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Present status of research on cancer chemoprevention in Japan.

The activities of the research group on cancer chemoprevention supported by the Ministry of Health and Welfare of Japan are summarized. Since the research on cancer chemoprevention is quite new in Japan, this paper describes the present status of the development of possible cancer preventive agents. Acyclic retinoid (E-5166) and sarcophytol A encourage further studies. New inhibitory compounds of experimental carcinogenesis, such as cryptoporic acid, oleanolic acid, mokko lactone and ursolic acid were isolated from plants. The importance of the keen collaboration between natural product chemists and clinicians for further meaningful progress in this research field has been emphasized.

Animals↗

[Qualitative analysis of Chinese drug xiakucao (Prunella)].

A high performance gas chromatographic procedure has been developed for the quantitation of ursolic acid and oleanolic acid in different parts of Xiakucao collected in different periods. The study provides a scientific foundation for increased availability and rational collecting periods of Xiakucao.

Chromatography, Gas↗

Protection of peroxynitrite-induced DNA damage by dietary antioxidants.

The present study was undertaken to test the hypothesis that dietary antioxidants protect DNA damage induced by peroxynitrite, a potent physiological inorganic toxin. The present study showed that dietary antioxidants such as (-)-epigallocatechin gallate, quercerin, rutin, resveratrol, and ursolic acid inhibit single strand breaks in supercoiled plasmid DNA induced by 3-morpholinosydnomine N-ethylcarbamide (SIN-1), a generator of peroxynitrite through the reaction between nitric oxide and superoxide anion. The formation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in calf thymus DNA by SIN-1 was also inhibited by dietary antioxidants. When U937 cells were incubated with 1 mM SIN-1 bolus, a significant increase of 8-OH-dG level was observed. However, oxidative DNA damage was significantly lower in the cells pre-treated with dietary antioxidants when cells were exposed to SIN-1.

8-Hydroxy-2'-Deoxyguanosine↗

DNA damaging activities of methanol extract of Ajuga postii and iridoid glucoside reptoside.

An iridoid glucoside reptoside (1) has been isolated as a DNA damaging active agent by bioassay-guided fractionation of the methanol extract of Ajuga postii. Furthermore, from the acetone extract of A. postii two known triterpenic compounds ursolic acid, alpha-amyrin and two steroidal compounds (24S)-24-ethylcholesta-5,25-dien-3beta-ol and beta-sitosterol were isolated. Their structures were elucidated based on 1D and 2D NMR techniques and mass data.

Ajuga↗

Antimicrobial activity of some pentacyclic triterpenes and their synthesized 3-O-lipophilic chains.

The major metabolites of Diopsyros melanoxylon viz. amyrins and ursolic acid and their lipophilic 3-O-fatty acid ester chains (C12-C18), which are synthesized now under mild esterification conditions in excellent yields (80-95%), were evaluated for their antimicrobial activity against a series of Gram positive and Gram negative bacteria. Significantly these compounds were found to exhibit potent activity against Gram negative bacteria Pseudomonas syringae (ATCC #13457) and fairly good activity against Gram positive bacteria, Bacillus sphaericus (ATCC #14577) and Bacillus subtilis (ATCC #6051).

Anti-Bacterial Agents↗

[Triterpenes from root of Rhaponticum uniflorum].

OBJECTIVE: To study the chemical constituents of the root of Rhaponticum uniflorum. METHOD: Separation and purification were performed on silica gel and Sephadex LH-20 column chromatography. Their structure were elucidated on the basis of physicochemical and spectral analysis. RESULT: Five triterpenoid compounds were isolated and identified as ursolic acid (1), 3-oxo-19alpha-hydroxyurs-12-en-28-oic acid (2), pomolic acid (3), arjunic acid (4) and tormentic acid (5), respectively. CONCLUSION: Compounds 1 approximately 5 were isolated from the genus Rhaponticum for the first time.

Leuzea↗

DNA polymerase beta inhibitors from Baeckea gunniana.

Crude plant extracts were surveyed for their ability to inhibit DNA polymerase beta. A methyl ethyl ketone extract prepared from Baeckea gunniana was identified as a potent inhibitor of the enzyme. Bioassay-guided fractionation of the extract, using an assay to monitor the inhibitory potential of individual fractions toward DNA polymerase beta, led to the isolation of four active ursane and oleanane triterpenoids (1-4). Inhibitory principle 1 is a new natural product, and 2 is a novel compound. Their structures were established as 3 beta-hydroxyrus-12,19(29)-dien-28-oic acid (1) and 3 beta-hydroxyrus-18,20(30)-dien-28-oic acid (2) by spectroscopic analysis and by comparison with the data for the structurally related compound ursolic acid (4). Also isolated as a DNA polymerase beta inhibitor was oleanolic acid (3). Compounds 1-4 had IC50 values of 5.3-8.5 microM as inhibitors of polymerase beta in the presence of bovine serum albumin (BSA) and 2.5-4.8 microM in the absence of BSA.

Animals↗

Antimutagens from Plumeria acuminata Ait.

Four isolates, A1, C1, D3, and F2, from the ethanol extract of the green leaves of Plumeria acuminata Ait, showed antimutagenic activity. The antimutagens were isolated from the bioactive hexane and carbon tetrachloride fractions following a bioactivity-directed fractionation scheme and using the micronucleus test to monitor the antimutagenic activities. Structure elucidation studies indicated that C1 is stigmast-7-enol[1], D3 is lupeol carboxylic acid [2] and F2 is ursolic acid [3]. The structure of A1 was not fully elucidated but MS data suggested that it contained a long hydrocarbon chain. At a dosage of 2 mg isolate/25 g mouse, A1 reduced the number of micronucleated polychromatic erythrocytes (MPCE) induced by the mutagen, mitomycin C, by 75%, C1 by 80%, D3 by 57%, and F2 by 76%. Compound A2 was also isolated but was found inactive. Its structure was identified to be lupeol acetate [4].

Animals↗

Bioactive constituents of Morus australis and Broussonetia papyrifera.

The biological activities of the active principles of two plants in the Moraceae have been investigated. A new prenylflavonoid, australone A (1), and a new triterpenoid, 3 beta-[(m-methoxybenzoyl)oxy]urs-12-en-28-ioc acid (2) were isolated from the root bark of Morus australis, and their structures determined by spectroscopic methods. Also isolated from this plant were seven known compounds, morusin (3), kuwanon C (4), betulinic acid, beta-amyrin, quercetin, ursolic acid, and compound A. Morusin (3) showed significant effects on arachidonic acid-, collagen-, and PAF-induced platelet aggregation, while kuwanon C (4) was active in the arachidonic acid- and PAF-induced platelet aggregation assays. In biological work on a second plant, Broussonetia papyrifera, broussoflavonols F (5) and G (6), broussoflavan A (7), and broussoaurone A (8) potently inhibited Fe(2+)-induced lipid oxidation in rat-brain homogenate. Compounds 5-7 also significantly inhibited the proliferation of rat vascular smooth muscle cells.

Animals↗

[Studies on the chemical constituents in herb of Ludwigia octovalvis].

OBJECTIVE: To study the chemical constituents of the herb of Ludwigia octovalvis. METHOD: Chemical constituents were isolated by the repeated silica gel column chromatography, and their structures were elucidated by the physicochemical properties and spectral analysis. RESULT: Thirteen compounds were obtained and determined as follows: beta-sitosterol (1), oleanolic acid (2), 2alpha-hydroxy ursolic acid (3), tormentic acid (4), daucosterol (5), maltol (6), luteolin (7), quercetin (8), apigenin (9), methyl brevifolincarboxylate (10), gallic acid (11), 3, 4, 8, 9, 10-pentahydroxydibenzo[b, d]pyran-6-one (12), and ellagic acid (13). CONCLUSION: Compounds 3, 4, 6-13 were isolated from the plant for the first time. And compounds 3, 4, 6, 10, 12 were obtained from the genus for the first time.

Molecular Structure↗

Cytotoxic triterpene acids from the Peruvian medicinal plant Polylepis racemosa.

Cytotoxicity-guided fractionation of the bark and stem extract of Polylepis racemosa led to the identification of ursolic acid, pomolic acid, 3-O-acetylpomolic acid, and 2-oxopomolic acid. Pomolic acid was the most cytotoxic component, and was specific for M-14 melanoma and ME180 cervical carcinoma, with GI50 values of 6.9 and 8.3 micrograms/mL respectively.

Animals↗

[Study on the chemical components of Rabdosia excisa].

Four kinds of components were extracted and separated from the stems and leaves of Rabdosia excisa. They had been identified as Kamebakaurin, Ursolic acid, Daucosterol and Liquiritin respectively by physical and chemical methods, as well as spectrum data. It was first time that Liquiritin was obtained from the Robdosia family, and its 13C-NMR data of Liquiritin had not been reported before.

Chromatography, Thin Layer↗

[Chemical constituents in the roots of Salvia przewalskii Maxim].

AIM: To investigate the chemical composition of the root of Salvia przewalskii Maxim. METHODS: Compounds were isolated by silica gel column chromatography. Structures of these compounds were elucidated by spectral analysis (EI-MS, FAB-MS, 1HNMR, 13CNMR, 1H-1H COSY, 1H-13C COSY, HMBC, NOESY) and phytochemical properties. RESULTS: Eight compounds were isolated and identified as: tanshinone II-A (I), crypotanshinone (II), przewaquinone A (III), sugiol (IV), ursolic acid (V), 2 alpha, 3 alpha-dihydroxy urs-12-ene-28-acid (VI), oleanolic acid (VII), and neo-przewaquinone A (VIII). CONCLUSION: Compound VIII is a new compound, and compound II, IV, V, VI and VII are isolated from this plant for the first time.

Heterocyclic Compounds, 4 or More Rings↗

Eclalbatin, a triterpene saponin from Eclipta alba.

From the whole plant of Eclipta alba, a new triterpene saponin, named eclalbatin, together with alpha-amyrin, ursolic acid and oleanolic acid were isolated. The structure of eclalbatin has been established as 3-O-beta-D-glucopyranosyl-3-beta-hydroxy-olean-12-en-28-oic acid, 28-O-beta-D-arabinopyranoside (1) on the basis of chemical and spectral data.

Asteraceae↗

Isolation of substances from glossy privet (Ligustrum lucidum Ait.) inhibiting the mutagenicity of benzo[a]pyrene in bacteria.

Methanol and hot-water extracts of glossy privet (Ligustrum lucidum Ait.) inhibited the mutagenic activity of benzo[a]pyrene in Salmonella typhimurium TA98 with S9 mix. The methanol extract was fractionated with ether and n-hexane. As the active components, oleanolic and ursolic acids were isolated, which were soluble in ether and insoluble in n-hexane. The hot-water extract was fractionated to water, 60% and 100% methanol fractions. Nuezhenide was isolated from the 60% methanol fraction as the active component.

Antimutagenic Agents↗

Determination of gambogic acid in human plasma by liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry.

A high-performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (HPLC-APCI-MS) method was established for the determination of gambogic acid (GA) in human plasma using ursolic acid as the internal standard (I.S.). Plasma samples were extracted with ethyl acetate and separated on a Hanbon Lichrospher 5-C18 column with a mobile phase of acetonitrile-tetrahydrofuran-water (70:23:7, v/v). Gambogic acid was determined by using atmospheric pressure chemical ionization (APCI) in a single quadrupole mass spectrometer. HPLC-APCI-MS was performed in the selected ion monitoring (SIM) mode using target ions at [M-H](-)m/z 627.4 for gambogic acid and [M-H](-)m/z 455.4 for the I.S. Calibration curve was linear over the range of 3.108-4144 microg/L. The lower limit of quantification was 3.108 microg/L. The intra- and inter-run precisions were less than 12.3 and 14.1%, respectively. The method has been successfully applied to study the pharmacokinetics of gambogic acid in patients with malignant tumour.

Atmospheric Pressure↗

Simultaneous quantification of three major bioactive triterpene acids in the leaves of Diospyros kaki by high-performance liquid chromatography method.

The leaf of Diospyros kaki, which is a traditional Chinese medicine, has been used for the treatment of various diseases. In order to improve the quality assurance of the leaves of D. kaki, derived extracts and phytomedicines, a simple, rapid and accurate high-performance liquid chromatography (HPLC) method was developed to simultaneously assess the three bioactive triterpene acids: barbinervic acid (BA) and its epimer, rotungenic acid (RA), along with 24-hydroxy ursolic acid (HA). This HPLC assay was performed on a reversed-phase C18 column with methanol and aqueous H3PO4 as the mobile phase and using a monitoring wavelength at 210 nm. This method was successfully applied to quantify these three bioactive triterpene acids in five different solvent extracts of the leaves of D. kaki and in the leaves from six different locations in China. The results demonstrated the total content and quantity of each of the main bioactive compounds were strongly dependent on the extraction solvents and locations, indicating that the quality control of the bioactive ingredients in the leaves of D. kaki, derived extracts and phytomedicines is critical to ensure its clinical benefits. The content of the total triterpenoids was also determined by the less selective colorimetric method, and the comparison with the HPLC method was given.

Calibration↗