4'-Substituted 4-halogenoacetamidodiphenyl ethers (B-type compounds) were synthesized by the application of p-chloronitrobenzene to the potassium salt of the corresponding para-substituted phenols to obtain 4'-substituted 4-nitrodiphenyl ethers (I to V), their reduction to 4-amino compounds (VI to X), and application of the corresponding halogeno (F, Cl, Br) acetyl chloride to obtain the 4'-substituted 4-halogenoacetamidodiphenyl ethers (XI to XXI). 4'-Substituted 4-iodoacetamidodiphenyl ethers (XXII to XXVI) were obtained by the application of patassium iodide to 4'-substituted 4-chloroacetamidodiphenyl ether. 4'-Substituted 4-halogenoacetamidodiphenyl sulfides (C-type compounds) were synthesized similarly from potassium salt of the corresponding para-substituted thiophenols, via 4'-substituted 4-nitrodiphenyl sulfides (XXVII to XXIX). 4'-Alkoxyl substituted compounds (XXX to XXXI) were obtained by alkylation of the sodium salt of the corresponding 4'-hydroxylated compounds with alkyl bromide. Reduction of the 4-nitro compounds (XXVII to XXXI) so obtained to 4-amino compounds (XXXII to XXXIX) and application of the corresponding halogenoacetyl chloride to them finally afforded the objective C-type compounds (XL to LXXXVII). Antibacterial tests of these B-and C-type compounds in vitro showed 4'-methyl-4-iodoacetamidodiphenyl ether (XXII) to be effective. Its secondary antibacterial tests and acute toxicity test(LD60), and skin stimulation tests gave a satisfactory result.
Thermal neutron activation analysis with measurement of 32P Cerenkov radiation by liquid scintillation spectrometer was used to determine phosphorus in cannabis. After irradiation of the sample, wet ashing was carried out with conc. nitric acid and 70% perchloric acid. The solution in 1M perchloric acid transferred to an inorganic ion-exchange column containing acid aluminum oxide and phosphorus was quantitatively eluted with 1M hydrofluoric acid. The 32P radioactivity of each fraction of the eluate was counted with Cerenkov radiation by a liquid scintillation spectrometer from 2 to 7 weeks after the irradiation. The activity curve decayed with 32P half-life. The isotope channel ratio technique was applied for the quench correction. The values for upper and middle leaves, and male flower of cannabis, in parts per million, were 0.55, 0.31, and 0.71, respectively, and a good agreement was found with the values from Molybdenum Blue method. The determination limit of phosphorus by this method was calculated to be 2.5×10-8g. The optimal experimental conditions for chemical separation of phosphorus and for measuring the 32P Cerenkov radiation were also examined.
Activation of adenylate cyclase of rat ascites hepatoma, AH-130, AH-272, AH-66, and AH-7974, by catecholamines, glucagon, and sodium fluoride, and effects of β-adrenergic blocking agents were studied. Basal adenylate cyclase activity of AH-7974 was higher than those of other three tumors as well as of the liver. Adenylate cyclase of AH-130 was activated by epinephrine and isoproterenol more than that of the liver, but those of other tumors showed lower responses to catecholamines. The response of tumor adenylate cyclase to glucagon was very low compared with that of the liver. Sodium fluoride activated all the tumor adenylate cyclases tested. The activation of AH-130 adenylate cyclase by epinephrine was inhibited by β-adrenergic blocking agents such as bufetolol, practolol, propranolol, and alprenolol, but was not blocked by α-adrenergic blocking agents like phentolamine and phenoxybenzamine, and by antitumor alkylating agents, bis(3-methylsulfonyloxypropyl) amine p-toluene sulfonate and nitrogen mustard. The effect of practolol, a selective β1-adrenergic blocking agent, was less than other three agents. These results suggest that AH-130 possesses β2-adrenergic receptor coupled with adenylate cyclase and may be useful as a tissue in place of liver for research on β-adrenergic blocking agents.
The structural elucidation of several unidentified metabolites of 1-butyryl-4-cin-namylpiperazine (I) in mice and guinea pigs was made by the ion cluster technique. A single dose of 100mg/kg of an equimolar mixture (I : I-d5) of 1-butyryl-4-(cinnamyl-arom.-d5)piperazine (I-d5) hydrochloride and I hydrochloride was injected subcutaneously into animals. After the administration of I : I-d5 the urine was collected for 24 hr. An aliquot of the urine sample was extracted with AcOEt at pH 11, the extract was subjected to thin-layer chromatography twice, and the isolated sample was introduced directly into the ion source, and its mass spectrum was recorded. The chemical structures of the metabolites in question were readily determined by the presence of doublet peaks separated by 3-5 mass units, and by the shift of the fragment ions. The possible structures of the unidentified metabolites were elucidated as 1-butyryl-4-(3-methoxy-4-hydroxycinnamyl)piperazine (V), 1-(3-hydroxybutyryl)-4-cinnamylpiperazine or 1-(4-hydroxybutyryl)-4-cinnamylpiperazine, and 1-(3-hydroxybutyryl)-4-(4-hydroxycinnamyl)piperazine or 1-(4-hydroxybutyryl)-4-(4-hydroxycinnamyl) piperazine. Unchanged I, 1-butyryl-4-(4-hydroxycinnamyl)piperazine (II), 1-cinnamylpiperazine (III), and 1-(4-hydroxycinnamyl) piperazine (IV), which had been identified by the inverse isotope dilution analysis, were also detected by this technique.
The effect of primary particle radius (γ0), addition of a polymer [po1y (vinylpyrrolidone), sodium carloxymethyl cellulose)], and centrifugal sedimentation on the sedimentation states of pyrimidine-penicillin G (PYG) were examined by the measurement of apparent aggregation degree (γm/γ0, where γm is the apparent secondary particle radius), porosity (ε) and tortuosity (q) of the sedimentation bed, and the amount of polymers adsorbed on particles. The following results were obtained. 1) γm/γ0 and ε did not change in parallel with each other while q and ε changed in the opposite direction with respect to the polymer concentration. 2) The effect of polymers on ε and difference in q with the direction of measurement were not found in the case of centrifugal sedimentation. 3) Saturated amount of adsorption X∞ of the polymers was higher than the theoretical value which was calculated by assuming monlayer adsorption of the monomers. 4) Plots of the data to see the relation between q and (1-ε) gave straight line. 5) qv/qH decreased with increase in ε at γ0= 1.8, 8.9, and 13.2 μmz, and almost constant against ε at 2.9 and 4.8 μm.
In vitro fertilization of mouse ova was examined with modified KRB buffer as a culture medium. Only the sperm from cauda epididymis and ductus deference was able to penetrate the zona pellucida of mouse ova. while the sperm from caput epididymis did not. At the final sperm concentration of 1.5×105 sperm/ml, ova were highly penetrated. In the high sperm concentration range, fertilization rate were decreased. It was suggested that this decrease was due to the anti-fertilization factor contained in the sperm suspension. Epididymal sperm required 40-50 min before penetration into the ova but preincubation of the sperm reduced this period. The difference of time before penetration between epididymal sperm and preincubated sperm shows the time need for capacitation (35-40min). Change of pH caused difference in the fertilization rate. Fertilization hardly occurred at pH 7.0, due to inhibition of penetration although the sperm were capacitated at pH 7.0.
Effect of ephedrine-isopropylantipyrine (EIA) on exudative inflammations and its physiological mechanisms were examined. When given to rats in a dose of 0.65 mmole/kg, EIA showed stronger inhibitory effect like phenylbutazone than isopropylantipyrine (IA) or ephedrine, on edema produced by carrageenin, formalin, and dextran, and increased vascular permeability induced by histamine and xylene. This compound greatly inhibited the primary edema by carrageenin in a rat, and its inhibitory action was blocked by reserpine and dibenamine. EIA also inhibited prostaglandin synthesis in vitro in 25 μM of ED50, equal to phenylbutazone and more than IA, in proportion to their inhibitory effect on exudative inflammation. Further, this compound promoted the excretion of noradrenaline into rat urine less than ephedrine and increased the level of cyclic AMP in lung and spleen of a rat. Anti-inflammatory action of EIA seems to result from inhibition of prostaglandin synthesis and moderate stimulation of sympathetic nerve system. EIA showed an antitussive effect and its ED50 was 23.1 mg/kg in guinea pigs, and improved the stimulated function of adrenal cortex system in a rat under stress by altera-tion of rhythm in temperature, but did not show a spontaneous motor activity like ephedrine and any effect on adrenal cortex function and blood glucose level of a normal rat.
Two new polyacetylenic compounds, named atractylodinol and acetylatractylodinol, were isolated from the rhizome of Atractylodes lancea DE CANDOLLE var.chinensis KITAMURA (Compositae), together with atractylodin. Their structures were determined from spectroscopic and chemical evidences. They were found to show piscicidal activity against Oryzias latipes TEMMINK et SCHLEGEL, the median tolerance limit (24 hr) of atractylodinol was 1.8 ppm and that of atractylodin, 3.0 ppm.
A fluorometric determination of phenylephrine hydrochloride (II) with formaldehyde, as described in the previous paper, was modified and a highly sensitive method for the determination of formaldehyde using II as an analytical reagent was established, and also the present method was applied to the determination of guaiacol glyceryl ether which produced formaldehyde by HIO4 oxidation. The recommended analytical procedure for formaldehyde was as follows : To 1 ml of HCHO solution (ca. 1μg/ml), 1ml of 0.1N NaOH and 1ml of 0.01% solution of II were added and the mixture was heated for 60 min at 80°, To this solution, 1ml of 2.5N NaOH and 1ml of 0.1% K3Fe(CN)6 were added, and the mixture was heated further for 5min at 80°. When cooled, 1ml of 0.1% ascorbic acid solution was added and the whole was diluted to 10ml with methanol. The fluorescence intensity of this solution was measured at 450nm, excited at 370nm.
Studies were made on the ultraviolet spectral change produced by the intermolecular interaction of acridines (acridine, 1-, 3-, 4-, and 9-aminoacridines, proflavine, Acridine Yellow, and Acridine Orange) and calf thymus deoxyribonucleic acid (DNA). The mixed systems of these acridines and DNA gave a hypochromic change in the ultraviolet region. The observed hypochromisn was quantified in terms of integrated intensity, H. The value of H run parallel with bacteriostatic index of the acridines. Protonation of acridines was favorable for the production of the hypochromic change. Examinations were also made on the mode of the intermolecular interaction from the point of flow dichroism of the interaction system, change in specific viscosity, and change in the melting temperature of DNA by this interaction. It was observed that polyguanylate and polyadenylate gave ultraviolet hypochromism on mixing with acridines, but no spectral change was observed with pyrimidine homopolynucleotides. On the basis of these informations, it was presumed that the modified intercalation model is applicable to the mode of interaction under the present experimental conditions.
The kinetics of the acid-catalyzed hydration of pentazocine (I) was examined in aqueous hydrochloric acid solutions from 3.9×10-2 to 5.0M. I undergoes hydration in aqueous acidic media to form an alcohol derivative corresponding to 1, 2, 3, 4, 5, 6-hexa-hydro-6, 11-dimethyl-3-(3-hydroxy-3-methylbutyl)-2, 6-methano-3-benzazocin-8-ol(II), and its reaction rate follows a pseudo-first-order reaction. Values of log kobs (hr-1 are far from directly proportional to acid concentration (log [HCl]) and instead nearly proportional to the acidity function (H0). Further, participation of water molecule in the transition state (as rate-determining step) was examined by Bunnett w-value, and a linear relation-ship was found between log kobs+H0 and log αH2O with the slope, w, of -1.28. These results suggest that the hydration transition state is I plus a proton, i.e., free carbonium ion. Accordingly, the hydration mechanism is free from firmly bound water molecules in the transition state. Arrhenius parameters were determined by plotting the hydration rate constants against temperature, and activation energy and activation entropy were calculated.
Essential oil of Lamium purpureum L. was investigated and main components of the odor of this plant were found to be alcohol and phenol. Thus far, 1-octen-3-ol, cis-3-hexen-1-ol, phenethyl alcohol, benzyl alcohol, phenol, 0-, m-, and p-cresols, guaiacol, eugenol, 15 kinds of fatty acid, and 21 kinds of paraffin were isolated and identified.
In order to introduce a carbon chain into the 4-position of 1-substituted 1H-pyrazolo-[3, 4-d]pyrimidine, substitution reactions were carried out with 4-chloro-1-methyl-(Im) and 4-chloro-1-phenyl-1H-pyrazolo[3, 4-d]pyrimidine (Ip) with a carbanion. The carbanion sources used were malononitrile, ethyl cyanoacetate, phenylacetonitrile, ethyl acetoace-tate, diethyl malonate, acetone, 2-butanone, 2-pentanone, acetophenone, propiophenone, cyclopentanone, cyclohexanone, and 1, 3-diphenyl-2-propanone. Reaction of I with malononitrile, in the presence of sodium amide, gave 1-methyl(or phenyl)-1H-pyrazolo[3, 4-d]pyrimidine-4-malononitrile (II-1), while the reaction of I with ethyl cyanoacetate, phenylacetonitrile, or diethyl malonate gave the corresponding derivatives (II-2, -3, -5). In the reaction of I with ethyl acetoacetate, the deacetylated product, ethyl 1-methyl(or phenyl)-1H-pyrazolo[3, 4-d]pyrimidine-4-acetate (II-4) was formed. Reaction of Ip with the ketones gave (1-phenyl-1H-pyrazolo[3, 4-d]pyrimidin-4-yl)methyl alkyl ketones (IIp-6 to-13). Tautomerism of the compounds (II) obtained by the present reaction was discussed from infrared, ultraviolet, and nuclear magnetic resonance spectral data.
Equilibria between copper (II) and acetate in N, N-dimethylformamide are written as follows : [Chemical formula] where Ac- denotes the acetate anion. Cupric acetate was found to exist primarily as dimeric species in the solution from electron spin resonance data. Equilibrium constants are calculated as log K=3.79, log K'=0.82 from paramagnetic susceptibilities of solutions determined by nuclear magnetic resonance spectra. From absorption at 704 nm, these are calculated as log K=3.48, log K'=0.31. Though an absorption band at 370 nm has been assigned to the dimeric copper(II) species, monomeric acetate-coordinated species was found to show a considerable absorbance in this region.
A strain of Scopulariopsis brevicaulis was cultured in a nutrient medium (primary culture) and the culture medium was changed with a medium consisting of uric acid and dipotassium hydrogen phosphate at various stages of growth (logarithmic, stationary, and autolytic) to continue cultivation (secondary culture). In the primary culture, maximum formation of uricase was found in the latter part of the logarithmic stage. In the secondary culture, almost constant uricase activity was seen after 24-48 hr of culture, irrespective of the amount of uricase present during the stage of fungal growth and before the start of secondary culture. In the cells of the logarithmic stage, this constant value of uricase activity was reached after 6 hr, indicating a significantly rapid adaptation of the cells in this stage to induction and formation of uricase, compared to cells in other stages of growth. Thus, difference in induction and formation of uricase according to different growth stages appeared markedly. The constant value of uricase activity found in the secondary culture was not greater than that of the maximum value in primary culture. It was concluded from the present series of experiments that, in practice, uricase production by this strain will be more efficient by the primary culture as reported previously4) than by the use of a secondary culture as in this work.
A simple quantitative determination of ethanol in five samples of tinctures was attempted by gas chromatography using the head-space technique. The apparatus used were two models of gas chromatograph, one of which was equipped with a hydrogen flame ionization detector and the other with a thermal conductivity detector. Column packings were 5% polyethyleneglycol 20M (PEG 20M)/Chamelite or 5% Reoplex 400/Chromosorb in case of the former detector and 5% PEG 20 M/Chamelite in the latter. Gas chromatograms derived from the untreated tinctures did not present any peak except that of ethanol using the former detector, while low level signals of air and moisture were observed with the latter detector. When propanol was used as an internal standard, ethanol content in tinctures was capable of being calculated by Eq. (3). The results obtained resembled the values determined by the conventional three methods of JP 9, alcoholmeter, and dichromate oxidation.
Application of sodium p-toluenesulfinate (II) to 6-chloro-9-phenyl-9H-purine (Ia), 4-chlorocinnoline (Ib), 4-chloroquinazoline (Ic), 2-chloroquinoline (Id), and 1-chloroiso-quinoline (Ie), in dimethyl sulfoxide or dimethylformamide, resulted in the formation of the corresponding sulfones in a good yield; 9-phenyl-6-(p-tolylsulfonyl)-9H-purine (IIIa), 4-(p-tolylsulfonyl)cinnoline (IIIb), 4-(p-tolylsulfonyl)quinazoline (IIIc), 2-(p-tolylsulfonyl)quinoline (IIId), and 1-(p-tolylsulfonyl)isoquinoline (IIIe). Application of potassium cyanide to IIIa and 6-(methylsulfonyl)-9-phenyl-9H-purine (III'a) in dimethyl sulfoxide gave 9-phenyl-9H-purine-6-carbonitrile (VIa) from both in a good yield. In order to simplify the reaction procedure, the above reaction was carried out and, without isolation of IIIa, IIIb, and IIIc, potassium cyanide was added to their reaction system, and VIa, 4-cinnolinecarbonitrile (VIb), and 4-quinazolinecarbonitrile were obtained. These reactions are considered to be applicable for the syntheses of sulfones and nitriles of other ring systems.
Alkaloidal constituents of Cephalotaxus wilsoniana HAY. collected in Formosa were examined, and 3-epi-schelhammericine (7), Base VI (8), and 3-epi-wilsonine (11) of homoerythrina alkaloids, and cephalotaxine (2), acetylcephalotaxine (12), isoharringtonine (3), and Alkaloid G(5) of the cephalotaxus alkaloids, were isolated and characterized.