Effect of various permeability modifying agents, such as bile salts, disodium ethylenediaminetetraacetate (EDTA-2Na), tetracycline, and sodium lauryl salfate, on the intestinal absorption of sulfanilic acid, normally poorly absorbable marker, was examined in rat small intestine in vivo. Presence of any one of these agents in the gut lumen markedly increased the transfer of sulfanilic acid to the thoracic duct lymph. Transfer to the tail vein was similarly increased except in the presence of EDTA-2Na. Good correlation was observed between this increase of transfer and the previously reported values of the apparent blood to gut lumen exsorption rate constant of the intravenously injected sulfanilic acid obtained under the concurrent perfusion of these agents in the small intestinel. In the case of lymphatic transfer, however, saturable tendency was noted at higher values of apparent exsorption rate constant.
In order to synthesize 2, 3-benzodiazocine and 2-benzazepine-type compounds, phenylpropylhydrazine and phenylpropylamine derivatives, possessing a cyclohexanol group, were condensed with formaldehyde and cyclohexanone by the Mannich reaction, and dibenzo[b, d]pyran derivatives were obtained unexpectedly.
Amino acid derivatives of Aminosidin (Paromomycin) were prepared. Eight kinds of amino acid were introduced into the amino group in 6-position of the 2, 6-diamino-2, 6-dideoxy-D-glucose moiety of Paromomycin by the active esterification method. All the derivatives prepared have an antimicribial activity but less than that of the original antibiotic. Acute toxicity of the derivatives of glycyl and DL-γ-amino-β-hydroxybutyl Paromomycin was also less than that of the original antibiotic.
The displacement effect of sulfonamides on warfarin-K binding to bovine serum albumin was studied by the technique of dynamic dialysis. Increasing concentrations of warfarin in outer solution of the dynamic dialysis apparatus was observed by the administration of seven kinds of sulfonamides, except sulfanilamide, in the warfarin-bovine serum albumin solution during all experimental periods. The area under the dynamic dialysis curve between 0 and 200 min was used for designating the displacement effect of sulfonamides and this was closely related to the magnitude of binding constant of sulfonamide to bovine serum albumin. The facilitating action of sulfanilamide on the warfarin-bovine serum albumin binding may be attributed to the interaction between two drugs (sulfanilamide and warfarin) and not to the disorganization of the serum albumin structure.
5-(3-tert-Butylamino-2-hydroxypropoxy)-8-hydroxy-3, 4-dihydrocarbostyril hydrochloride, the main metabolite of the new adrenergic β-blocking agent, 5-(3-tert-butylamino-2-hydroxypropoxy)-3, 4-dihydrocarbostyril hydrochloride (Carteolol hydrochloride), was synthesized and its antagonism against the positive chronotropic and hypotensive actions of isoproterenol was examined in anesthetized dogs. This compound was found to be a little more active than Carteolol hydrochloride.
Thirty-one kinds of pyrimido[5, 4-d]pyrimidine derivatives possessing methylsulfonyl or sulfonamide group at 2-position were prepared by the reaction of 4, 8-disubstituted 6-chloro-2-methylsulfonylpyrimido[5, 4-d]pyrimidine (Va-c) or 4, 8-disubstituted 6-chloropyrimido[5.4-d]pyrimidine-2-sulfonyl chloride (IXa-b) and various amines. Reaction of N, N-bis(2-hydroxyethyl)-6-chloro-4, 8-dipiperidinopyrimido[5, 4-d]pyrimidine-2-sulfonamide (Xa-10) with an excess of diethanolamine by heating to gave dipyridamole (XIIIa) with elimination of the SO2 group. N-(2-Hydroxyethyl)-N-methyl-sulfonamide derivative (Xa-11) also underwent a similar elimination of the SO2 group. The relationship between the structure of these derivatives and their activity as coronary vasodilators and inhibitor of platelet aggregation was examined. 6-Bis(2-hydroxyethyl)amino-2-methylsulfonyl-4, 8-dipiperidinopyrimido[5, 4-d]pyrimidine (VIa) exhibited a strong coronary vasodilation, and sulfonamide derivatives (XIa-10, 12, 14, and XIIa) exhibited a comparatively strong inhibition of platelet aggregation.
In order to examine the relation between the structure and the activity of coronary vasodilation and inhibition of platelet aggregation, 9 kinds of 2-methylsulfonylpyrimido-[5, 4-d]pyrimidine derivatives possessing a sulfonamide group at 6-position were synthesized. Reaction of N, N-bis(2-hydroxyethyl)-2-methylsulfonyl-4, 8-dipiperidinopyrimido-[5, 4-d]pyrimidine-6-sulfonamide (Va) with an excess of sodium hydroxide solution or an excess of diethanolamine at low temperature gave 6-[2-(2-hydroxyethylamino)ethoxy]-2-methylsulfonyl-4, 8-dipiperidinopyrimido[5, 4-d]pyrimidine (VIIa) in a quantitative yield. When a mixture of Va and triethylamine was refluxed in butanol, 6-[bis(2-hydroxyethyl)-amino]-2-methylsulfonyl-4, 8-dipiperidinopyrimido[5, 4-d]pyrimidine (IXa) was obtained in a good yield, and IXa was obtained by heating VIIa in butanol. Analogous sulfonamide derivatives (Vb, c, j) also underwent a similar reaction.
Application of phosphorus pentachloride to 4, 8-dihydroxy-2, 6-dimercaptopyrimido-[5, 4-d]pyrimidine (I) gave 4, 8-dichloropyrimido[5, 4-d]pyrimidine-2, 6-disulfenyl dichloride (II) in a good yield, and application of piperidine to this chloride (II) afforded 4, 8-dipiperidinopyrimido[5, 4-d]pyrimidine-2, 6-disulfenopiperidide (V). Oxidation of V with chlorine in 40%, acetic acid gave 4, 8-dipiperidinopyrimido[5, 4-d]pyrimidine-2, 6-disulfonyl dichloride (VI) in a good yield. The reaction of this VI with diethanolamine at low temperature resulted in the formation of N, N, N', N'-tetrakis-(2-hydroxyethyl)-4, 8-dipiperidinopyrimido[5, 4-d]pyrimidine-2, 6-disulfonamide (VIIa). Heating of VI and diethanolamine at 110 -120°for 72 hr resulted in the formation of dipyridamole (VIII) in a good yield.
The mechanism of dipyridamole (V) formation from 4, 8-dipiperidinopyrimido[5, 4-d]-pyrimidine-2, 6-disulfonyl dichloride (IV) and diethanolamine was examined : Reaction of 14Cdisu1fonamide (VI) with cold diethanolamine at 110-120°gave V. This reaction was thought to be an intramolecular rearrangement. Four compounds (VII, VIII, IX, X) were isolated as intermediates in the formation of V. The reaction of these compounds with diethanolamine gave V in a good yield. The reaction of sulfonamide compounds (VI, VII, IX) with a string base, such as sodium hydroxide solution or diethanolamine, gave aminoether compounds (VIII, X) by intramolecular rearrangement, and the effect of acid catalysis for the rate of intramolecular rearrangement of aminoether compounds was proved.
Browning reaction of L-ascorbic acid and its related compounds in the presence of amines, as previously reported, was examined in more detail. Colorimetric and gas chromatographic determination of ascorbic acid indicated its presence in the reaction system of dehydroascorbic acid-p-toluidine. Reducing power of 2, 6-dichlorophenol-indophenol and 1, 1-diphenylpicrylhydrazyl was determined by colorimetry, and the resulting difference (FR values) might be due to the quantities of electron-donating compounds such as free radicals. The FR values also suggested the participation of the free radicals in the reaction of dehydroascorbic acid-p-toluidine. The free radicals were detected by electron spin resonance (ESR) spectra in the ascorbic acid-p-toluidine and dehydroascorbic acid-p-toluidine reaction. The spectra of ESR showed a broad triplet two weeks later and, in the later stage of the browning, it changed to a broad singlet which was stable for a long period of time. These results, suggested that ascorbic acid generated in dehydroascorbic acid-p-toluidine system was produced together with dehydroascorbic acid from 2 mol of monodehydroascorbic acid which acted as an intermediate in the browning reactions of both ascorbic acid-7-toluidine and dehydroascorbic acid-p-toluidine. The monodehydroascorbic acid radical reacted with p-toluidine to give p-tolylaminyl radical which accelerated the browning and decomposition reactions as a radical carrier in these reaction systems.
The effect of bufetolol, a β-adrenergic blocking agent, on the activities of the adenyl cyclase of rat heart (β1) and liver (β2) was examined. Bufetolol did not activate adenyl cyclase of the heart at concentrations from 10-5 to 10-10M, but showed a slight inhibition at high concentrations. Bufetolol inhibited the activation of heart adenyl cyclase by 10-5M epinephrine or isoproterenol at above 10-9 or 10-6M, respectively, but did not inhibit the activation by 3μg/ml of glucagon or 10-3M of NaF. The inhibition of Bufetolol against isoproterenol was shown to be competetive from Lineweaver-Burk analysis. Bufetolol did not affect the basal activity of the liver adenyl cyclase. With the activation by 10-5M epinephrine, bufetolol inhibited it at 10-6M or higher concentrations. The activities of phosphodiesterase and Mg2+, Na+, K+-adenosine triphosphatase of rat heart or protein kinase of rabbit skeletal muscle were not affected by bufetolol. These results suggest that the β-adrenergic blocking activity of Bufetolol is shown by the inhibition of the activation of adenyl cyclase in competition with catecholamines at β-receptor sites.
In the course of screening studies on new substances inhibiting gastric juice secretion among metabolites of Streptomyces, Streptomyces bottropensis F4708 was found to produce a new principle in its culture filtrate inhibiting gastric juice secretion. A crude active fraction was obtained as an isoelectric precipitate by adjusting the culture filtrate to pH 3.0 and then defatted. The lipid-free active fraction was purified by chromatography on hydroxylapatite column and gel filtration on Sephadex G-100 column. The purified preparation was named gastric juice inhibitory substance. From the result if chemical analysis, this substance was considered to be a kind of glycoprotein. At the dose of 1.0 mg/kg (i-p.), this substance markedly decreased gastric juice volume, total acid output, and total peptic activity in pylorus-ligated rats. Furthermore, it showed an inhibitory effect on the gastric acid secretion stimulated by tetragastrin or histamine in the perfused stomach preparation of rats, and also inhibited ulceration developed in the forestomach of pylorus-ligated rats. However, antagonism between this substance and acetylcholine was not recognized on the contraction of guinea-pig ileum.
Dihydrolycorine, which is the sole product from the hydrogenation of the alkaloid lycorine of the Amaryllidaceae, has now been synthesized by the following sequence of reactions. Methanolysis of the tetrahydrophthalic anhydride (3) yie1ded a mixture of a pair of half-esters (4a and 4b). This mixture was subjected to the Friedel-Crafts cyclization to give the tetrahydrofluorenone (5) and the tetralone (6). The keto-alcohol, derived from 5 by treatment with LiAlH4 followed by MnO2, underwent the Schmidt rearrangement, giving, after hydrolysis, two lactams (11 and 14), one (11) of which was found to be an abnormal rearrangement product. The lactam (14) has been prepared alternatively in a better yield. The mixture of the half-esters (4a and 4b) was treated with ethyl chloroformate followed by NaN3 to give the azide, which, after heating, gave the lactam-ester (16) as a sole isolable product by treatment with trifluoroacetic acid. Hydrolysis of 16, conversion of the resulting acid (17) into the mixed anhydride (18), and its reduction with NaBH4 yielded the lactam-alcohol (14) which was converted to the lactam-homoacid (21) through the cyanide (20). The stereochemistry of 21 was established by its conversion into γ-lycorane. Halolactonization if 21 gave the iodo-lactone (36) which was transformed into the iodo-acetoxy-imide (38) with acetic anhydride and acetic acid. Treatment of 38 with LiCl in dimethylformamide gave the olefin (41) which was oxidized to the epoxide (42) with m-chloroperbenzoic acid. Reductive cleavage of 42 with LiAlH4 in the presence of ZnCl2 gave dl-dihydrolycorine identical with that from natural sources.
Two kinds of carboxypeptidases, CPase Sa and CPase Sb, were separated from germinating soybeans (Glycine Max. AKIYOSHI) in a highly purified form. The enzymes liberated neutral, acidic, and basic amino acids including proline from the C-termini of N-substituted dipeptides at varying rates. Of the substrates tested, Z-Gly-Pro, Z-Pro-Pro, Z-Gin-Pro, Bz-Gly-Lys, and Bz-Gly-Arg were hydrolyzed very slowly. The specific activities of purified CPase Sa and Sb were 0.11 and 3.68 units/mg of protein, respectively, for Z-Glu-Phe. The enzyme also released C-terminal amino acid residues from peptide and protein substrates. They possessed weak esterase activity, but were free of endopeptidase and aminopeptidase activities. They had a pH optimum at 5.5 for Z-Glu-Phe. CPase Sb was strongly inhibited by diisopropylfluorophosphate (DFP) and HgCl2, whereas CPase Sa was only partially inhibited. Other metal ions, anions, ethylenediaminetetra-acetic acid (EDTA), o-phenanthroline, N-ethylmaleimide (NEM), p-chloromercuribenzoic acid (PCMB), dithiothreitol (DTT). and monoiodoacetic acid (MIA) showed no significant effect on the enzymic activity. The molecular weight of CPase Sa was 99000 (s20, w=6.9S, D20, w=6.4×10-7cm2/sec) and that of CPase Sb, 106000 (s20, w=6.1S, D20, w=5.4×10-7cm2/sec).
Tannic acid has an inhibitory effect against the proteolytic activity of pepsin in vitro, but its activity was nonspecific. In pylorus-ligated rat, tannic acid inhibited not only peptic activity but also gastric acid secretion. However, the volume of gastric juice was not influenced by tannic acid. Tannic acid showed antiulcer activity by its oral administration (50 mg/kg) in Shay's ulcer. By the use of artificial gastric juice perfusion technique, formation of ulcer was delayed by the addition of tannic acid in the perfusate. The acetic acid ulcer was slightly healed by the oral administration of tannic acid (200 mg/kg) for 10 days without any effect on the growth curve of rats. Tannic acid formed a precipitate with starch in water and this complex has the same activities. From these results, it is suggested that tannic acid has ideal properties as an antiulcer agent.
6-Chloro-5-cyclohexyl-1-indancarboxylic acid (Ib), a potent nonsteroidal analgesic, antipyretic, and antiinflammatory agent, and related compounds were synthesized by the new routes via cyanation of 1-chloro-5-cyclohexylindan (IXa), which was obtained from 5-cyclohexyl-1-indanol (VIa), or cyanation of 1, 6-dichloro-5-cyclohexylindan (IXb), which was obtained from VIa or IXa.
From the heartwood of Magnolia obovata THUNB. (Magnoliaceae), l-N-acetyl-anonaine (I) and a mixture (II) of ω-O-feruloyl-ω-hydroxy fatty acids were isolated besides liriodenine. II was hydrolyzed with alkali to give trans-ferulic acid (IIa) and a mixture of ω-hydroxy-fatty acids (HO(CH2)n COOH, IIb). Treatment of IIb with acetic anhydride and pyridine followed by diammethane gave a mixture of ω-acetoxy-fatty acid methyl esters. Gas chromatography and mass spectroscopy of the mixture revealed the chain length of the fatty acids in IIb to be n=21-27.
Reaction of 1-and 2-naphthols with 2-chloro-4-halonitrobenzene in diglyme, in the presence of sodium hydride, gave nitro compounds (I to VI) which were reduced with stannous chloride and hydrochloric acid to the amino compounds (VII to XII). These amino compounds were reacted with p-chlorophenyl isocyanate in ether and carbanilides (XIII to XVIII) were obtained. Other carbanilides (XIX to XXV) were obtained by reacting p-chlorophenyl isocyanate with 4, 4'-diamino-4-methyldiphenylamine, 4-amino-4'-chlorophenyl sulfide, 2-amino-4, 4'-dichlorodiphenylamine, 2-amino-4, 4'-dichlorodi-phenyl sulfide, and 2-amino-4, 4'-difluorodiphenyl ether, in the same way as above. Examination of antibacterial activity of these compounds showed that XV to XVII and XXII to XXIV had a strong antibacterial activity against gram-positive bacteria.
In an attempt to develop a more specific and convenient method for the determination of vitamin A in preparations, high-speed liquid chromatographic method was investigated. Samples were dissolved, directly or after saponification, in a mixture of methanol and chloroform, and injected into a Permaphase "ODS" column. Vitamin A was separated from other substances by employing a mixture of methanol and water as the mobile phase, and assayed using chrysene or benzo[a]pyrene as internal standard. This method showed a substantial improvement over the Carr-Price method which had been used officially.
In order to study the microbial transformation of lysergic acid diethylamide, several amide derivatives of norlysergic acid were prepared from the corresponding amide derivatives of Iysergic acid by treatment with cyanogen bromide followed by hydrolysis with 1.4% HCl-dioxan at 100°. N6-Allyl-, -propyl-, and-ethyl-norlysergic acid diethylamide were also prepared to examine their pharmacological activities.
The content and accumulation of gentiopicroside (I) in the fresh root or Gentiana triflora var. japonica and Gentiana scabra var. buergeri were examined by the thin-layer chromatography-densitometer method. It was found that the content of I in the fresh root of G. triflora var. japonica was 10-13% and hardly varied it growing of 1-3 years after planting but the content showed a decrease in the sample from this plant after 4 years' growth. The content of I in various parts of G. scabra var. buergeri decreased in the order of roots, flowers, stems, and leaves.