P388 Murine Leukemia Cells Research Articles

Kumemicinones A–G, Cytotoxic Angucyclinones from a Deep Sea-Derived Actinomycete of the Genus Actinomadura

Chemical investigation of the fermentation products of a deep sea water-derived actinomycete, Actinomadura sp. KD439, identified seven new angucyclinones, designated as kumemicinones A-G (1-7), together with the known SF2315B and miaosporone E. NMR and MS spectroscopic analyses, combined with X-ray crystallography and quantum chemical calculations of NMR chemical shifts and electronic circular dichroism (ECD) spectra, uncovered the structures of new angucyclinones as regioisomers of SF2315B at the allyl alcohol unit (1 and 2), an epoxy ring-opened γ-hydroxy enone isomer (3), a B/C-ring-rearranged product (4), or dimers with a new mode of bridging (5-7), adding new structural variation to this antibiotic group. The absolute configuration of SF2315B was also determined by comparison of ECD spectra with those of 1 and 2. All the angucyclinones exhibited cytotoxicity against P388 murine leukemia cells, with IC50 values ranging from 1.8 to 53 μM.

Tenacibactins K-M, cytotoxic siderophores from a coral-associated gliding bacterium of the genus Tenacibaculum.

HPLC/DAD-based chemical investigation of a coral-associated gliding bacterium of the genus Tenacibaculum yielded three desferrioxamine-class siderophores, designated tenacibactins K (1), L (2), and M (3). Their chemical structures, comprising repeated cadaverine–succinic acid motifs terminated by a hydroxamic acid functionality, were elucidated by NMR and negative MS/MS experiments. Compounds 1–3 were inactive against bacteria and a yeast but displayed cytotoxicity against 3Y1 rat embryonic fibroblasts and P388 murine leukemia cells at GI50 in submicromolar to micromolar ranges. Their iron-chelating activity was comparable to deferoxamine mesylate.

Characterization and Anticancer Activity from Gaharu (Aquilaria malaccensis) Stem Bark Extract

Aquilaria malaccensis is a plant species from the Thymeleaceae family. A. malaccensis contains secondary metabolites of phenolic groups, namely benzophenone, chromone, coumarin, xanthone, and flavonoids with various biological activities such as antioxidants and anticancer. This study aims to determine the potential of A. malaccensis stem bark extract as an anticancer and to determine the characteristics of secondary metabolite compounds contained in A. malaccensis stem bark extract. Extraction of the stem bark of A. malaccensis by maceration method in stages using n -hexane, ethyl acetate and methanol as solvents. The anticancer activity test used the microculture tetrazolium (MTT) assay against Murine Leukemia P-388 cells. Compound separation using thin layer chromatograph (TLC) and gravity column chromatography (CC) methods. Determination of chemical compounds using liquid chromatography mass spectroscopy/mass spectroscopy (LCMS/MS). The extracts of n -hexane, ethyl acetate, and methanol each had anticancer activity which was classified as active with an IC 50 value of 20.64, 26.61, and 14.49 µg/mL. Based on the results of separation and identification of compounds in methanol extract with TLC, CC, and LCMS/MS, methanol extract contains acetaminophen, aminosamptothesin, isophorone, diethyl phthalate, 3-hydroxy-6H-benzochromen-6-one, isolongifolenic, 2-(2-phenethyl) chromone, 4′,7-dihydroxy-2′,5-dimethoxyisoflavone, 4-hydroxy-3-(1-phenylpropyl)-2H-chromone-2-one; 6-methylchromone-2-carboxylic acid; 3,4-dimethyl-2- phenylmorpholine; and β-calacoren compounds.

Nomimicins B-D, new tetronate-class polyketides from a marine-derived actinomycete of the genus Actinomadura.

Three new tetronate-class polyketides, nomimicins B, C, and D, along with nomimicin, hereafter named nomimicin A, were isolated from the culture extract of Actinomadura sp. AKA43 collected from floating particles in the deep-sea water of Sagami Bay, Japan. The structures of nomimicins B, C, and D were elucidated through the interpretation of NMR and MS analytical data, and the absolute configuration was determined by combination of NOESY/ROESY and ECD analyses. Nomimicins B, C, and D showed antimicrobial activity against Gram-positive bacteria, Kocuria rhizophila and Bacillus subtilis, with MIC values in the range of 6.5 to 12.5 μg/mL. Nomimicins B and C also displayed cytotoxicity against P388 murine leukemia cells with IC50 values of 33 and 89 μM, respectively.

Antiproliferative Activity of Triterpenoid and Steroid Compounds from Ethyl Acetate Extract of Calotropis gigantea Root Bark against P388 Murine Leukemia Cell Lines

Calotropis gigantea has been known to produce bioactive secondary metabolites with antiproliferative activities against cancer cells. Herein, we extracted the secondary metabolites using ethyl acetate from its root bark and further tested its antiproliferative activities against P388 murine leukemia cell lines. The subfractions from the ethyl acetate extract was obtained from Vacuum Liquid Column Chromatography (VLCC), and followed by Gravity Column Chromatography (GCC). The subfraction C2 and D1 were identified to contain triterpenoids and steroids with the most potent cytotoxicity against Brine Shrimp Lethality Test (BSLT). A 3-(4,5-dimethylthiazol-2-yl) -2-5 diphenyl tetrazolium bromide (MTT) assay suggested that ethyl acetate extract has the highest antiproliferative activities against P388 murine leukemia cell lines (IC50 = 21.79 μg/mL), as opposed to subfraction C2 (IC50 = 50.64 µg/mL) and subfraction D1 (IC50 = 49.33 µg/mL). The compound identified in subfraction C2 and D1 are taraxerol acetate and calotropone, respectively. Though taraxerol acetate and calotropone were active in inhibiting the leukemic cell lines, their IC50s were lower than the ethyl acetate extract, which is probably due to the synergism of the secondary metabolites.

Synthesis, Cytotoxic Evaluation and Molecular Docking of Bromo-Substituted 1,3,6-Trihydroxyxanthone as Protein Tyrosine Kinase Inhibitor

A series of bromo-substituted hydroxyxanthone were synthesized using bromine in glacial acetic acid as a solvent, producing compounds 3 and 5 with 82.1% and 84.2% yield, respectively. In vitro analyses of these two compounds using MMT assay against murine leukemia P388 cell line resulted in IC50 of 2,550 and 3,455 μg/mL and selectivity index of 43.21 and 74.40, respectively. These results indicated that the bromo-substituted hydroxyxanthone compounds could potentially be developed as selective and sensitive anti-cancer agents. Furthermore, through the molecular docking conducted using Discovery Studio, compound 3 exhibited interactions between amino acid residues and protein tyrosine kinase (1T46.pdb). This article provides supporting data that xanthone derivatives can be used as drugs for cancer chemotherapy through the mechanism of how this compound can inhibit protein tyrosine kinase.

TMKS8A, an antibacterial and cytotoxic chlorinated α-lapachone, from a sea slug-derived actinomycete of the genus Streptomyces.

TMKS8A (1), a new chlorinated α-lapachone derivative, along with five known related metabolites, A80915 C (2), SF2415B1 (3), chlorinated dihydroquinone 3 (4), SF2415B3 (5), and A80915 C (6), were identified from the culture extract of Streptomyces sp. TMKS8, which was isolated from a sea slug, Paromoionchis tumidus. The structure of 1 was determined by the analysis of NMR and MS spectral data, assisted by NMR chemical shift prediction using DFT-based calculation. The absolute configuration was determined to be R by comparison of experimental and calculated ECD spectra. Compound 1 displayed antimicrobial activity against Gram-positive bacteria with MIC values ranging from 6.25 to 12.5 μg ml-1 and cytotoxicity against murine leukemia P388 cells with IC50 9.8 μM.

Nyuzenamides A and B: Bicyclic Peptides with Antifungal and Cytotoxic Activity from a Marine-Derived Streptomyces sp.

Two bicyclic peptides, nyuzenamides A (1) and B (2), were discovered from Streptomyces isolated from suspended matter in deep sea water collected in the Sea of Japan. Their structures were determined through nuclear magnetic resonance and mass spectrometry analyses in combination with X-ray crystallography and the chiral-phase gas chromatography-mass spectrometry method to comprise ten amino acid residues containing four unusual amino acids along with aromatic acyl units. Both compounds displayed antifungal activity against pathogenic fungi and cytotoxicity against P388 murine leukemia cells.

Cytotoxicity Assay of Water, Ethanol N-Hexane Extract of Dates Fruit (Phoenix dactylifera) Against Murine Leukemia Cancer Cell P388

The World Health Organization (WHO) states, in 2015, there are an estimated 9 million people who die of cancer and in 2030 there are an estimated 11.4 million deaths from cancer. Blood cancer reaches 3% of all cancers found in humans and is most common in children. The cancer drug that is available today has quite severe side effects, therefore in this study an antileukemia activity test will be carried out on water, 70% ethanol, and n-hexane extract of dates (Phoenix dactylifera). This study, the extracts from dates will be tested for cytotoxicity on Murine Leukemia P388 cells using the MTT Assay method. The IC50 value of extract of dates is used as an assessment of the activity of palm fruit extract as antileukemia. Citotoxicity assay against Murine Leukemia P388 cells by MTT assay, showed IC50 value is more than 100 µg/mL for all extracts. This result informed that the water, ethanol, and n-hexane extract of dates fruit are less potencial as a antileukemia agent.

Small-Scale Preparation of Fluorescently Labeled Chemical Probes from Marine Cyclic Peptides, Kapakahines A and F.

A number of bioactive marine natural products have been isolated so far, but it is still difficult to disclose their modes of action. In this study, we prepared fluorescently labeled chemical probes from the cytotoxic marine cyclic peptides kapakahines A (1) and F (2) to visualize their localization as the first step of the study of their modes of action. We used fluorescent dyes 3a or 3a/b (a 1:1 mixture of 3a and 3b) whose terminal N-hydroxysuccinimide (NHS) group can react with the free amino groups of kapakahines. The fluorescently labeled kapakahine A (Kap A-5-FL, 5a) stained P388 murine leukemia cells and HeLa human cervical cancer cells, while cells treated with fluorescently labeled kapakahine F (Kap F-5-FL, 6a) only weakly stained them. Further analysis of the confocal images of the stained cells with higher magnification (×100) indicated the localization of Kap A-5-FL (5a) in the cells. In this paper, we report the small-scale preparation and a new delivery method of fluorescent probes, as well as the application of these procedures to cell staining.

Identification of marine bacteria HPP.4A and HPP.T13 and its anticancer activity against P388 murine leukaemia cell

Sea cucumber has been widely studied as a source of bioactive compounds with various biological activities such as antibacterial, antifungal, antioxidant, and anticancer. However, there are a few studies have reported on the biological activity of its associated bacteria. The purpose of this study were to determine the potential of sea cucumber associated bacteria from Panjang Island, Jepara, Central Java, Indonesia as a natural source of anticancer compounds and identify the prospective isolates through DNA barcoding. Bacteria HPP.4A and HPP.T13 were isolated from the gut of sea cucumber Holoturia atra. The bacteria were cultivated in three different media (A3, A11, and A16) then extracted using 1-butanol with maceration method. Cytotoxic assay of each extract was conducted against P388 murine leukaemia cell. Bacteria HPP.4A and HPP.T13 were identified through molecular approach as Sallinicoccus roseus and Sphingobium yanoikuyae with 99.73% similarity. The strongest anticancer activity was showed by Sallinicoccus roseus extract which cultivated in A11 medium while Sphingobium yanoikuyae extract in A3 medium.

Study of biomedical properties of Rhizophora mucronata fruit from Rembang, Central Java

Indonesian in coastal region utilize mangrove ecosystem for aquaculture, fishing ground, firewood source, tourism, nevertheless it is rarely reported as a source of biomedical substances. Rhizophora mucronata is one of the most common mangrove species in Rembang, Central Java. However, study of biomedical prospect of its fruit is neglected. This study aimed to investigate the antimicrobial activity of R. mucronata fruit extract to inhibit human pathogens (E. coli, S. aureus, M. luteus, T. rubrum, and C. albicans), as well as its anticancer activity against P388 murine leukemia cells. Metabolite profile was characterized through phytochemical test and HPLC. The result showed there was no antimicrobial activity of R. mucronata fruit extract against all human pathogens. Cytotoxic assay indicated a moderate anticancer potential with IC50 value of 398 μg/mL. Phytochemical test indicated the presence of alkaloid, flavonoid, and steroid/triterpenoid in the crude extract.

Antibacterial, cytotoxicity and metabolite profiling of crude methanolic extract from andaliman (Zanthoxylum acanthopodium) fruit

Abstract. Sibero MT, Siswanto AP, Frederick EH, Wijaya AP, Syafitri E, Farabi K, Murwani R, Saito S, Igaras Y. 2020. Antibacterial, cytotoxicity and metabolite profiling of crude methanolic extract from andaliman (Zanthoxylum acanthopodium) fruit. Biodiversitas 21: 4147-4154. The local community in North Sumatra has utilized andaliman fruit (Zanthoxylum acanthopodium) as spices for traditional cuisines because it has a unique flavor. Information on the antimicrobial activity of Z. acanthopodium fruit against aquaculture pathogens and its bioactivity against leukemia cell lines are limited. The purposes of this study were to evaluate the antimicrobial activity of Z. acanthopodium fruit against Tenacibaculum maritimum, Vibrio alginolyticus, V. anguillarum, V. harveyi that are known as pathogens in aquaculture; to determine cytotoxic property against murine P388 leukemia cells; and to characterize its metabolites profile. The sample was extracted using methanol by the maceration method. Antibacterial assay was conducted by Kirby-Bauer disc diffusion method; while cytotoxicity assay using the XTT method. Proximate analysis showed that Z. acanthopodium fruit contained 63.41% of moisture, 24.73% of crude fiber, 9.81% of crude protein, 6.90% of ash, and 2.55% of crude fat. Several phytochemical components were detected, such as alkaloid, flavonoid, tannin, triterpenoid, and steroid. The GC/MS analysis indicated the presence of various compounds from terpenoid and terpenes derivatives. This study indicated that Z. acanthopodium fruit was not potential as antibacterial agents against the aquaculture pathogens; however, the methanol extract showed cytotoxic potential with IC50 19.14 µg/mL against murine P388 leukemia cells.

Compounds 7-Hydroxy-6-Metoxic Cytotoxic Cytotoxic Stem Leather From Chisochetonmacrophyllus (Meliaceae) Bark

A phenyl propanoid compound, 7-hydroxy-6-methoxycoumarin has been isolated from the bark of Chisocheton macrophyllus (Meliaceae). The chemical structure of isolated compound was identified on the basis of spectroscopic data including UV, IR, 1D-NMR, 2D-NMR and mass along with comparison with those spectral data previously reported. 7-hydroxy-6-methoxycoumarin compound showed cytotoxic effect against P-388 murine leukemia cell with IC50 value of 16.5 mg/mL. 7-hydroxy-6-methoxycoumarin compound was reported for the first time from genus of Chisocheton.

Compounds 7-Hydroxy-6-Metoxic Cytotoxic Cytotoxic Stem Leather From Chisochetonmacrophyllus (Meliaceae) Bark

A phenyl propanoid compound, 7-hydroxy-6-methoxycoumarin has been isolated from the bark of Chisocheton macrophyllus (Meliaceae). The chemical structure of isolated compound was identified on the basis of spectroscopic data including UV, IR, 1D-NMR, 2D-NMR and mass along with comparison with those spectral data previously reported. 7-hydroxy-6-methoxycoumarin compound showed cytotoxic effect against P-388 murine leukemia cell with IC50 value of 16.5 mg/mL. 7-hydroxy-6-methoxycoumarin compound was reported for the first time from genus of Chisocheton.

A cyclopeptide and three oligomycin-class polyketides produced by an underexplored actinomycete of the genus Pseudosporangium.

Aside from the well-studied conventional actinomycetes such as Streptomyces, the less investigated genera of actinomycetes also represent a promising source of natural products. Genome mining indicated that members of the underexplored genus Pseudosporangium, from which no secondary metabolites have been reported to date, may harbor the biosynthetic machinery for the formation of novel natural products. The strain RD062863, that is available at a public culture collection, was obtained and subjected to metabolite analysis, which resulted in the discovery of a novel cyclopeptide, pseudosporamide (1), along with three new oligomycin-class polyketides, pseudosporamicins A–C (2–4). The unusual structure of compound 1, featured by a biaryl-bond bridging across a tripeptide scaffold, N-acetyl-ʟ-Tyr-ʟ-Pro-ʟ-Trp, was determined by a combination of spectroscopic analyses, chemical derivatization, ECD calculation, and DFT-based theoretical chemical shift calculation, revealing the presence of an (Sa)-axial chirality around the biaryl bond. Compounds 2–4 lacked hydroxylation on the side chain of the spiroacetal rings, which showed clear contrast to other oligomycin congeners and related polyketides with ring-truncation or expansion. The new macrolides 2–4 displayed potent antimicrobial activity against the Gram-positive bacterium Kocuria rhizohpila and the plant pathogenic fungus Glomerella cingulata. All compounds showed moderate cytotoxicity against P388 murine leukemia cells with IC50 values in the micromolar to submicromolar ranges. These results exemplified the validity of phylogeny-focused strain selection combined with biosynthetic gene-directed genome mining for the efficient discovery of new natural products.

The effect of drying treatment to metabolite profile and cytotoxic potential of Rhizophora apiculata leaves

Abstract. Sibero MT, Siswanto AP, Pribadi R, Sabdono A, Radjasa OK, Trianto A, Frederick EH, Wijaya AP, Haryanti D, Triningsih DW, Hayuningrat SJ, Igarashi Y. 2020. The effect of drying treatment to metabolite profile and cytotoxic potential of Rhizophora apiculata leaves. Biodiversitas 21: 2180-2187. Coastal communities in Indonesia have utilized Rhizophora spp. leaves as a traditional medicine for many years. The previous studies have succeeded in extracting bioactive compounds from this mangrove after drying treatment, but there is a possibility of the compound decomposition or breakdown. This study aimed to determine the effect of drying treatment on the metabolite profile of R. apiculata leaves which were taken from mangrove forests in Rembang, Central Java. The effect of pre-drying treatment was examined by comparing the metabolites profiles of fresh, oven-dried, and sun-dried leaves crude extracts. Extraction was carried out using maceration method with agitation (110 r.p.m.) for 24 hours in methanol. The metabolite profile was analyzed using high-performance liquid chromatography (HPLC) with diode array detector (DAD) and thin layer chromatography (TLC), while secondary metabolites were studied by phytochemical test. The phytochemical results showed that there were no differences in metabolites in dried and fresh R. apiculata leaves. Crude extract of fresh and oven-dried gave 10 spots on the TLC, while sun-dried crude extract had 9 spots. The one missing spot (Rf value of 0.79) in the sun-dried crude extract might have unstable compounds that are easily degraded or damaged by the sunlight. Moreover, HPLC chromatogram indicated the pre-drying treatment gave alteration to the R. apiculata metabolites that only detected at 400 nm. Cytotoxic assay against P388 murine leukemia cell indicated that oven-dried treatment gave the best anticancer activity with IC50 value of 0.0323 mg/mL.

Bulbimidazoles A-C, Antimicrobial and Cytotoxic Alkanoyl Imidazoles from a Marine Gammaproteobacterium Microbulbifer Species.

Three new alkanoyl imidazoles, designated bulbimidazoles A-C (1-3), were found from the culture extract of the gammaproteobacterium Microbulbifer sp. DC3-6 isolated from a stony coral of the genus Tubastraea. The absolute configuration of the anteiso-methyl substitution in 1 was established to be a mixture of (R)- and (S)-configurations in a ratio of 9:91 by applying the Ohrui-Akasaka method. Compounds 1-3 displayed unique broad-spectrum antimicrobial activity against Gram-positive and -negative bacteria and fungi with MICs ranging from 0.78 to 12.5 μg/mL. They also exhibited cytotoxicity toward P388 murine leukemia cells with IC50 in the micromolar range.

Flavonoid Derivatives from The Leaves of Muntingiacalabura L.

The Isolation and Purification of ethyl acetate fraction from Muntingiacalabura leaves have resulted three flavonoid derivatives. UV-Vis, 1H-NMR dan13C-NMR data showed that isolate compounds are 2’,4’-dihydroxychalcone (1), 3,7-dimetoxy-5-hydroxyflavone (2), and 3,5,7-trihydroxy-8-methoxyflavone (3). The antioxidant evaluation against 2,2-diphenyl-1-picrylhydrazyl (DPPH), compound (3), showedto be a very high activity (IC503,7 × 109 ppm). The isolated compounds were also evaluated for their cytotoxic activities against murine leukemia P-388 cells resulted that all isolated compounds were inactive.

Kurzichalcolactone A and B, two chalcone derivatives from Indonesian Cryptocarya

Kurzichalcolactone A (1) and B (2), two chalcone derivatives were isolated from two species of Indonesian Cryptocarya: Cryptocarya phoebeopsis and Cryptocarya morotaiense. The structures of the isolated compounds were elucidated based on the analysis of spectroscopic data, including UV, IR, NMR, and mass spectra. On biological evaluation of these compounds for their cytotoxic activity, it was found that (1) exhibited moderate inhibition activities against murine leukemia P388 cells while (2) was inactive.