Anti-biofilm, nitric oxide inhibition and wound healing potential of purpurin-18 phytyl ester isolated from Clinacanthus nutans leaves.

AIMS: Clinacanthus nutans (C. nutans) has demonstrated anti-inflammatory activity, however, the active compound generating this activity remains unknown. The aim of this study was to identify the bioactive compound in C. nutans responsible for its anti-inflammatory, in-vitro wound healing, and anti-biofilm activities. MAIN METHODS: A pure compound was isolated from the chloroform extract (CE) of C. nutans leaves by chromatographic techniques and bioassay-guided fractionation. This compound's structure was determined by spectroscopic analyses (FTIR/NMR/HRES-MS). Biological activities were evaluated using cytotoxicity, nitric oxide (NO), wound scratch, anti-microbial activity, and anti-biofilm assays; and the compound's bactericidal depth into the biofilm was visualized by confocal laser scanning microscopy. KEY FINDINGS: CE and its pure isolated compound, purpurin-18 phytyl ester (P18PE), significantly inhibited lipopolysaccharide (LPS)-induced NO production in RAW 264.7 cells at concentrations of 100 µg/ml and 10-100 µg/ml, respectively. These concentrations significantly induced wound closure by human gingival fibroblasts. CE (100-1000µg/ml) and P18PE (1-500 µg/ml) did not inhibit Streptococcus (S.) mutans growth. However, these concentrations significantly reduced S. mutans biofilm formation below 50% at 250 µg/ml for CE, and 25 µg/ml for P18PE (p<0.05). SIGNIFICANCE: C. nutans contains a bioactive compound, P18PE, which exhibits anti-inflammatory, in-vitro wound healing, and anti-biofilm activities.

Preclinical Study of Antineoplastic Sinoporphyrin Sodium-PDT via In Vitro and In Vivo Models.

Photodynamic therapy (PDT) investigations have seen stable increases and the development of new photosensitizers is a heated topic. Sinoporphyrin sodium is a new photosensitizer isolated from Photofrin. This article evaluated its anticancer effects by clonogenic assays, MTT assays and xenograft experiments in comparison to Photofrin. The clonogenicity inhibition rates of sinoporphyrin sodium-PDT towards four human cancer cell lines ranged from 85.5% to 94.2% at 0.5 µg/mL under 630 nm irradiation of 30 mW/cm² for 180 s. For MTT assays, the IC50 ranges of Photofrin-PDT and sinoporphyrin sodium-PDT towards human cancer cells were 0.3 µg/mL to 5.5 µg/mL and 0.1 µg/mL to 0.8 µg/mL under the same irradiation conditions, respectively. The IC50 values of Photofrin-PDT and sinoporphyrin sodium-PDT towards human skin cells, HaCaT, were 10 µg/mL and 1.0 µg/mL, respectively. Esophagus carcinoma and hepatoma xenograft models were established to evaluate the in vivo antineoplastic efficacy. A control group, Photofrin-PDT group (20 mg/kg) and sinoporphyrin sodium group at three doses, 0.5 mg/kg, 1 mg/kg and 2 mg/kg, were set. Mice were injected with photosensitizers 24 h before 60 J 630 nm laser irradiation. The tumor weight inhibition ratio of 2 mg/kg sinoporphyrin sodium-PDT reached approximately 90%. Besides, the tumor growths were significantly slowed down by 2 mg/kg sinoporphyrin sodium-PDT, which was equivalent to 20 mg/kg Photofrin-PDT. In sum, sinoporphyrin sodium-PDT showed great anticancer efficacy and with a smaller dose compared with Photofrin. Further investigations are warranted.

Antiproliferative effect of alkaloids via cell cycle arrest from Pseuduvaria rugosa.

CONTEXT: Pseuduvaria rugosa (Blume) Merr. (Annonacaea) grows widely in the south and southeast regions of Thailand. Preliminary screening for biological activities revealed that crude hexane, ethyl acetate, and acetone extracts from mixtures of leaves and twigs of P. rugosa showed cytotoxicity. OBJECTIVE: Chemical constituents and their antiproliferative activity in K562, U937, and HL-60 human leukemic cell lines from P. rugosa were performed for the first time. MATERIALS AND METHODS: The isolated compounds were obtained from chromatographic separation. The structures were established by spectroscopic techniques including IR, UV, NMR together with 2D NMR (HMBC, COSY, and NOE) and MS. The K562, U937, and HL-60 cell lines were treated with isolated aporphine alkaloids (0-100 µg/mL) and cell viability was measured with the MTT assay. Cell cycle analysis was performed using propidium iodide (PI) based staining methods. RESULTS: Two known aporphine alkaloids, 1,2,3-trimethoxy-5-oxonoraporphine (1) and ouregidione (2) were isolated. Treatment of the cells with compounds 1 and 2 at a concentration of 100 µg/mL for 72 h reduced the viability of K562, U937, and HL-60 cell lines to 63 and 64, 38 and 66, and 49 and 64%, respectively. In addition, compounds 1 and 2, at a concentration of 100 µg/mL, exposed to U937 and HL-60 cell lines showed cell cycle arrest. The U937 cell line treated with compounds 1 and 2 increased significantly the proportion of the cell in S phase, whereas the HL-60 cell line-induced G2/M and G1 phase, respectively. DISCUSSION AND CONCLUSION: The results showed that 1,2,3-trimethoxy-5-oxonoraporphine and ouregidione-induced cytotoxicity with HL-60, U937, and K562 cells where 1,2,3-trimethoxy-5-oxonoraporphine was more active than ouregidione.

A novel porphyrin photosensitizer from bamboo leaves that induces apoptosis in cancer cell lines.

BACKGROUND: In the screening of new anticancer agents, we found that a methanol extract of bamboo leaves induced rapid apoptosis in the human leukemia CMK-7 cell line. MATERIALS AND METHODS: The active compounds in the extract were isolated by chromatographic methods and their structures were determined by NMR and mass spectroscopy. Apoptosis by the compounds were evaluated in CMK-7 and human colon adenocarcinoma Colo320 DM cells by monitoring the caspase-3 activation and DNA cleavage. RESULTS: The active compounds are 201-hydroxypurpurin-7 delta-lactone ethyl methyl diester (1) and the corresponding methyl phytyl diester (2). The apoptosis by compound 1 (0.3 to 0.1 microM for CMK-7 cells) was enhanced when the culture was briefly irradiated with a fluorescent lamp. This photodynamic induction of apoptosis by compound 1 was much stronger than that by a known photosensitizer, pyropheophorbidemethyl. Compound 2 was a weaker inducer of apoptosis than compound 1, but the apoptosis occurred after light irradiation. CONCLUSION: The 201-hydroxypurpurin-7 delta-lactone esters are promising lead compounds as photosensitizers for photodynamic therapy of cancer.

Naturally occurring aristolactams, aristolochic acids and dioxoaporphines and their biological activities.

Aristolactams, having a phenanthrene chromophore are a small group of compounds mainly found in the Aristolochiaceae together with the aristolochic acids and 4,5-dioxoaporphines. In this report, these three important classes of natural products are reviewed and classified on the basis of their oxygenation pattern. In addition the biological activities of these compounds and their general chemistry are discussed.

The alkaloids of Artabotrys uncinatus.

A novel type of alpha,beta-butenolide alkaloid, uncinine (1), two novel oxoaporphines, artabonatine C (2) and artabonatine D (3), a new oxazoloaporphine, artabonatine E (4), and a new 7,7'-bisdehydroaporphine, artabonatine F (5), along with 25 known alkaloids, were isolated from Artabotrys uncinatus. The structures of 1-5 were determined using NMR and mass spectral data. Atherospermidine and squamolone exhibited cytotoxicity against hepatocarcinoma cancer cell lines (Hep G(2) and 2,2,15), and the activity of some of the alkaloids in an antithrombin assay is also discussed.

Cytotoxic oxoisoaporphine alkaloids from Menispermum dauricum.

Four new oxoisoaporphine alkaloids, daurioxoisoporphines A-D (1-4), were isolated from the rhizomes of Menispermum dauricum. The structures of these alkaloids were established by spectroscopic methods. The cytotoxic evaluation of 1 and 2 is reported against four cancer cell lines.

[A-ring formylated flavonoids and oxoaporphinoid alkaloid from Dasymaschalon rostratum Merr. et Chun].

OBJECTIVE: To study the chemical components in the stem of Dasymaschalon rostratum. METHOD: The components were extracted with solvent, separated and purified with chromatographic methods, identified by NMR, MS, UV, IR and physicol-chemical constants. RESULTS: Three A-ring-formylated flavonoids and one oxoaporphinnoid aikaloid were isolated and identified as lawinal, unonal, isounonal and 7-oxodehydroasimilobine. CONCLUSION: All the four compounds were isolated for the first time from the genus Dasymaschalon. According to all the phytochemistry papers on Annonaceae, A-ring formylated flavonoids in this family were isolated from the genus Desmos for the first time. Thus, it is an interesting discovery in chemotaxonomy which reveals the close relationship between the two genera Desmos and Dasymaschalon.

Constituents from the leaves of Aristolochia elegans.

One new biphenyl ether, aristogin C (1), and two new porphyrins, aristophylls A (2) and B (3), as well as 11 known compounds, were isolated from the leaves of Aristolochia elegans. Their structures were elucidated according to the spectroscopic (NMR and MS) analyses or by comparison with literature values.

5-Oxonoraporphines from Mitrephora cf. maingayi.

Two new 5-oxonoraporphines, 1 and 2, together with three known compounds, ouregidione, 3-methoxycepharadione B, and isoelemicin, have been isolated from the bark of Mitrephora cf. maingayi. Structures of 1 and 2 were determined to be 1,2,3-trimethoxy-5-oxonoraporphine and 1,2-dimethoxy-3-hydroxy-5-oxonoraporphine on the basis of NMR and MS studies.

Computerised gas chromatographic-mass spectrometric analysis of complex mixtures of alkyl porphyrins.

Computerised capillary gas chromatography-mass spectrometry (GC-MS) analysis of complex mixtures of alkyl porphyrins, as their bis-(trimethylsiloxy)silicon(IV) and bis(tert.-butyldimethylsiloxy)silicon(IV) derivatives, is described. The latter derivative is more suitable for routine GC-MS analysis. This computerised GC-MS approach, when applied to the alkyl porphyrins of two geological samples, a bitumen (Gilsonite, Eocene age, UT, U.S.A.) and a crude oil (Boscan, Cretaceous age, West Venezuela), has revealed the highly complex compositions of these fractions. Computer-aided data processing, using relative retention index (RRI) calculations, facilitated the classification of the chromatographic peaks according to structural type and membership of pseudo-homologous series. Computerised GC-MS is compared with, and contrasted to high-performance liquid chromatography as a means of petroporphyrin analysis.

Identification of porphyrin present in apo-cytochrome C oxidase of copper-deficient yeast cells.

Heme a was not detected either in mitochondria isolated from copper-deficient yeast or in the intact cells. Nevertheless, the intracellular concentration of free porphyrins indicated that the pathway of porphyrin and heme synthesis was not impaired in copper-deficient cells. The immunoprecipitated apo-oxidase from copper-deficient cells revealed an absorption spectrum with maxima at 645, 592, 559, 519 and 423 nm, similar to that of purified porphyrin a. When solubilized mitochondria from [3H]leucine and delta-amino[14C]levulinic acid-labeled copper-deficient yeast cells were incubated with rabbit antiserum against cytochrome c oxidase, a precipitate was obtained. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of this immunoprecipitate showed [3H]leucine associated with six bands and delta-amino[14C]levulinic acid resolved in a single band. HCl fractionation of copper-deficient mitochondria labeled with delta amino[14C]levulinic acid showed a high specific radioactivity in the fraction extracted by 20% HCl, a solvent which extracts porphyrin a. Thin-layer chromatography of the radioactivity found in 20% HCl showed an RF value identical to that of purified porphyrin a. When delta-amino[3H]levulinic acid-labeled, copper-deficient yeast cells are grown in copper-supplemented medium, the porphyrin a accumulated in copper-deficient cells was converted into heme a, and this conversion was prevented by cycloheximidine. These observations suggest that porphyrin a is present in the apo-oxidase of copper-deficient cells, but that the conversion to heme a does not occur. This conversion reaction appears to be a point in the biosynthetic pathway of cytochrome c oxidase which is blocked by copper deficiency.