P388 Murine Leukemia Cells Research Articles

Herbidomicins, two pairs of polyketide tautomers produced by an actinomycete of the genus Herbidospora.

Herbidospora is one of the underexplored actinomycete genera from which only a limited number of secondary metabolites are reported. In our continuing investigation on less explored actinomycetes, a liquid culture of Herbidospora sp. RD 11066 was found to contain unknown metabolites that had no match in our in-house UV database. Chromatographic separation and following structural analysis using NMR and MS identified these metabolites to be chromanone and chromene derivatives, which were respectively composed of an inseparable mixture of two isomeric forms. The former polyketides, designated to be herbidomicins A1 (1) and A2 (2), are positional isomers in terms of a methyl substituent on an aromatic ring that mutually interconvert by acetal exchange by two phenolic hydroxy groups. The latter pair, herbidomicins B1 (3) and B2 (4), is Z/E-isomers regarding an enol ether double bond. Herbidomicins 1-4 were weakly antifungal against a dermatophytic fungus Trichophyton rubrum and were moderately cytotoxic against murine leukemia P388 cells.

Wychimicins E and F from a rare actinomycete of the genus Cryptosporangium.

Two new spirotetronate class compounds designated wychimicins E (1) and F (2) were isolated from the culture extract of an actinomycete Cryptosporangium sp. RD061707. Their structures were determined through extensive NMR analysis in comparison with wychimicin C. Both compounds lack one hydroxy group in the decalin moiety, and the relative configuration of the remaining hydroxy group was assigned by NMR analysis of triacetylwychimicin E (3). Compounds 1 and 2 showed potent antimicrobial activity against Kocuria rhizophila and Staphylococcus aureus. These compounds were also modestly cytotoxic against P388 murine leukemia cells.

Sporangimicins A-D, acylated maltose derivatives from a rare actinomycete of the genus Pseudosporangium.

Sporangimicins A-D (1-4), four anomeric pairs of diacyl disaccharides that represent a new metabolite class, were discovered from the culture extract of an actinomycete Pseudosporangium sp. RD061809. Compounds 1-4 caused peak separation in the HPLC chromatogram and partial duplication of the NMR resonances by anomeric interconversion of a maltose core modified at the two sugar 6-positions with an isobutanoyl and a methyl-branched long-chain dienoyl groups. A highlight of the structure elucidation was application of Ohrui-Akasaka's method to a chromatographically inseparable mixture of 3 and 4, which proved the composition ratio of 3 and 4 to be 82:18 and the R/S ratio at the anteiso-methyl bearing chiral center in 3 to be 66:34. Compounds 1-4 showed antimicrobial activity against Gram-positive bacteria and modest cytotoxicity toward P388 murine leukemia cells.

Cladoic Acid, an Anti-Trypanosoma cruzi Polyketide from Cladosporium sp. TP-F2020.

A new polyketide, cladoic acid, was isolated from a fungus of the genus Cladosporium. The structure of the highly oxygenated trans-decalin ring with an all-E triene side chain was elucidated by extensive spectroscopic analysis. The unique chair/twist-boat conformation of the trans-decalin core and the flexibility of the B-ring were demonstrated by computer-aided conformational analysis. Cladoic acid was active against Trypanosoma cruzi and inhibited the proliferation of amastigotes and epimastigotes with IC50 values of 27 and 46 μM, respectively, but it did not show any appreciable activity against P388 murine leukemia cells, bacteria, or fungi, indicating it is a potential candidate for drug development against Chagas disease.

Structure revision of tricholomenyn B, an antimitotic geranylcyclohexenone rediscovered as a bitter and antibacterial substance, from a basidiomycete Tricholoma japonicum (Shiro-shimeji).

Tricholomenyn B, an antimitotic geranylcyclohexenone originally discovered from a basidiomycete Tricholoma acerbum, was isolated as a bitter and antibacterial constituent from fruiting bodies of T. japonicum. Careful comparison of NMR, MS, and other physicochemical properties of the isolated substance with the literature values revised a previously proposed macrolide structure 1 to a macrodiolide 2. Compound 2 was perceived bitter at a minimum dose of 37.5 μg, showed weak antimicrobial activity against Kocuria rhizophila and Staphylococcus aureus, and was marginally cytotoxic (IC50 2.6 µM) against P388 murine leukemia cells.

Ciliatonoids D–M, highly modified limonoids from the twigs of Toona ciliata

A new ring-intact limonoid, ciliatonoid D (1), five new 7,8-seco limonoids, ciliatonoids E–I (2–6), three new 7,8-seco-29-nor limonoids, ciliatonoids J–L (7–9), and a new 7,8:9,11-diseco limonoid, ciliatonoid M (10), together with eleven known limonoids were isolated from the twigs of Toona ciliata. Their structures were established by comprehensive analyses of spectroscopic, electronic circular dichroism, single-crystal X-ray diffraction data. Compound 6 bears an uncommon 1,30-oxygen bridge. Compound 10 represents a rare example of a structure with 9,11-seco limonoid from Meliaceae. The orientation of the 14,15-epoxy moiety in a known limonoid, toonayunnanae C (12), was revised as in a β-orientation by quantum NMR calculations with ML-J-DP4 probability analysis. Compound 9 and a known analogue, toonanoronoid C (11), were found to show modest in vitro cytotoxicity against HL-60 (human premyelocytic leukemia) and P-388 (murine leukemia) cell lines.

Morushalunin D, a tri-O-bridged Diels-Alder type adduct from Morus alba var. shalun root cultures and the related precursors

Three Diels-Alder type adducts (1-3) along with their precursors, including one 2-arylbenzofuran (4) and one stilbene (5), were isolated from the MeOH extract of M. alba var. shalun root cultures. Among them, 1 is a new Diels-Alder type adduct named morushalunin D. The molecular structures of 1-5 were elucidated based on spectroscopic data and comparison with the literatures. Cytotoxic properties of compounds 1-5 were evaluated against murine leukemia P-388 cells. Morushalunin D (1), mulberrofuran T (2), sorocein A (3), moracin M (4), and oxyresveratrol (5) were active, significantly inhibiting the growth of P-388 cells with IC50 values of 0.5, 1.0, 0.6, 2.0, and 3.3 μg/ml, respectively.

Botryorhodines K and L, two new cytotoxic depsidones from a fungus of the genus Arcopilus.

Botryorhodines K (1) and L (2), two new depsidone derivatives, along with one known metabolite, 4-O-demethylbarbatic acid (3), were isolated from the culture extract of a fungus of the genus Arcopilus. The structures of 1‒3 were determined by the analysis of NMR and MS spectral data and the absolute configuration of 1 was established by single-crystal X-ray diffraction analysis. Compounds 1 and 2 showed antimicrobial activity against Gram-positive bacteria and cytotoxicity against murine leukemia P388 cells.

Nortopsentins as Leads from Marine Organisms for Anticancer and Anti-Inflammatory Agent Development.

The marine environment is an excellent source of molecules that have a wide structural diversity and a variety of biological activities. Many marine natural products (MNPs) have been established as leads for anticancer drug discovery. Most of these compounds are alkaloids, including several chemical subclasses. In this review, we focus on the bis-indolyl alkaloid Nortopsentins and their derivatives with antiproliferative properties. Nortopsentins A-C were found to exhibit in vitro cytotoxicity against the P388 murine leukaemia cell line. Their structural manipulation provided a wide range of derivatives with significant anti-tumour activity against human cell lines derived from different cancer types (bladder, colon, gastric, CNS, liver, lung, breast, melanoma, ovarian, pancreatic, prostate, pleural mesothelioma, renal, sarcoma, and uterus). In vivo assays on animal models also proved that Nortopsentins and related bis-indolyl compounds have potent anti-inflammatory activity. These remarks set the foundation for future investigations into the development of new Nortopsentin derivatives as new anticancer and anti-inflammatory agents.

Iseoic acids and bisiseoate: three new naphthohydroquinone/naphthoquinone-class metabolites from a coral-derived Streptomyces.

Two new naphthohydroquinone derivatives designated iseoic acids A (1) and B (2) and a new symmetrical glycerol bisester of naphthoquinonepropanoic acid designated bisiseoate (3) were isolated from the culture extract of a marine-derived actinomycete Streptomyces sp. DC4-5. The structures of 1-3 were determined by analyzing one- and two-dimensional NMR data and MS analytical data. The absolute configurations were determined by NOESY analysis and the phenylglycine methyl ester (PGME) method for 1 and by considering the structural similarity and biosynthesis for 2 and 3. Compound 3 exhibited modest cytotoxicity against P388 murine leukemia cells with an IC50 value of 19 μM.

Coumarin Derivative from the Unripe Fruit of Mengkudu (Morinda citrifolia Linn) and Cytotoxic Activity

A coumarin derivative namely scopoletin (7-hydroxy-6-methoxychromen-2-one)had been isolated from the methanol extract of unripe noni fruits (Morinda citrifolia Linn.). The extraction was conducted by maceration, continuing with separation and purification using several techniques of chromatography. The yellowish isolated compound showed a melting point at 200 – 202 C. The compound structure was analyzed based on spectral data 1H-NMR, 13C-NMR, and NOE 1D. The inhibition of methanol extract of noni fruits and scopoletin against murine leukemia P388 cells using MTT assay showed IC50 were > 100 μg/mL and 65.69 μg/mL respectively. These showed that scopoletin was more cytotoxic than methanol extract but weaker than artonin E as a positive control IC50 0.48 μg/mL).

Anticancer Activity Study of Modified Artocarpin Compound from Pudau Plant (Artocarpus kemando Miq.)

This research is a continuation of the successful isolation of artocarpin from the root of Artocarpus kemando Miq reported in our previous study. In the previous study, the artocarpin was characterized with UV-Vis and FTIR techniques. In this follow-up investigation, the artocarpin was subjected to a transesterification reaction using acetic anhydride and pyridine as catalysts, and the product of the reaction was specified as compound 1. The compound 1 was further characterized with different techniques to gain more complete data and then tested for anticancer activity test against P-388 murine leukemia cells. Characterizations of the compound 1 using 1H-NMR and 13C-NMR techniques suggest that the modification reaction resulted in the conversion of the -OH groups at C2' and 4' at the artocarpin molecule to -OOCH3, and based on the MS analysis, the compound was proposed to have the molecular formula of C30H32O8. Another important feature of compound 1 that should be noted is the significant improvement in stability compared to the unmodified artocarpin. Anticancer activity tests against P-388 murine leukemia cells revealed that compound 1 has an IC50of 2.35 g/mL, confirming that the compound is categorized as an active anticancer agent and suggesting that the compound has promising potential that deserves further investigations. Doi: 10.28991/ESJ-2023-07-03-05 Full Text: PDF

Species-specific secondary metabolism by actinomycetes of the genus Phytohabitans and discovery of new pyranonaphthoquinones and isatin derivatives.

To further exploit secondary metabolic potential of a minor actinomycete genus Phytohabitans within the family Micromonosporaceae, metabolite profiling by HPLC-UV analysis, combined with 16S rDNA sequence-based phylotyping were attempted on seven Phytohabitans strains available at the public culture collection. The strains were grouped into three clades and each exhibited unique and distinct metabolite profiles, which were highly conserved among strains within the same clade. These results were consistent with previous observations on two other actinomycetes genera, reconfirming species-specificity of secondary metabolite production, which were conventionally thought to be strain-specific. A strain RD003215, belonging to the P. suffuscus clade, produced multiple metabolites, some of which were presumed to be naphthoquinones. Liquid fermentation followed by chromatographic separation of the broth extract led to the discovery of three new pyranonaphthoquinones, designated habipyranoquinones A-C (1-3), and one new isatin derivative, (R)-N-methyl-3-hydroxy-5,6-dimethoxyoxindole (4), along with three known synthetic compounds, 6,8-dihydroxydehydro-α-lapachone (5), N-methyl-5,6-dimethoxyisatin (6), and 5,6-dimethoxyisatin (7). Structures of 1-4 were unequivocally elucidated by NMR, MS, and CD spectral analysis, with assistance of density functional theory-based NMR chemical shift prediction and ECD spectral calculation. Compound 2 displayed antibacterial activity against Kocuria rhizophila and Staphylococcus aureus with MIC 50 µg/mL and cytotoxicity against P388 murine leukemia cells with an IC50 value of 34 µM. Compounds 1 and 4 also showed cytotoxicity against P388 cells with IC50 values of 29 and 14 µM, respectively.

Bulbiferamide, an Antitrypanosomal Hexapeptide Cyclized via an N-Acylindole Linkage from a Marine Obligate Microbulbifer.

UV absorption spectroscopy-guided fractionation of the culture extract of a marine obligate bacterium of the genus Microbulbifer yielded a novel cyclic hexapeptide, bulbiferamide (1). NMR spectroscopic and mass spectrometric analyses revealed the structure of 1 to be a cyclic tetrapeptide appending a ureido-bridged two amino acid unit. Notably, Trp is a junction residue, forming on one hand a very rare N-aminoacylated indole linkage for cyclization and on the other hand connecting the ureido-containing tail structure, which is an unprecedented way of configuring peptides. The component amino acids were determined to be l by the advanced Marfey's method. Compound 1 displayed growth inhibitory activity against Trypanosoma cruzi epimastigotes with an IC50 value of 4.1 μM, comparable to the currently approved drug benznidazole, while it was not cytotoxic to P388 murine leukemia cells at 100 μM.

Ethnomedicinal bioprospecting of Rhizophora apiculata leaves through in silico and in vitro approaches as antioxidant, ?-glucosidase inhibitor and anticancer

Abstract. Sibero MT, Pribadi R, Ambariyanto A, Haryanti D, Kharisma VD, Dewi AS, Patantis G, Zilda DS, Murwani R. 2022. Ethnomedicinal bioprospecting of Rhizophora apiculata leaves through in silico and in vitro approaches as antioxidant, a-glucosidase inhibitor and anticancer. Biodiversitas 23: 6437-6447. Investigating marine natural products for biopharmaceutical development leads to a massive study of mangrove metabolites. Rhizophora apiculata is utilized as a traditional medicine by the local community in Indonesia. However, only a few studies reported the lead compounds. The aim of this study was to discover the biological properties of R. apiculata metabolites from the leaves as an antioxidant, a-glucosidase inhibitor, and anticancer agent through in vitro and in silico approaches. The leaves of R. apiculata were extracted and then fractionated using the silica-OCC method. All fractions were screened for antioxidant, a-glucosidase inhibitors, and cytotoxicity assays. Then the bioactive compounds in prospective fractions were identified using LC-HRMS. The selected compounds with ppm error <10 were applied for in silico analysis. The result of biological properties screening indicated Fr. 5 and Fr. 6 as the most potent sources of antioxidants, a-glucosidase inhibitors, and cytotoxic compounds. In total, 19 compounds were selected from two prospective fractions. The drug-likeness and bioavailability prediction results indicated that all selected compounds act as drug-like molecules. In addition, 17 were predicted to have antioxidant activity, and 15 compounds had antidiabetic activity. Moreover, 15 compounds had a more negative affinity binding than cytarabine. Molecular docking analysis showed that the mechanism of cytotoxicity against P388 Murine Leukaemia Cells was through the interaction of apigenin with protein tyrosine kinase (c-Kit) with a stronger inhibitory activity than the control drug (Cytarabine) and had the same binding site as the control (Cytarabine).

Krasilnikolides A and B and Detalosylkrasilnikolide A, Cytotoxic 20-Membered Macrolides from the Genus Krasilnikovia: Assignment of Anomeric Configuration by J-Based Configuration Analysis.

A chemical investigation of strain RD003821, belonging to the underexplored actinomycetes genus Krasilnikovia, led to the discovery of three novel polyketides: two 20-membered glycomacrolides, krasilnikolides A (1) and B (2), and an aglycone of 1, detalosylkrasilnikolide A (3). A major challenge in the structure elucidation of 1 was to determine the anomeric configuration of the α-l-6-deoxytalose (6dTal) unit, which was achieved by J-based configuration analysis (JBCA) that incorporated anomeric carbon- and proton-specific two-bond 13C-1H spin-spin coupling constants as diagnostic parameters. The updated criteria for the conformation/configuration assignment facilitated discrimination of three out of four stereochemical variants at the anomeric and the adjacent C2 positions, which expanded the scope of the JBCA method to determination of the anomeric configuration of aldohexopyranoses. Compounds 1 and 2 are the first macrolides decorated by 6dTal. Compounds 1-3 exhibited cytotoxicity against P388 murine leukemia cells with IC50 values of 14, 8.4, and 3.9 μM, respectively. In addition, 1-3 were antibacterial against the Gram-positive bacterium Kocuria rhizophila with MIC values of 25, 50, and 100 μg/mL. 1 was inhibitory against Staphylococcus aureus with an MIC of 50 μg/mL.

Cyclic enaminones and a 4-quinazolinone from an unidentified actinomycete of the family Micromonosporaceae

Four novel cyclic enaminones, designated RD4123A-D (1-4), and a new 4-quinazolinone metabolite, RD4123E (5), were isolated from the culture extract of an unidentified actinomycete strain RD004123, which belongs to the family Micromonosporaceae. Structures of 1-5 were determined by spectroscopic analyses using NMR, MS, and electronic circular dichroism (ECD), combined with quantum chemical calculations of ECD and NMR chemical shifts and biosynthetic consideration. Compounds 1-5 showed weak to modest cytotoxicity against murine leukemia P388 cells, while being inactive against bacteria and fungi.

Derivatization of Inseparable Ursolic and Oleanolic Acids of Fagraea fragrans Fruits to Enhance Their Anticancer Activity

Inseparable Ursolic acid and its isomeric oleanolic acid are the major compounds in Fagraea fragrans fruits. The white solid crystals , 3.1 % of these triterpenic acids are easyly isolated from alcoholic extracts of these dried fruits. They are well known in both cosmeticeutical and medicinal industries. Therefore the objective of this work is to derivative those to be the inseparable ethyl ursolate 3-ethyl ether [its isomeric ethyl oleanolate 3-ethyl ether], and to evaluate their anticancer activity against P-388 murine leukemia cells. The results are 71% of the derivatives have been successfully made from the inseparable ursolic [its isomeric oleanolic] acids by in situ reaction between those triterpenic acids with thionyl chloride and ethanol in benzene. The anticancer activity of ethyl ursolate 3-ethyl ether [its isomeric ethyl oleanolate 3-ethyl ether] against P-388 murine leukemia cells with IC50 value of 31.36 µg/mL is twofold (1.7) more potent then their mother compounds, the inseparable ursolic [its isomeric oleanolic] acids with IC50 value of 53.5 µg/mL.

Marinoquinolones and Marinobactoic Acid: Antimicrobial and Cytotoxic ortho-Dialkylbenzene-Class Metabolites Produced by a Marine Obligate Gammaproteobacterium of the Genus Marinobacterium

Chemical investigation of the culture extract of a marine obligate proteobacterium, Marinobacterium sp. C17-8, isolated from scleractinian coral Euphyllia sp., led to the discovery of three new o-dialkylbenzene-class metabolites, designated marinoquinolones A (1) and B (2) and marinobactoic acid (3). Spectroscopic analysis using MS and NMR revealed the structures of 1 and 2 to be 4-quinolones with an o-dialkylbenzene-containing side chain at C3 and 3 to be a fatty acid bearing an o-dialkylbenzene substructure. The 4-quinolone form of 1 and 2 was unequivocally determined by comparison of the 1H, 13C, and 15N chemical shifts of 1 with those predicted for 2-methyl-4-quinolone A and its tautomer 2-methyl-4-quinolinol B by quantum chemical calculation. Compound 1 was proven to be racemic by X-ray crystallographic analysis and chiral-phase HPLC analysis of its chemical degradation product. Compounds 1-3 exhibited antimicrobial activity against bacteria and filamentous fungi at MIC of 6.3-50 μg/mL. In addition, all compounds showed cytotoxicity against P388 murine leukemia cells at micromolar ranges.

The Extraction of Curcuminoids From Ethanol Extract of Yellow Turmeric (Curcuma longa L) and Activity Test on P-388 Murine Leukemia Cells

Abstract: Research on the Extraction of Curcuminoids from Ethanol Extract of Yellow Turmeric (Curcuma longa L) and its Activity Test on P-388 Murine Leukemia Cells has been carried out. This study aims to determine the per cent yield, solvent concentration, activity of P-388 murine leukemia cells and pH of the number of curcuminoids produced in ethanol extract. The curcuminoids were obtained by the extraction process through the soxhlet method using 70 and 96% ethanol solvent with a ratio of sample to solvent of 1:8 with variations in operating time for 3, 6, 9 and 12 hours. The absorbance of curcuminoid extract was measured to determine the concentration of curcumin. The cytotoxic test on murine leukemia P-388 cells used the Microculture Tetrazolium Technique (MTT) method. The results obtained were yellow turmeric curcuminoid compounds using 96% ethanol solvent at an extraction operating time of 9 hours. The largest extract yield was 21.15% with a concentration of 1121.10 ppm, which has activity against murine leukemia P-388 cells with an IC50 of 6.15 g/mL have the potential as anticancer leukemia, and dye analysis at acidic pH (2,4,6) is yellow, while alkaline pH (9,12) is brownish red as a natural food coloring.Abstrak: Penelitian ekstraksi kurkuminoid dari ekstrak etanol kunyit kuning (Curcuma longa L) dan uji aktivitasnya terhadap sel murine leukimia P-388 telah dilakukan. Penelitian ini bertujuan untuk mengetahui rendemen, konsentrasi pelarut, aktivitas sel murine leukemia P-388 dan pH terhadap jumlah kurkuminoid yang dihasilkan pada ekstrak etanol. Kurkuminoid diperoleh dengan proses ekstraksi dengan metode sokletasi menggunakan pelarut etanol 70 dan 96% dengan perbandingan sampel dan pelarut 1:8 variasi waktu operasi selama 3, 6, 9 dan 12 jam. Ekstrak kurkuminoid diukur absorbansi untuk mengetahui konsentrasi kurkumin. Pengujian sitotoksik terhadap sel murine leukimia P-388 dengan metode Microculture Tetrazolium Technique (MTT). Hasil yang diperoleh senyawa kurkuminoid kunyit kuning menggunakan pelarut etanol 96% pada waktu operasi ekstraksi 9 jam rendemen ekstrak terbesar 21,15% dengan konsentrasi 1121,10 ppm, memiliki aktivitas terhadap sel murine leukimia P-388 dengan IC50 sebesar 6,15 μg/mL berpotensi sebagai antikanker leukemia, dan analisis zat warna pada pH asam (2,4,6) berwarna kuning, pada pH basa (9,12 ) berwarna merah kecoklatan sebagai pewarna alami makananan.