Hui-Ming Wu

Antioxidant and Anticancer Aporphine Alkaloids from the Leaves of Nelumbo nucifera Gaertn. cv. Rosa-plena

Fifteen compounds were extracted and purified from the leaves of Nelumbo nucifera Gaertn. cv. Rosa-plena. These compounds include liriodenine (1), lysicamine (2), (−)-anonaine (3), (−)-asimilobine (4), (−)-caaverine (5), (−)-N-methylasimilobine (6), (−)-nuciferine (7), (−)-nornuciferine (8), (−)-roemerine (9), 7-hydroxydehydronuciferine (10) cepharadione B (11), β-sitostenone (12), stigmasta-4,22-dien-3-one (13) and two chlorophylls: pheophytin-a (14) and aristophyll-C (15). The anti-oxidation activity of the compounds was examined by antiradical scavenging, metal chelating and ferric reducing power assays. The results have shown that these compounds have antioxidative activity. The study has also examined the antiproliferation activity of the isolated compounds against human melanoma, prostate and gastric cancer cells. The results shown that 7-hydroxydehydronuciferine (10) significantly inhibited the proliferation of melanoma, prostate and gastric cancer cells. Together, these findings suggest that leaves of Nelumbo nucifera Gaertn. cv. Rosa-plena are a good resource for obtaining the biologically active substances with antioxidant properties.

The Pharmacological Activities of (−)-Anonaine

Several species of Magnoliaceae and Annonaceae are used in Traditional Chinese Medicine. (−)-Anonaine, isolated from several species of Magnoliaceae and Annonaceae, presents antiplasmodial, antibacterial, antifungal, antioxidation, anticancer, antidepression, and vasorelaxant activity. This article provides an overview of the pharmacological functions of (−)-anonaine.

The Effect of Butanolides from Cinnamomum tenuifolium on Platelet Aggregation

This study investigated the effects of isotenuifolide and tenuifolide B from the stems of Cinnamomum tenuifolium on adenosine diphosphate (ADP)-induced human platelet aggregation. Treatment of human platelet-rich plasma with isotenuifolide (1 and 2 μg/μL) and tenuifolide B (1, 2 and 4 μg/μL) did not have any significant effect on human platelet aggregation in vitro, however, treatment of human platelet-rich plasma with isotenuifolide (4 μg/μL) resulted in an inhibitory effect on platelet aggregation, suggesting the potential of this compound as an anti-atherosclerogenic agent in humans. Isotenuifolide is a new butanolide compound, whose structure was characterized by spectral analyses.

Chemical Constituents of the Stems of Michelia champaca

Champacaine (1), (–)-anonaine (2), (–)-norushinsunine (3), (–)-ushinsunine (4), (–)-N-acetylanonaine (5), (–)-roemerine (6), (–)-asimilobine (7), (–)-anolobine (8), (–)-isocorydine (9), liriodenine (10), atherospermidine (11), O-methylmoschatoline (12), (+)-syringaresinol (13), N-trans-feruloyltyramine (14), 4-hydroxybenzaldehyde (15), vanillin (16), vanillic acid (17), syringic acid (18), 3,4-dimethoxybenzoic acid (19), coniferyl aldehyde (20), syringin (21), scopoletin (22), 4-acetonyl-3,5-dimethoxy-p-quinol (23), β-sitostenone (24), and stigmasta-4,22-dien-3-one (25) were isolated from the stems of Michelia champaca L. (Magnoliaceae). Among them, 1 is a new 4,5-dioxoaporphine compound. The structures of these compounds were characterized and identified by spectral analyses.

Secondary Metabolites from the Leaves of Ilex Cornuta

Ilex (Aquifoliaceae) species are found throughout China, and some are used extensively in traditional Chinese medicine. For example, I. rotunda is antipyretic and used for the treatment of the common cold, tonsillitis, and stomach and intestinal ulcers. I. pubescens is used for the treatment of coronary disease, myocardial infarction, dysentery, and erysipelas, and I. cornuta and I. latifolia are used for the treatment of headaches, toothaches, bloodshot eyes, and tinnitus [1]. Previous studies on the Ilex genus have resulted in the isolation of essential oils [2] triterpenes, triterpene glycosides [3–7], hemiterpene glycosides [8], and phenolic compounds [9, 10]. In the present study, 12 compounds were isolated from the leaves of I. cornuta. A MeOH extract of the I. cornuta leaves was concentrated to obtain a residue, which was partitioned between CH2Cl2 and H2O. The aqueous layer was extracted with n-BuOH, and then the n-BuOH layer was concentrated and analyzed by chromatography. As a result, 12 compounds, including eight flavonoids and four phenolic carboxylic acids, were isolated. The flavonoids were quercetin [11], quercetin 3-O-D-glucopyranoside [12], quercetin 3-O-D-galactopyranoside [13], quercetin 3-O-D-glucopyranosyl-(1 6)-D-glucopyranoside [14], isorhamnetin [15], isorhamnetin 3-O-Dglucopyranoside [15], isorhamnetin 3-O-D-galactopyranoside [15], and isorhamnetin 3-O-D-glucopyranosyl-(1 6)D-glucopyranoside [15]. The phenolic carboxylic acids were caffeic acid [16], p-hydroxybenzoic acid [17], protocatechuic acid [11], and vanillic acid [16]. This is the first report of the isolation of these compounds from this source. Leaves from I. cornuta Lindl. & Paxton were collected in Kaohsiung, Taiwan, in October 2006. Plant material was identified by Dr. Fu-Yuan Lu (Department of Forestry and Natural Resources, College of Agriculture, National Chiayi University, Chiayi City, Taiwan). A voucher specimen was deposited in the School of Medical and Health Sciences, Fooyin University, Kaohsiung, Taiwan. The air-dried leaves of I. cornuta (4.2 kg) were extracted with MeOH (5 10 L) at room temperature, and each MeOH fraction was concentrated under reduced pressure. The MeOH fractions were concentrated by evaporation and then partitioned to yield CH2Cl2 and aqueous extracts. The aqueous extracts were concentrated by evaporation and extracted with n-BuOH to give a n-BuOH layer and an aqueous layer. The n-BuOH layer (178.6 g) was subjected to silica gel chromatography (500 g, 70–230 mesh) with EtOAc–MeOH elution. Ten fractions were obtained. Part of fraction 2 (20.3 g) was subjected to silica gel chromatography with n-hexane–EtOAc (1:1, v/v). The volume fraction of EtOAc was gradually increased to yield five fractions (2-1–2-5). Fraction 2-2 (2.2 g) was further purified on a silica gel column using an n-hexane– CH2Cl2 mixture to obtain caffeic acid (7 mg), vanillic acid (3 mg), and p-hydroxybenzoic acid (5 mg). Fraction 2-4 (5.7 g) was further purified on a silica gel column with n-hexane–EtOAc elution to obtain isorhamnetin (12 mg). Part of fraction 3 (12.7 g) was subjected to silica gel chromatography with n-hexane–EtOAc (1:8, v/v). The volume fraction of EtOAc was gradually increased to yield quercetin (20 mg) and protocatechuic acid (34 mg). Part of fraction 4 (32.5 g) was subjected to silica gel chromatography with EtOAc–MeOH (20:1, v/v). The volume fraction of MeOH was gradually increased to yield isorhamnetin 3-O-D-glucopyranoside (67 mg) and isorhamnetin 3-O-D-galactopyranoside quercetin (42 mg). Part of fraction 5 (17.3 g) was subjected to silica gel chromatography with EtOAc–MeOH (20:1). The volume fraction of MeOH was gradually increased to obtain five further fractions (5-1–5-5). Fraction 5-3 (5.8 g) was further purified on a silica gel column with EtOAc–MeOH to obtain quercetin 3-O-D-glucopyranoside (78 mg) and quercetin 3-O-D-galactopyranoside (113 mg). Fraction 6-8 (22.7 g) was eluted with EtOAc–MeOH (8:1) and was repeatedly subjected to silica gel chromatography to give quercetin 3-O-D-glucopyranosyl-(1 6)-D-glucopyranoside (56 mg) and isorhamnetin 3-O-D-glucopyranosyl-(1 6)-D-glucopyranoside (24 mg).

Secondary Metabolites from the Stems of Capsicum annuum var. conoides

Red pepper, Capsicum annuum (Solanaceae), is used as a spice all over the world. Red pepper is studied actively because its pungent principal component, capsaicin, has a dietary effect, analgesic activity, and antioxidant activity [1]. The pungent principal component of red peppers is a group of acid amides of vanillylamine and C8 to C13 fatty acids, which are known generally as capsaicin [2]. More than 16 other capsaicinoids have been found as minor components [3]. Numerous studies have been done on the red pepper fruit, but there are few studies on stems [2–4]. The chemical constituents of stems of C. annuum L. var. conoides Irish have not yet been reported. This study has shown the isolation of secondary metabolites from the stems of C. annuum var. conoides. To further understand the chemotaxonomy and to continue our search for biologically and chemically novel agents from Solanaceous plants, the stems of C. annuum collected from Chiayi County, Taiwan in March, 2011 were chosen for further phytochemical investigation. The MeOH extract of its plants were subjected to solvent partitioning and chromatographic separation to afford 19 pure substances. The chemical constituents in the plant of C. annuum var. conoides were separated by column chromatography. Investigation of the MeOH extract of the plants has led to the isolation of 21 compounds, 10 amides 1–10, four steroids 11–14, one lignan 15, and six benzenoids 16–21. These compounds were obtained and characterized by comparison of their physical and spectral data (UV, IR, NMR, and MS) with values in the literature. All of these compounds were found for the first time from this species. The specimen of C. annuum var. conoides Irish was collected from Chiayi County, Taiwan in March, 2011. A voucher specimen was characterized by Dr. Jin-Cherng Huang of the Department of Forest Products Science and Furniture Engineering, National Chiayi University, Chiayi, Taiwan and deposited in the School of Medical and Health Sciences, Fooyin University, Kaohsiung, Taiwan. The air-dried stems of C. annuum var. conoides (6.2 kg) were extracted with MeOH (10 L 4) at room temperature, and an MeOH extract (64.3 g) was obtained upon concentration under reduced pressure. The MeOH extract was chromatographed over silica gel using CH2Cl2–MeOH as eluent to produce 10 fractions. Part of fraction 2 (6.52 g) was subjected to silica gel column chromatography by eluting with n-hexane–acetone (50:1) to furnish seven fractions (2-1–2-7). Fraction 2-2 (2.52 g) was further purified by another silica gel column using n-hexane–acetone (10:1) to obtain p-hydroxybenzoic acid (16) (2 mg) and p-hydroxybenzaldehyde (17) [5] (5 mg). Fraction 2-3 (1.86 g) was further purified by another silica gel column using n-hexane–acetone (9:1) to obtain vanillic acid (18) [5] (2 mg) and isovanillic acid (19) [5] (3 mg). Part of fraction 3 (5.12 g) was subjected to silica gel column chromatography by eluting with n-hexane–acetone (40:1), then enriched with acetone to furnish eight fractions (3-1–3-8). Fraction 3-1 (1.24 g) eluted with n-hexane–EtOAc (30:1) was further separated using silica gel column chromatography and preparative TLC (n-hexane–EtOAc, 10:1) to give a mixture of -sitosterol (11) [6] and stigmasterol (12) [6] (37 mg). Fraction 3-2 (1.41 g) was further purified on a silica gel column using the CH2Cl2–MeOH system to obtain (+)-syringaresinol (15) [7] (5 mg), ferulic acid (20) [8] (3 mg), and hydroferulic acid (21) [9] (2 mg).

Cinnapine, a New Pyridine Alkaloid from Cinnamomum Philippinense

Cinnapine (1), a new pyridine alkaloid, has been isolated from the roots of Cinnamomum philippinense (Lauraceae). The structure was characterized and identified by spectral analysis.

Secondary Metabolites from the Leaves of Aquilaria sinensis

Lignum Aquilariae Resinatum has been widely used in the treatment of various kinds of pain, cough, and anaphylaxis for hundreds of years in Asia, especially in China, Vietnam, and Indonesia. The genus Aquilaria (Thymelaeaceae) is widely distributed in Asia. Aquilaria sinensis Gilg is of particular interest economically because it is the principal source of agarwood, one of the most highly valued forest products currently traded internationally. The leaves of A. sinensis Gilg, which is widely cultivated in Guangdong, Hainan, and Taiwan Provinces in China, are reported to be used locally in trauma-related diseases such as fracture, bruise, etc. [1]. Previous phytochemical investigations on Chinese eaglewood have revealed characteristic sesquiterpenes and chromone derivatives [2–15] but few on the chemical constituents of the leaves [16, 17]. The analgesic and anti-inflammatory activities of the ethanol extract of A. sinensis leaves were observed in various experimental models related to nociception and inflammation, thus providing some evidence for its traditional use. In the investigations of interrelated studies, it was found that agarwood has significant anticancer activities [18], analgesic and anti-inflammatory activities [19], and anti-depressant activities [20, 21]. These observations provide useful information for potential chemopreventive drug design. The MeOH extract of its leaves were subjected to solvent partitioning and chromatographic separation to afford 12 pure substances. Twelve compounds, including three flavonoids, 5-hydroxy-4 ,7-dimethoxyflavonoid (1) [22], luteolin-7,3 ,4 -trimethyl ether (2) [23], and 5,3 -dihydroxy-7,4 -dimethoxyflavone (3) [24], five benzenoids, methylparaben (4) [25], vanillic acid (5) [26], p-hydroxybenzoic acid (6) [27], syringic acid (7) [28], and isovanillic acid (8) [28], and four steroids, -sitosterol (9) [29], stigmasterol (10) [29], -sitostenone (11) [29], and stigmasta-4,22-dien-3-one (12) [29], were isolated from the leaves of A. sinensis. All of these compounds (1–12) were obtained for the first time from the leaves of this plant. The specimen of A. sinensis was collected from Guansi Township, Hsinchu County, Taiwan in May, 2007. A voucher specimen was identified by Prof. Fu-Yuan Lu (Department of Forestry and Natural Resources, College of Agriculture, National Chiayi University) and was deposited in the School of Medical and Health Sciences, Fooyin University, Kaohsiung, Taiwan. The leaves (1.2 kg) of A. sinensis were air dried and extracted repeatedly with MeOH (6 L 8) at room temperature. The combined MeOH extracts (31.3 g) were then evaporated and further separated into four fractions by column chromatography on silica gel (4.3 kg, 70–230 mesh) with gradients of n-hexane–CH2Cl2–acetone–MeOH. Part of fraction 1 (11.6 g) was subjected to silica gel chromatography by eluting with n-hexane–acetone (50:1), enriched with acetone, to furnish three further fractions (1-1–1-3). Fraction 1-1 (2.8 g) was further purified on a silica gel column using n-hexane–acetone mixtures to obtain 5-hydroxy-4 ,7-dimethoxyflavonoid (1) (4.8 mg). Part of fraction 1-2 (2.5 g) was subjected to silica gel chromatography by eluting with n-hexane–acetone (50:1), enriched gradually with acetone, to furnish two fractions (1-2-1–1-2-2). Fraction 1-2-1 (0.5 g) was further purified on a silica gel column using n-hexane–acetone mixtures to yield luteolin-7, 3 ,4 -trimethyl ether (2) (3.4 mg) and 5,3 -dihydroxy-7,4 -dimethoxyflavone (3) (3.2 mg). Fraction 1-2-2 (0.5 g) was further purified on a

Secondary Metabolites from the Stems of Capsicum annuum var. longum

Red pepper, Capsicum annuum (Solanaceae), is used as a spice all over the world. Red pepper is studied actively because its pungent principal component, capsaicin, has a dietary effect and analgesic and antioxidant activity [1]. The pungent principal component of red peppers is a group of acid amides of vanillylamine and C8 to C13 fatty acids, which are known generally as capsaicin [2]. More than 16 other capsaicinoids have been found as minor components [3]. Numerous studies have been done on the red pepper fruit, but there are few studies on stems [2–4]. Earlier investigations on the chemical constituents of stems of C. annuum L. dealt with a new amide, 7 -(4 -hydroxyphenyl)-N-[(4-methoxyphenyl)-ethyl]propenamide, along with 13 known compounds [2]. The chemical constituents of stems of C. annuum L. var. longum Sendtn. cv. have not yet been reported, and this study shows their isolation. To further understand the chemotaxonomy and to continue searching for biologically and chemically novel agents from Solanaceous plants, the stems of C. annuum L. var. longum Sendtn. cv. were chosen for further phytochemical investigation. The MeOH extract of its plants was subjected to solvent partitioning and chromatographic separation to afford 19 pure substances. The chemical constituents in the plant of C. annuum L. var. longum Sendtn. cv. were separated with column chromatography. Investigation of the MeOH extract of the plants has led to the isolation of 19 compounds, including eight amides: N-trans-feruloyltyramine (1) [5], N-cis-feruloyltyramine (2) [6], N-trans-caffeoyltyramine (3) [5], N-cis-caffeoyltyramine (4) [7], N-p-trans-coumaroyltyramine (5) [8], N-p-cis-coumaroyltyramine (6) [8], cinnaretamine (7) [9], and cinnabutamine (8) [10]; four steroids: -sitosterol (9) [5], stigmasterol (10) [5], -sitostenone (11) [5], and stigmasta-4,22-dien-3-one (12) [5]; three lignans: (+)-syringaresinol (13) [11], (+)-dia-syringaresinol (14) [12], and threo-2,3-bis-(4-hydroxy-3-methoxyphenyl)3-methoxypropanol (15) [13]; three benzenoids: p-hydroxybenzoic acid (16) [14], p-hydroxybenzaldehyde (17) [14], and vanillic acid (18) [14]; and one ionone: (+)-abscisic acid (19) [15]. These compounds were obtained and characterized by comparison of their physical and spectral data (UV, IR, NMR, and MS) with values obtained in the literature. All of these compounds were found for the first time from this species. The specimen of C. annuum L. var. longum Sendtn. cv. was collected from Chiayi County, Taiwan in April, 2010. A voucher specimen was characterized by Dr. Jin-Cherng Huang of the Department of Forest Products Science and Furniture Engineering, National Chiayi University, Chiayi, Taiwan and deposited in the School of Medical and Health Sciences, Fooyin University, Kaohsiung County, Taiwan. The air-dried stems of C. annuum L. var. longum Sendtn. cv. (12.5 kg) were extracted with MeOH (30 L 6) at room temperature, and the MeOH extract (302.7 g) was obtained upon concentration under reduced pressure. The MeOH extract was chromatographed over silica gel using CH2Cl2–MeOH as eluent to produce 13 fractions. Part of fraction 3 (12.23 g) was subjected to silica gel column chromatography by eluting with n-hexane–acetone (60:1) to furnish 10 fractions (3-1–3-10). Fraction 3-3 (5.83 g) was further purified by another silica gel column using n-hexane–acetone (10:1) to obtain 16 (5 mg) and 17 (7 mg). Part of fraction 4 (14.56 g) was subjected to silica gel column chromatography by eluting with n-hexane–acetone (30:1), then enriched with acetone to furnish six fractions (4-1–4-6). Fraction 4-1 (5.45 g) eluted with n-hexane–EtOAc (30:1) was further separated using silica gel column chromatography and preparative TLC (n-hexane–EtOAc,

Secondary Metabolites from the Stems of Nelumbo nucifera cv. Rosa-plena

Nelumbo nucifera Gaertn. cv. Rosa-plena is a perennial aquatic crop grown and consumed throughout Asia. All parts of N. nucifera have been used for various medicinal purposes in oriental medicine. In particular, the leaves are known for diuretic and astringent properties and are used to treat fever, sweating, and strangury, and as a styptic. [1]. To further understand the chemotaxonomy of the Nelumbo species [1, 2], Nelumbo nucifera Gaertn. cv. Rosa-plena were chosen for phytochemical investigation. There is only one paper describing the constituents of this scientific name [3]. Previously, we have isolated seven aporphines, two oxoaporphines, one dioxoaporphine, one dehydroaporphine, two steroids, seven flavonoids, and two chlorophylls from this plant [3, 4]. As part of our continuing investigation of the phytochemical and bioactive compounds of Nelumbo plants, five aporphines, (–)-nuciferine [2], (–)-nornuciferine [5], (–)-asimilobine [6], (–)-anonaine [5] and (–)-roemerine [2]; two oxoaporphines, lysicamine [5], and liriodenine [5]; two isoquinolones, northalifoline (1) [7] and thalifoline (2) [7]; and two steroids, -sitostenone [6] and stigmasta-4,22-dien-3-one [6], were obtained by systematic extraction and isolation from the stems of Nelumbo nucifera Gaertn. cv. Rosa-plena. Compounds 1 and 2 were isolated for the first time from Nelumbo nucifera Gaertn. cv. Rosa-plena. These compounds were obtained and characterized by comparison of their physical and spectral data with values obtained in the literature [2, 5–7]. The stems of Nelumbo nucifera Gaertn. cv. Rosa-plena were collected from Tainan County, Taiwan, November 2008. Plant material was identified by Dr. Fu-Yuan Lu (Department of Forestry and Natural Resources, College of Agriculture, National Chiayi University). A voucher specimen (Nelumbo nucifera Gaertn. cv. Rosa-plena) was deposited in the School of Medical and Health Sciences, Fooyin University, Kaohsiung, Taiwan. The air-dried stems of N. nucifera Gaertn. cv. Rosa-plena (0.7 kg) were extracted with MeOH (5 L 4) at room temperature, and a MeOH extract (24.3 g) was obtained upon concentration under reduced pressure. The MeOH extract was chromatographed over silica gel (580 g, 70–230 mesh) using n-hexane–EtOAc–MeOH mixtures as eluents to produce four fractions. Part of fraction 1 (3.56 g) was subjected to silica gel chromatography by eluting with n-hexane–acetone (10:1) enriched gradually with acetone to furnish two fractions (1-1–1-5). Fraction 1-2 (1.25 g) was further purified on a silica gel column using n-hexane–acetone mixtures to obtain -sitostenone and stigmasta-4,22-dien-3-one (12 mg). Part of fraction 2 (6.21 g) was subjected to silica gel chromatography by eluting with n-hexane–acetone (6:1) enriched with acetone to furnish three further fractions (2-1–2-6). Fraction 2-1 (1.24 g) was further purified on a silica gel column using n-hexane–acetone mixtures to obtain liriodenine (4 mg), lysicamine (2 mg), and (–)-nornuciferine (5 mg). Fraction 2-2 (1.06 g) was further purified on a silica gel column using n-hexane–acetone mixtures to obtain (–)-roemerine (2 mg), (–)-nuciferine (5 mg), and (–)-anonaine (9 mg). Fraction 2-5 (1.57 g) was further purified on a silica gel column using n-hexane–acetone mixtures to obtain (–)-asimilobine (2 mg). Part of fraction 3 (5.78 g) was subjected to silica gel chromatography by eluting with n-hexane–EtOAc (60:1) enriched with EtOAc to furnish six further fractions (3-1–3-4). Fraction 3-1 (1.54 g) was further purified on a silica gel column using CH2Cl2–MeOH mixtures to obtain northalifoline (1) (2 mg) and thalifoline (2) (4 mg).