Several proteolytic enzymes and dehydrogenases of microbial origin were studied with special regard to structure-activity relationship. Enzyme genes of Zn-proteases, subtilisin and pyroglutamyl aminopeptidase from genus Bacillus, prolyl endopeptidases from Flavobacterium and Aeromonas, and of protease II from E. coli were cloned, sequenced and overproduced in E. coli, their active site structures being elucidated by chemical modification as well as by site-directed mutagenesis. Homology analysis revealed that there is a prolyl endopeptidase family as a new family of serine endopeptidases. In addition, enzymatic properties and the primary structures of glutathione-independent formaldehyde dehydrogenase of Pseudomonas putida and 7α-hydroxysteroid dehydrogenase from E. coli were elucidated. Amino acid sequences deduced from the nucleotide sequences of their genes indicated that the former enzyme should be classified into a long-chain metallo alcohol dehydrogenase family, and the latter belongs to a member of the short-chain nonmetallo-alcohol dehydrogenase family.
To understand the previous result of higher bioavailability of rectal salbutamol (SB) compared with oral SB, in situ experiments using rabbit duodenal and rectal loop were carried out. After the intravenous (i.v.) and intraportal (i.p.) dosing of SB, fraction of dose which avoids the hepatic first-pass-effect (Fh) was calculated from the areas under the blood concentration-time curve (AUC). The Fh was about 10% and unchanged significantly with increasing i.p. dose (5-20 mg). Intraduodenal (i.d.) or intrarectal (i.r.) dosing of SB was made after the i.v. and i.p. dosing, and the AUC's and the residual amount in the loop were obtained to estimate the parameters. The results of the i.d. and i.r. dosing were as follows ; for the extent of bioavailability (EBA), 7.7±1.5% and 14.5±2.3%, for the fraction of dose absorbed (fa), 93.9±3.7% and 33.8±3.3%, and for the fraction of absorbed SB which avoids first-pass-effects (F), 8.4±1.9% and 43.0±6.0% (mean±S.E., n=4). Consequently, SB dosed i.d. was absorbed completely, and received first-pass-metabolism in the mucosa (about 20%) and then in the liver (about 90%), which caused the low bioavailability. While, in i.r. dosing, SB absorption was poor. However, higher bioavailability was obtained owing to about 40% of rectal venous blood flow which bypasses the liver and negligible first-pass-metabolism in the mucosa (about 4%).
We developed a new subdermal silicone elastomer implants containing chlormadinone acetate (CMA), which was effective for prevention of estrus in bitches. The implants gave a continuous CMA release over a year. The CMA release from the implants was dependent on the CMA loading level and surface area of the implants. The CMA release patterns both in vitro and in vivo system fitted well with the Higuchi square root low. The dissolution test using organic solvents was found to be useful for the brief estimation of CMA release from the implants.
As a continuing study on the chemical evaluation of the utility of the farinfrared drying method for crude drugs, the efficiency in the drying process of Zingiberis Rhizoma was examined in comparison with the conventional drying methods such as an air drying and a hot-air drying. Furthermore, the chemical fluctuation of the constituents during the drying process has been investigated by means of HPLC quantitative analyses for pungent principles and galanolactone, and GLC quantitative analyses for bisabolane-type sesquiterpenes. It has been found that, by use of the far-infrared drying method (oven temperature 60°C), the rhizome of Zingiber officinale was dried efficiently without reducing the contents of bioactive constituents.
Effects of 70% ethanolic extract from leaves of Ginkgo biloba (GBE) on the hair regrowth in normal and high butter diet-pretreated C3H strain mice which posterior hair we shaved were investigated. GBE showed a promoting effect on the hair regrowth. GBE had the inhibitory effects on blood platelet aggregation, thrombin activity and fibrinolysis. GBE inhibited the increase of serum the triglyceride level in high cholesterol diet-treated rats. These results suggested that GBE promotes the hair regrowth and could be used as a hair tonic.
The stability test in storage of "Rokumi-gan" powders was carried out under accelerated (40°C, 75%RH, in the dark) and low-temperature conditions (10°C, in the dark). Morroniside (1), loganin (2), paeoniflorin (3), paeonol (4), γ-aminobutyric acid (5) and 50% (v/v) ethanol extract (Extract) were selected as quality indicators. The powder preparation was confirmed to be stable for six months in tight storage under 10°C because the contents of 1, 2, 3, 5 and Extract did not change and that of 4 decreased by 7%. During six-month tight storage under the accelerated condition, the contents of 2, 3 and Extract did not change and that of 4 decreased by 12%. Those of 1 and 5 decreased according to apparent one and two-stage firstorder disappearance, respectively. In open storage, the contents of 1-5 and Extract decreased faster than in tight storage according to apparent two-stage first-order disappearance. These results indicated that temperature was one of the critical factors for good storage and that the stability of the powder preparation in storage under room temperature was not validated.
A new type of nonionic surface-active agents containing sugars as the hydrophilic group were synthesized and evaluated for their antimicrobial activity. Methyl 6-deoxy-3-O-dodecyl-6-halo-D-glucopyranoside derivatives showed powerful antimicrobial activity, but no significant differences were observed on the activity with regard to the kinds of anomers as well as halogen atoms.