The Alkaloid-Enriched Fraction of Pachysandra terminalis (Buxaceae) Shows Prominent Activity against Trypanosoma brucei rhodesiense.

In the course of our studies on antiprotozoal natural products and following our recent discovery that certain aminosteroids and aminocycloartanoid compounds from Holarrhena africana A. DC. (Apocynaceae) and Buxus sempervirens L. (Buxaceae), respectively, are strong and selective antitrypanosomal agents, we have extended these studies to another plant, related to the latter-namely, Pachysandra terminalis Sieb. and Zucc. (Buxaceae). This species is known to contain aminosteroids similar to those of Holarrhena and structurally related to the aminocycloartanoids of Buxus. The dicholoromethane extract obtained from aerial parts of P. terminalis and, in particular, its alkaloid fraction obtained by acid-base partitioning showed prominent activity against Trypanosoma brucei rhodesiense (Tbr). Activity-guided fractionation along with extended UHPLC-(+)ESI QTOF MS analyses coupled with partial least squares (PLS) regression modelling relating the analytical profiles of various fractions with their bioactivity against Tbr highlighted eighteen constituents likely responsible for the antitrypanosomal activity. Detailed analysis of their (+)ESI mass spectral fragmentation allowed identification of four known constituents of P. terminalis as well as structural characterization of ten further amino-/amidosteroids not previously reported from this plant.

Two new pregnane alkaloids from Pachysandra terminalis Sieb. et Zucc.

Two new pregnane alkaloids, (20S)-20α-cinnamoylamino-3ß-dimethylamino-5-en-pregnane (1) and (20S)-20α-cinnamoylamino-3ß-dimethylamino-pregnane (2), and four known alkaloids (+)-(20S)-20-(dimethylamino)-3-(3'R-isopropyl)-lactam-5α-pregn-2-en-4-one (3), axillaridine A (4), pachysamine M (5) and 20α-dimethylamino-16ß-hydroxy-3ß-senecioylamino-pregn-5-ene (6) were obtained from the whole herb of Pachysandra terminalis Sieb. et Zucc. Their structures were determined by various spectral techniques and computed electronic circular dichroism (ECD) data. Compounds 1-4 were tested for cytotoxicity against three human tumor cell lines and a human umbilical vein endothelial cell (HUVEC) line. Compound 4 exhibited moderate cytotoxicity against MCF-7, U251 and A549 cells with IC50 values of 15.01 ± 0.47 µM, 20.13 ± 1.34 µM and 20.04 ± 1.16 µM, respectively; compounds 1-3 showed weak cytotoxic activity against three tumor cells.

The synthesis and anti-metastatic effects of optical isomers of ionone alkaloid 9-(N,N-dimethyl)-4,7-megastigmedien-3-one.

Ionone alkaloid 9-(N,N-dimethyl)-4,7-megastigmedien-3-one (compound 1) is a new anti-metastatic natural product. However, it was previously reported as optical isomers mixture. Herein, the optical isomers (6a-6d) of compound 1 were synthesized. The absolute configurations of 6a-6d were determined by ECD experiments and calculated spectra with time-dependent density functional theory (TDDFT). The anti-metastatic effects of the optical isomers were examined by transwell assay. These results revealed that compound 6a had potential anti-metastatic activity with an IC50 value of 0.512 ±â€¯0.093 µM.

A novel pregnane-type alkaloid from Pachysandra terminalis inhibits methicillin-resistant Staphylococcus aureus in vitro and in vivo.

A new kind of pregnane-type alkaloid, 20α-dimethylamino-3ß-senecioylamino-16ß-hydroxy-pregn-5-ene (K-6), was isolated from Pachysandra terminalis Sieb. et Zucc., and its antibacterial activity against MRSA and MRSE was evaluated. We found that K-6 showed antibacterial effects against MRSA and MRSE with minimum inhibitory concentration values (25 mg/L), but did not induce antibiotic resistance in bacteria easily. The administration of K-6 dose-dependently improved the animal survival rate of mice infected with MRSA, with survival rates of 36.34% and 66.67% in the low-dose and high-dose groups, respectively. The protective effects were associated with the reduction of the bacterial titers in the blood and with the morphological amelioration of infected tissues. Scanning and transmission electron microscopy analyses indicated that the cytoplasm shrink of bacterial cells led to noticeable gaps between the cell membrane and cell cytoplasm, and the severely damaged cell membrane resulted in leakage of intracellular content, which ultimately caused the lethal effect of K-6 on bacteria. These findings demonstrated that K-6 is a potential agent against MRSA and MRSE.

Synthesis and antimetastatic effects of E-salignone amide derivatives.

The preparation of novel E-salignone derivatives and their biological evaluation as potential antimetastatic agents is described. The E-salignone amide derivatives were prepared from epiandrosterone and androsterone, and characterized by analytical (1) H NMR, (13) C NMR, and mass spectrometry. The derivatives were evaluated for antimetastatic activity in MDA-MB-231 cells by using a transwell assay. Comparing with the positive control, LY294002, compounds 19b, 19d, and 19e exhibited significant inhibitory effects on the EGF-induced invasion of MB-MDA-231 cells. Moreover, compound 19b also had antimigration effects in wound-healing assay. Compound 19b may represent a novel antimetastatic agent for treating breast cancer.

Antitumor metastasis pregnane alkaloids from Pachysandra terminalis.

Three new pregnane alkaloids, named terminamines H-J (1-3), together with two known alkaloids (4 and 5), were isolated from the ethanol extract of Pachysandra terminalis. The structures of isolated compounds were elucidated by spectroscopic methods, including (1)H and (13)C NMR, 2D NMR, and HR-ESI-MS. Compounds 1, 4, and 5 revealed significant anti-metastasis activities. In addition, compound 1 inhibited the expression of p-PKCζ in MDA-MB-231 cells, and compound 4 inhibited the expressions of p-PKCζ in MDA-MB-231 and A549 cells.

[Chemical constituents from Pachysandra terminalis].

To study chemical constituents from Pachysandra terminalis. By repeated column chromatography, including silica gel, Toyopearl HW-40, and preparative HPLC, four new (14) and one known (5) compounds were isolated and purified. On the basis of spectral data analysis, the structure of isolated compounds were elucidated as follow: 2-methyl-3-methylenepentane-1, 2, 5-triol (1), 4-methyl-3-methylenepentane-1, 2, 5-triol (2), 4-methyl-3-methylenepentane-1, 2, 5-triol-5-O-beta-D-glucopyranoside (3), 4-methyl-3-methylenep- entane-1, 2, 5-triol-1-O-beta-D-glucopyranoside (4), (7S, 8R, 8' R)-(+)-lariciresinol-9-O-beta-D-glucopyrano-side (5). Compound 5 was isolated from this genus for the first time.

Alkaloids from Pachysandra terminalis inhibit breast cancer invasion and have potential for development as antimetastasis therapeutic agents.

The aim of the present study was to identify potentially useful natural compounds for the development of novel therapeutic agents to inhibit metastasis. A phytochemical investigation of Pachysandra terminalis resulted in the isolation of seven new pregnane alkaloids, terminamines A-G (1-7), and seven known alkaloids (8-14). The structures of 1-7 were elucidated by 1D- and 2D-NMR spectroscopic and mass spectrometric methods. Compounds 1-5 and 8-14 inhibited the migration of MB-MDA-231 breast cancer cells induced by the chemokine epithelial growth factor. In addition, compound 1 inhibited phosphorylation of integrin ß(1), which plays an important role in MB-MDA-231 cell adhesion and metastasis.

[Phenol constituents of Pachysandra terminalis and their antioxidant activity].

OBJECTIVE: To study the phenol constituents from Pachysandra terminalis and their antioxidant activities. METHOD: Constituent isolation and purification was carried by repeated column chromatography (silica gel, Toyopearl HW-40 and preparative HPLC), and their structures were elucidated on the basis of spectral data analysis. DPPH method was used to evaluate the free radical scavenging activity of the isolated compounds. RESULT: Nine phenol compounds (1-9) were isolated and their structures were identified as follow: p-hydroxybenzaldehyde (1), vanillin (2), 1-(4-hydroxy-3-methoxyphenyl) -ethanone (3), syringaldehyde (4), salicylic acid (5), p-hydroxybenzoic acid (6), ferulic acid (7), 2,3,4-trihydroxybenzoic acid (8), 3,4-dihydroxybenzoic acid (9). The isolated compounds showed obviously antioxidant activity. At the concentration of 50 micromol x L(-1), compounds 7-9 revealed DPPH free radical scavenging rates were 87.8%, 97.8% and 92%, respectively. CONCLUSION: Compounds 1-9 were isolated from this genus for the first time. They showed the significant antioxidant activity.

[Studies on the chemical constituents of Pachysandra terminalis and their antioxidant activity].

OBJECTIVE: To study the chemical constituents from Pachysandra terminalis and their antioxidant activity. METHODS: Chemical constituents were isolated by repeated column chromatography (silica gel, Toyopearl HW-40C and preparative HPLC). The structures of isolated compounds were elucidated on the basis of spectral data analysis. DPPH method was used to evaluate the free radical scavenging activity of the isolated compounds. RESULTS: Six compounds were isolated and their structures were identified as follow: 2-Phenylethyl-beta-D-glucopyranoside (1), (+)Pinoresinol-4'-O-beta-D-glucopyranoside (2), Pinoresinol (3), cis-Syringin (4), 4-hydroxy-4-(3-oxo-l-butenyl)-3,5,5-trimethylcyclohex-2-en-l-one (5), 3alpha-hydroxy-5,6-epoxy-7-megastigmen-9-one (6). Compounds 2, 4, 5 showed obviously antioxidant activity, their DPPH free radical scavenging rates were 82.50%, 87.36%, and 84.56% on the concentration of 50 micromol/L, respectively. CONCLUSION: Compounds 1-6 are isolated from this genus for the first time. Compounds 2, 4, and 5 showed significant antioxidant activities.

Pregnane alkaloids from Pachysandra axillaris.

Nine new pregnane alkaloids, pachysamines J-R (1-9), together with seven known ones, were isolated from Pachysandra axillaris. The chemical structures of the new alkaloids were elucidated by spectroscopic methods. All the compounds were evaluated for their inhibitory activities against HL-60, SMMC-7721, A-549, SK-BR-3, and PANC-1 cell lines. Compound 15 possessed moderate activities against A-549, SK-BR-3, and PANC-1 cells, with the IC(50) values of 11.17, 4.17, and 10.76 microM, respectively. Besides, compound 11 showed cytotoxicities against A-549 cell, with the IC(50) values as 24.94 microM.

Two new pregnanone derivatives with strong cytotoxic activity from Pachysandra axillaris.

Two new, bioactive, pregnane-based natural products, pachysanonin (= 3beta,11alpha,12beta)-12-acetoxy-3-(dimethylamino)-11-[(3,4-dimethylpent-3-enoyl)oxy]pregnan-20-one; 1) and pachysanone (= (11alpha,12beta)-12-acetoxy-11-[(3,4-dimethylpent-3-enoyl)oxy]pregnan-3,20-dion; 2) have been isolated from Pachysandra axillaris. Their structures were determined by spectroscopic methods, and, in the case of 2, by single-crystal X-ray crystallography (Figure). Compound 2 showed significant antitumor activity against Lewis lung carcinoma (LCC) tumor cells, with an IC50 value of 0.020+/-0.006 microg/ml, which is equal or even lower than those of the well-known natural antitumor agents harringtonine (0.02), homoharringtonine (0.15), and adriamycin (0.06 microg/ml; positive control).