ABSTRACT
Chemical investigation of the methanol extract of the roots of Baphia bancoensis led to the isolation and characterization of three new isoflavonoid glycosides (1-3). Their structures were determined on the basis of spectroscopic studies andchemical evidence. Antibacterial activity of isolated compounds was evaluated against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa.
Subject(s)
Anti-Bacterial Agents/isolation & purification , Fabaceae/chemistry , Glycosides/isolation & purification , Isoflavones/isolation & purification , Isoflavones/pharmacology , Plants, Medicinal/chemistry , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Enterococcus faecalis/drug effects , Escherichia coli/drug effects , Glycosides/chemistry , Glycosides/pharmacology , Isoflavones/chemistry , Microbial Sensitivity Tests , Molecular Structure , Plant Roots/chemistry , Pseudomonas aeruginosa/drug effects , Staphylococcus aureus/drug effectsABSTRACT
The pH-zone refining centrifugal partition chromatography technique was used to separate the two acetylcholinesterase inhibitors huperzines A and B from a crude alkaloid extract of the club moss Huperzia serrata. Complete co-elution of huperzines A and B was initially observed with the well-known methyl tert-butyl ether-acetonitrile-water (4:1:5, v/v/v) solvent system with triethylamine (8mM) as the displacer and methane sulfonic acid (6mM) as the retainer. An efficient biphasic system was designed on the basis of solvent association that provided selectivity in the elution mode: n-heptane/ethyl acetate/n-propanol/water (5:15:35:45, v/v/v/v). Lowering the bridge solvent content (n-propanol) of this system increased the polarity difference between the two phases thus adapting it to the pH-zone refining mode. Thus, the purification of these compounds was achieved using the biphasic system n-heptane/ethyl acetate/n-propanol/water (10:30:15:45, v/v/v/v) with triethylamine (8mM) as the displacer and methane sulfonic acid (6mM) as the retainer.
Subject(s)
Alkaloids/isolation & purification , Centrifugation/methods , Countercurrent Distribution/methods , Huperzia/chemistry , Sesquiterpenes/isolation & purification , Hydrogen-Ion ConcentrationABSTRACT
Nowadays, centrifugal partition chromatography (CPC) separations can be routinely achieved at the laboratory scale. The solvent system selection has been made easy, as generic sets of solvent systems are described in publications and books. This approach, however, generally reduces the scope of optimization strategies for two important parameters: selectivity and sample solubility. This can be very limiting for the preparative separation of structurally similar compounds. Multiple dual-mode (MDM) CPC has been developed to provide an easy-to-use alternative technique to circumvent this problem. A MDM separation consists of a succession of dual-mode runs (i.e. multiple inversion of stationary and mobile phase) that can only be achieved because both chromatographic phases are liquids. This original elution mode is thus a semi-continuous process with a classical sample injection and which only requires a single CPC column. Underlying mechanisms of MDM were studied using a model mixture of acenaphthylene and naphthalene. A mixture of two synthetic pairs of diastereomers was then successfully submitted to MDM CPC, in the framework of the synthesis of biologically active compounds.
Subject(s)
Centrifugation/methods , Countercurrent Distribution/methods , Acenaphthenes/isolation & purification , Chromatography, High Pressure Liquid , Naphthalenes/isolation & purification , StereoisomerismABSTRACT
Fagaronine, a benzophenanthridine alkaloid from Fagara zanthoxyloides Lam. (Rutaceae), has been tested on the erythroleukemic cell line K562 in order to explain some previous results on cell differentiation. In this study we showed that fagaronine induces a significant hemoglobinization of the human erythroleukemic cell line K562. This hemoglobin synthesis was accompanied by a strong increase of erythroid mRNA expression such as gamma- and alpha-globin, and PBGD, an enzyme of heme synthesis. In addition, the Epo-R transcripts were also stimulated indicating that cells are engaged in a maturation process. Both transcription factors GATA-1 and NF-E2, which play an important role in the regulation of genes involved in the erythroid differentiation, were also transcriptionally up-regulated. To elucidate the possible role of GATA-1 in the FAG-induced differentiation of K562 cells, we transfected reporter constructs containing regulatory regions of erythroid genes encompassing GATA-1 binding sites. After 48 hours of treatment, FAG stimulated the EPO-R and gamma-globin promoters by 2- to 3-fold and the promoter/enhancer region of GATA-1 gene by 3.2-fold. A mutation within the GATA-1 binding sites strongly decreased the promoter activation induced by FAG. Taken together, our results represent a demonstration that FAG exerts its differentiating activity by a specific activation of the regulating GATA-1 regions of genes involved in the erythroid phenotype expression.
Subject(s)
Alkaloids/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Phenanthridines/pharmacology , Phytotherapy , Benzophenanthridines , Cell Differentiation/drug effects , DNA Primers , DNA-Binding Proteins/genetics , Erythroid-Specific DNA-Binding Factors , Erythropoiesis/drug effects , GATA1 Transcription Factor , Gene Expression , Humans , K562 Cells/drug effects , NF-E2 Transcription Factor , NF-E2 Transcription Factor, p45 Subunit , Plant Roots , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction , Transcription Factors/genetics , ZanthoxylumABSTRACT
Ion-exchange displacement chromatography has been adapted to centrifugal partition chromatography. The use of an ionic liquid, benzalkonium chloride, as a strong anion-exchanger has proven to be efficient for the preparative separation of phenolic acid regioisomers. Multigram quantities of a mixture of three hydroxycinnamic acid isomers were separated using iodide as a displacer. The displacement process was characterized by a trapezoidal profile of analyte concentration in the eluate with narrow transition zones. By taking advantage of the partition rules involved in support-free liquid-liquid chromatography, a numerical separation model is proposed as a tool for preliminary process validation and further optimization.
ABSTRACT
Five dammarane-type saponins were isolated by means of centrifugal partition chromatography from the leaves of Zizyphus lotus. Their structures were elucidated using a combination of 1D and 2D 1H and 13C NMR spectra and mass spectroscopy. One of these glycosides is the known jujuboside B (5). Three are new jujubogenin glycosides, identified as 3-O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranosyljujubogenin-20-O-(2,3,4-O-triacetyl)-alpha-L-rhamnopyranoside (1), 3-O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranosyljujubogenin-20-O-alpha-L-rhamnopyranoside (2), and 3-O-alpha-L-rhamnopyranosyl-(1-->2)-[(4-sulfo)-beta-D-glucopyranosyl-(1-->3)]-alpha-L-arabinopyranosyljujubogenin (3). The last is a new sulfated derivative of jujubasaponine IV, identified as 3-O-alpha-L-rhamnopyranosyl-(1-->2)-[(4-sulfo)-beta-D-glucopyranosyl-(1-->3)]-beta-D-galactopyranosyl-(20R,22R)-16beta,22:16alpha,30-diepoxydammar-24-ene-3beta,20-diol (4).
Subject(s)
Plants, Medicinal/chemistry , Saponins/isolation & purification , Triterpenes/isolation & purification , Ziziphus/chemistry , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Plant Leaves/chemistry , Saponins/chemistry , Stereoisomerism , Triterpenes/chemistry , Tunisia , DammaranesABSTRACT
A new derivative of strychnopentamine was isolated from the leaves of Strychnos usambarensis. This compound, named chrysopentamine, was identified by detailed spectroscopic methods (UV, IR, HR-ESI-MS, 1D and 2D NMR). Chrysopentamine presented an original hydroxy substitution on C-14 and an aromatization of the ring D of strychnopentamine leading to anhydronium base properties and exhibited strong antiplasmodial properties (IC (50) less than 1 microM).
Subject(s)
Antimalarials/pharmacology , Heterocyclic Compounds, 4 or More Rings/pharmacology , Phytotherapy , Plasmodium falciparum/drug effects , Pyrrolidines/pharmacology , Strychnos , Animals , Antimalarials/chemistry , Heterocyclic Compounds, 4 or More Rings/chemistry , Inhibitory Concentration 50 , Magnetic Resonance Spectroscopy , Parasitic Sensitivity Tests , Plant Leaves , Pyrrolidines/chemistryABSTRACT
In the course of our search for new antiplasmodial alkaloids from Strychnos icaja, we have isolated five alkaloids: three monomers, protostrychnine and genostrychnine, previously described in Strychnos nux-vomica, pseudostrychnine, already found in the leaves of the plant, a new bisindolic alkaloid, named strychnogucine C, and the first naturally occurring trimeric indolomonoterpenic alkaloid: strychnohexamine. This latter trimeric alkaloid presented an antiplasmodial activity against the FCA Plasmodium falciparum line near 1 microM.
Subject(s)
Antimalarials/isolation & purification , Carbazoles/isolation & purification , Indole Alkaloids/isolation & purification , Monoterpenes/isolation & purification , Plasmodium falciparum/drug effects , Strychnine/isolation & purification , Strychnos/chemistry , Animals , Antimalarials/chemistry , Antimalarials/pharmacology , Carbazoles/chemistry , Carbazoles/pharmacology , Democratic Republic of the Congo , Indole Alkaloids/chemistry , Indole Alkaloids/pharmacology , Inhibitory Concentration 50 , Magnetic Resonance Spectroscopy , Molecular Structure , Monoterpenes/chemistry , Monoterpenes/pharmacology , Plant Roots/chemistry , Stereoisomerism , Strychnine/analogs & derivatives , Strychnine/chemistry , Strychnine/pharmacologyABSTRACT
The in vitro antiplasmodial activities of 69 alkaloids from various Strychnos species were evaluated against chloroquine-resistant and chloroquine-sensitive lines of Plasmodium falciparum. The compounds, comprising mainly indolomonoterpenoid alkaloids, exhibited a wide range of biological potencies in the antiplasmodial assays. The most active alkaloids were also tested for cytotoxicity against HCT-116 colon cancer cells to determine their antiplasmodial selectivity. As a result of these studies, structure-activity relationships for these alkaloids have begun to emerge. Alkaloids presenting four types of bisindole skeleton exhibited potent and selective activities against Plasmodium. They were sungucine-type (IC(50) values ranging from 80 nM to 10 microM), longicaudatine-type (IC(50) values ranging from 0.5 to 10 microM), matopensine-type (IC(50) values ranging from 150 nM to 10 microM), and usambarine-type alkaloids. Within the last structural type, isostrychnopentamine (49) and ochrolifuanine A (46) were found to be active against chloroquine-sensitive and -resistant strains (IC(50) values of 100-150 and 100-500 nM, respectively), and dihydrousambarensine (51) exhibited a 30-fold higher activity against the chloroquine-resistant strain (IC(50) = 32 nM) than against the chloroquine-sensitive one.
Subject(s)
Alkaloids/pharmacology , Antimalarials/pharmacology , Plasmodium falciparum/drug effects , Strychnos/chemistry , Alkaloids/chemistry , Alkaloids/isolation & purification , Animals , Antimalarials/chemistry , Antimalarials/isolation & purification , Drug Evaluation, Preclinical , Structure-Activity RelationshipABSTRACT
Ten triterpenes and phytosterols beta-amyrin, lupeol, lupeol acetate, ursolic acid, friedelin, canophyllol, 29-hydroxy-friedelan-3-one, beta-sitosterol, 3- O-beta- D-glucopyranosyl-beta-sitosterol, 3-O-(6'- O-palmitoyl)-beta-D-glucopyranosyl-beta-sitosterol, were evaluated as potential inhibitors of human leucocyte elastase (HLE). In this series, lupeol, ursolic acid and canophyllol showed marked HLE inhibitory activity with IC(50) values at 1.9 microM, 4.4 microM, and 2.5 microM, respectively. It appeared that HLE inhibition depended on the presence and the orientation of two reactive groups in the tested molecules, distant from 10-12 A, reacting with Arg-217 in S(4) -S(5) subsites of the extended substrate-binding domain of HLE, and S(3), respectively.
Subject(s)
Leukocyte Elastase/antagonists & inhibitors , Leukocyte Elastase/drug effects , Phytosterols/pharmacology , Phytotherapy , Plants, Medicinal , Triterpenes/pharmacology , Bignoniaceae , Caesalpinia , Humans , Inhibitory Concentration 50 , Moraceae , Phytosterols/administration & dosage , Triterpenes/administration & dosageABSTRACT
One new diterpene, 2 alpha-hydroxy-12beta-hydroxy-isopimara-8(14), 15-diene, and six known compounds as triterpenes, sterols and fatty acid, were isolated from the stem bark of Tanaecium jaroba (Bignoniaceae), a Bolivian plant used in traditional medicine. Their structures were established mainly by 1D and 2D NMR (COSY, HMQC, HMBC, ROESY) and their antiplasmodial activities were evaluated in vitro against Plasmodium falciparum.
Subject(s)
Bignoniaceae , Diterpenes/pharmacology , Animals , Antimalarials/pharmacology , Bolivia , Diterpenes/chemistry , Diterpenes/isolation & purification , Inhibitory Concentration 50 , Magnetic Resonance Spectroscopy , Medicine, Traditional , Molecular Structure , Plant Bark/chemistry , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Extracts/pharmacology , Plant Stems/chemistry , Plasmodium falciparum/drug effectsABSTRACT
In addition to six cyclopeptide alkaloids lotusines A-F, a new compound lotusine G, was isolated from the root bark of Zizyphus lotus by centrifugal partition chromatography.