In order to obtain ganglionic blocking agent, dialkylaminoalkyl isoketopinates (I to XIII) were prepared and derived to their quaternary ammonium salts (XIV to XXVII). N-Isoketopinoyl-ω-dialkylaminoalkylamines (XXVIII to XXXII) were also prepared and derived to their quaternary ammonium salts (XXXIII to XXXVII). Of these compounds, hypotensive effect was found in (III), (XIV) to (XXI), (XXXVI), and (XXXVII), and the intensity and duration of the compounds (XVII) to (XXI) were comparable to those of Hexamethonium.
In order to obtain ganglionic blocking agent, 9-oxocamphor oxime was reduced to 9-aminocamphor which was condensed with dialkylaminoalkyl chloride to form d-9-(dialkylaminoalkylamino) camphors (I to IV), which were further derived to their respective quaternary ammonium salts (VIII to XIII). 9-Aminocamphor was derived to 9-(chloroacetamido) camphor and 9-(chloropropionamido) camphor, which were reacted with secondary amines to form d-9-(dialkylaminoacylamino) camphors (XIV to XVIII). Quaternary ammonium salts (XIX to XXIII) of these compounds were also synthesized. Hypotensive effect was found in compounds (X) to (XII), (XXI), and (XXII).
In order to obtain ganglionic blocking agent, 9-aminocamphor was condensed with monochloroacetic acid ester to form ethyl 9-camphorylaminoacetate (I) and with acrylic acid ester to form ethyl 3-(9-camphorylamino) propionate (II). (I) and (II) were respectively methylated to form ethyl ω-[N-(9-camphoryl) methylamino] alkanecarboxylates (III and IV) which were reacted with dialkylaminoalcohol to effect ester exchange and dialkylaminoalkyl d-9-camphorylaminoalkanecarboxylates (V to XIV). These were derived to their quaternary ammonium salts. Of these compounds synthesized, (XV) to (XX), and (XXII) to (XXIV) were found to have hypotensive action.
In order to obtain ganglionic blocking agent, d-9-aminocamphor was methylated to form d-9-dimethylaminocamphor (I) or reacted with 1, 4-dibromobutane to form 1-(d-9-camphoryl) pyrrolidine (II). The reaction of d-9-bromocamphor and secondary amines afforded 9-dialkylaminocamphor (III to V), while condensation of (I), (II), and (III) respectively with 2-dialkylaminoethyl chloride afforded d-3-(2-dialkylaminoethyl)-9-dialkylaminocamphor (VI to IX), which were derived to their quaternary ammonium salts (X to XIII) by reaction with methyl iodide. Hypotensive effect was found in the compounds (X) to (XIII).
As a means of utilizing 3, 3′-dihydroxydiphenylamine in the distillation residue of m-aminophenol, the amine or its methylated 3, 3′-dimethoxydiphenylamine was derived to 2, 8-dihydroxyphenothiazine or 2, 8-dimethoxyphenothiazine by fusion reaction with sulfur and anthelmintic action of these compounds was compared with that of phenothiazine and piperazine phosphate. 2, 8-Dihydroxyphenothiazine was so unstable that it was derived to its triacetate. The amino group in these diphenylamines was reacted with alkylaminoalkyl chloride with sodium amide or sodium hydride as the dehydrochlorination agent and 3, 3′-dimethoxydiphenylamine and 2, 8-dimethoxyphenothiazine, substituted with diethylaminoethyl, dimethylaminoethyl, diethylaminopropyl, and 1-methyl-3-piperidylmethyl groups were prepared. Antihistamine action of these derivatives was compared with that of chlorpromazine and Anergen.
Some of 2-alkoxy-4-aminobenzoic acid, 4-alkylaminosalicylic acid, and 2-alkoxy-4-nitrobenzoic acid (alkyl=methyl, butyl, octyl, dodecyl, hexadecyl) were prepared and their bacteriostatic action against tubercle bacilli (human type) was tested in Kirchner medium. In these three compound series, it was found, as in other alkyl compounds already reported, that antibacterial activity increased with the number of carbon atoms in the alkyl, reached the maximum at around C12, and decreased again over that number. The dodecyl derivatives of not only PAS but also of 4-nitrosalicylic acid, which by itself showed no remarkable activity, were as strongly antituberculous as PAS. This fact seems to suggest that, in some cases, even non-active compound may aquire strong antitubercular activity by introduction of alkyl group into it. The order of activity was: 4-alkylaminosalicylic acid>2-alkoxy-4-aminobenzoic acid>2-alkoxy-4-nitrobenzoic acid.
Tetracycline-metal chelates were prepared with Th4+, Zr4+, Fe3+, Cu2+, Al3+, Co2+, Mn2+, and UO22+. Their properties, especially their solubility, and method of preparation are described. Tetracycline-Al chelate has a tendency to change rapidly into anhydrotetracycline-Al chelate when it is wet. This change is delayed on the storage of tetracycline-Al chelate shut off from air oxygen. The reason is not fully understood. The chelates isolated are as follows: Tetracycline-Th4+, yellow-orange powder; tetracycline-Zr4+, yellow-orange vitreous mass; tetracycline-Al3+, yellow-orange powder which gradually changes to red; tetracycline-Cu2+, yellow-brown powder; tetracycline-Co2+, yellow-brown powder; tetracycline-Mn2+, brown-yellow powder; tetracycline-UO22+, red vitreous mass; tetracycline-Fe3+, black-brown powder. Infrared spectra of tetracycline-Th and -Zr indicated disappearence of carbonyl of C-11 in the tetracycline molecule, and the presence of metal chelate in C=O of C-11, which previously had been suggested by the present anthors.
Some experiments were carried out to make chemical confirmation of the proposed structures for thalicberine (I) and O-methylthalicberine (II). Primary cleavage reaction of (I) with sodium in liquid ammonia afforded a dauricine-type base (VIII) which was ethylated and second cleavage reaction of this O-ethylated base (IX) with sodium in liquid ammonia finally afforded the two cleaved bases, d-1-(4-ethoxybenzyl)-2-methyl-6, 7-dimethoxy-1, 2, 3, 4-tetrahydroisoquinoline (X) and d-1-(4-hydroxybenzyl)-2-methyl-6-ethoxy-7-methoxy-1, 2, 3, 4-tetrahydroisoquinoline (XII), in almost quantitative yield. These reactions prove the presence of an ether-oxygen binding the two isoquinoline system in a diphenyl-ether linkage and the linkage is in 6-position of the isoquinoline ring. The steric configuration (optical rotation) of the two asymmetric centers in this molecule is (+, +). This has given basis for the assumption that the chemical structure of thalicberine and O-methylthalicberine is a new, specific type, represented respectively by (I) and (II), differing from the oxyacanthine-berbamine type of biscoclaurine-type bases in the linkage of ether-oxygen in the isoquinoline portion.
In continuation of earlier works, amino acids constituting the homogeneous callicrein obtained and purified from hog pancreas were examined through paper chromatography. Hydrolysis of callicrein (Product L, 570U/mg.) with hydrochloric acid afforded 15 kinds of amino acid, viz. cystine, aspartic acid, glutamic acid, serine, glycine, threonine, tyrosine, alanine, proline, valine, leucine, phenylalanine, arginine, histidine, and lysine. Determination of these amino acids was effected by hydrolysis of callicrein with hydrochloric acid, followed by the dinitrophenylation method of Sanger, Schroeder, and Levy, and a two-dimensional paper chromatography. For the determination of tryptophan, which is likely to be destroyed by acid hydrolysis, colorimetry with p-dimethylaminobenzaldehyde by the method of Spies and Chamber was carried out. Content of each amino acid is listed in Table I. For the detection of amine-terminal amino acid, dinitrophenylation and two-dimensional paper chromatography were carried out and it was identified as phenylalanine while the carbonyl-terminal amino acid was found to be alanine alone through Akabori's hydrazone decomposition method.
Callicrein contains sugar and amino sugar, comprising 9.2% of galactose and 2.9% of glucosamine, does not contain phosphorus, and its elementary analyses gave the values of C, 47.79, H, 7.00, N, 10.93, S, 0.69, and O, 33.59%. It is a kind of saccharoprotein.
The substance which accelerates heart contraction was extracted from bovine and whale myocardium, as reported previously, and its purification by column chromatography over alumina was carried out. The fraction (AL) of highest unit, 2700U/mg., was thereby obtained and this showed absorption maximum at 260mμ and minimum at 228mμ in its ultraviolet spectrum.
An amorphous or crystalline precipitate separates out almost immediately upon addition of ethanolic or dioxane solution of diiodo-β-resorcylic acid, diiodosalicylic acid, or methyl diiodo-β-resorcylate to aqueous or organic solvent solution of organic bases with nitrogen-containing heterocyclic system. This reaction was examined with respect to 76 substances of 28 kinds of heterocyclic nitrogenous bases and those containing quinoline, isoquinoline, quinoline and quinuclidine, hydroindolizine and hydrocarbazole, acridine, and phenazine ring showed detectable reaction. Sulpyrine, which has a pyrazole ring, also formed a precipitate by reaction with methyl diiodo-β-resorcylate in dilute dioxane solution. Optimal conditions and detectable limit of the reaction were also examined.
3-Oxo-2, 3-dihydro-1H-pyrido[3, 2, 1-k, l]phenothiazine derivatives obtained by cyclization of 3-(10-phenothiazinyl) propionic acid derivatives and were converted to 3-amino-2, 3-dihydro-1H-pyrido[3, 2, 1-k, l]phenothiazine derivatives. In order to determine the position of substituents in these derivatives, infrared absorption spectra of phenothiazine and pyrido-phenothiazine derivatives in the region of 700-900cm-1 were examined and it was found that the wave number of CH out-of-plane vibration absorption was useful enough in assuming the position of a substituent. The absorption of adjacent four, three, and two hydrogens and isolated hydrogen appears respectively at 764-732, 792-760 and 746-718, 833-797, and 891-825cm-1. By the use of the wave number of this CH out-of-plane deformation vibration, structure of pyrido-phenothiazine derivatives was determined. Consequently, it was found that in 3-(2-chloro-10-phenothiazinyl) propionic acid, in which a strong electron-attracting chlorine atom is substituted, cyclization occurs chiefly in the non-substituted benzene ring while in the methoxyl derivative in which a strong electron-releasing methoxyl group is substituted, the cyclization occurs in the substituted benzene ring carrying the methoxyl.
3-Oxo-2, 3-dihydro-1H-pyrido[3, 2, 1-k, l]phenothiazine was derived to 3-alkyl (or aryl)-3-hydroxy-2, 3-dihydro-1H-pyrido[3, 2, 1-k, l]phenothiazine and further to their dimethylcarbamate. Heating of these 3-alkyl (or aryl) derivatives with hydrochloric acid in glacial acetic acid results in dehydrogenation reaction with dehydration and phenothiazinium salts were obtained. This pyrido-phenothiazinium salts produced by the action of alkali a kind of pseudo base, 1-hydroxy-3-alkyl (or aryl)-1H-pyrido-[3, 2, 1-k, l]phenothiazines and the pseudo base easily formed the ether by the action of alcohol, and regenerated the pyrido-phenothiazinium saits by the action of acid.
Pearl barley decoction has long been used as a home remedy for tuberculosis, hemoptysis, and tumor, and is well known as being effective for removing wart, its clinical effect having been proved by Sakurane. However, no detailed examinations have been made on its pharmacological action and, therefore, experiments were made on the effect of pearl barley oil on relaxation of muscles, blood sugar and serum calcium. Pearl barley oil and saturated fatty acids of 10-18 carbons act on skeletal muscles and damage their contractibility, while oleic acid, one of unsaturated fatty acids, has no effect. The amount of blood sugar and serum calcium decreases by the effect of pearl barley oil and fatty acids of over 12 carbon atoms, but not by ethyl caprate. The lowering of blood sugar is antagonized by pyruvic acid. There was no significant change in the amount of inorganic phosphate in the serum.
Process for preparation of 6α-methyl-4-androstene-3, 17-dione from 5α, 6-epoxyandro-stane-3β, 17β-diol was reexamined and a substance (XI) of m.p. 145-147°, [α]D26: -55°, was obtained, differing from 6β-methylandrostane-3β, 5α, 17β-triol (XI) obtained by Madaeva, Campbell, Ringold, and Ackroyd. [M]D value of this substance agreed with the assumed value and it was proved to be (XI) from analytical values of its 3, 17-diacetate (XII) and of 6β-methyl-5α-hydroxyandrostane-3, 17-dione (XIII) obtained by oxidation of (XI). 6α-Methyl-17α-hydroxyprogesterone (I) was prepared from 17α-hydroxypregnenolone (III). (III) was derived to 20-ethylene ketal (IV) by ethylene glycol, then to 5α, 6-epoxide (V) by the action of perphthalic acid, and (V) was cleaved with methyl magnesium bromide to 6β-methyl-3β, 5α, 17α-trihydroxypregnan-20-one 20-ethylene ketal (VI). Oxidation of (VI) with chromium trioxide-pyridine gave 6β-methyl-5α, 17α-dihydroxypregnane-3, 20-dione 20-ethylene ketal (VII) whose hydrolysis with dil. sulfuric acid finally afforded 6β-methyl-5α, 17α-dihydroxypregnane-3, 20-dione (VIII) which was found to be identical with the substance prepared by the process of Babcock with modification of Bowers and others.
In 1954, Junkmann found that in spite of the fact that 17α-hydroxyprogesterone (I) had almost no progestational activity, its ester showed such activity, but why such activity should appear by acylation had remained obscure to date. The present writer had revealed in the preceding paper of this series that acylation of 17α-OH in (I) had rendered 20-CO inactive and resistant to reduction. It was assumed that, if CO group in 20-position of (I), having free OH in 17α-position, is more easily reduced in the body than that of progesterone (II), then this might be the reason why (I) does not show progestational activity. Attempt was therefore made to prove this by reduction of (I) and (II) under the same conditions and to determine the amount of unreacted material from the ultraviolet absorbancy coefficient of 20-CO, but the measurement of 20-CO was difficult due to the strong effect of the absorption of α, β-unsaturated ketone. Therefore, 17α-hydroxypregnenolone (V) and pregnenolone (VI), which are in the same relationship with respect to the 17-position as (I) and (II), were reduced with sodium borohydride under identical conditions and it was proved that (V) is more easily reduced than (VI), as had been assumed. (I) and its caproate (IV) were administered to normal female rabbit and determination of pregnanetriol (III) excreted in the urine suggested that hydrolysis of (IV) in vivo hardly took place and that (IV) has progestational activity in the state of an ester.
The substance which retards heart function, contained in the aqueous extract of bovine myocardium, contains 6.5% of nitrogen, is positive to the ninhydrin, Dragendorff, and diazo reactions, and negative to biuret, orcinol, and nitroprusside reactions. It colors in the Jaffe reaction when heated, fluorescences under ultraviolet rays, but shows no absorption maximum (in water) in its ultraviolet spectrum. It was found that the composite effective principle of this substance is composed of three substances, anserine, a substance which forms a picrate of m.p. 220-223° (decomp.), and a substance which is adsorbed on talc column and colors pink to ninhydrin.
A polypeptide composed of 12 kinds of amino acid and thought to be a unity in ultracentrifugal property was isolated from bovine myocardium. This substance depresses blood pressure and increases the amplitude of heart contraction. Its isoelectric point is at pH 5.5 and it has ultraviolet absorption maximum at 275mμ. Its sedimentation constant, S, is 2.615×10-13, it contains a saccharide, and is analyzed to 14.64% nitrogen and 1.04% phosphorus. This substance is considered to be a callicrein-like substance contained in the bovine myocardium. A similar substance was also obtained from hog myocardium.
Administration of the substance which retards heart function, isolated from myocardium, and anserine, one of its components, to rat decreased incorporation of 32P into myocardium from labeled phosphorus compound administered at the same time. On the other hand, administration of heart callicrein, which accelerates heart function, increases incorporation of 32P. In this case, effect of various substances extracted from myocardium on incorporation of 32P into rat myocardium is observed by percentage incorporation or specific activity, but is detected the best by analysis of covariance of percentage incorporation and heart weight. This is also significantly affected by the period after administration and rat weight. However, primary regression of dose-response curve of the effective substance was not recognized under the present conditions.
In order to examine whether dibasic organic acids bonded to an aglycone would play the same role as the sugar moiety in cardiotonic glycosides, 3-O-succinoyldigitoxigenin (III), 16-O-succinoylgitoxigenin (VII), 3-O-succinoyloleandrigenin (VIII) 3-O-phthaloyldigitoxigenin (IV) and 3-O-phthaloyloleandrigenin (IX) were prepared. Considering the lethal dose tested with pigeons, succinoylation failed to increase accumulation and toxicity. Phthaloylation resulted in decrease of solubility and marked decrease in lethal dose.
Reaction conditions for the formation of monomethylol (II) and dimethylol (III) compounds by heating 2, 5-lutidine (I) and formaldehyde were examined. The monomethylol compound (II) was derived to the dimethylol compound (III), which formed a monoacetate (IV) (yield, 78.1%) and a diacetate (V) (yield, 6.3%) on heating with acetic anhydride. The acetate (IV) was reacted with ethyl malonate in the presence of sodium ethoxide and the product was catalytically reduced to the malonate derivative (VIII). On the other hand, the ester (IX) was reduced with lithium aluminium hydride, chlorinated with thionyl chloride, and reacted with ethyl malonate in the presence of sodium ethoxide to form the objective (VIII). The picrates of the ester (VIII) obtained by both routes were found by mixed fusion to be identical.
3, 3-Dimethylvaleric acid, homologous to (+)-3-methylvaleric acid, which is considered to be one of the aromatic components of tobacco leaf, and 12 of its esters were prepared. Some of lower esters of this series were found to have an aroma and the odor tended to weaken in higher esters. The boiling point and refractive index of these esters became higher with increasing molecular weight but esters with branched-chain alkyl group did not follow this rule. These compounds and the 3, 3-dimethylated, branched fatty acids (C7H14O2 to C23H46O2) reported previously were tested in Dubos medium for growth-inhibitory action against human-type tubercle bacilli, H37RV strain. Higher fatty acids above C17H34O2 (3, 3-dimethyl-pentadecanoic acid) showed such activity in 320, 000 dilution and approximately the same result was obtained with bovine-type tubercle bacilli, B15 strain. In Kirchner medium, none of the compounds showed any antibacterial activity. Derivatives of 3, 3, 6, 6-tetramethylsuberic acid and 3, 3-dimethylisooctanoic acid were prepared.
Oxidation of trachelogenin with potassium permanganate affords methyl 3-methoxy-4-hydroxyphenylacetate and an acid substance of m.p. 163°, which was identified with 3-methoxy-4-(3, 4-dimethoxyphenyl) butyric acid synthesized in the present series of experiments. The proposed structure of trachelogenin enabled rational explanation to be made of the formation of the above two substances and the structure was further clarified.
Physical constants of the purified and homogeneous callicrein (Product L, 570U/mg.) extracted from hog pancreas were measured in order to find its molecular weight. Sedimentation constant by ultracentrifugal analysis was 2.85×10-13and diffusion constant was 8.39×10-7cm2/sec. from which the molecular weight was calculated as 33, 000. Absorption maximum of this Product L was found to be at 281mμ, same as that in the Product G (220U/mg.). Amine-terminal amino acid was examined through phenyl isothiocyanate by the dinitrophenylation method and was found to be phenyl-alanine. The fact that this callicrein product did not lose potency by trypsin treatment was reported earlier and the variation in ultraviolet absorption, paper electrophoretic migration, and paper chromatographic movement were examined.
In order to examine relationship between the structure and Rf value of steroids, various steroids were separated by the reverse-phase chromatography using liquid paraffin as the stationary phase and developing with a polar solvent. It was thereby found from Rf values of stanols (Table I) and stanones (Table II) that the effect of the A/B ring (configuration of H at C-5) of a steroid ring on the polarity of the molecule was greater in the coprostane type (A/B cis) than in cholestane type (A/B trans). It was also found from Rf values of 3β-stenols [cholestenols (Table III) and ergostenols (Table IV)] that the effect of a double bond in a steroid ring on the polarity of a molecule decreases in the order of the position of the double bond at 14, 8 (14), 4 or 7, and 5.
Triterpenoids were submitted to various kinds of paper chromatography and relationship between Rf value and chemical structure was examined. 1) Triterpenoids were separated by paper chromatography with water absorbed in the filter paper as the stationary phase and following conclusions were drawn from their Rf values (Table I): (a) General effect of oxygen-containing functional groups on polarity of the molecule decreased in the order of -COOH, -OH, -CH2OH, =O, lactone, -OCOCH3, and -COOCH3. (b) The effect of various types triterpenoid ring on the polarity was greater in ursane and lupane than oleanane. 2) Triterpenoids were separated by reversed-phase paper chromatography with liquid paraffin as the stationary phase and following conclusions were drawn from their Rf values (Tables III and IV): (a) General effect of various oxygen-containing functional groups on the polarity of molecule decreased in the order of -CH2OH or -COOH, =O, lactone, -OCOCH3 and -COOCH3. (b) Effect of various types of triterpenoid ring on the polarity was in the decreasing order of oleanane, ursane, and lupane. 3) The same result as above (2) was obtained from Rf values (Table VI) of triterpenoids separated by reversed-phase paper chromatography using methyl stearate as the stationary phase. Some structurally unknown triterpenoids. were also submitted to paper chromatography by the foregoing three methods and a satisfactory result was obtained.
2-Salicylidenehydrazono-4-thiazolidinone (I) and its 5-methylated derivative (II) possess strong anti-tubercular activity but have small solubility. Its derivative (III) substituted with a methyl in the position alpha to salicylidene group, prepared in order to increase the solubility, was found to have lost the anti-tubercular action. In the present series of work, derivatives of (III) possessing hydroxyl or methoxyl in the benzene ring were prepared in the form of 2-(2, 4(or 5)-dihydroxy-α-methyl-benzylidenehydrazono)-4-thiazolidinone and its 5-methyl derivative, and 2-(2-hydroxy-4(or 5)methoxy-α-methylbenzylidenehydrazono)-4-thiazolidinone and its 5-methyl derivative were prepared and submitted to antibacterial tests. It was thereby found that 4- or 5-methylated derivatives of the latter possessed antibacterial activity close to that of (I) or (II). The preparation of these derivatives was made by the reaction of dihydroxyacetophenone and its monomethyl ether with thiosemicarbazide to form the corresponding thiosemicarbazones which were condensed with ethyl monochloroacetate or α-bromo-propionate, in the presence of sodium acetate.
In order to examine the effect of the presence or absence of and kind of substituent in 3-position of rhodanine on anti-tubercular action, 3-substituted rhodanines having various alkyl or aryl groups were prepared and their in vitro antibacterial activity was examined. The derivatives containing isonicotinoyl, α-naphthyl, p-tolyl, and p-methoxyphenyl groupo were found to have a fair amount of such activity. The preparation of these derivatives follow the method of usinig ammonium alkylor aryl-dithiocarbamate and sodium monochloroacetate and that using bis (carboxymethyl) trithiocarbonate and amines. The latter process gave better yield when using aromatic sulfanilamides and carbowylic acid hydrazides as the starting material.
Following isolation of the quaternary base, magnoflorine, and aristolochic acid from the root of Aristolochia debilis SIEB. et ZUCC., examinations were made with larger amount of the plant material and two kinds of acid substance and one neutral substance were newly isolated. These substances were found to be identical with the aristolochic acid-B and -C, and aristololactam (I), isolated from Tao-Fang-chi and reported in the preceding paper. Components of the crude drug sold on Formosan market as Fang-chi were examined. Its outward appearance was quite similar to Tao-Fang-chi and the components were also similar, isolating magnoflorine, aristolochic acid, aristolochic acid-B and -C, and aristololactam (I). It was thereby found that this crude drug is a root of the plant of Aristolochiaceae family and the same or closely related to that of Tao-Fang-chi.