P388 Murine Leukemia Cells Research Articles

Antioxidant, antibacterial and cytotoxic activities of essential oils and ethanol extracts of selected South East Asian herbs

Antioxidant, antibacterial and cytotoxic activities of essential oils and ethanol extracts of four South East Asian traditional herbs were investigated. Their essential oils were analyzed byGas chromatography mass spectrometry (GC-MS) and a total of 57 types of volatile organics were identified. Artemisia argyi, Centella asiatica, Cosmos caudatus, and Polygonum hydropiper contained 30, 20, 19 and 21 types of volatiles, respectively. The major volatile hydrocarbon for A. argyi, C. asiatica and C. caudatus was a-cadinene while dodecanal was the major component for P. hydropiper. Total phenolic content of ethanol extracts analyzed by Folin-Ciocalteu method were in the range of 31.58±3.08 to 84.03±8.15 mg GAE/100 g. Free radical-scavenging activity of the four ethanol extracts were in the range of 654.43±17.22 to 5857.54±164.13 mg AA/100 g; while ferric-reducing antioxidant power were in the range of 26.58±1.10 to 50.08±0.71 mg AAE/g. Essential oils of all four species were inactive for both these assays. Ethanol extracts of these herbs were active against 60 to 80% of ten strains of human pathogenic bacteria tested, compared to 20 to 50% inhibition for essential oils. Only ethanol extracts of A. argyi and C. caudatus showed activity against P388 murine leukemia cell line, while all the essential oils were inactive. Key words: Antimicrobial, antioxidant, cytotoxicity, ethanol extracts, essential oils, total phenolic content.

Novel Acrylonitrile Derivatives, YHO-13177 and YHO-13351, Reverse BCRP/ABCG2-Mediated Drug ResistanceIn VitroandIn Vivo

Breast cancer resistance protein (BCRP/ABCG2) confers resistance to anticancer drugs such as 7-ethyl-10-hydroxycamptothecin (SN-38, an active metabolite of irinotecan), mitoxantrone, and topotecan. In this study, we examined the reversing effects of YHO-13177, a novel acrylonitrile derivative, and its water-soluble diethylaminoacetate prodrug YHO-13351 on the BCRP-mediated drug resistance. YHO-13177 potentiated the cytotoxicity of SN-38, mitoxantrone, and topotecan in both BCRP-transduced human colon cancer HCT116 (HCT116/BCRP) cells and SN-38-resistant human lung cancer A549 (A549/SN4) cells that express BCRP, but had little effect in the parental cells. In addition, YHO-13177 potentiated the cytotoxicity of SN-38 in human lung cancer NCI-H460 and NCI-H23, myeloma RPMI-8226, and pancreatic cancer AsPC-1 cells that intrinsically expressed BCRP. In contrast, it had no effect on P-glycoprotein-mediated paclitaxel resistance in MDR1-transduced human leukemia K562 cells and multidrug resistance-related protein 1-mediated doxorubicin resistance in MRP1-transfected human epidermoid cancer KB-3-1 cells. YHO-13177 increased the intracellular accumulation of Hoechst 33342, a substrate of BCRP, at 30 minutes and partially suppressed the expression of BCRP protein at more than 24 hours after its treatment in both HCT116/BCRP and A549/SN4 cells. In mice, YHO-13351 was rapidly converted into YHO-13177 after its oral or intravenous administration. Coadministration of irinotecan with YHO-13351 significantly increased the survival time of mice inoculated with BCRP-transduced murine leukemia P388 cells and suppressed the tumor growth in an HCT116/BCRP xenograft model, whereas irinotecan alone had little effect in these tumor models. These findings suggest that YHO-13351, a prodrug of YHO-13177, could be clinically useful for reversing BCRP-mediated drug resistance in cancer chemotherapy.

Cytotoxic Tetramic Acid Derivative Produced by a Plant Type-III Polyketide Synthase

The tetramic acid (2,4-pyrrolidinedione) scaffold has been recognized as an important structural feature because of its mycotoxic, antibacterial, antiviral, and antioxidant activities. This important class of natural products is reportedly produced by the type-I polyketide synthase/nonribosomal peptide synthetase (PKS/NRPS) hybrid megaenzyme systems. In contrast, the benzalacetone synthase (BAS) from Rheum palmatum is a structurally simple, plant-specific type-III PKS that catalyzes the one-step decarboxylative condensation of malonyl-CoA with 4-coumaroyl-CoA. The type-III PKS exhibits unusually broad substrate specificity and notable catalytic versatility. Here we report that R. palmatum BAS efficiently produces a series of unnatural, novel tetramic acid derivatives by the condensation of malonyl-CoA with aminoacyl-CoA thioesters chemically synthesized from L- and D-amino acids. Remarkably, the novel tetramic acid dimer D-5 formed from D-phenylalanoyl-CoA showed moderate antiproliferative activity against murine leukemia P388 cells.

Phytochemical Study of Fagraea spp. Uncovers a New Terpene Alkaloid with Anti-Inflammatory Properties

Phytochemical investigation of the stem bark of F. racemosa JACK ex WALL (Loganiaceae) from East Java, Indonesia, has resulted in the isolation of a new alkaloid fagraeoside along with the iridoid glycoside secologanoside. Fagraeoside may be derived from the condensation of secologanin with L-asparagine, and represents a rare example of a terpene alkaloid in which the amino acid component is non-aromatic. Investigation of three additional species of Fagraea provided known lignans, iridoid or secoiridoid glycosides, and flavanol-6-C-glucosides, thus it is likely that iridoid and secoiridoid glucosides are chemotaxonomic markers for the Fagraea genus. Fagraeoside inhibited the production of prostaglandin E2 in 3T3 murine fibroblasts (IC50 ~5.1 µM), and was not cytotoxic to this cell line or to a P388 murine leukaemia cell line. Selected isolated compounds, including fagraeoside, showed low to moderate activity in anti-acetylcholinesterase screening.

Pyrinodemins E and F, new 3-alkylpyridine alkaloids from sponge Amphimedon sp.

Two new 3-alkylpyridine alkaloids, pyrinodemins E (1) and F (2), were isolated from an Okinawan marine sponge Amphimedon sp. and the structures of 1 and 2 were elucidated on the basis of spectroscopic data. Pyrinodemins E (1) and F (2) were novel 3-alkylpyridine alkaloids possessing a 4-(methoxyamino)piperidinone moiety and an indol-3-glyoxylamide moiety, respectively. Pyrinodemin E (1) showed cytotoxicity against P388 and L1210 murine leukemia cells.

Lagunamides A and B: Cytotoxic and Antimalarial Cyclodepsipeptides from the Marine Cyanobacterium Lyngbya majuscula

Lagunamides A (1) and B (2) are new cyclic depsipeptides isolated from the marine cyanobacterium Lyngbya majuscula obtained from Pulau Hantu Besar, Singapore. The planar structural characterization of these molecules was achieved by extensive spectroscopic analysis, including 2D NMR experiments. In addition to Marfey's method and (3)J(H-H) coupling constant values, a modified method based on Mosher's reagents and analysis using LC-MS was deployed for the determination of the absolute configuration. Lagunamides A and B displayed significant antimalarial properties, with IC(50) values of 0.19 and 0.91 μM, respectively, when tested against Plasmodium falciparum. Lagunamides A and B also possessed potent cytotoxic activity against P388 murine leukemia cell lines, with IC(50) values of 6.4 and 20.5 nM, respectively. Furthermore, these cyanobacterial compounds exhibited moderate antiswarming activities when tested against Pseudomonas aeruginosa PA01.

Cytotoxic 3,4-seco-Atisane Diterpenoids from Croton barorum and Croton goudotii

In a screening program directed to the discovery of new anticancer agents from Madagascan plants, ethyl acetate extracts of Croton barorum and C. goudotii showed strong cytotoxic activity, with 100% inhibition at 10 μg/mL in a primary screen using the murine lymphocytic leukemia P388 cell line. Bioassay-guided fractionation led to the isolation of two new 3,4-seco-atisane diterpenoids, crotobarin (1) and crotogoudin (2). Their structures were elucidated by spectroscopic data interpretation. Compounds 1 and 2 produced a net progression in the number of cells arrested at the G2/M growth stage in the cell cycle of the K562 human leukemia cell line at 4 μM.

Geranylated Flavonols from Macaranga Rhizinoides

Two geranylated and methylated flavonol derivatives, macarhizinoidins A (1) and B (2), along with a known phenolic compound methyl 4-isoprenyloxycinnamate (3), have been isolated from the methanol extract of the leaves M. rhizinoides. The structures of these compounds were identified based on their spectroscopic data. On cytotoxic evaluation against murine leukemia P-388 cells, compounds 1-2 showed IC50 values of 11.4 and 13.9 microM, respectively, while compound 3 was inactive.

Molecular Docking of Lunacridine from Lunasia amara to DNA : Its Inhibition And Interaction Study Correlated With The Cytotoxic Activity on P388 Murine Leukemia Cells

DNA Topoisomerase II inhibitors are a type of anticancer drugs. These drugs perform their biological activity either by forming a DNA-intercalator-topoisomerase II ternary complex or by inhibiting other enzymes and/or transcription factors that act on DNA. The strong interactions with DNA play a crucial role for their pharmacological properties. Lunacridine, the active principle from Lunasia amara, was known as DNA intercalating Topoisomerase II inhibitor. With the aims to explore the affinity and molecular interaction of lunacridine compound isolated from Lunasia amara with DNA, molecular docking study has been carried out with DNA model using Autodock 4.0 software. Cytotoxicity test on P388 murine leukemia cells was done also using 100, 30, 10, 3 and 1 μg/ml series of lunacridine concentration. The docking result shows that Lunacridine itself could to dock intercalatively between base pairs of DNA and the possibility interaction with adenine, thymine and cytosine by dipole-dipole interaction. The lowest predicted binding energy of lunacridine is –6,22 kcal/mol, whereas original ligand bis thiazole is -16,37 kcal/mol. Lunacridine compound itself has less cytotoxic activity on P388 murine leukemia cells with the IC50 value of 39,52 μg/ml or 129,41 μM. The binding energy of lunacridine on DNA higher than original ligand show that the interaction of lunacridine with DNA is not stable afford the less cytotoxic activity. However, based on the IC50 value, lunacridine could be depeloved as anticancer.Key words: docking, lunacridine, Lunasia amara, cytotoxic, P388 murine leukimia cells.

SYNTHESIS OF UK-3A ANALOGUE AND ASSAY ON P 388 MURINE LEUKEMIA CELLS

UK-3A is secondary metabolite of Streptomyces sp.517-02 which has IC50 38 µg/mL against P388 Murine Leukemia cells. An analogue of UK-3A was synthesized from L-methyl serine as the starting material by amidation and esterification. An analogue UK-3A was analyzed and identified by TLC, FT-IR, LC-MS and NMR spectrometer. It was found to have IC50 15.4µg/mL against the same Leukemia cells. The overall yield was 87.10%. Keywords: UK-3A, Streptomyces sp. 517-02, Anticancer, P388 Murine Leukeumia cell

RESVERATROL DIMERS FROM STEM BARK OF <i>Hopea gregaria</i> AND THEIR CYTOTOXIC PROPERTIES

Six resveratrol dimers have been isolated from the stem bark of H. gregaria, ampelopsin A (1), balanocarpol (2), ε-viniferin (3), hopeafuran (4), heimiol A (5), and parviflorol (6). The structures of these compounds were determined based on spectroscopic evidence such as UV, IR, 1-D, 2-D NMR and comparison with the reported data. These compound inventions are strengthen conclusion that Hopea tends to produce resveratrol dimers. Biological activity of those compounds against murine leukemia P-388 cells showed that ε-viniferin (3) is the most active compound with IC50 value 5.1 ± 0.3 μg/mL. Keywords: H. gregaria, resveratrol dimers, murine leukemia P-388 cells

Antineoplastic agents 582. Part 1: Isolation and structure of a cyclobutane-type sesquiterpene cancer cell growth inhibitor from Coprinus cinereus (Coprinaceae)

Bioassay-guided (murine P388 lymphocytic leukemia and human cancer cell lines) separation of an ethyl acetate extract prepared from the inky cap fungus Coprinus cinereus led to the isolation of three new sesquiterpenes, 7,7a-diepicoprinastatin 1 (1), 14-hydroxy-5-desoxy-2S,3S,9R-illudosin (2), and 4,5-dehydro-5-deoxyarmillol (3), together with the known armillol (4). The structure and relative configuration of 1 was determined by single-crystal X-ray diffraction experiments. The structures of compounds 2, 3, and 4 were each deduced by a combination of HRMS and 1D and 2D NMR techniques. Cyclobutane 2 led to modest inhibition of the murine P388 leukemia cell line.

Vanchrobactin and anguibactin siderophores produced by Vibrio sp. DS40M4.

The marine bacterium Vibrio sp. DS40M4 has been found to produce a new triscatechol amide siderophore, trivanchrobactin (1), a related new biscatecholamide compound, divanchrobactin (2), and the previously reported siderophores vanchrobactin (3) and anguibactin (4). Vanchrobactin is comprised of l-serine, d-arginine, and 2,3-dihydroxybenzoic acid, while trivanchrobactin is a linear trimer of vanchrobactin joined by two serine ester linkages. The cyclic trivanchrobactin product was not detected. In addition to siderophore production, extracts of Vibrio sp. DS40M4 were screened for biologically active molecules; anguibactin was found to be cytotoxic against the P388 murine leukemia cell line (IC(50) < 15 microM).

Phenolic compounds from Cryptocarya konishii: their cytotoxic and tyrosine kinase inhibitory properties

Two chalcone derivatives, 2'-hydroxychalcone (1) and desmethylinfectocaryone (2), together with five known phenolic compounds infectocaryone (3), cryptocaryone (4), kurzichalcolactone A (5), pinocembrin (6) and trans-N-feruloyltyramine (7), were isolated from the methanol extract of the wood of Cryptocarya konishii. The structures of the new compounds were determined based on the analysis of spectroscopic data, including UV, IR, 1D and 2D NMR, and mass spectra. Evaluation of the cytotoxic and tyrosine kinase inhibitory activities of compounds 1-7 showed that compounds 2-4 strongly inhibited the growth of murine leukemia P-388 cells, whereas compound 4 significantly inhibited the enzyme.

A Role for Hydrogen Peroxide in the Pro‐apoptotic Effects of Photodynamic Therapy

Although the first reactive oxygen species (ROS) formed during irradiation of photosensitized cells is almost invariably singlet molecular oxygen (1O2), other ROS have been implicated in the phototoxic effects of photodynamic therapy (PDT). Among these are superoxide anion radical (*O2(-)), hydrogen peroxide (H2O2) and hydroxyl radical (*OH). In this study, we investigated the role of H2O2 in the pro-apoptotic response to PDT in murine leukemia P388 cells. A primary route for detoxification of cellular H2O2 involves the peroxisomal enzyme catalase. Inhibition of catalase activity by 3-amino-1,2,4-triazole led to an increased apoptotic response. PDT-induced apoptosis was impaired by addition of an exogenous recombinant catalase analog (CAT-skl) that was specifically designed to enter cells and more efficiently localize in peroxisomes. A similar effect was observed upon addition of 2,2'-bipyridine, a reagent that can chelate Fe+2, a co-factor in the Fenton reaction that results in the conversion of H2O2 to *OH. These results provide evidence that formation of H2O2 during irradiation of photosensitized cells contributes to PDT efficacy.

(+)-N-(2-Hydroxypropyl)lindcarpine: A New Cytotoxic Aporphine Isolated from Actinodaphne pruinosa Nees

One new alkaloid; (+)-N-(2-hydroxypropyl)lindcarpine (1), together with four known aporphine alkaloids, (+)-boldine (2) (+)-norboldine (3), (+)-lindcarpine (4) and (+)-methyllindcarpine (5) were isolated from the stem bark of Actinodaphne pruinosa Nees (Lauraceae). (+)-N-(2-Hydroxypropyl)lindcarpine (1) exhibited cytotoxic activity against P-388 murine leukemia cells with an IC50 value of 3.9 μg/mL. Structural elucidation of all the compounds were performed by spectral methods such as 1D- and 2D- NMR, IR, UV, and HRESIMS.

Phenylpropanoid triesters from Daucus glaber

Three new phenylpropanoid triesters, glaberins A–C ( 1– 3), were isolated from the leaves and stems of Daucus glaber (Forssk.) Thell. (Apiaceae). Their gross structures were determined on the basis of 2D NMR experiments, and the absolute structure of 1 by X-ray crystallography and chemical methods. Glaberins A ( 1) and B ( 2) were evaluated for cytotoxicity against P-388 murine lymphocytic leukemia cells.

Prenylated Flavones from Artocarpus Lanceifolius and their Cytotoxic Properties against P-388 Cells

New prenylated flavones, artoindonesianins Z-4 and Z-5, together with four known prenylated flavones, artonin E, 12-hydroxyartonin E, artobiloxanthone, and cycloartobiloxanthone, have been isolated from the methanol extract of the tree bark of Artocarpus lanceifolius. The structures of these compounds were determined on the basis of spectroscopic data, including UV, IR, 1D and 2D NMR, and mass spectra. The cytotoxic effect of the isolated compounds against murine leukemia P-388 cells is described.

Synthetic Studies of the Zoanthamine Alkaloids: The Total Syntheses of Norzoanthamine and Zoanthamine

The zoanthamine alkaloids, a type of heptacyclic marine alkaloid isolated from colonial zoanthids of the genus Zoanthus sp., have distinctive biological and pharmacological properties in addition to their unique chemical structures with stereochemical complexity. Namely, norzoanthamine (1) can suppress the loss of bone weight and strength in ovariectomized mice and has been expected as a promising candidate for a new type of antiosteoporotic drug, while zoanthamine (2) has exhibited potent inhibitory activity toward phorbol myristate-induced inflammation in addition to powerful analgesic effects. Recently, norzoanthamine derivatives were demonstrated to inhibit strongly the growth of P-388 murine leukemia cell lines, in addition to their potent antiplatelet activities on human platelet aggregation. Their distinctive biological properties, combined with novel chemical structures, make this family of alkaloids extremely attractive targets for chemical synthesis. However, the chemical synthesis of the zoanthamine alkaloids has been impeded owing to their densely functionalized complex stereostructures. In this paper, we report the first and highly efficient total syntheses of norzoanthamine (1) and zoanthamine (2) in full detail, which involve stereoselective synthesis of the requisite triene (18) for an intramolecular Diels-Alder reaction via the sequential three-component coupling reactions, the key intramolecular Diels-Alder reaction, and subsequent crucial bis-aminoacetalization as the key steps. Ultimately, we achieved the total synthesis of norzoanthamine (1) in 41 steps with an overall yield of 3.5 % (an average of 92 % yield each step) and that of zoanthamine (2) in 43 steps with an overall yield of 2.2 % (an average of 91 % yield each step) starting from (R)-5-methylcyclohexenone (3), respectively.

Ophiodilactones A and B, Cytotoxic Tetrameric Phenylpropanoids, from the Ophiuroid Ophiocoma scolopendrina

Two tetrameric phenylpropanoids, ophiodilactones A (1) and B (2), were isolated from the ophiuroid Ophiocoma scolopendrina. Their structures were determined on the basis of spectroscopic data. Ophiodilactone B has a novel carbon skeleton. They exhibit moderate cytotoxic activity against P388 murine leukemia cells.