacuo, and DPH, PHB, CBZ, and PRM were simultaneously determined by a high-performance liquid chromatography. While, isoamylacetate was added to the other aliqout of the extract, and chloroform was evaporated in vacuo, then VPA and ES were simultaneously determined by a gas-liquid chromatography. These relatively simple procedures gave enough accuracy to determine the serum levels of these drugs in epileptic patients with multipledrug administration. Some cases of the antiepileptic medication were also described in this report. In an epileptic patient, the ratio of cerebrospinal/serum levels of PHB was increased by coadministration of acetazolamide.
Physical dependence on orally administered loperamide hydrochloride (loperamide) was investigated in comparison with codeine phosphate (codeine) in mice and rats. Jumpings were produced in 50% or more of mice used by an intraperitoneal injection of nalorphine after the repeated administration of 60 mg/kg/2 d or more of loperamide and of 399 mg/kg/2 d or more of codeine, but in 10% after 10.5 to 45 mg/kg/2 d of loperamide. Dosing of 50 mg/kg/d of loperamide resulted in a partial substitution for morphine in morphine physically dependent rats, but dosing of 5 and 10 mg/kg/d did not produce significant activity. Codeine (s.c.) showed complete substitution activity for morphine. Significant withdrawal signs were not observed after abrupt withdrawal of loperamide or after the administration of narcotic antagonist instead of loperamide in the rat given chronic administration of loperamide at maintenance doses of 2.5 and 5 mg/kg×2/d, though the high dose (10 mg/kg×2/d) of loperamide as well as codeine (40 mg/kg×2/d) caused the definite withdrawal signs. Its withdrawal signs, however, were partly different from that in the rat given with codeine. Serum loperamide levels were 1336 and 155 ng/ml after repeated dosing of 60 mg/kg/2 d to mice and 10 mg/kg×2/d to rats, respectively. These levels in mice and rats were approximately 136 and 263 times, respectively, that after the single oral administration of the dose (0.1 and 1 mg/kg) effective in preventing diarrhea induced by castor oil. From these results, it was suggested that loperamide produced definite physical dependence similar to codeine if its high dose was given repeatedly to mice and rats, and that there was a large difference between the doses producing physical dependence and anti-diarrheal action.
A new secoiridoid glucoside has been isolated together with oleuropein from the fruits of Ligustrum obtusifolium SIEB. et ZUCC. Its structure was elucidated to be 10-hydroxyoleuropein on the basis of the spectroscopic and chemical evidence. p-Hydroxyphenethyl β-D-glucoside and mannitol were also identified from the methanolic extract.
From dried aerial parts of Youngia denticulata, paraffins (A), higher fatty acid esters of hexacosanol, β-amyrin, α-amyrin and taraxasterol (B), acetic acid esters of germanicol, lupeol and taraxasterol (C), and hexacosanol (D) were proved to be found. Fractions B and C were supposed to be main components of milky juice.
A high performance liquid chromatographic method for the determination of lnsulin in preparations is described. Insulin was eluted on Nucleosil 5CN as packing material with acetonitrile-water-acetic acid (33 : 67 : 2) containing sodium octanesulfonate as mobile phase. The assay results of 7 preparations was 37.9-41.2 IU/ml for these indicated potency of 40 IU/ml and coefficient of variations were 0.85-2.10% (n=3).
Pentafluorobenzyloxylamine hydrochloride (PFBOA) is an excellent reagent used for the syntheses of some derivatives in gas chromatography of water-soluble low molecular weight carbonyl compounds. To a sample solution (3.0 ml) containing 5-50μg of guaiacol glyceryl ether (GGE) 0.5 ml of 0.01 M NaIO4 was added and the solution was oxidized on standing at room temperature for 10 minutes. After decomposing excess NaIO4 by 0.1 N Na2S2O3, HCHO produced reacted with 0.5 ml of PFBOA aqueous solution (40 mg/ml) on standing at room temperature for 60 minutes. One drop of 18 N sulfuric acid saturated with sodium chloride was further added to this solution, which was extracted with 0.3 ml of hexane containing an internal standard (IS) and applied to gas chromatography. Indobenzene (30μg/ml) and allethrin (25μg/ml) were well suited as internal standards for the determination of HCHO and methoxyphen-oxyacetaldehyde (MPA), respectively, which were both synthesized by the oxidative reaction. Gas chromatography was carried out on a glass column of 3% XE-60, 2.0 m, FID and the column temperatures for HCHO and MPA were 80°C and 190°C, respectively.
Saponins from Panax ginseng were studied by thin layer chromatography (TLC) using an NH2-chemically bonded stationary phase, prepared from pre-coated silica gel plate treated with benzene solution containing 3-aminopropyltriethoxysilane (3APTS). TLC plate treated with 3APTS was applied to the evaluation of the commercial "Ginseng Extract". After developement, 3APTS-treated TLC plates were dried, sprayed with sulfuric acid solution, and followed by heating. Each saponin spot on the chromatogram was measured with a TLC densitometer equipped with a dual-wave length TLC scanner at λs 525 nm (sample) and λR 760 nm (reference).