P388 Leukemia Cells Research Articles

Antioxidant and cytotoxic agent from the rhizomes of Kaempferia pandurata

ObjectiveTo determine antioxidant and cytotoxic activity of two flavanones, pinocembrin (1) and pinostrobin (2) from the rhizomes of Kaempferia pandurata. The chemical structures of both compounds were determined based on spectroscopic data, including UV, IR, MS and NMR spectra. MethodsThe antioxidant activities of pinocembrin (1) and pinostrobin (2) were assayed by using 2,2-diphenyl-1-picrylhydrazyl. Cytotoxic assay was done by using brine shrimp lethality test, and cytotoxic properties was tested against murine leukemia P-388 cells. ResultsCompounds 1-2 were evaluated for their antioxidant properties against DPPH, showing their IC50 were 5 816 and 6 268 μmol/L; brine shrimp lethality test: LC50 23.3 and 60.5 μg/mL; murine leukemia P-388: IC50 176.3 and 218.5 μmol/L. ConclutionsThe results indicated that pinocembrin (1) was slightly more active than pinostrobin (2).

Cycloforskamide, a Cytotoxic Macrocyclic Peptide from the Sea Slug Pleurobranchus forskalii

A macrocylic dodecapeptide, cycloforskamide, was isolated from the sea slug Pleurobranchus forskalii, collected off Ishigaki Island, Japan. Its planar structure was deduced by extensive NMR analyses and was further confirmed by MS/MS fragmentation analyses. Finally, the absolute configuration was determined by total hydrolysis and chiral-phase gas chromatographic analysis. This novel dodecapeptide contains three d-amino acids and three thiazoline heterocycles and exhibits cytotoxicity against murine leukemia P388 cells, with an IC50 of 5.8 μM.

Andirolides Q–V from the flower of andiroba (Carapa guianensis, Meliaceae)

Two new gedunins, an andirobin, two mexicanolides, and a phragmalin-type limonoid, named Andirolides Q (1), R (2), S (3), T (4), U (5), and V (6), were isolated from an oil of the flower of Carapa guianensis AUBLET (Meliaceae). Their structures have been elucidated on the basis of spectroscopic analyses using 1D and 2D NMR spectra and FABMS. Andirolide S (3) and Andirolide T (4) showed significant cytotoxic activity against the murine P388 leukemia cell line (IC50 of 1.4μM for 3; 1.8μM for 4) and the human HL-60 leukemia cell line (IC50 of 1.3μM for 3 and 4).

Synthesis, Structure and Cytotoxic Activity of New Acetylenic Derivatives of Betulin

A new series of betulin derivatives containing one or two pharmacophores bearing an acetylenic and carbonyl function at the C-3 and/or C-28 positions has been synthesized and characterized by 1H- and 13C-NMR, IR, MS and elemental analyses. The crystal structure of 28-O-propynoylbetulin was determined by X-ray structural analysis. All new compounds, as well as betulin, were tested in vitro for their antiproliferative activity against human SW707 colorectal, CCRF/CEM leukemia, T47D breast cancer, and against murine P388 leukemia and Balb3T3 normal fibroblasts cell lines. Most of the compounds showed better cytotoxicity than betulin and cisplatin used as reference agent. 28-O-Propynoylbetulin was the most potent derivative, being over 500 times more potent than betulin and about 100 times more cytotoxic than cisplatin against the human leukemia (CCRF/CEM) cell line, with an ID50 value of 0.02 μg/mL.

Phytochemical of Indonesian Macaranga and their Cytotoxic Properties Against P388 Cells

Plants as a source of bioactive chemicals have been known since very long time, and thus it is a rational strategy to develop phytochemical studies on the Indonesian plants and tested the isolated chemicals on a various biological systems. Since 1999, the author, as a part the Group of Natural Products Chemistry, Institut Teknologi Bandung, has actively wotk on a number of Indonesian plant families, including Lauraceae, Moraceae, Dipterocarpaceae, Zingiberaceae, and Euphorbiaceae, aiming to find new biologically active chemicals from nature. In this presentation, studies on Macaranga species (Euphorbiaceae) will be presented, hightlighting the results from six species of the genus, namely M. gigantea, M. lowii, M. pruinosa, M. recurvata, M. rhizinoides, and M. trichocarpa. Samples of the plants to be extracted were the leaves, and extraction was done either with acetone or methanol. Fractionation was carried out by vacuum liquid chromatography eluted by a gradient solvent system of n-hexaneEtOAc. Purification of the metabolites was done mainly with radial chromatography eluted with various solvent systems. Twenty seven flavonoid and stibene derivatives were isolated and identified, seventeen of which were new compounds. These compounds are substituted with isoprenyl, geranyl, farnesyl, and an iregular sesquiterpenyl group with a cyclobutane skeleton. In the presentation, several selected NMR spectra of the compounds will be described. On biological evaluation against murine leukemia P-388 cells by MTT assay these compounds showed varying cytotoxicities, with the highest active compounds was an isoprenylated dihydroflavanone isolated from M. recurvata. In conclusion, Macaranga is a unique source of flavonoid and stilbene derivatives, some of them have potential for biological activities. Keeping that Indonesia is one of the center of Macaranga’s diversity, pytochemical and biological research on this plant should be continued.

Cytotoxic Activity of Secondary Metabolites from the Bark of Dipterocarpus Confertus Sloot

Four compounds, namely β-sitosterol (1), betulinic acid (2), cinnamic acid (3), and α-viniferin (4) have been successfully isolated from the bark methanol extract of Dipterocarpus confertus Sloot. The structures of the isolated compounds have been established on the basis spectroscopic data evidence, and comparison with the published data. In the cytotoxicity study, cinnamic acid (3) and betulinic acid (2) have been found to be very strong active against murine leukemia P388 and vero cells lines with the IC50 values of 2.25 and 5.10 µg/mL, respectively, while the other compounds were not active.

Fluorinated and rearranged gedunin derivatives

When treated with (diethylamino)sulfur trifluoride (DAST), 11α-hydroxygedunin gave 11β-fluorogedunin and 9,11-didehydrogedunin, whereas deacetylgedunin afforded two skeletal rearranged products 6 and 7, in which the Me-30 had shifted from position 8 to position 7. Of those products, 11β-fluorogedunin and 6 were shown to be more cytotoxic than gedunin on P-388 leukemia cells.

Abstract 3733: Combined effect of linalool as component of Humulus lupulus on doxorubicin-induced antitumor activity

Abstract [Purpose] Cancer chemotherapy has been applied to advanced tumors and intractable tumors, playing an important role in their treatments. However, there are many problems including the appearance of adverse reactions and acquirement of drug resistance. The increased numbers of medications resulted in a decreased quality of life for the patients. However, food and food components can be used to increase the activity of antitumor agents without increasing the number of medicines. In this study, the effects of some components in Humulus lupulus (Hop) were examined on the doxorubicin (DOX) induced antitumor activity in vitro and in vivo. [Methods] The effects of Hop components (linalool, myrcene, α-humulene, α-caryophyllene) on DOX permeability and DOX cytotoxicity in P388 leukemia cells in vitro were examined. P388 leukemia cells (5 x 105 cells per animal) were transplanted onto the backs of BDF1 mice, and DOX (2.0mg/kg/day x 4 days, i.p.) and linalool (1.0mg/kg/day x 4 days, p.o.) were administered. The solid tumor and normal tissues were removed and measured DOX concentrations in each tissue. [Results and Discussion] In vitro, myrcene, α-humulene and α-caryophyllene did not have beneficial effects on DOX permeability in tumor cells. In contrast, the addition of linalool was shown to significantly promote on DOX influx and increased DOX concentration in tumor cells. The combination of DOX with linalool (0.1 μM) showed 3.5-fold (P<0.05) cytotoxicity, compared to treatment with DOX alone. Therefore, the combined linalool with DOX enhanced DOX cytotoxicity by the increase of DOX concentration in tumor cells. In vivo, the tumor weight in the DOX alone group decreased to 77% of the control level, whereas the combination of linalool with DOX decreased the tumor weight by 53% (P<0.05) of that in DOX alone group. In the tumor, the DOX concentration in the linalool combined group increased by 1.8-fold (P<0.05) compared to DOX alone group. In the normal tissue, the DOX concentration after combined linalool and DOX treatment tended to decrease. In particular, the DOX concentration in the heart decreased by 49% (P<0.05) of that in DOX alone group, expecting that linalool may decrease DOX-induced cardiotoxicity. Thus, the combination of linalool with DOX increased DOX concentration in the tumor, consequently enhanced antitumor activity of DOX. In conclusion, linalool is expected effectiveness as DOX modulator. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3733. doi:1538-7445.AM2012-3733

TERPENOIDS FROM THE STEM BARK OF JATROPHA PLANTS AND THEIR BIOLOGICAL ACTIVITIES

Three terpenoids, including two diterpenes (curcusone B and jatrophone) and a triterpene (stigmasterol) have been isolated from the stem bark of Jatropha plants. Curcusone B and stigmasterol were isolated from J. curcas, meanwhile jatrophone and stigmasterol were from J. gossypifolia. The biological activities of these compounds have been evaluated toward bacteria, fungi and tumour cells. Isolation was carried out in vacuum liqiud cromatography (VLC) technique with silica gel as an adsorben and some solvents as eluents. The compound structures were determined by spectroscopic methodes i.e. UV-vis, FTIR, NMR (1-D, 2-D) and were then compared based on their spectroscopic data with similiar data from literatures. The biological properties of these compounds were evaluated against four strains of bacteria (Acetobacter sp., Eschericia coli, Staphylococcus aureus, and Streptococcus sp.), 4 strains of fungi (Aspergilus niger, Penicillium sp. (grey), Penicillium sp. (white) and Rhizopus sp.) and murine leukemia P-388 cells. The results showed that cytotoxic property of curcusone B towards murine leukemia P-388 cells is better than jatrophone and stigmasterol which are IC50 = 0.57 μg/mL (1.93 μM) for curcusone B and IC50 > 100 μg/mL for jatrophone and stigmasterol. Meanwhile, activities against bacteria, jatrophone is better than curcusone B and stigmasterol. Jatrophone is the most active against S. aureus (bacteria) with growth inhibition zone 36 mm and A.niger (fungi) is 44 mm. Further study indicated that jatrophone was bacteriostatic against S. aureus.

Cytogenetic and antineoplastic effects by newly synthesised steroidal alkylators in lymphocytic leukaemia P388 cells in vivo

New compounds with potential antitumour activity were synthesised by combining nitrogen mustard with the steroidal skeleton, in an effort to improve specificity and at the same time reduce systemic toxicity. The steroidal part is aimed to serve as a biological platform enabling the alkylating moiety to approach its site of action by altering its physicochemical properties. The purpose of the present investigation was to evaluate these compounds for anti-neoplastic activity. The compounds tested have as alkylators either para-NN-bis(2-chloroethyl)-aminophenyl-butyrate (CHL) or para-N,N-bis(2-chloroethyl)-aminophenyl-acetate (PHE) esterified with a differently modified steroidal nucleus. The eight newly synthesised compounds were compared on a molar basis with respect to their ability to induce sister chromatid exchanges (SCEs) and to modify proliferation rate indices (PRI) in lymphocytic leukaemia P388 cells in mice in vivo. The life span of BDF1 mice inoculated with P388 leukaemia cells was also estimated (anti-leukaemic activity). The compounds that were effective in inducing cytogenetic effects in lymphocytic leukaemia cells in vivo were also effective in inducing antineoplastic effects in BDF1 mice inoculated with P388 leukaemia cells.These results suggest that the in vivo cytogenetic effects in conjunction with the antineoplastic activity of modified steroidal alkylators depend on the configuration of the whole molecule and on the appropriate combination of the alkylator with the steroidal molecule: a pronounced cytogenetic and anti-neoplastic action was demonstrated by the compounds that contain either PHE or CHL as alkylators and are esterified with either a steroidal nucleus that carries a cholesten group in the 17 position of the D-ring, or with a steroidal nucleus having an exocyclic NHCO-group in the D-ring. In contrast, a ketone group or an NHCO-group in the D-ring inserted endocyclically in the steroidal nucleus esterified with either CHL or PHE failed to induce cytogenetic or anti-neoplastic effects.

Regulation by survivin of cancer cell death induced by F14512, a polyamine-containing inhibitor of DNA topoisomerase II

F14512, an epipodophyllotoxin derivative equipped with a spermine moiety, is selectively taken up by the polyamine transport system over-active in tumor cells. F14512 was identified as a selective anticancer agent with a broad spectrum of antitumor activities and is currently undergoing phase I clinical trial in onco-hematology. However, the mechanism by which F14512 exerts its selective effects on neoplastic cells remains poorly understood. In this study, using mainly P388 leukemia cells, we showed that activation of the DNA damage response by F14512 did not induce immediate apoptosis but resulted in an early growth arrest. F14512-induced G2 arrest was accompanied by the appearance of a senescence-like phenotype (characterized by an increased β-galactosidase staining) with up-regulation of the cyclin-dependent kinase inhibitor p16, and cyclin D1. The early senescence-based cell cycle block was characterized by a marked increase of the level of the IAP protein survivin, but not cIAP2, in P388 cells as well as in three other leukemia and melanoma cell types. The Thr(34)-phosphorylated form of survivin was observed within 4 h after F14512 exposure. Inhibition of survivin by siRNA resulted in a switch from senescence-like growth arrest to apoptosis. Compared with the parental drug etoposide, F14512-induced DNA damage signaling pathway resulted in greater senescence like-growth arrest and delayed apoptosis. Collectively, our data show that senescence arrest and subsequent apoptosis are powerful mechanisms mediating the chemotherapeutic effects of F14512 and identify survivin as the molecular determinant responsible for a qualitative shift in cell fate from senescence to apoptosis upon treatment with F14512.

Induction of apoptosis in murine leukemia by diarylheptanoids from Curcuma comosa Roxb.

Diarylheptanoids, isolated from the rhizome of Curcuma comosa Roxb., have several biological activities including anti-oxidant and anti-inflammation. The present study investigated the effect of five diarylheptanoids isolated from C. comosa rhizome on the proliferation of murine P388 leukemic cells. Compound-092, (3S)-1-(3,4-dihydroxyphenyl)-7-phenyl-(6E)-6-hepten-3-ol, bearing a catechol moiety, was the most potent diarylheptanoid (IC(50) of 4μM) in inhibiting P388 leukemic cell viability by causing DNA breakage and inducing apoptosis. Apoptotic cell death was characterized by the presence of chromatin condensation, formation of apoptotic bodies, DNA fragmentation, and externalization of plasma membrane phosphatidylserine. This compound increased caspase-3 activity about fivefold above the untreated control, decreased the intracellular reduced glutathione level, and impaired mitochondrial transmembrane potential. In the presence of Cu(II) ion, the compound exhibited a pro-oxidant activity causing DNA strand breakage and enhancing the anti-proliferative activity. The results provide evidence for the pro-oxidant activity of the diarylheptanoid bearing a catechol moiety in the induction of apoptosis in murine P388 leukemia.

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.

Recent Advances in the Studies on Luotonins

Luotonins are alkaloids from the aerial parts of Peganum nigellastrum Bunge. that display three major skeleton types. Luotonins A, B, and E are pyrroloquinazolino-quinoline alkaloids, luotonins C and D are canthin-6-one alkaloids, and luotonin F is a 4(3H)-quinazolinone alkaloid. All six luotonins have shown promising cytotoxicities towards selected human cancer cell lines, especially against leukemia P-388 cells. Luotonin A is the most active one, with its activity stemming from topoisomerase I-dependent DNA-cleavage. Such intriguing biological activities and unique structures have led not only to the development of synthetic methods for the efficient synthesis of these compounds, but also to interest in structural modifications for improving the biological properties. Recent progress in the study of luotonins is covered.

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.

Per-N-methylated analogues of an antitumor bicyclic hexapeptide RA-VII

Penta- N-methyl and hexa- N-methyl analogues of RA-VII, an antitumor bicyclic hexapeptide of plant origin, were prepared. In the former, the nitrogens of d-Ala-1 and Ala-4 and in the latter, those of d-Ala-1, Ala-2, and Ala-4 were methylated under the phase-transfer catalysis conditions. Their solution structures were established by NOESY experiments and the crystal structures by X-ray crystallography. Those two methylated analogues showed much weaker cytotoxicity against P-388 leukemia cells than the parent RA-VII.

Plant Derived Compounds Having Activity against P388 and L1210 Leukemia Cells

Cancer is a major cause of death and the number of new cases as well as the number of individuals living with cancer is expanding continuously. Due to the enormous propensity of plants that synthesize mixture of structurally derived bioactive compounds, the plant kingdom is potentially diverse source of chemical constituents with murine leukemia activity. Plant derived compounds have played an important role in the development of several clinically useful anticancer agents having activity against murine leukemia. These include vinca alkaloids, lignans, terpenoids, quinines and phytochemicals such as flavones, flavonoids, flavonol. The active compounds which have been isolated from various plant sources are listed, classified and discussed in terms of their activity against murine leukemia cells. Structure-activity relationship has been studied for further development of compounds which show activity against leukemia cells.

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.

Synthesis and biological evaluation of 2-amino-1-thiazolyl imidazoles as orally active anticancer agents

Designed from a high throughput screened hit compound, novel 2-amino-1-thiazolyl imidazoles were synthesized and demonstrated cytotoxicity against human cancer cells. 1-(4-Phenylthiazol-2-yl)-4-(thiophen-2-yl)-1H-imidazol-2-amine (compound 2), a 2-amino-1-thiazolyl imidazole, inhibited tubulin polymerization, interacted with the colchicine-binding sites of tubulins, and caused cell cycle arrest at the G(2)/M phase in human gastric cancer cells. Disruption of the microtubule structure in cancer cells by compound 2 was also observed. Compound 2 concentration-dependently inhibited the proliferation of cancer cells in histocultured human gastric and colorectal tumors. Given orally, compound 2 prolonged the lifespans of leukemia mice intraperitoneally inoculated with the murine P388 leukemic cells. We report 2-amino-1-thiazolyl imidazoles as a novel class of orally active microtubule-destabilizing anticancer agents.

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.