Boonchoo Sritularak

Phenolics with antiviral activity from Millettia Erythrocalyx and Artocarpus Lakoocha

From the leaves of Millettia erythrocalyx, a new flavone named 3′,5′-dimethoxy-[2′′,3′′ : 7,8]-furanoflavone and three known compounds were isolated. Assays for anti-herpes simplex virus activity (HSV-1 and HSV-2) were performed on 24 phenolic compounds obtained from M. erythrocalyx and Artocarpus lakoocha. It was found that the flavones ovalifolin, pongol methyl ether and millettocalyxin A, and the stilbene oxyresveratrol possessed moderate activity against both types of HSV. In addition, oxyresveratrol was evaluated for potential anti-HIV activity against a wild-type human immunodeficiency virus type 1 (HIV-1/LAI) isolate and was found to be a modest inhibitor of HIV (EC50 28.2 µM), showing no toxicity in PBM, CEM and Vero cells at 100 µM. The heartwood of A. lakoocha, which contains a large amount of oxyresveratrol, could be considered as a source of starting material for the development of new natural product-based anti-HSV and anti-HIV agents.

Gigantol, a Bibenzyl from Dendrobium draconis, Inhibits the Migratory Behavior of Non-Small Cell Lung Cancer Cells

Lung cancer is one of the most common causes of cancer death due to its high metastasis potential. The process of cancer migration is an early step that is required for successful metastasis. The discovery and development of natural compounds for cancer therapy have garnered increasing attention in recent years. Gigantol (1) is a bibenzyl compound derived from the Thai orchid, Dendrobium draconis. It exhibits significant cytotoxic activity against several cancer cell lines; however, until recently, the role of 1 on tumor metastasis has not been characterized. This study demonstrates that 1 suppresses the migratory behavior of non-small cell lung cancer H460 cells. Western blot analysis reveals that 1 down-regulates caveolin-1 (Cav-1), activates ATP-dependent tyrosine kinase (phosphorylated Akt at Ser 473), and cell division cycle 42 (Cdc42), thereby suppressing filopodia formation. The inhibitory effect of 1 on cell movement is also exhibited in another lung cancer cell line, H292, but not in normal human keratinocytes (HaCat). The inhibitory activity of 1 on lung cancer migration suggests that this compound may be suitable for further development for the treatment of cancer metastasis.

Quantitative analysis of ginsenosides Rb1, Rg1, and Re in American ginseng berry and flower samples by ELISA using monoclonal antibodies

Quantitative analysis of ginsenosides Rb1, Rg1, and Re in American ginseng berry and flower samples, which were collected in various months throughout the year, was performed by enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies. The American ginseng flower had the highest content of ginsenosides Rb1, Rg1, and Re (GRb1, GRg1, and GRe). The content of GRb1, GRg1, and GRe decreased in going from young through to ripe berry.

Moscatilin Inhibits Lung Cancer Cell Motility and Invasion via Suppression of Endogenous Reactive Oxygen Species

Lung cancer is the leading cause of death among cancer patients worldwide, and most of them have died from metastasis. Migration and invasion are prerequisite processes associated with high metastasis potential in cancers. Moscatilin, a bibenzyl derivative isolated from the Thai orchid Dendrobium pulchellum, has been shown to have anticancer effect against numerous cancer cell lines. However, little is known regarding the effect of moscatilin on cancer cell migration and invasion. The present study demonstrates that nontoxic concentrations of moscatilin were able to inhibit human nonsmall cell lung cancer H23 cell migration and invasion. The inhibitory effect of moscatilin was associated with an attenuation of endogenous reactive oxygen species (ROS), in which hydroxyl radical (OH∙) was identified as a dominant species in the suppression of filopodia formation. Western blot analysis also revealed that moscatilin downregulated activated focal adhesion kinase (phosphorylated FAK, Tyr 397) and activated ATP-dependent tyrosine kinase (phosphorylated Akt, Ser 473), whereas their parental counterparts were not detectable changed. In conclusion, our results indicate the novel molecular basis of moscalitin-inhibiting lung cancer cell motility and invasion and demonstrate a promising antimetastatic potential of such an agent for lung cancer therapy.

A new phenanthrenequinone from Dendrobium draconis

A number of Dendrobium species (Orchidaceae) have been used as health foods. In Thailand, the tea prepared from the stems of Dendrobium draconis Rchb.f. (Orchidaceae) has been used as a blood tonic. Our phytochemical investigation of this plant led to the isolation of a new compound namely 5-methoxy-7-hydroxy-9,10-dihydro-1,4-phenanthrenequinone (1), along with four known stilbenoids including hircinol (2), gigantol (3), batatasin III (4), and 7-methoxy-9,10-dihydrophenanthrene-2,4,5-triol (5). The structures of these compounds were determined through extensive spectroscopic studies, including 1H and 13C NMR, DEPT, COSY, NOESY, HMQC, HMBC, ESI-MS, and HR-ESI-MS experiments. In the DPPH-free radical assay, these stilbene derivatives showed appreciable antioxidant activity.

Development of a combined technique using a rapid one-step immunochromatographic assay and indirect competitive ELISA for the rapid detection of baicalin.

A colloidal gold conjugated anti-baicalin monoclonal antibody (anti-BA MAb) was prepared and used in an immunochromatographic assay (ICA) for BA in Scutellariae Radix and Kampo medicines. This competitive ICA uses an anti-BA MAb which shows a high specificity for BA and baicalein. Its advantages include a short assay time (15 min), no dependence on any instrumental systems, and it can detect BA in plant materials and Kampo medicines. The limit of detection for the ICA was found to be around 0.6 μg mL(-1)of baicalin. Moreover, the usefulness of the combination of indirect competitive ELISA and the ICA using anti-BA MAb as a quality control method was confirmed for analysis of BA in Scutellariae Radix and Kampo medicines with a sufficient sensitivity (200 ng mL(-1) to 2 μg mL(-1)), obtainable in an easy and timely manner.

Topical cream-based oxyresveratrol in the treatment of cutaneous HSV-1 infection in mice

Anti-herpes simplex virus (HSV) activities of oxyresveratrol in vitro and topical administration in cutaneous HSV-1 infection in mice were examined. The inhibitory concentrations for 50% plaque formation (IC(50)) of oxyresveratrol against HSV-1 clinical isolates and HSV-2 clinical isolates were 20.9-29.5 and 22.2-27.5 μg/ml, respectively. In topical administration in cutaneous HSV-1 infection in mice, 2.5%, 5%, 10% and 20% oxyresveratrol in cream vehicle applied three times daily for 7 days after infection were evaluated and 10% and 20% oxyresveratrol cream were significantly effective in delaying the development of skin lesions and protection from death (P < 0.01). The concentration of 10% oxyresveratrol in cream was significantly more effective than that of 30% oxyresveratrol in vaseline applied three times daily (P < 0.01). Oxyresveratrol cream at 20% was as effective as 5% ACV cream applied three times daily (P < 0.01). Both 10% and 20% oxyresveratrol cream were as effective as that of 5% ACV cream applied two times daily (P > 0.05). Therapeutic efficacy of oxyresveratrol in cream vehicle was dose-dependent and the maximum efficacy observed on day 6 after infection was shown at 10% oxyresveratrol in cream applied three times daily. The frequency of application of 10% oxyresveratrol cream at three, four and five times daily was as effective as that of 5% ACV cream applied five times daily (P > 0.05). These results demonstrated that topical administration of oxyresveratrol in novel cream vehicle reduced the concentration of oxyresveratrol to 10% and was suitable for cutaneous HSV infection.

Preparation of a specific monoclonal antibody to asiaticoside for the development of an enzyme-linked immunosorbent assay

Asiaticoside (AS), the major active component of Centella asiatica (L.) Urban, is used as a memory enhancer and for wound healing. We have successfully prepared monoclonal antibodies (MAbs) against AS, and developed an enzyme-linked immunosorbent assay (ELISA) system for its determination. AS was conjugated to the carrier protein bovine serum albumin (BSA), which acted as an immunogen. In order to confirm its immunogenicity, the ratio of hapten in the AS-BSA conjugate was determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). After immunization, hybridomas secreting MAbs against AS were produced by fusing splenocytes with the mouse myeloma cell line, SP2/0-Ag14. After the screening, anti-asiaticoside MAb 2B4 was obtained. Weak cross-reactivities occurred with madecassoside (7.08%), but no cross-reactivities were observed with other related triterpenoid glycosides (<0.01%). The assay was suitable for quantitating AS in the range of 0.78 to 50 µg mL(-1). A good correlation of AS concentrations in crude extracts of C. asiatica between ELISA and HPLC methods was obtained (r(2) = 0.999). The contents of AS in various cultivated C. asiatica samples were assayed by the newly established ELISA. The recovery rates of AS in the samples were in the range of 95-103% with coefficients of variation of <10%. The intra- and inter-assay variations were 3.9 and 4.5%, respectively. The ELISA method described should prove useful as an analytical tool for quality control and standardization of medicinal plants and pharmaceutical products containing AS.

Phenolics with Anti-HSV and Anti-HIV Activities from Artocarpus gomezianus., Mallotus pallidus., and Triphasia trifolia.

Abstract A total of 25 phenolic compounds were studied for their inhibitory effects against herpes simplex virus (HSV)-1, HSV-2, and human immunodeficiency virus (HIV)-1. These include five flavonoids (1–5) and two dimeric stilbenes (6,7) from Artocarpus gomezianus. Wall. ex tréc., five phloroglucinol derivatives (8–12) from Mallotus pallidus. (Airy Shaw) Airy Shaw, and 13 courmarins (13–25) from Triphasia trifolia. (Burm.f.) P. Wilson. The data obtained in this study suggest the bis-hydroxyphenyl structure as a potential target for anti-HSV and HIV drugs development.

Flavones with free radical scavenging activity from Goniothalamus tenuifolius.

From the leaves ofGoniothalamus tenuifolius, a new natural product namely 3′-hydroxy-3,5,7,4′-tetramethoxyflavone (1) was isolated, along with seven other known compounds (2–8). Each of these isolates was evaluated for free radical scavenging activity on the DPPH decoloration test. The data obtained in this study suggested that theortho 3′,4′-diphenolic structure was essential for the activity of these flavonol derivatives.