Synthesis,Crystal Structure and Cytotoxic Activities of Oxazolidin-2-one Derivatives

Four cytotoxic oxazolidin-2-one derivatives were prepared from alkynyl alcohol and isocynate with high yields of 83～95%, and their structures were characterized by IR, H-RESI-MS and NMR analysis. Meanwhile, the crystal of （Z）-4-benzylidene-3-ethyl-1-oxa-3-azaspiro[4.4] nonan-2-one （5a） was obtained and determined by X-ray single-crystal diffraction. Crystal data： monoclinic system, space group P121/c1, a = 10.9284（2）, b = 9.47510（10）, c = 14.2510（2） ？, β = 111.917（2）o, V = 1369.01（3） ？3, Z = 4, F（000） = 552.0, Dc = 1.248 Mg/m3, μ = 0.652 mm-1, R = 0.0473 and wR = 0.1207 for 2699 independent reflections （Rint = 0.0206） and 2581 observed ones （I 〉 2σ（I））.

Synthesis,Crystal Structure and Cytotoxic Activities of 1-(Prop-2-yn-1-yl)-7,8-dihydro-1H-benzo[d][1,3]-thiazine-2,5(4H,6H)-dione Derivatives

The important synthetic precursor（Ⅲ）, 1-（prop-2-yn-1-yl）-7,8-dihydro-1Hbenzo[d][1,3]thiazine-2,5（4H,6H）-dione（C（11）H（11）NO2S）, was prepared through a three-component reaction, which was further transferred into cytotoxic triazoles by alkylation and ＂click＂ synthesis in satisfactory yields of 87%^95%. Their structures were characterized by IR, H-RESI-MS and NMR analysis. Meanwhile, the crystal of Ⅲ was obtained and determined by X-ray single-crystal diffraction. Crystal data： orthorhombic system, space group P212121, a = 5.189（4）, b = 8.661（6）, c = 23.498（17） A, V = 1056.2（13） A^3, Z = 4, F（000） = 464, Dc = 1.392 g/cm^3, μ =0.284 mm^-1, R = 0.0637 and wR = 0.1668 for 8182 independent reflections（R（int） = 0.1580） and 2166 observed ones（I 〉 2σ（I））.

New Secodaphnane-Type Alkaloids with Cytotoxic Activities from Daphniphyllum angustifolium Hutch

One new Daphniphyllum alkaloid,daphnioldhanol A(1),together with three known ones,were isolated from the stem part of Daphniphyllum angustifolium Hutch.Their structures were elucidated by spectroscopic methods and comparing with the literature data.Compound 2 is a new natural product,but known by synthesis as a racemate.Compound 1 exhibited week cytotoxic activity against Hela cell line with IC50 of 31.9μM.

In-vitro Anti-food Poisoning Bacteria and Cytotoxic Activities of Bioactive Compounds Produced by Streptomyces sp.A10 Isolated from Fermented Food Running Title:Anti Bacteria and Cytotoxic Activities of Bioactive Compounds

Streptomyces sp.A10 was isolated from fermented food,PLA-SOM,collected from Nakhon Si Thammarat province,Thailand and identified by means of MALDI Biotyper,designated as Streptomyces violaceoruber.It could produce bioactive compounds(BCs)that showed spectrum of antibacterial activity against representative of Gram-positive bacteria(Staphylococcus aureus TISTR 517 and Bacillus cereus ATCC 11778)and Gram-negative bacteria(Pseudomonas aeruginosa TISTR 1467 and Salmonella typhimurium TISTR 292)by using cross streak method.Moreover,it could excrete the BCs into medium broth(half-formula of Luria Bertani)within 4 days of incubation period at 30°C in shaking incubator 200 rounds per minute(RPM).The excreted BCs from cell-free supernatant(CFS)was determined the protein concentration by means of Bradford assay and investigated the antibacterial activity and cytotoxicity against brine shrimp nauplii by using broth microdilution method and brine shrimp lethality bioassay,respectively.Moreover,cell cytotoxicity activity of CFS was also investigated using MTT assay in HepG2 cells.The results demonstrated that the BCs which had protein concentration equal to 254μg protein/ml showed strong activity against representative of Gram-positive bacteria,moderate activity against representative of Gram-negative bacteria,but Ps.aeruginosa TISTR 1467 is resist to these BCs.The MIC,MBC and LC50 of BCs were in the range of 0.49-63.50,0.49-127.00,and 131.58μg protein/ml,respectively.Moreover,CFS(10-80%)had no cytotoxic effect on HepG2 cells.It implied that BCs may contain the important polypeptide substances that can be used as model for development of novel polypeptide antibiotic or food preservative agents.

Antimicrobial and cytotoxic activities of endophytic fungus Colletotrichum gloeosporioides isolated from endemic tree Cinnamomum malabatrum

In a survey of endophytic fungi associated with endemic plant Cinnamomum malabatrum leaves harbored a bioactive endophytic isolate CMS 3 was identified as Colletotrichum gloeosporioides through morphological and phylogenetic analysis based on ITS-rDNA.The ethyl acetate extract of fermentation broth of Colletotrichum gloeosporioides CMS 3 displayed antimicrobial activity against gram positive and gram negative bacteria as well as the fungal pathogen,Candida albicans.The ethyl acetate crude extract showed in vitro cytotoxicity against the HeLa,MCF-7 and MG63 cancer cell lines with the IC50 values of 94.2μg/ml,84.3μg/ml and 162μg/ml respectively.Gas chromatography and Mass Spectrophotometry(GC-MS)analysis of crude extract confirmed that CMS 3 was a prolific producer of secondary metabolites,in which nearly 74%of the metabolites not listed in the NIST database.Major compounds were phenol 3,5-dimethoxy acetate(11.82%),4'-isopropylidene-bis-(2-cyclohexyl)phenol,N-Didehydrohexacarboxyl-2,4,5-trimethylpiperazine and 1,2,4-Triazolium ylide.These metabolites may be responsible for its antimicrobial and cytotoxic activities.

Distribution and diversity of endophytic fungi in Gentiana rigescens and cytotoxic activities

Objective:In the present study,Gentiana rigescens was screened for fungi communities to clarify their diversity and community assemblage in hosts.Meanwhile,the identification and activity assays of the strains were also conducted.Methods:By culture-dependent(endophytic fungi isolations from plant sections)and culture-independent(metagenomic library and cloning from plant sections)techniques,fungi communities were studied.The metagenomic library was generated using direct DNA isolation of whole plants,plant radixes,plant stems,plant leaves,plant flowers and soils around the plant.Meanwhile,endophytes were isolated from all parts ofG.rigescens plants.After fermentation of the fungi isolations,all the isolates were evaluated for their cytotoxicity against four kinds of human cancer cell lines(HCT116,BEL7404,A549,MDA-MB-231).Results:Eventually,200 strains were isolated and 103 strains were further identified through the internal transcribed spacer(ITS,ITS1 and ITS2 regions)sequence by using the universal primers ITS5 and ITS4.A total of 59,106 fungal sequences corresponding to 374 putative operational taxonomic units(OTU)were identified by 454 pyrosequencing.Through 454 pyrosequencing,the main fungal genera were Sebacina,Botrytis,Mycosphaerella,Boletus and Gibberella,and the major fungal genera which were directly isolated were Aspergillus,Fusarium,Penicillium and Alternaria.Activity assays showed strains 5-26(Aspergillus sp.)and 6-2(Fusarium avenaceum)had the outstanding cytotoxicity to all the tested cell lines with IC5o values<5μg/mL.Conclusion:This study revealed the abundance of endogenetic fungal resources and a variety of genetic information in G.rigescens by high-throughput 454 sequencing technology and fungi isolation methods.Activity assays indicated that endophytes were a promising natural source of potential anticancer agents.

Cytotoxic Activities of Total Saponins from Plena Clematis on Human Tumor Cell Lines In Vitro

Objective： To investigate the anti-proliferative effects of saponins prepared from Plena Clematis （PC） cultured in Fujian Province, China on 4 human tumor cell lines and its possible anti-tumor mechanism. Methods： The growth inhibition assays of saponins on human esophageal squamous carcinoma cell line （EC9706）, human hepatoma cell line （HepG-2）, human oral cancer cell line （KB） and human gastric cancer cell line （BGC-823） were evaluated in vitro by thiazolyl blue （MTT） method. The inhibitory effects on EC9706 treated with different concentrations of saponins （15.62, 31.25, 62.50, 125, 250 and 500 μg/mL） were performed in vitro by MTT method. The morphology and nuclear staining with acridine orange/ethidium bromide of EC9706 calls treated with saponins were illustrated under an inverted phase fluorescence microscope. The apoptotic effects of saponins were further evaluated by annexin-V/propidium iodide dual staining experiment to examine the occurrence of phosphatidylserine externalization onto the call surface by a flow cytometer. Results： MTT assay showed that the saponins could inhibit the proliferation of 4 tumor cell lines. Among them, the maximum inhibition rate of 73.1% was detected in EC9706 cells at the saponins concentration of 250 μg/mL for 24 h. Further investigation indicated that the saponins induced EC9706 cells apoposis. The EC9706 cells presented apoptotic characteristics when treated with saponins, including that the morphologies of EC9706 cells were appeared round-shaped with higher refraction, and the cell nuclear stained orange with EB after 250 μ g/mL saponins exposure. The flow cytometry analysis results showed that the induction of cell cycle arrest in apoptotic system may participate in the anti-proliferative activity of saponins on EC9706 cells. Conclusion： The saponins from PC exhibited significant cytotoxicity against human EC9706, KB, BGC-823, and HepG-2 cells and might be beneficial to development of ethnic pharmaceutical plant for potential anti-tumor drugs.

Synthesis and cytotoxic activities of E-resveratrol derivatives

The present study was designed to synthesize derivatives of E-resveratrol and evaluate their cytotoxic activity in vitro.Different functional groups were conjugated with the phenolic hydroxyl group of E-resveratrol,and the double bond of E-resveratrol was reduced.The in vitro cytotoxicity of the synthetic derivatives was evaluated against three tumor cell lines(A549,LAC,and He La) using the MTT assay.Twenty-six E-resveratrol derivatives were synthesized and their structures were confirmed by 1H NMR,MS,IR,and elemental analyses.Compounds 1-6,12,15-21,and 23-26 were reported for the first time.Among them,Compounds 1,2,4,5,and 9-11,showed significant cytotoxicity against tumor cells;especially,Compound 1 showed an IC50 value of 4.38 μmol·L-1 in the A549 cells which was 15-fold more active than E-resveratrol;Compound 9 showed an IC50 value of 1.41 μmol·L-1 in the He La cell line which was 90-fold more active than E-resveratrol,and close to adriamycin.The structure–activity relationships were also investigated.Compounds 1,2 and 9-11 may serve as potential lead compounds for the discovery of new anticancer drugs.

Synthesis and Characterization of Novel Azo-containing or Azoxy-containing Schiff Bases and Their Antiproliferative and Cytotoxic Activities

A series of novel Schiff base derivatives was designed and synthesized from 3,3＇-azobenzaldehyde and 3,3＇-azoxybenzaldehyde. The newly synthesized compounds were characterized by FTIR, ^1H NMR, MS and elemental analysis and tested for their in vitro antiproliferative activities against HeLa cell lines. At the same time, the impact on the antitumor activity of the sulfanilamido, benzamido, phenolic hydroxyl or thiourea groups was investigated and discussed. Compounds 4, 6 and 10 were found to exhibit strong cytotoxic activity.

Two cardenolide glycosides from the seed fairs of Asclepias curassavica and their cytotoxic activities

Two cardenolide glycosides,corotoxigenin 3-0-[β-D-glucopyranosyl-(l→4)-6-deoxy-β-D-glucopyranoside](1)and coroglaucigenin 3-0-[β-D-glucopyranosyl-(l→4)-6-deoxy-β-D-glucopyranoside](2),were isolated from the seed fairs of Asclepias curassavica.The structures of 1-2 were determined based on the combination of the analysis of their MS,NMR spectroscopic data and acid hydrolysis.The inhibitory effects of compounds 1 and 2 on human colorectal carcinoma cells(HCT116),non-small cell lung carcinoma cells(A549)and hepatic cancer cells(SMMC-7721)were evaluated.The results showed that both compounds 1 and 2 significantly inhibited the viability,proliferation,and migration of A549,HCT116 and SMMC-7721 cells,suggesting that compounds 1 and 2 can be applied in the treatment of lung,colon and liver cancers in clinical practice.This study may not only provide a scientific basis for clarifying the active ingredients in A.curassavica,but also help to understand its antitumor activity,which can promote the application of A.curassavica in clinical treatment of various cancers.

Synthesis and Cytotoxic Activities of Spin-labeled Derivatives of Cinobufagin

Two series of novel spin-labeled derivatives of Cinobufagin（compounds 5 and 8a--8f in series 1 with five-membered ring nitroxyl free radical and compounds 6 and 9a-9f in series 2 with six-membered ring nitroxyl free radical） were synthesized. The cytotoxic activities in vitro against two tumor cell lines（HepG2 and HeLa） were evaluated, and the results indicate that all compounds display potent cytotoxicity against HepG2 and HeLa cells, and most compounds show better activities on HeLa cells than on HepG2 cells except for compounds 8a and 9d. Gene- rally, the compounds in series 2 have more potent cytotoxic activity against HepG2 than the compounds in series 1. Especially, compounds 6 and 9f in series 2 exhibit even more potent activities against the two tumor cell lines than Cinobufagin. Thus incorPoration of different L-amino acids as the linker changed the cytotoxic profile of the spin-labeled Cinobufagin. In addition, the representative compound 9f significantly changed the cell cycle distribution and led to HeLa cell cycle arrested at G2/M phase.

Synthesis, Crystal Structures, and Binding Behaviors with CT-DNA/BSA, Antimicrobial and Cytotoxic Activities of Three New Pyrazine Acylhydrazones

Three new acylhydrazones containing pyrazine ring(C12H11N5O2·CH3COOH,1;C13H13N5O·2CH3COOH,2;C13H13N5O3·C13H13N5O3,3)were synthesized and fully characterized.The single-crystal XRD indicated that both 1 and 2 crystallized in monoclinic,P21/c space group but 3 belonged to monoclinic,C2/c space group.The temperature of the maximum thermal decomposition peaks measured by thermogravimetry for 1~3 is 284,289 and 276℃respectively,all showing better thermal stabilities.The interactions of 1~3 with calf thymus DNA(CT-DNA)and bovine serum albumin(BSA)were studied by UV-Vis absorption spectroscopy and fluorescence spectroscopy,respectively,presenting that 1~3 could bind to CT-DNA via groove binding mode and quench the fluorescence of BSA through static process.Moreover,molecular docking studies of the interactions between 1~3 with DNA/BSA were in good agreement with experimental results.From antimicrobial activities of 1~3 and gentamycin sulfate against Staphylococcus aureus,Escherichia coli and Salmonella typhimurium,it was inferred that 3 had generally stronger antibacterial activity than 1 and 2 and is more active against Staphylococcus aureus than gentamycin sulfate.The cytotoxic tests of 1~3 and etoposide on human lung cancer cells(A549)were carried out by using the MTT method.

Synthesis,Crystal Structures and Cytotoxic Activities of Two New Pyrimidine Derivatives of Ursolic Acid

The title compounds(5a and 5b)were synthesized from ursolic acid and their structures were characterized by spectroscopic methods including ESI-MS,^(1)H-NMR,^(13)C-NMR and elemental analysis.The crystal structures of compounds 5a and 5b were determined by single-crystal X-ray diffraction analysis.Compound 5a crystallizes in monoclinic system,P21 space group with a=12.258(3),b=10.396(2),c=15.570(3)Å,b=107.21(3)°,Z=2,V=1895.3(7)Å^(3),Mr=659.90,Dc=1.156 Mg/m^(3),S=1.003,µ=0.076 mm^(−1),F(000)=716,the final R=0.0686 and wR=0.1430 for 1859 observed reflections(I>2σ(I)).Compound 5b crystallizes in monoclinic system,P21 space group with a=12.371(3),b=10.647(2),c=15.722(3)Å,b=109.44(3)°,Z=2,V=1952.8(8)Å^(3),M_(r)=655.93,D_(c)=1.116 Mg/m^(3),S=1.002,µ=0.069 mm^(−1),F(000)=716,the final R=0.0686 and wR=0.1882 for 2574 observed reflections(I>2σ(I)).The preliminary cytotoxic assay indicated that compound 5b exhibited notable cytotoxic activity against MCF-7 and HeLa cells with the IC_(50)values of 10.71±0.23 and 12.63±0.31μM,respectively.

Antioxidant and Cytotoxic Activities of A Novel Isomeric Molecule(PF5)Obtained from Methanolic Extract of Pleurotus Florida Mushroom

The Pleurotus florida is recognized as a medicinal and edible mushroom and the present study intends to reveal the active isomeric molecules from this mushroom.The P.florida was cultivated using different nutrient supplements:groundnut husk,maize powder,horse gram powder and coconut oil-cake powder.Horse gram supplement showed the higher mushroom yield and henceit was used for the cultivation of P.florida.Methanolic extract of P.florida was found to be efficient in antioxidant activity among ethanol,aqueous,ethyl acetate and hexane extracts.The bioactive fraction 3-methoxy-4-hydroxy cinnamic acid(PF5)was isolated and purified from the methanolic extract of P.florida by column chromatography,thin layer chromatography(TLC)and gas chromatography-mass spectrum(GC-MS)and further it was characterized by Nuclear magnetic resonance(NMR).The PF5 was tested for its DPPH and reducing power assays,and the IC_(50) values were found to be 21.7μg/mL and 105μg/mL,respectively.We found that the cytotoxic effect of 3-methoxy-4-hydroxy cinnamic acid was tested against the lung cancer cell line using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT),sulphorodamine B(SRB)and trypan blue assays which exhibited a higher cytotoxic effect(CTC_(50),645μg/mL).These results suggested that 3-methoxy-4-hydroxy cinnamic acid from P.florida could be explored as a novel and potent natural antioxidant and cancer preventive agent,alternative to existing synthetic molecules.

Cytotoxic triterpenoid saponins from Ardisia pusilla

A new triterpenoid saponin, 3-O-{ β-D-xylopyranosyl-（1 → 2）-β-D-glucopyranosyl-（1 → 4）-[β-D-glucopyranosyl-（1 → 2）]-α- L-arabinopyranosyl}-3β,16α,28α-trihydroxy-13β,28-epoxy-oleanan-30-al （ardipusilloside Ⅲ, 1）, together with two known saponins, ardisiacrispins A （2） and B （3）, were isolated from the whole plants of Ardisia pusilla A. DC. Their structures were elucidated by extensive spectral analysis and chemical evidences. Saponins 1 and 3 exhibited significant cytotoxicity against human glioblastoma U251MG cells.

Synthesis, Crystal Structure and Cytotoxic Activity of a Novel Nickel(II) Complex with Schiff Base Derived from Salicylhydrazide

A novel Schiff base complex with π-conjugated system, [Ni（L1）2（py）2] 1 （L1 =（E）-N′-（2,4-dichlorobenzylidene）-2-oxidobenzohydraizide）, was synthesized and characterized by elemental analysis and single-crystal X-ray determination. Complex 1 crystallizes in the monoclinic system, space group P21/n with a = 12.8286（10）, b = 16.3573（13）, c = 19.0206（14） A, β = 108.2920（10）°, V = 3789.6（5）A^3, Z = 4, Mr = 833.17, Dc = 1.460 g/cm^3,/t = 0.843 mm^-1, F（000） = 1704, the final R = 0.0537 and wR = 0.0640 for 3836 observed reflections with I 〉 2σ（I）. In the molecular structure of 1, the Ni^Ⅱ atoms are six-coordinated by two N and two O atoms from two Schiff base ligands （LI） and two N atoms from two pyridine solvent molecules to form a distorted octahedral geometry. The cytotoxic activities of complex 1 have been experimentally studied against a human HeLa cell in vitro.

Antioxidant and Cytotoxic Phenolic Compounds of Areca Nut(Areca catechu)

Areca catechu L.（Palmae）, commonly known as an important economical seed crop, is widely culti- vated in tropical and subtropical areas, including India, Southeast Asia, East Africa and New Guinea. Areca nut（frequently known as betel nut） is the ripe fruit of the tree A. catechu. Areca nut can be chewed and it is a common masticatory in tropical and subtropical countries. It was estimated in the early 1990s that 10% to 20% of the world＇s population chewed betel quid daily. Areca nut is commonly used in folklore medicine for treatment of various diseases such as dyspep sia, constipation, beriberi and oedema.

Diterpenoid alkaloids from a Tibetan medicinal plant Aconitum richardsonianum var. pseudosessiliflorum and their cytotoxic activity

The chemical constituents from Aconitum richardsonianum var.pseudosessiliflorum were investigated.The roots of this plant were extracted three times with 90% EtOH at the room temperature.The ethanol extracts were combined and concentrated under reduced pressure to yield residue,which was suspended in water and successively partitioned with chloroform.The chloroform extraction was isolated and purified by silica gel and Sephadex LH-20 column chromatography.Six compounds were isolated and elucidated as delelatine（1）,isodelpheline（2）,3-acetylaconitine（3）,isoatisine（4）,nordhagenine A（5）and yunaconitine（6）.Compounds 1-5 were obtained from Aconitum Brunneum for the first time.Compound（1）showed significant cytotoxic activities（IC50=4.36 μM）against the human tumor cell line P388.

Cytotoxic Cycloartane Triterpenoid Saponins from the Rhizomes of Cimicifuga foetida

To enrich the bioactive cycloartane triterpenoid glycoside named actein and find out more cytotoxic cycloartane triterpenes,a phytochemical study of Cimicifuga foetida was conducted.113 g(0.17%)actein was purified by recrystallization while eight cycloartane-type triterpenes(1-8)were isolated from the mother liquid.The chemical structures of new compounds(1-4)were elucidated by 1D and 2D NMR and HRESIMS spectroscopic analyses.Moreover,new compounds showed moderate and broad-spectrum cytotoxicity against 5 human cancer cell lines with IC_(50) values ranging from 4.02 to 15.80μM.

A Cytotoxic Saponin with Two Monoterpenoids from Albizia julibrissin

A new cytotoxic saponin(1). Julibrosides J(27), was isolated from the stem barks of Alibizia. julibrissin by chromatography, and the structure was elucidated as 3-O-beta-D-xylopyranosyl-(1-->2)-beta -D- fucopyranosyl - (1-->6) -beta -D-glucopyranosyl - 21-O-[(6S)-2 -trans-2-hydroxymethyl-6-methyl-6-O- [4-O-((6S)-2-trans-2-hydroxylmethy 6- methyl - 6- hydroxy)-2,7-octadienoyl-beta-D-quinovopy- -ranosyl]-2.7-octadienoyl}- acacic acid- 28 -O-beta-D-glucopranosyl-(1-->3)-[(alpha-L-arabinofuranosyl-(1-->4)]-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyrnosyl ester based on spectral and chemical evidences.