This paper reviews the researches on powdered preparations from the point of view of molecular pharmaceutics. Molecular behaviours of medicinals in the preparations are discussed in relating with physicochemical properties and bioavailability. Polymorphism, crystallinity including disorder parameter, and inclusion compounds are quoted as an example. Two methods that give the molecules high activity-grinding with microcrystalline cellulose or cyclodextrins and the mixing with porous powders-are explicated. The characteristic molecular behaviours in the mixtures are explained on the basis of X-ray diffraction, thermal, and infrared spectroscopic measurements.
Two-step laser excitation (TSLE) time-resolved fluorescence has been developed as one of the recent developments of laser spectroscopy for the investigations on the kinetics and dynamics of unstable molecules in the photochemical processes. Applications of the TSLE fluorescence to the excited-state proton transfer and isomerization in the several inter- and intramolecular hydrogen bonding systems have been reviewed. The TSLE fluorescence consists of formation of unstable species by the first laser excitation. which were observed first in the excited state proton transfer by us, and of the fluorescence observation by the second laser excitation of unstable species. Further, variable delay technique provides us with the time-resolved spectra and decays of several unstable species having different lifetimes and spectra both in the ground and excited states. A possibility of three dimensional time-resolved fluorescence spectroscopy is suggested.
The solid state reactions between 1-methyl-4-oxo-cis-2, 6-diphenylthianium tetrafluoroborate (1) and several alkali halides were ascertained by the infrared absorption spectrometry and powder X-ray diffraction. Various facts concerning these reactions indicate that an ion exchange occurs after molecules of 1 diffuse into alkali halides. In the cases of KCl and KBr, elimination reaction proceeds by withdrawal of hydrogen from 3-position of oxothianium ring as a result that chloride and bromide ions produced by the ion exchange act as a base. In this process, it turned out that the reaction proceeds by an anion mechanism. In the case of KI, however, an iodide ion acts as nucleophilic reagent to give demethylated product (4-oxo-cis-2, 6-diphenylthiane).
In connection with our studies on the chemical evaluation of Cucurbitaceae plants, isolation and identification of saponins of fruits and herb of Luffa cyclindrica ROEM. were carried out. From fruits, nine saponins 2-5, 6, 10, 11, 14, 15 were isolated. The saponins 6, 10, 11, 14 and 15 were identified as lucyosides-A (6), -E (10), -F (11), 3-O-β-D-glucopyranosyl hederagenin (14), 3-O-β-D-glucopyranosyl oleanolic acid (15), respectively, the three former of which have already been isolated from herb of this plant. The structures of new saponins, named lucyosides-J (2), -K (3), -L (4) and -M (5) were established as 3, 28-O-bis-β-D-glucopyranosyl 21β-hydroxygypsogenin, 3-O-β-D-glucopyranosyl gypsogenin, 3-O-β-sophorosyl-28-O-β-D-glucopyranosyl gypsogenin and 3-O-(6'-acetoxy-β-O-D-glucopyranosyl)-28-O-β-D-glucopyranosyl gypsogenin, respectively. From herb, a new saponin, named lucyoside-I (1) was isolated as minor saponin and established as 3-O-β-D-glucopyranosyl arjunolic acid.
Bis (methoxy-2-benzofuranyl) ketones (IIa-e) were synthesized by the reaction of 1, 3-dichloro-2-propanone with methoxysalicylaldehydes (Ia-e) in the presence of sodium hydroxide. Bis (hydroxy-2-benzofuranyl) ketones (IIIa-e) were prepared from IIa-e by treatment with alminium chloride. Bis (7-hydroxy-2-benzofuranyl) methane (IV) was obtained by reduction of bis (7-hydroxy-2-benzofuranyl) ketone (IIIa) with 80% hydrazine monohydrate in the presence of potassium hydroxide. Bis (7-hydroxy-2-benzofuranyl) methanol (V) was prepared by reduction of IIIa with sodium borohydride. Some of these compounds showed an antiviral activity against influenza virus in vitro. Bis (6-hydroxy-2-benzofuranyl) ketone (IIIb) was most active in the screening of in vitro, which was also effective in vivo.
The anti-inflammatory and analgesic effects of 17 crude drugs used for the relief of arthritic symptoms in traditional Chinese medicine have been investigated by utilizing carrageenan-induced edema in rats and acetic acid-induced writhing in mice. In 13 crude drugs anti-inflammatory activity on carrageenan edema was observed with 250 mg/kg i.p. administration and 16 crude drugs (1000 mg/kg s.c.) inhibited the number of writhings induced by acetic acid in mice.
Simple, rapid and sensitive methods for separation and quantification of caffeine and/or dimethylxanthines in biological fluids and tissues were developed by high performance liquid chromatography. A 0.1-ml of the serum was denatured and precipitated with 0.1 ml of acetonitrile. A 0.01-ml of the supernatant was injected into the chromatograph with a reversed-phase Zorbax ODS column and ultraviolet (UV) detector set at 273 nm and 0.01a.u.f.s. The flow rate of the mobile phase of 0.2 M acetate buffer-acetonitrile (85 : 15, v/v) was 0.5 ml/min. The limit of detection was 0.2 μg/ml for caffeine. On analyses of biological fluids, caffeine and its metabolites were extracted from 0.2 ml of the plasma or saliva by using 2.5 ml of chloroform-isopropanol solution (75 : 25, v/v). On analyses of tissues, the liver, placenta or fetal cerebrum was homogenated by saline, adjusted to 10% solution, and caffeine and its metabolites were extracted from 0.5 ml of 10% tissue homogenates by using 5.0 ml of chloroform-isopropanol solution. The reversed-phase μBondapak C18 column was used, connected with UV detector set at 280 nm and 0.01 a.u.f.s. The flow rate of mobile phase of 0.005 M acetate buffer-methanol-acetonitrile-tetrahydrofuran (92.5 : 3.0 : 2.8 : 1.7, v/v) was 1.5 ml/min. The limits of detection of caffeine and dimethylxanthines on analyses of biological fluids and tissues were 0.1-0.2 μg/ml and 1.0-2.0 μg/g wet weight, respectively. The practicability and utility of the method were demonstrated in human and rat studies.
The degradation of fluorescent substances, N-substituted-1, 4-dihydropyridine-3, 5-dialdehyde derivatives, derived from the reaction of malondialdehyde (MDA) which is one of end -degradative products during lipid peroxidation with various amino compounds, was studied in rat liver microsomal fractions. The fluorescent substances from the reaction of MDA with 1-amonopentane, 1-aminoheptane, 1-aminodecane and phenylethylamine (PEA) rapidly changed into water soluble compounds. On the other hand, the fluorescent compounds from short-length amino compounds such as methyl or ethylamine had a little or no change. The degradation system required nicotineamide adenine dinucleotide phosphate, and was inhibited by CO. Furthermore, in microsomal fractions from phenobarbital-pretreated rats, the rate of degradation increased. The degradative compounds of the fluorescent substance from MDA with 14C-phenylethylamine were separated by high performance liquid chromatography. Two major water soluble fluorescent compounds, 4-methyl-1, 4-dihydropyridine-3, 5-dialdehyde (WM-1) and 1-phenylethyl-4-hydroxy-4-methyl-1, 4-dihydropyridine-3, 5-dialdehyde (WM-2), and two minor fat soluble fluorescent compounds were isolated. All of these isolated degradative compounds retained 1, 4-dihydropyridine structure, and exhibited also the same maximum excitation and emission spectra at 392 and 448 nm, respectively, as those of native fluorescent substance. The present results suggested that the microsomal degradation of fluorescent substances related to MDA was dependent on the structure of N-alkyl sidechain of amino compounds.
Capacity of 7 immunomodulating agents to induce cytotoxic factor in the sera was examined in mice. These agents were injected intravenously to Bacillus Calmatte-Guerinsensitized C3H/He mice and the sera were collected 2 h later. Cytotoxic activity of the sera was tested against L929 cells in the presence of actinomycin D. Poly (I)-poly (C), concanavalin A and a Streptococcus preparation OK 432 induced cytotoxic factors as well as bacterial lipopolysaccharide. However, dextran sulfate, lentinan and a linear β-1→3-glucan, TAK did not. These agents have been classified according to the extent to which they increase LB, a serum protein in mice. In comparison with the classification, these results were discussed.
When cysteamnie was administered before the treatment with carbon tetrachloride (CCl4), changes in glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and alkaline phosphatase in the serum and those in glucose-6-phosphatase and triglyceride in the liver of rats with CCl4-induced hepatic injury were observed to be significantly prevented. Under conditions described in this report, cystamine was recognized to have almost the same effect with cysteamine and thiopronin had a weak effect. However, little effect was observed in cysteine and WR-2721. These results suggested the possibility of screening test of drugs preventing CCl4-induced hepatic injury under conditions described in this report.
Survival effects of Shimotsu-toh and its four kinds of constituents (Tohki ; Angelicae Radix, Senkyu ; Cnidii Rhizoma, Shakuyaku : Paeoniae Radix, Jioh : Rehmanniae Radix) on mice irradiated with a lethal dose of soft X-ray were investigated. It was observed that significant protective effects exist in both MeOH and H2O extracts of Shimotsu-toh and that most of these effects were dependent on Senkyu.
Morphological studies on the effects of ten guanidine, three urea, two thiourea derivatives and others including anticancer agents on chick embryos were examined on the 14th day of incubation, when each drug was injected into the albumen of fertile eggs of the 5th day ofincubation. Aminoguanidine sulfate (AG) produced retardation of the growth of liver and hypoplasia of gallbladder of chick embryos, but did not cause any external abnormality. Hydroxyurea, semicarbazide, ethionine, mitomycin C (MMC) and vinblastine induced severe external anomalies. Hypoplasia of gallbladder and hepatic lesion were also observed after the injection of these drugs. Anomalies produced by MMC on the gallbladder and liver were similar to those by AG regardless of structural similality. On the other hand, guanidine derivatives produced neither AG-like abnormalities nor other marked anomalies. Consequently, the effect of AG on the liver and gallbladder of chick embryo was thought to be due to a peculiarity of its structure.
Effects of protein compositions of Persicae semen on inflammation, allergy and blood coagulation were investigated in experimental animals. Tohnin sen is lyophilized powder of boiling water extracts of Persicae semen, and PR-A and PR-B (mol. wt.=about 300000 and 10000, respectively) are proteins purified from the water extracts. (1) PR-A and PR-B showed strong inhibitoty activity on hind paw edema induced by carrageenin in rats, and their potency (ED50=1.08 and 0.100 mg/kg, i.v., respectively) was superior to or comparable to that of indomethacin and much stronger than that of Tohnin sen. (2) PR-A (2.5 mg/kg, i.p.), PR-B (0.5 mg/kg, i.p.) and Tohnin sen (300 mg/kg, p.o.) produced a significant inhibition of granuloma formation induced by felt-pellet implantation in rats ; the activity of PR-B was approximately comparable to that of indomethacin (1-2 mg/kg, p.o.). (3) When injected into the carboxymethyl cellulose pouch, PR-B (0.1 to 10 mg/pouch) enhanced exudation of serum protein ; PR-B but not PR-A, unlike indomethacin, enhanced leucocyte emigration. (4) PR-A, PR-B or Tohnin sen did not show any significant inhibitory activity on vascular permeability and on adjuvant arthritis ; and also PR-B did not potentiate the anti-arthritic action of prednisolone. (5) PR-A and PR-B (10 mg/kg, i.p. each), like indomethacin, did not show any inhibitory activity on passive cutaneous anaphylaxis and on delayed type hypersensitivity in mice. (6) Tohnin sen (100 mg/kg, p.o.) and PR-B (20 mg/kg, i.p.) did not influence on the blood coagulation time and on erythrocyte absorption from the tissue with internal hemorrhage, respectively, in rats. (7) PR-A and PR-B showed anti-writhing activity at doses of until 5 and 0.1 mg/kg, i.v., respectively, in mice, and the anti-writhing ED50-value of Tohnin sen was 43.3 mg/kg, p.o. These results demonstrate that PR-A, PR-B and Tohnin sen have anti-edema, antigranulation and anti-writhing activities, and that their pharmacological profile was dissimilar to that of non-steroidal anti-inflammatory drugs such as indomethacin.
PR-B is a protein composition of Persicae semen (mol. wt.=about 10000) ; and its antiinflammatory activity is comparable to that of indomethacin in the carrageenin-induced hind paw edema test. In order to explore the mode of anti-inflammatory action of PR-B, inhibitory activities of PR-B on kinin-release by kallikrein and superoxide anion production were investigated in vitro. PR-B produced a dose-related inhibition of kinin-release by kallikrein at concentrations of 10-5-10-3M, and its potency (IC50=1.4×10-4M) was comparable to that of soybean trypsin inhibitor (SBTI) ; superoxide dismutase (SOD) and N-α-p-tosyl-L-lysine-chrolomethylketone, a synthetic trypsin inhibitor were inactive. The potency of PR-B as a trypsin inhibitor (IC50=3.2×10-4 M or more than 104- M) was much weaker than that of SBTI (IC50=7.2 or 7.4×10-8M) ; SOD was inactive. In contrast to those activities, PR-B had comparatively strong SOD-like activity ; i.e., PR-B inhibited dose-relatedly the superoxide anion production by peritoneal macrophages of guinea pigs at concentrations of 10-6-5×10-5M, and its potency (IC50=5.7×10-6M) was approximately equivalent to that of SBTI while much weaker than that of SOD (IC50=5.7×10-9M). Lysozyme and egg albumin as reference proteins lacked the SOD-like activity. The anti-edema activity of PR-B given intravenously was stronger than that of SBTI and SOD in the carrageenininduced hind paw edema test in rats. These results were discussed in connection with the mode of anti-inflammatory action of PR-B.
A high sensitive determination of ketoprofen (KP) in the human serum after cutaneous administration of KP was developed by using electron capture detector gas chromatography (GC-ECD). The extracted KP was derivatized with pentafluorobenzyl bromide, and the reactant purified with Sep-Pak C18 cartridge. The KP derivative was separated by high performance liquid chromatographic technique, and determined by using GC-ECD. This procedure provided linear calibration curve ranging 2 to 80 ng/ml and the limit of detection was approximately 0.5 ng/ml in the human serum. This method was applicable to pharmacokinetic studies of this drug.