Yasuhiro Tezuka

Comparative analysis of the constituents in Saposhnikoviae Radix and Glehniae Radix cum Rhizoma by monitoring inhibitory activity of nitric oxide production.

During the development of natural herbal medicines in Japan, Glehniae Radix cum Rhizoma (Hamabofu in Japanese) has been used as a substitute for Saposhnikoviae Radix (Bofu). Bofu and Hamabofu are blended differently in several Kampo formulae. For example, Bofu is included in Jumihaidokuto by a manufacturer, whereas Hamabofu is included instead of Bofu in the same formula by other manufacturers. Although both Bofu and Hamabofu are used for their expected anti-inflammatory effects, differences in their medicinal properties are not well characterized. In addition, there have been very few reports comparing the pharmacological activities of the constituents in Bofu and Hamabofu. In the present study, we investigated the anti-inflammatory effects of the extracts of Bofu and Hamabofu by monitoring levels of the inflammatory mediator nitric oxide (NO) produced in rat hepatocytes. Moreover, the chemical constituents responsible for the activity were investigated. Our results showed that ethyl acetate fractions of Bofu and Hamabofu extracts contain different compounds, although both fractions suppressed NO production in rat hepatocytes. The linear dihydropyranochromones from the Bofu extract (i.e., 3′-O-angeloylhamaudol, ledebouriellol and hamaudol) suppressed NO production, whereas the coumarins from the Hamabofu extract (i.e., umbelliferone and scopoletin) also suppressed NO production. These results suggest that linear dihydropyranochromones and coumarins are responsible for the anti-inflammatory effects of Bofu and Hamabofu. It is plausible that Bofu and Hamabofu are blended differently in several Kampo formulae due to many constituents with as yet unidentified pharmacological activity.

A new polyoxygenated cyclohexane and other constituents from Kaempferia rotunda and their cytotoxic activity

A new polyoxygenated cyclohexane and other constituents from Kaempferia rotunda and their cytotoxic activity Subehan Lallo, Sullim Lee, Dya Fita Dibwe, Yasuhiro Tezuka & Hiroyuki Morita a Faculty of Pharmacy, Biofarmaka Research Center, Hasanuddin University, Jl. Perintis Kemerdekaan Km 10, Makassar, 90245, Indonesia b Institute of Natural Medicine, University of Toyama, 2630Sugitani, Toyama, 930-0194, Japan c Faculty of Pharmaceutical Sciences, Hokuriku University, Ho-3 Kanagawa-machi, Kanazawa, 920-1181, Japan Published online: 11 Aug 2014.The isolation of secondary metabolites from a methanolic extract of Kaempferia rotunda yielded 12 compounds (1–12), including a new polyoxygenated cyclohexane compound, (–)-3-acetyl-4-benzoyl-1-benzoyloxymethyl-1,6-diepoxycyclohexan-2,3,4,5-tetrol (1). The structures of the isolated compounds were determined based on their spectroscopic data and comparison with references. All of the isolated compounds were tested for their cytotoxic activity against pancreatic (PSN-1) and breast (MDA-MB231) cancer cell lines. Compound 12 showed moderate cytotoxic activity against PSN-1 and MDA-MB231 without showing any cytotoxicity against the normal cell line, TIG-3.

The Potent Inhibitors of Protein Tyrosine Phosphatase 1B from the Fruits of Melaleuca leucadendron.

Background:Melaleuca leucadendron (Myrtaceae) is a kind of fruit used as Indonesian medicinal component and recorded in Jamu (tonic made of medical herbs) prescription records for the diabetes treatment. Its methanol extract exhibited a strong inhibitory activity with the half maximal inhibitory concentration (IC50) value of 2.05 μg/mL, while it is the same value with positive control RK-682. Objective: To isolate the chemical constituents of M. leucadendron and to evaluate their activity against protein tyrosine phosphatase 1B (PTP1B). Further, determine their toxicity potential against T-cell protein tyrosine phosphatase (TCPTP). Materials and Methods: Methanol extract was fractionated using silica column chromatography, and the obtained fraction was purified using Sephadex 20-LH. The structure of isolated compounds was identified based on 1H and 13Nuclear Magnetic Resonance Spectrometry. Furthermore, the compounds were examined against PTP1B and TCPTP. Results: Methanol extract of M. leucadendron (Myrtaceae) afforded two triterpenes: Betulinic acid and ursolic acid in high quantities. Both compounds exhibited a strong inhibitory activity against PTP1B inhibition with IC50value of 1.5 and 2.3 μg/mL, respectively (positive control RK-682, IC50 =2.05 μg/mL). Their activity toward TCPTP, on the other hand, were at 2.4 and 3.1 μg/mL, respectively. Based on this purification work, betulinic acid and ursolic acid presented 7.6% and 2.4%, respectively, as markedly M. leucadendron most potential for betulinic acid source among Indonesian plants. The result should have demonstrated that the antidiabetes of M. dendron could be through the inhibition of PTP1B.

Oviposition inhibitor in umbelliferous medicinal plants for the common yellow swallowtail ( Papilio machaon )

Umbelliferous medicinal plants, such as Angelica acutiloba Kitagawa and Angelica dahurica Bentham et Hooker filius ex Franchet et Savatier, account for a large percentage of crude drug consumption in Japan. The most serious problem in the cultivation of umbelliferous medicinal plants is the feeding damage caused by the common yellow swallowtail (Papilio machaon hippocrates C. & R. Felder, 1864). When we compared the numbers of eggs laid by P. machaon on six umbelliferous medicinal plants, the eggs on A. acutiloba, A. dahurica, and Glehnia littoralis Fr. Schmidt ex Miquel were the most numerous, those on Saposhnikovia divaricata Schischkin and Cnidium officinale Makino were rare, and Bupleurum falcatum Linné was not oviposited at all. To identify oviposition inhibitors for P. machaon in B. falcatum, S. divaricata, and C. officinale, the volatile chemical constituents of these umbelliferous medicinal plants were compared with GC–MS. We carried out multivariate analysis of gas chromatographic data and concluded that germacrene d, α-humulene, and trans-caryophyllene play important roles in protecting plants from oviposition by P. machaon. Their oviposition repellent activity was confirmed by the fact that the number of eggs laid on the leaves around a repellent device containing a mixture of germacrene d, α-humulene, and trans-caryophyllene was reduced by 40% compared to a control.

[Dedicated to Prof. T. Okada and Prof. T. Nishioka: data science in chemistry]Analysis of Volatile Constituents in Fermented Brown Rice and Rice Bran by Aspergillus oryzae (FBRA)

In recent years, consumers’ interest in health foods has increased significantly. Among these health foods, fermented foods are used traditionally in Japanese food culture and have contributed to the maintenance of peoples health. Recently, the biological effects of fermented brown rice and rice bran by Aspergillus oryzae (FBRA) have been comprehensively studied, and inhibitory effects on carcinogenesis have been reported. Regarding the bioactive chemical constituents in FBRA, the involvement of ferulic acid on the biological activity has been reported. In this study, we quantitatively investigated the dependence on fermentation time of the production of ferulic acid and related compounds in FBRA. In addition, we analyzed the generation of aroma-active compounds by fermentation.

Preferential cytotoxicity of crude drugs used in Japanese Kampo medicines against human pancreatic cancer PANC-1 and PSN-1 cells

Pancreatic cancer is the most aggressive cancer of all and has an exceptionally high global mortality rate, ranking seventh as the most frequent cause of cancer death. In order to identify an anti‐pancreatic cancer drug, we screened 24 crude drugs used in Kampo medicine for preferential cytotoxicity based on anti‐austerity strategy.