There are two different techniques for the X-ray structure analyses of materials ; that for small molecules and that for macromolecules. The latter technique, is called"protein crystallography."The characteristics of protein crystallography as opposed to ordinary crystallography for small molecules are described. Some of the achievements of protein crystallography and their impact on other scientific fields are discussed, and perspectives for the possible contribution of protein crystallography to molecular pharmacology are given.
Effects of Dai-Saiko-Toh, Sho-Saiko-Toh and Saiko-Keishi-Toh on body weight and weights of various organs of rats were observed, and histological changes in various organs were examined. While no difference in the rate of body weight increase was noted among normal rats and those treated with Dai-Saiko-Toh or Saiko-Keishi-Toh, Sho-Saiko-Toh treated rats gained weight faster than normal. A weight increase of the adrenal glands and atrophy of the thymus were observed in rats treated with Dai-Saiko-Toh or Sho-Saiko-Toh. Histologically, the adrenal cortex showed lipid accumulation in the lower zona fasciculata in Dai-Saiko-Toh and Sho-Saiko-Toh treated rats. In the liver, no change in weight or histology was noted in any group.
Changes in mitochondrial and microsomal enzyme activities and in hepatic ultrastructure were examined in rats treated with Dai-Saiko-Toh, Sho-Saiko-Toh or Saiko-Keishi-Toh. In liver homogenates of rats treated with Dai-Saiko-Toh, the activities of NADPH-cyt. c reductase and G-6-Pase were markedly decreased ; a similar, but weaker trend was noted in rats treated with Sho-Saiko-Toh. Succinate-cyt. c reductase activity was also decreased in the rats treated with Dai-Saiko-Toh or Sho-Saiko-Toh. Electronmicroscopy of liver tissue showed that mitochondria in the liver of rats treated with Dai-Saiko-Toh or Sho-Saiko-Toh were very small and that the volume density of mitochondria was also lower than in the control.
Cannabinoids (CN) of CBDA stock, Tochishi No. 1 stock and crossed stock were determined on a high-pressure liquid chromatograph equipped with a LiChrosorb RP-18 column (25 cm×4 mm) by elution with a mixture of methanol and 0.02 N sulphuric acid (4 : 1). The CBDA stock was found to contain cannabidiolic acid (CBDA) and cannabichromenic acid (CBCA). Tochishi No. 1 stock was detected to have thtrahydrocannabinolic acid (THCA), CBCA and a small amount of CBDA. F1 was obtained by crossing CBDA stock (female) and Tochishi No. 1 stock (male). All 79 F1 specimens were found to contain CBDA, THCA and CBCA. Therefore F1 was designated the median type. F2 was then obtained by crossing F1 and out of 199 F2 specimens, 58, 40 and 101 stocks were designated as CBDA stock type, Tochishi No. 1 stock type and the median type by CN analysis, respectively. F3 was also gained by crossing CBDA stock type of F2 and all 166 F3 specimens were CBDA stock type. F4, which was obtained by crossing F3, was also CBDA stock type. In addition, the fibers of F4 were found to be better than those of the original CBDA stock, and F4 contained a negligible amount of THCA.
The tumor-inhibiting effect of yeast cell wall (YCW) against a transplanted 3-methyl-cholanthrene-induced fibrosarcoma in C57BL/6 mice was examined alone and in combination with mitomycin-C, methotrexate or 5-fluorouracil. These drugs and/or YCW were administered intraperitoneally 3 or 5 times after tumor inoculation. While the tumorinhibiting effect of YCW alone was weak, in combination with mitomycin-C or methotrexate, it was stronger than with YCW or either of these drugs alone. Injection of 5-fluorouracil alone was as effective as in combination with YCW. These results suggest that some antitumor drugs can be augmented in their antitumor activity by combination with YCW. To investigate the effect of YCW on the immune response of mice during tumor progression, the change in mitogenic response (3H-thymidine uptake) was examined in spleen cells from tumor-bearing mice that had received YCW. The mitogenic response to bacterial lipopolysaccharide (a B-cell mitogen) was enhanced following intraperitoneal injection of YCW. The response of spleen cells from mice with tumors over 3 g to concanavalin A (a T-cell mitogen) was decreased markedly in spite of YCW-injection, while YCW augmented this response in animals with 1 g tumors.
Metabolism of Sodium 7-[D (-)-α-(4-ethyl-2, 3-dioxo-1-piperazinecarboxamido)-α-(4-hydroxyphenyl) acetamido]-3-[(1-methyl-1H-tetrazol-5-yl) thiomethyl]-3-cephem-4-carboxylate (T-1551), a new β-lactam antibiotic, was studied in vivo, in situ and in vitro. Only unchanged T-1551 was detected by bioautography in the urine of human and animals receiving T-1551 intramuscularly. When 3H, 14C double-labeled T-1551 (3H, 14C-T-1551) was intramuscularly administered to rats, in urine and bile, small amounts of two metabolites, 3H and 14C radioactive 7-[D (-)-α-3-[2-(N-ethyl-N-oxaloamino) ethyl] ureido-α-(4-hydroxyphenyl) acetamido]-3-[(1-methyl-1H-tetrazol-5-yl) thiomethyl]-3-cephem-4-carboxylic acid (T-1551 A) and 3H radioactive 5-mercapto-1-methyl-1H-tetrazole (T-1551F), and in feces, 14C radioactive N-formylmethyl D (-)-α-(4-ethyl-2, 3-dioxo-1-piperazinecar-boxamido)-α-(4-hydroxyphenyl) acetamide (T-1551D), were detected by radio thin layer chromatography. Similarly, when T-1551 was administered to monkeys, small amounts of two metabolites, T-1551A and T-1551F, were detected in urine and bile by means of high pressure liquid chromatography (HPLC). Furthermore, quantitative analysis of each metabolite was carried out by HPLC (for separation) and radioactive measurement (for determination). In situ and in vitro, 3H, 14C-T-1551 was stable in various tissue homogenates, but seemed to change into 14C-T-1551D and 3H-T-1551F by β-lactamase produced from intestinal flora. In rats, 14C-T-1551D was hardly absorbed from the gastro-intestinal tracts. On the other hand, when T-1551 was intramuscularly administered to human, small amounts of urinary T-1551A and T-1551F were detected by HPLC similar to the findings in animal studies. We suggest that the metabolism of T-1551 in human was the same as in the animals examined.
A series of decahydrocinnoline derivatives (II), (VIII), (IX), (X), (XI), (XII), (XIII), (XVI), (XVII) and (XVIII), possessing a 3-substituted phenyl group at the 4a-position were synthesized and evaluated for their analgesic activity.
Derivatives of 3-[4-(1, 3-dihydro-1-oxo-2H-isoindol-2-yl) phenyl]-2-oxobutanamide (1), exhibiting potent anti-inflammatory and analgesic activity, were obtained from 3-(4-aminophenyl)-2-oxo-butanamide (2) and 3-[4-(1, 3-dihydro-1-oxo-2H-isoindol-2-yl) phenyl]-2-oxobutanoic acid (25) as intermediates. Furthermore, the structures of E and Z isomers of 2-butenoic acid and oxiranecarboxylic acid derivatives as synthetic intermediates of compound (2) were studied. Among the derivatives, 3-[4-(2, 5-dihydro-1H-pyrrol-1-yl) phenyl]-2-oxobutanamide (6), 2-oxo-3-[4-(1-piperidinyl) phenyl] butanamide (8), 3-[4-(3-carboxypropanoylamino)-phenyl]-2-oxobutanamide (11), 3-[4-(2, 5-dihydro-2, 5-dioxo-1H-pyrrol-1-yl)-phenyl]-2-oxobutanamide (14), and N-benzyl-3-[4-(1, 3-dihydro-1-oxo-2H-isoindol-2-yl) phenyl]-2-oxobutanamide (16) showed potent anti-inflammatory activity comparable to phenylbutazone at the first screening with carragenin-induced rat paw edema.
6, 8-Dimethoxyflavone (I) and 2', 3', 5'-trimethoxyflavone (II) were synthesized in good yield from 2-bromo-6-methoxy-1, 4-benzoquinone (VIII) as a starting material. The flavone isolated from Alnus glutinosa and presumed to be 5, 4'-dihydroxy-6, 8-dimethoxyflavone (IV) was suggested to be 5, 4'-dihydroxy-6, 7-dimethoxyflavone by comparison with the melting points of synthetic flavone derivatives. The 13C-NMR spectra of the flavones are also discussed.
The anti-inflammatory actions of water extracts of snake's moulted skin were investigated to determine profile. The extract was found to have anti-edematous action on carrageenin edema, mustard edema and inhibitory action against leucocyte emigration. However, the extract showed somewhat weaker inhibitory action against increased vascular permeability. In in vitro tests, the extract showed marked inhibitory action on heat-induced hemolysis of rat erythrocytes.
A search for catalysts effective in the demethylation of p-methoxybenzonitrile was undertaken in the liquid-phase. When the reaction was carried out using anhydrous aluminum chloride or Raney nickel as a catalyst and piperidine or pyridine as a solvent, a successful result was obtained. Especially when anhydrous aluminum chloride was used as a catalyst, and piperidine as a solvent, p-hydroxybenzonitrile was obtained in a yield of 85% by heating at 240° for 15 hours. The above reaction systems were also effective for the demethylation of m-methoxybenzonitrile (maximum yield of m-hydroxy-benzonitrile : 77% at 230°), but not as effective for the demethylation of o-methoxybenzonitrile (maximum yield of o-hydroxybenzonitrile : 18% at 250°) by heating for 15 hours in liquid-phase.
A method for the determination of serum bromvalerylurea (BVU) was established, using high-speed liquid chromatography. BVU was extracted from serum with ethanol containing 3-nitro-p-anisidine as an internal standard, and chromatographed on a reversephase column (LiChrosorb RP-18, 5 μm) with a mobile phase of acetonitrile/water (1/3 (v/v)). This method facilifated BVU determination within 1.16 % error over the serum level range from 10 to 500 μg/ml, with the limit of detection being 10 μg/ml.
A rapid method for the determination of gentiopicroside, the bitter principle of the herb of Swertia japonica, was established by the use of high-speed liquid chromatography. Gentiopicroside in Swertiae Herba was separated on a 25 cm PC8, column, (Shimadzu-Du Pont Model LC-2), using 12% acetonitrile as desorption solution ; separation was completed within 7 min. Gentiopicroside in the crude fraction was extracted with water and the extract was injected directly into the column. The content of gentiopicroside was calculated from the calibration curve previously prepared using a standard. This method is considered to be useful for the evaluation of Swertiae Herba.
A new 8-epi-eremophilenolide derivative, 3β-angeloyloxy-9-en-8-epi-eremophilenolide and 3β-angeloyloxy-8-epi-eremophilenolide were isolated from the rhizome of Farfugium japonicum (L.) KITAM. (Ligularia tussilaginea MAKINO).