Purification and Characterization of Cytotoxins from Agkistrodon acutus Venom and Their Anticancer Activity

Aim To investigate the anticancer activity of two new cytotoxins from thevenom of Agkistrodon acutus. Methods The venom was isolated by FPLC column chromatography consistingof DEAE Sepharose FF and Source 30S. The cytotoxic activity on tumor cells was detected by MITmethod. Purity and molecular weight were determined by SDS-PAGE (silver staining). Their stabilitiesto temperature and pH were also detected. Results Two pure cytotoxins named ACTX-6 and ACTX-8 wereobtained. Their molecular weights are 98 kDa and 27 kDa, respectively. ACTX-6 consists of twosubunits bonded together by disulfide bonds. Conclusion ACTX-6 and ATCX-8 have highest inhibitoryactivity on lung cancer cell A549. ACTX-6 is stable to heat while ACTX-8 not. ACTX-6 is stablebetween pH 7-9 and ACTX-8 between pH 6 - 9.

Total synthesis and anticancer activity studies of the stereoisomers of asperphenamate and patriscabratine

All stereoisomers of asperphenamate 1a and patriscabratine 2a were achieved with a high yield,and total synthesis of 2a is firstly described here.The absolute configuration of patriscabratine was determined as(S,S).The compounds 1a-d and 2a-d have been tested by MTT assay in T47D,MDA-MB231,HL60,Hela and SGC-7901 cell lines in vitro.Among them,the(R,S) stereoisomer shows the strongest anticancer effects,while the(S,R) shows the weakest one.

Sulforaphene in Raphanus sativus L. var. caudatus Alef increased in late-bolting stage as well as anticancer activity

Objectives: To evaluate the concentration differences of sulforaphene and sulforaphane at various ages and in different parts of Raphanus sativus L. var. caudatus with respect to their potential cancer preventive effect on HCT116 colon cancer cells.Methods: FTIR–ATR and GC–MS were used to characterize the isothiocyanates in the plant extracts followed by HPLC for quantification. Antiproliferation and apoptosis induction were determined by using MTT assay and flow cytometry, respectively.Results: The respective rank of anticancer activity of Raphanus sativus were as follows:vegetative(3 week) < older rosette(4 week) < early-bolting(5 week) < senescence(7 week) < late-bolting(6 week). The low to high concentration of sulforaphene and sulforaphane occurred in the same stage order.Conclusions: The reproductive parts(flower, pod, and dry seed) of Raphanus sativus have the greatest isothiocyanate concentration, evidenced by a sulforaphene concentration higher than the sulforaphane. This result should inform the selection of the most appropriate harvesting stage and plant part for use as a potential chemopreventive agent.

Comparative Molecular Field Analysis(CoMFA) of Curcumin-related Compounds for Anticancer Activity

Three-dimensional （3D） quantitative structure-activity relationship （QSAR） studies of 44 curcumin-related compounds have been carried out based on our previously reported result for their anticancer activity against pancreas cancer Panc-I cells and colon cancer HT-29 cells. The established 3D-QSAR models from the comparative molecular field analysis （CoMFA） in training set showed not only significant statistical quality, but also satisfying predictive ability, with high correlation coefficient values （R12= 0.911, R22= 0.985） and cross-validation coefficient values （q2= 0.580, q22= 0.722）. Based on the CoMFA contour maps, some key structural factors responsible for anticancer activity of these series of compounds were revealed. The results provide some useful theoretical references for understanding the mechanism of action, designing new curcumin-related compounds with anticancer activity and predicting their activities prior to synthesis.

Anticancer activity of crude acetone and water extracts of Tulbaghia violacea on human oral cancer cells

Objective: To evaluate the anticancer activity of crude acetone and water leaf extracts of Tulbaghia violacea on a human oral cancer cell line(KB).Methods: The antioxidant activity of the leaf extracts was evaluated by using the DPPH assay while the anti-proliferative activity was assessed by using the MTT assay.The morphological characteristics of apoptotic cells were examined by using the dual acridine orange/ethidium bromide staining.Flow cytometry was used to evaluate the induction of multi-caspase activity and changes in the cell cycle.Results: The acetone and water extracts exhibited antioxidant activity in a concentration dependent manner.The extracts inhibited the growth of the KB cell line with IC_(50) values of 0.2 mg/mL and 1 mg/mL, respectively for acetone and water.Morphological changes such as cell shrinkage, rounding and formation of membrane blebs were observed in the treated cells.In acridine orange/ethidium bromide staining, the number of apoptotic cells increased as the concentration of the extracts increased.The activation of multi-caspase activity in KB cells treated with Tulbaghia violacea extracts was concentration dependent, leading to cell death by apoptosis and cell cycle arrest at the G_2/M phase.Conclusions: The acetone and water extracts of Tulbaghia violacea appear to have anti-cancer activity against human oral cancer cells and need to be investigated further.

Potential anticancer activity analysis of piscidin 5-like from Larimichthys crocea

Antitumor activity is one characteristic function of some certain antimicrobial peptides(AMPs)found in recent years.In the present study,we attempted to detect potential anticancer activity of a recombinant piscidin 5-like from Larimichthys crocea(r Lc-P5L)which owned widely antibacterial and strong antiparasitic activity in vitro.The light microscope observation indicated r Lc-P5L was of antitumor activity to He La cells,293 T cells and L929 cells.MTT assay showed the toxic sensitivity of r Lc-P5L to three tumor cell strains was 293 T>L929>He La.Scanning electron microscope(SEM)results showed r Lc-P5L behaved like a lytic peptide to cause damage on cells membrane of L929 cells by forming globular clusters,even pores at 60μmol/L,or degrading membrane to make it completely lose cytoskeleton structure at 80μmol/L;r Lc-P5L treatment also resulted in DNA degradation.Fluorescence observation results indicated r Lc-P5L could cause L929 cells at least two obvious changes:one is nucleus,nuclear chromatin condensed in the margin,nuclear volume became smaller and shrank to be out of shape,or lysed to be debris;the other is cytoskeleton,they became disordered and polarized to make cells atrophic shapes,or even lysed to be debris.In summary,r Lc-P5L owned potential anticancer activity causing membrane structure damage and genome DNA degradation.Interestingly,treatment with different concentration of r Lc-P5L seemingly caused the similar but different changes,whether it indeed gave rise to cancer cells diverse death way,the further studies should be performed,and the detailed mechanisms were still need further explored.

Thymoquinone enhances the antioxidant and anticancer activity of Lebanese propolis

BACKGROUND Reactive oxygen species(ROS)are produced by multiple cellular processes and are maintained at optimal levels in normal cells by endogenous antioxidants.In recent years,the search for potential exogenous antioxidants from dietary sources has gained considerable attention to eliminate excess ROS that is associated with oxidative stress related diseases including cancer.Propolis,a resinous honeybee product,has been shown to have protective effects against oxidative stress and anticancer effects against several types of neoplasms.AIM To investigate the antioxidant and anticancer potential of Lebanese propolis when applied alone or in combination with the promising anticancer compound Thymoquinone(TQ)the main constituent of Nigella sativa essential oil.METHODS Crude extracts of Lebanese propolis collected from two locations,Rashaya and Akkar-Danniyeh,were prepared in methanol and the total phenolic content was determined by Folin–Ciocalteu method.The antioxidant activity was assessed by the ability to scavenge 2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical and to inhibit H2O2-induced oxidative hemolysis of human erythrocytes.The anticancer activity was evaluated by[3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide]MTT assay against HCT-116 human colorectal cancer cells and MDAMB-231 human breast cancer cells.RESULTS The total phenolic content of propolis extract from Rashaya and Akkar-Danniyeh were 56.81μg and 83.503μg of gallic acid equivalent/mg of propolis,respectively.Both natural agents exhibited strong antioxidant activities as evidenced by their ability to scavenge DPPH free radical and to protect erythrocytes against H2O2-induced hemolysis.They also dose-dependently decreased the viability of both cancer cell lines.The IC50 value of each of propolis extract from Rashaya and Akkar-Danniyeh or TQ was 22.3,61.7,40.44μg/mL for breast cancer cells at 72 h and 33.3,50.9,33.5μg/mL for colorectal cancer cells at the same time point,respectively.Importantly,the inhibitory effects of propolis on DPPH radicals and cancer cell viability were achieved at half its concentration when combined with TQ.CONCLUSION Our results indicate that Lebanese propolis extract has antioxidant and anticancer potential and its combination with TQ could possibly prevent ROS-mediated diseases.

Thymoquinone anticancer activity is enhanced when combined with royal jelly in human breast cancer

BACKGROUND Breast cancer is the most common cause of the majority of cancer-related deaths in women,among which triple-negative breast cancer is the most aggressive type of breast cancer diagnosed with limited treatment options.Thymoquinone(TQ),the main bioactive constituent of Nigella sativa,has been extensively studied as a potent anticancer molecule against various types of cancers.Honeybee products such as the royal jelly(RJ),the nutritive secretion fed to honeybee queens,exhibit a variety of biological activities besides its anticancer effect.However,the anticancer activity of the combination of TQ and RJ against breast cancer is still unknown.AIM To investigate cytotoxicity of RJ in FHs 74 Int cells and the anticancer effects of TQ,RJ,and their combinations in the MDA-MB-231 cell line.METHODS Cells were treated with TQ,RJ,and their combinations for 24 h.Using 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay,we determined the half-maximal inhibitory concentration of TQ.Trypan blue and 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were then performed to assess the cell viability in response to different treatment conditions.Cell death and cycle regulation were investigated using propidium iodide deoxyribonucleic acid staining followed by flow cytometry in response to a single dose of TQ,RJ,and their combination.Immunostaining for cleaved caspase 3 and Ki67 expression was used to determine apoptosis induction and changes in cell proliferation.RESULTS TQ alone inhibited cell viability in a dose-dependent manner at concentrations below and above the half-maximal inhibitory concentration.RJ exhibited relatively nontoxic effects against MDA-MB-231 cells and FHs 74 Int small intestinal cells at concentrations below 5μg/mL.High doses of RJ(200μg/mL)had greater toxicity against MDA-MB-231 cells.Interestingly,the inhibition of cell viability was most pronounced in response to 15μmol/L TQ and 5μg/mL RJ.A dose of 15μmol/L TQ caused a significant increase in the PreG1 population,while a more pronounced effect on cell viability inhibition and PreG1 increase was observed in response to TQ and RJ combinations.TQ was the main inducer of caspase 3-dependent apoptosis when applied alone and in combination with RJ.In contrast,no significant regulation of Ki67 expression was observed,indicating that the decrease in cell viability was due to apoptosis induction rather than to inhibition of cell proliferation.CONCLUSION This study is the first to report enhanced anticancer effects of TQ and RJ combination against MDA-MB-231 breast cancer cells,which could confer an advantage for cancer therapy.

PROGRESS OF RESEARCH IN THE PREPARATION OF CHINESE TRADITIONAL AND HERBAL DRUGS WITH ANTICANCER ACTIVITY

Research into anticancer substances madefrom Chinese herbal drugs and their clinicalapplication is gaining international attention bythe medical profession of the more than 20analogues of camptothecine isolated from Camp-totheca tree in China, most exhibited anticanceractivity. Among them, 10-hydroxycamptothe-cine has a wide anticancer spectrum and is lesstoxic. In suspension, it exhibits some therapeu-tic effects on primary hepatic cancer, gastriccarcinoma, cancer of the urinary bladder andleukemia.

Synthesis and Anticancer Activity of 2,3,4-Trimethoxyacetophenoxime Ester Containing Benzothiazole Moiety

A series of 2,3,4-trimethoxyacetophenoxime esters containing benzothiazole moiety were synthesized by the reaction of oxime with acyl chloride in alkaline medium. Their structures were established by elemental analysis, IR, and ^1H NMR spectra. The bioassay tests showed that these title compounds exhibit moderate anticancer activity in vitro by MTT method and compounds 6c and 6d could inhibit ERK phosphorylation in NIH 3T3 cell induced by PDGF.

Lobelia chinensis: chemical constituents and anticancer activity perspective

Research has demonstrated that many chemical constituents dominated by piperidine alkaloids and flavonoids, such as lobelanidine, lobeline, and lobelanine, have been obtained from Lobelia chinensis Lour. Experimental studies and clinical applications have also indicated that L. chinensis possesses a number of pharmacological activities(e.g., diuretic, choleretic, breathing excitement, anti-venom, anti-bacterial, and anticancer). This paper focuses on the properties, chemical constituents, and anticancer activity of L. chinensis to clarify the connection among them, and identify the active anticancer compounds. This work serves as the foundation for further research and development of L. chinensis.

Facile Synthesis of 7-epi-Taxane and Its Derivatives and Preliminary Evaluation of Anticancer Activity

7-epi-Taxane has been achieved efficiently in gram scale from natural taxane via inversion of the 7-hydroxyl group simply using Ag20 as catalyst and DMF as solvent. The catalyst could he quantitatively recovered by filtra- tion without loss of catalytic activity. This condition is also applicable to the direct epimerization of taxane derivatives （e.g., docetaxel and paclitaxel） to 7-epi-taxane derivatives （e.g., 7-epi-docetaxel and 7-epi-paclitaxel）. Furthermore, 33 ester derivatives of 7-epi-taxane with different amino acid moieties at the position of C-13 were successfully synthesized via esterification without protecting C-7-OH. Bioassay results revealed that compounds 13 and 18 have good selectivity against prostatic cancer cell line DU145, with ICs0 value as low as 15.9 nmol/L for 18.

QSAR Studies on a Series of 7,8-Dialkyl-1,3-diaminopyrrolo-[3,2-f]quinazolines with Anticancer Activity

The quantitative structure-activity relationship （QSAR） studies on a series of 7,8-dialkyl-1,3-diaminopyrrolo-[3,2-f]quinazolines, dihydrofolate reductase （DHFR） inhibitors as potential anticancer agents, have been carried out by using the density functional theory （DFT） method, molecular mechanics method （MM＋） and statistical method. Some QSAR models based on their lipophilic and steric parameters were built up via a stepwise regression analysis. It is very interesting to find that the established optimal QSAR equation involves only two descriptors： lipophilicity indexes Clog P and Clog P^2 However, such descriptors can quite well describe a significant statistic quality and have a remarkable predictive activity according to the square of adjusted correlation coefficient （RA^2 = 0.937） and the square of cross-validation coefficient （q^2=0.911） of this equation. The results show that the lipophilicity is a main factor affecting the anticancer activity of this series of antimetastatic agents, and the obtained equation describes a parabolic correlation between pIC50 and Clog P, and indicates a suitable range of Clog P （around 4.43） being very important for optimal pIC50 values. These QSAR studies can offer some useful references for understanding the action mechanism and performing the molecular design or modification of this series of antimetastatic agents.

Improvement of Stability and Anticancer Activity of ChlorambuciI-Tetrapeptide Conjugate Vesicles

Chlorambucil is a classic nitrogen mustard drug that has been used in the treatment of cancers. It may induce neutropenia, thrombocytopenia and other side effects because of its short lifetime and off-target effect. In this report, chlorambucil-tetrapeptide （AAAK, A3K） conjugate vesicles were developed to improve the stability and bioactivity of chlorambucil. First of all, chlorambucil-A3K conjugate was synthesized by solid phase synthesis strategy. Sec- ondly, the chlorambucil-A3K conjugate was assembled and characterized by critical aggregation concentration, cir- cular dichroism, dynamic light scattering and transmission electron microscopy. The results indicated that the chlo- rambucil-A3K conjugate can be assembled to form spherical vesicles with an average diameter of 390.5 nm, and high drug loading about 47.1% is reached. Surprisingly, the preliminary biological evaluation of the chlorambu- cil-A3K conjugate vesicles revealed the best in vitro anticancer activity against HeLa, HepG-2 and MCF-7 cell lines compared with chlorambucil and chlorambucil-A3K conjugate free drugs. Furthermore, conjugate vesicles showed excellent in vivo antitumoral activity. It can be partly attributed to their vesicular structure which isolates chloram- bucil active moiety from aqueous solution to retard degradation before killing cancer cells. Therefore, chlorambu- cil-peptide （A3K） conjugate vesicles may be an alternative delivery system of chlorambucil.

Two Cu(Ⅱ)and Co(Ⅱ)-based Coordination Polymers Based on the N,S-donor Ligand:Crystal Structures and Anticancer Activity Evaluation

Two new Cu(II)and Co(II)-based coordination polymers with the chemical formulae of{Cu(L)2]·DMF}n(1)and{[Co(L)2]·2DMF}n(2)have been successfully constructed from a pyridine-substituted N-heterocyclic thioamide ligand,namely[3,4?-bipyridine]-2?-thiol(HL).Complex 1 is composed of two identical coordination networks with dia topology interpenetrating into each other,and shows 1-D microporous channels along the a axis.Complex 2 is assembled by 2-D square-grid layers stacked in an eclipsed fashion to give 1-D large square channels along the b axis.CCK-8 was used for the detection of cancer cells.Results indicated that compound 1 exhibited stronger anti-cancer activity than compound 2 on the H1650 cancer cells.The Annexin V-FITC/PI was recommended for cancer cell apoptosis,and the western blot was applied for the detection of PI3K/AKT pathway activation.

Design, Synthesis and Anticancer Activity of Novel 6-（Aminophenyl）-2,4-bismorpholino-1,3,5-triazine Derivatives Bearing Arylmethylene Hydrazine Moiety

In an attempt to develop potent and selective anticancer agents,we designed and synthesized a series of novel bis（morpholino-1,3,5-triazine） derivatives beating aylmethylene hydrazine moiety and evaluated their cytotoxicity,in vitro,against H460（non-small-cell lung cancer）,HT-29（human colorectal cancer） and MDA-MB-231（human breast cancer） cell lines.The pharmacological results indicate that all the compounds exhibit enhanced cytotoxicity than BMCL-200908069-1,and six target compounds（7e,7h,7j,9a,9b,9c） were superior to PAC-1 against all the tested cancer cell lines.The most active compound 7j,with IC50（inhibitory concentration 50％）values of 0.75,0.34 and 0.60 μ mol/L against HT-29,H460 and MDA-MB-231 cancer cell lines,was 39-,28-,and 60-fold more potent than BMCL-200908069-1 （29.24,9.52 and 36.21 μmol/L）,respectively.

Synthesis and anticancer activity containing benzyl piperidone study of curcumin-related compounds

Sixteen curcumin-related compounds containing benzyl piperidone were synthesized and evaluated for their anticancer activity by the MTT assay towards cultured prostate cancer （PC-3）, pancreatic cancer （BxPC-3）, colon cancer （HT-29）, and lung cancer （H1299） cells. Compounds A1 and B3 exhibited potent growth inhibitory effects against these cells in culture. The IC50 values of these compounds were lower than 1 μM in all four cell lines.

In vivo anticancer activity of vanillin,benzophenone and acetophenone thiosemicarbazones on Swiss albino mice

Objective:To study the anticancer activities of three schiff bases viz.vanillin thiosemicarbazone,benzophenone thiosemicarbazone and acetophenone thiosemicarbazone against Ehrlich ascites carcinoma(EAC)cells in Swiss albino mice.Methods:Synthesized compounds have administrated into the intraperitoneal cavity of the EAC inoculated mice at two doses.The anticancer activities have studied by monitoring the parameters such as cell growth inhibition,tumor weight measurement,survival time of EAC bearing mice as well as the changes in depleted hematological parameters due to tumorgenesis.All such data have been compared with those of a known standard drug bleomycin at the dose of 0.3 mg/kg(i.p.).Results:It has been found that these bases enhanced life span,reduced average tumor weight and inhibited tumor cell growth of EAC cell bearing mice remarkably.The results were similar in potency to those obtained with bleomycin.It was also found that the depleted hematological parameters(red blood count,white blood count and haemoglobin content)were found to be restored gradually towards normal within few weeks after ceasing the treatment.Conclusions:The compounds can be primarily considered more or less as potent anticancer agents.

Anticancer Activities of Substituted Cinnamic Acid Phenethyl Esters on Human Cancer Cell Lines

Caffeic acid phenethyl ester (CAPE) and sixteen substituted cinnamic acid phenethyl esters were prepared via conventional procedures in order to test their in vitro anticancer activities by either MTT assay or SRB assay on six different human cancer cell lines. The results indicated that in the concentration of 10 μmol·L -1 the lead compound CAPE possessed anticancer activities against human HL 60, Bel 7402, and Hela cell lines, and two other compounds possessed potent anticancer activities against Bel 7402 and Hela cell lines.

Cantharidin and Its Analogues:Anticancer and Ser/Thr Protein Phosphatase Inhibitory Activities

This paper mainly describes the anticancer activities and Ser/Thr protein phosphatase inhibitory activities of cantharidin and its analogues.