Promising Antiproliferative Effects Research Articles

Discovery of novel thienoquinoline-2-carboxamide chalcone derivatives as antiproliferative EGFR tyrosine kinase inhibitors.

Novel thienoquinoline carboxamide-chalcone derivatives were prepared via the cyclization of acylated chalcones and 2-mercaptoquinoline-3-carbaldehyde in DMF with K2CO3. Thienoquinolines 9a-f, h exhibited promising antiproliferative effect against all the tested cell lines and gave a significant activity as EGFR inhibitors, with IC50 values ranging from 0.5 and 3.2 µM, and compounds 9e and 9f being the most active of the series. They also showed better activity than Erlotinib against melanoma cancer cell line A375. Moreover, compound 9f influenced pre G1 apoptosis and cell cycle arrest at G2/M phase. The binding mode of the best EGFR inhibitor 9e in the EGFR active site revealed that the thienoquinoline ring occupied the ATP-binding site while the chalcone moiety is located in the allosteric site and is responsible for the enhanced activity of these compounds.

A new class of prophylactic metallo-antibiotic possessing potent anti-cancer and anti-microbial properties.

Immunocompromised cancer patients are often at high risk of developing infections. Standard infection control measures are required to prevent the onset of infection but, under some circumstances, antimicrobial prophylaxis is necessary. We have developed a family of innovative metallo-antibiotics of general formula [Cu(N,N)(CipA)Cl] where N,N represents a phenanthrene ligand and CipA stands for a derivative of the clinically used fluoroquinolone antibiotic ciprofloxacin. The X-ray crystal structure of one member from this family, [Cu(phen)(CipA)Cl] (where phen is 1,10-phenanthroline), is also reported. These complexes combine into one drug entity a Cu-N,N-framework with DNA binding and DNA oxidant properties and an antibiotic derivative with known anti-proliferative and anti-microbial activities. The complexes were all found to exhibit excellent DNA recognition with binding affinity of lead agents in the order of ∼107 M(bp)-1. Biophysical studies involving calf thymus DNA indicate the complexes intercalate or semi-intercalate DNA via the minor groove. All complexes exhibited excellent nuclease activity with DNA strand scission being mediated predominantly via superoxide and hydroxyl radicals. The complexes were found to have promising anti-proliferative effects against a human breast adenocarcinoma cell line (MCF-7) and a human prostate carcinoma cell line (DU145) with low micromolar and, in some cases, nanomolar cytotoxicities observed. Selective targeting of Gram positive bacteria was also identified by this complex class with one lead compound having an order of magnitude greater potency against Methicillin-resistant S. aureus (MRSA) as compared to the CipA ligand. Importantly, from a clinical stand point, these complexes were also found to be well tolerated in an in vivo Galleria mellonella larvae model, which has both functional and structural similarities to that of the innate immune system of mammals.

Innovative DNA-Targeted Metallo-prodrug Strategy Combining Histone Deacetylase Inhibition with Oxidative Stress.

Cancer remains a global health challenge. There is an urgent need to develop innovative therapeutics that can overcome the shortcomings of existing cancer therapies. DNA enzymes involved in nucleic acid compaction and organization are an attractive cancer drug target for therapeutic exploitation. In this work, a family of Cu(II) prodrugs containing suberoylanilide hydroxamic acid (SAHA), a well-established histone deacetylase inhibitor (HDACi) and clinically approved cancer drug, and phenanthrene ligands as DNA intercalative components have been rationally developed. The complexes, of general formula [Cu(SAHA-1H)( N, N'-phenanthrene)]+, exhibit excellent DNA recognition with binding affinity of lead agents in the order of ∼107 M(bp)-1. Biophysical studies involving nucleic acid polymers indicate intercalative binding at both adenine-thymine (A-T) and guanine-cytosine (G-C) rich sequences but thermodynamically stable interactions are favored in G-C tracts. The complexes mediate DNA damage by producing reactive oxygen species (ROS) with spin trapping experiments showing that superoxide, the hydroxyl radical, and hydrogen peroxide play critical roles in strand scission. The agents were found to have promising antiproliferative effects against a panel of epithelial cancers, and in two representative cell lines possessing mutated p53 (SK-OV-3 and DU145), enhanced cytotoxicity was observed. Significantly, mechanistic experiments with the most promising candidates revealed HDAC inhibition activity was achieved over a shorter time frame as compared to clinical standards with DNA damage-response markers identifying upregulation of both DNA synthesis and nucleotide excision repair (NER) pathways. Finally, confocal imaging and gene expression analysis show this metallodrug class exerts cytotoxic activity predominantly through an apoptotic pathway.

Novel anticancer and antibacterial organometallic polymer based on ferrocene as a building block and xanthone bioactive scaffolds: Synthesis, characterization, and biological study

Poly(azine ferrocene‐triazole‐xanthone amide) (PAFTXA) was successfully synthesized from azine ferrocene bisacetylene (AFcB) and 2,7‐(N‐2‐azidoacetamide) xanthone (NAzX) by using click chemistry protocol. In this paper, innovative AFcB moiety incorporated in this procedure as a monomer. This monomer was prepared in three steps from ferrocene scaffold, and the synthetic processes were discussed in details. Characterization of the synthesized novel intermediate materials, monomer, and the resulting polymer was performed by nuclear magnetic resonance, Fourier transform infrared, thermal gravimetric analysis, differential scanning calorimetry, and ultraviolet‐visible spectrophotometer. In addition, the in vitro antibacterial activity of the polymer was appraised against Gram‐positive and Gram‐negative bacteria by using minimum inhibitory concentration method. Moreover; antitumoral effects of synthesized polymer were evaluated on Hela, MCF‐7, and Saos cancer cell lines and compared with normal fibroblast cell line by means of 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay. Promising antiproliferative effects were detected on both epitheloid and mesenchymal cancer cell lines depending on PAFTXA concentration and time. Survey on its characteristic features revealed the proficiencies of our prepared biomaterial such as its high thermal stability as well as valuable antitumoral and antibacterial capabilities that make it an appreciable candidate for further bioapplications.

Abstract 5430: Evaluating a novel phytobiologic mixture against breast cancer cell lines: Effect on glycolytic enzymes and EMT gene expression profile

Abstract Background: Recently, there has been a growing evidence proposed regarding role of phytobiological compounds against various malignancies. One of the novel phytobiological mixture (PB), which is a mixture composed of the following: quercetin, resveratrol, cruciferex, epigallocatechin-3-gallate and curcumin at different proportions. To the best of our knowledge, this is the first study to evaluate the effects of a PB on breast cancer cell lines and its impact on the expression profile of these cells. Aim: Herein, we investigated the anti-proliferative effects of PB against two breast cancer cell lines: MDA-MB-231 and MCF-7, in addition, to investigating whether PB may alter the expression profile of glycolytic enzymes and epithelial mesenchymal transition (EMT) genes in these cell lines. Method: The anti-proliferative effect of PB was evaluated in vitro against MCF-7 and MDA-MB-231 cell lines using SRB assay. The effect of PB on the relative expression profile of some of the glycolytic enzymes and EMT genes was conducted using TaqMan gene expression assays via reverse transcription-quantitative PCR (RT-qPCR). Results: At 72 hrs, treatment with PB showed cytotoxic effects against MDA-MB-231 and MCF-7 cells with an IC50's of 25.9±3 µg/ml and 29.5± 0.9 µg/ml, respectively. At 24 hrs post PB treatment, there was a significant increase in the gene expression of most glycolytic enzymes such as; Phosphofructokinase muscle type (PFKM) (1.4± 0.1, p<0.01) and Pyruvate dehydrogenase kinase isoenzyme 1 (PDK-1) (1.6± 0.2 fold, p< 0.001). PB did not seem to alter the EMT gene expression profile with both cadherin 1 and 2 showing no significant changes. Interestingly, however, a significant up-regulation was seen with ZEB2 member of the zinc finger E-box binding homeobox 2, post PB treatment. Conclusion: In conclusion PB showed a promising anti-proliferative effect against two breast cancer cell lines. However, we postulate that the drug could be modulating glycolytic enzymes expression, which ultimately can affect key molecules in cell differentiation. Further studies are needed to elucidate the exact underlying mechanism and to better understand the most beneficial drug combinations that can work synergistically with PB to interfere with different tumor hallmarks. Citation Format: Aliaa A. Alamoudi, Afnan A. Alqarni, Ghada Ajabnoor, Aleksandra Niedwiecki, Matthias Rath, Steve M. Harakeh, Ahmed M. Al-Abd. Evaluating a novel phytobiologic mixture against breast cancer cell lines: Effect on glycolytic enzymes and EMT gene expression profile [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5430.

2325 Targeting isoform-specific PI3 kinase signaling for treatment of cutaneous T cell lymphoma

OBJECTIVES/SPECIFIC AIMS: (1) Determine the anti-proliferative effect of Copanlisib (α/δ PI3 kinase inhibitor) in CTCL cell lines and synergy with other anti-tumor drugs such as HDAC inhibitors. (2) Determine the effect of Copanlisib treatment on downstream targets of the PI3K/AKT/mTOR pathway. METHODS/STUDY POPULATION: We will test the anti-proliferative effect of Copanlisib on the cell lines H9 and HH, which are well characterized for evaluating new therapies for CTCL (Netchiporouk et al., 2017). We will also test the anti-proliferative effect of Copanlisib in combination with HDAC inhibitors on CTCL cell lines. Cell Titer Glo (Promega) will be used for the proliferation assay. Briefly, Cell-Titler Glo will be added after 24, 48, and 72 hours and luminescence will be measured as a % of maximal growth. Inhibitory effect will be determined by comparison to % growth of the control cultures without Copanlisib treatment. Our next objective is to determine the effect of Copanlisib treatment on downstream targets of the PI3K/AKT/mTOR pathway using Western blot analysis. In brief, 30 μg of each lysate will be subjected to 4%–12% gradient SDS-PAGE gel eletrophoresis. All primary antibodies were purchased from Cell Signaling Technology. Membranes will be washed with TBST and incubated with 1:10,000 dilution of IRDye-conjugated secondary antibody (Licor) for 1 hour. Results will be expressed as relative intensity: the intensity of each band adjusted to that of GAPDH. Experiments will be done in triplicate and 1-way analysis of variance followed by multiple comparison test will be applied to compare the cell proliferation between different treatment groups. RESULTS/ANTICIPATED RESULTS: Previous results from the Ai lab have demonstrated the importance of the PI3 kinase signaling cascade using high-throughput proliferation assays and siRNA knockdown of individual and double PI3 kinase isoforms (unpublished data). So far I have successfully established culture of HH and H9 cells in 96 well plates (100,000 cells/200 μL). We are titrating Copanlisib at doses from 20 nm to 20 μM. Initial results suggest a promising anti-proliferative effect and currently we are optimizing the most effective pharmacologic dose. Thus we anticipate that Copanlisib will exhibit a potent anti-proliferation activity in CTCL cell lines. H9 and HH cell lysates have been collected and preserved for Western blot analysis. We anticipate that Copanlisib treatment will significantly decrease the phosphorylation of AKT and 4EB-P1, both downstream targets of PI3 kinase signaling. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings will elucidate the importance of the PI3 kinase/AKT/mTOR pathway in tumor proliferation in CTCL. Identifying the importance of specific isoforms of PI3 kinase in CTCL will allow for more targeted selection of treatment. Copanlisib targets α and δ isoforms of PI3K and is newly approved by the FDA for low grade B cell lymphoma. Our results seek to quantify the anti-proliferative effect of Copanlisib and determine an on-target mechanism of action by investigating the drug’s effects on downstream signaling molecules of the PI3 kinase pathway. This project will elucidate the disease process of CTCL and provide important insight for its management.

Modular synthesis of new C-aryl-nucleosides and their anti-CML activity

The C-aryl-ribosyles are of utmost interest for the development of antiviral and anticancer agents. Even if several synthetic pathways have been disclosed for the preparation of these nucleosides, a direct, few steps and modular approaches are still lacking. In line with our previous efforts, we report herein a one step - eco-friendly β-ribosylation of aryles and heteroaryles through a direct Friedel-Craft ribosylation mediated by bismuth triflate, Bi(OTf)3. The resulting carbohydrates have been functionalized by cross-coupling reactions, leading to a series of new C-aryl-nucleosides (32 compounds). Among them, we observed that 5d exerts promising anti-proliferative effects against two human Chronic Myeloid Leukemia (CML) cell lines, both sensitive (K562-S) or resistant (K562-R) to imatinib, the “gold standard of care” used in this pathology. Moreover, we demonstrated that 5d kills CML cells by a non-conventional mechanism of cell death.

Anti-proliferative activity of earthworm coelomic fluid using oral squamous carcinoma KB 3-1 cells: An In vitro study with serine protease analysis

Background: Naturally available extracts have gained importance as adjunctive pharmacologic modalities. The earthworm coelomic fluid (ECF) has shown promising in vitro cytotoxic effect in hepatocellular, colorectal, and breast cancers. Few in vitro studies have been performed pertaining to oral cancer. Objective: The objective of this study was to evaluate the time- and dose-dependent antiproliferative effect of ECF of Eudrilus eugeniae (EE), Eisenia fetida (EF), and Perionyx excavatus (PE) on oral cancer cell line KB 3–1. The chemical nature of the ECF was analyzed as well. Materials and Methods: The ECF was collected employing the cold shock method. Lowry's protein estimation was used for determining the total protein content. ECF of EE, EF, and PE at concentration of 6.25, 12.5, 25, and 50 μg/mL was tested on KB 3–1 cells in vitro. The 3-(4, 5-dimethylthiazol-2-Yl)-2, 5-diphenyltetrazolium bromide antiproliferative assay was used to determine the time- and dose-dependent antiproliferative effect. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed to separate and observe prominent protein bands in the three extracted fluids. The chemical nature of the band was analyzed through an enzyme zymography. Results: ECF of PE showed a maximum inhibitory percentage of 94.29% at 50 μg/mL, followed by ECF of EE and EF with values of 89.23% and 79.44%, respectively. ECF of EE showed a superior IC50 value of 3.70 μg/mL as compared to EF and PE at 48 h. Conclusion: ECF of EE has a promising antiproliferative effect in a time- and dose-dependent manner on oral squamous carcinoma KB 3–1 cells in vitro. The serine protease presence in ECF of EE was demonstrated. Abbreviations used: EFE: Earthworm fibrinolytic enzyme; BSA: Bovine serum albumin; DMSO: Dimethyl sulphoxide; ECF: Earthworm coelomic fluid; EE: Eudrilus eugeniae; EF: Eisenia fetida; PE: Perionyx excavatus; ELISA: Enzyme-linked immunosorbent assay; FBS: Fetal bovine serum; IC: Inhibitory concentration; MEM: Minimal essential medium; MTT: 3-(4, 5-dimethylthiazol-2-Yl)-2, 5-diphenyltetrazolium bromide; NCCS: National Centre for Cell Science; NCI: National Cancer Institute; OD: Optical density; PBS: Phosphate-buffered saline; SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TEMED: Tetramethylethylenediamine.

Development of novel anti-filarial agents using carbamo(dithioperoxo)thioate derivatives

A series of novel carbamo(dithioperoxo)thioate derivatives have been prepared in excellent yield using a significantly fast, one-pot three component reaction and experimented for their potential as anti-filarial agents against model filarial nematode Setaria cervi. Among 23 compounds (4a-w) evaluated for the anti-filarial activities, five compounds (4a, 4b, 4c, 4d and 4h) have shown promising anti-proliferative effects on the juvenile stage microfilariae (mf) as well as in adults in a time and dose dependent manner. Compound 4a was found most active against oocytes, mf and adult nematods as well as non-cytotoxic to the normal cells. It has been established that the anti-filarial activity of the compounds were observed due to the involvement of reactive oxygen species (ROS) and apoptosis. Several biochemical and microscopic experiments have been carried out to establish the fact that both intrinsic and extrinsic pathways of apoptosis contribute to the compound 4a mediated death phenomenon of the filarial nematodes.

Anti-cancer effects of F16: A novel vascular endothelial growth factor receptor-specific inhibitor.

Vascular endothelial growth factor receptor-2 is a dynamic target for therapeutic intervention in various types of cancers. This study was aimed to explore the anti-angiogenic activity of a novel vascular endothelial growth factor receptor-specific inhibitor named F16 in both in vitro and in vivo experimental models. This compound effectively reduced cell proliferation, tube formation, and migration of human umbilical vein endothelial cells in a concentration-dependent manner by directly inhibiting vascular endothelial growth factor binding and subsequent vascular endothelial growth factor receptor-2 phosphorylation. The F16 was also able to inhibit the phosphoinositide 3-kinase/protein kinase B-mediated survival and migration pathways in cancer in addition to inhibiting the focal adhesion kinase and mitogen-activated protein kinases-mediated signaling in GI-101A cancer cells. The chorioallantoic membrane assay followed by tumor growth inhibition measurements with GI-101A breast cancer xenograft implanted athymic nude mice confirmed the in vivo tumor reductive effects of F16. It was interesting to observe a decrease in tumor burden after F16 treatment which correlated very well with the decrease in the plasma levels of mucin-1 (MUC-1). Our studies so far have confirmed that F16 is a specific inhibitor of angiogenesis in both in vitro and in vivo models. The F16 also works very efficiently with Taxol in combination by limiting the tumor growth that is better than the monotherapy with any one of the drugs that were tested individually. Thus, F16 offers a promising anti-proliferative and anti-angiogenic effects with better specificity than some of the existing multi-kinase inhibitors.

Abstract 4208: Inhibition of proliferation and induction of apoptosis by a novel phytobiological mixture against breast cancer cell lines

Abstract Breast cancer remains a leading cause of women’s death worldwide with progressive emergence of resistance towards approved chemotherapies. For this reason, new complementarytreatments have been sought. One of those includethe use of a phytobiological mixture against different types of malignancies, including but not limited to breast cancer. Herein, we investigated the anti-proliferative effects of a novel phytobiological mixture (PB)against two breast cancer cell lines :MDA-MB-231 and MCF-7.The PB mixture is composed of the following: quercetin, resveratrol, cruciferex, epigallocatechin-3-gallate and curcumin at different proportions. Theanti-proliferative and pro-apoptotic effects of PB were evaluated at non-cytotoxic concentrations as determined usingthe Sulfa-Rhodamine B assay (SRB).The effects of PB on the induction of apoptosis were valuated using Annexin-V and flow cytometry to measure the resultingDNA fragments. The results showed that at 72hrs, PB had cytotoxic effects against MDA-MB-231 and MCF-7 cells with an IC50’s of 25.8±5.4 µg/ml and 56.9±4.2 µg/ml, respectively. Exposure of MDA-MB-231 to PB for 72hrs resulted in the induction of apoptosis in a dose-dependent manner. PB concentrations of 10 µg/ml and 30 µg/ml, resulted in early programmed cell death of 9.1±1.5% and 22.1±1.8% respectively. Exposure to PB (10 µg/ml for 24 hrs) resulted in mild cell cycle arrest at the G2/M phase20.3±0.3% ;while higher dosesshowed a significant cell cyclearrest at theS-phaseas compared to the control untreated cells (p<0.001). However, exposure to PB for 48 hrs at both doses,10 µg/ml and30 µg/ml,resulted in a significant cell cycle arrest at the S-phase 25.3±1.6% and 20.5± 0.4%respectivelyas compared to the control untreated cells 12.4±1.5% (p<0.001). In conclusion, PB was found to have a promising anti-proliferative and pro-apoptotic effect in breast cancer cell lines. Further studies are needed to address the underlying mechanism of action of PB against breast cancer cell lines. Citation Format: Afnan A. Alqarni, Aliaa A. Alamoudi, Ghada M. Ajabnoor, Ahmed M. Al-Abd, Aleksandra Niedwiecki, Matthias Rath, Steve Harakeh. Inhibition of proliferation and induction of apoptosis by a novel phytobiological mixture against breast cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4208. doi:10.1158/1538-7445.AM2017-4208

Synthesis and anti-proliferative activity of a small library of 7-substituted 5H-pyrrole [1,2-a][3,1]benzoxazin-5-one derivatives

In this study, we investigate the anti-proliferative activity of a small library of 7-substituted 5H-pyrrolo[1,2-a][3,1]benzoxazin-5-one derivatives, against a panel of human cancer cell lines. We reported the synthesis of these compounds in a previous work. 7-Bromo-5H-benzo[d]pyrrolo[2,1-b][1,3]oxazin-5-one showed a promising anti-proliferative effect. As starting material for Suzuki-Miyaura cross coupling reaction, it was selected for the design and the synthesis of six further derivatives, with the aim to better define structure-activity relationships. The anti-proliferative MTT assay revealed a dose-dependent reduction of cell viability, especially for 7-([1,1′-biphenyl]-4-yl)-5H-benzo[d]pyrrolo[2,1-b][1,3]oxazin-5-one. Cell cycle and western blotting analysis suggested apoptosis as possible mechanism for its anti-proliferative activity. These preliminary results encourage our interest for further optimizations.

In vitro Antiproliferative Effect of Earthworm Coelomic Fluid of Eudrilus Eugeniae, Eisenia Foetida, and Perionyx Excavatus on Squamous Cell Carcinoma-9 Cell Line: A Pilot Study.

Introduction:The earthworm coelomic fluid (ECF) has shown proven antiproliferative effect against breast, liver, gastrointestinal, and brain cancer, but it is least explored in oral cancer. The present in vitro study is an attempt to investigate the antiproliferative activity of ECF on oral cancer cell line squamous cell carcinoma (SCC)-9.Materials and Methods:ECF was collected from the species Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) stored at −80°C. Percentage inhibition of ECF on squamous cell carcinoma-9 cells in vitro was recorded at 24 h. Protein estimation was done using Bradford protein assay validated by the biuret method. Cytotoxicity was tested at 2.5, 5, 10, 20, 40, and 80 μg/ml concentrations by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay in SCC-9 cells in vitro. GraphPad Prism 7.0 software was used to calculate the inhibitory concentration (IC50). Chi-square test was used to analyze the difference between samples.Results:The test samples EE, EF, and PE inhibited the growth of SCC-9 cells significantly in a dose-dependent manner, and the IC50 values were found to be 4.6, 44.69, and 5.27 μg/ml, respectively. The antiproliferative effect was found to be variable among the three earthworm species with EE showing the most promising effect followed by PE and EF.Conclusion:Establishing the antiproliferative effect of ECF on oral cancer cells could be an initial step toward drug development and future anticancer research. The preliminary investigation has shown that ECF has a promising antiproliferative effect on oral cancer cells in vitro.SUMMARYThe present pilot study evaluated the in vitro antiproliferative effect of earthworm coelomic fluid (ECF) of Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) on squamous cell carcinoma-9 cell line. The ECF inhibitory activity was promising at inhibitory concentration values of 4.6, 44.69, and 5.27 μg/ml, respectively. Further studies pertaining to antiproliferative mechanism of EE, EF, and PE have been planned.Abbreviations Used: ECF: Earthworm coelomic fluid, EE: Eudrilus eugeniae, EF: Eisenia foetida, PE: Perionyx excavatus, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, SCC: Squamous cell carcinoma, BSA: Bovine serum albumin, PBS: Phosphored buffered saline, ATCC: American Type Culture Collection

In Vitro Anticancer Effect of Gedunin on Human Teratocarcinomal (NTERA-2) Cancer Stem-Like Cells.

Gedunin is one of the major compounds found in the neem tree (Azadirachta indica). In the present study, antiproliferative potential of gedunin was evaluated in human embryonal carcinoma cells (NTERA-2, a cancer stem cell model) and peripheral blood mononuclear cells (PBMCs), using Sulforhodamine (SRB) and WST-1 assays, respectively. The effects of gedunin on expression of heat shock protein 90 (HSP90), its cochaperone Cdc37, and HSP client proteins (AKT, ErbB2, and HSF1) were evaluated by real-time PCR. Effects of gedunin on apoptosis were evaluated by (a) apoptosis associated morphological changes, (b) caspase 3/7 expression, (c) DNA fragmentation, (d) TUNEL assay, and (e) real-time PCR of apoptosis related genes (Bax, p53, and survivin). Gedunin showed a promising antiproliferative effect in NTERA-2 cells with IC50 values of 14.59, 8.49, and 6.55 μg/mL at 24, 48, and 72 h after incubations, respectively, while exerting a minimal effect on PBMCs. Expression of HSP90, its client proteins, and survivin was inhibited and Bax and p53 were upregulated by gedunin. Apoptosis related morphological changes, DNA fragmentation, and increased caspase 3/7 activities confirmed the proapoptotic effects of gedunin. Collectively, results indicate that gedunin may be a good drug lead for treatment of chemo and radiotherapy resistant cancer stem cells.

Phenolic composition, antioxidant and anti-proliferative activities of edible and medicinal plants from the Peruvian Amazon

Among 23 extracts of medicinal and edible plants tested, Mauritia flexuosa L.f., Arecaceae, showed significant antioxidant ability (DPPH and ORAC=1062.9 and 645.9±51.4μg TE/mg extract, respectively), while Annona montana Macfad., Annonaceae, demonstrated the most promising anti-proliferative effect (IC50 for Hep-G2 and HT-29=2.7 and 9.0μg/ml, respectively). However, combinatory antioxidant/anti-proliferative effect was only detected in Oenocarpus bataua Mart., Arecaceae (DPPH=903.8 and ORAC=1024μg TE/mg extract; IC50 for Hep-G2 and HT-29 at 102.6 and 38.8μg/ml, respectively) and Inga edulis Mart., Fabaceae (DPPH=337.0 and ORAC=795.7μg TE/mg extract; IC50 for Hep-G2 and HT-29 at 36.3 and 57.9μg/ml, respectively). Phenolic content was positively correlated with antioxidant potential, however not with anti-proliferative effect. None of these extracts possessed toxicity towards normal foetal lung cells, suggesting their possible use in development of novel plant-based agents with preventive and/or therapeutic action against oxidative stress-related diseases.

A Polyphenol-Enriched Fraction of Rose Oil Distillation Wastewater Inhibits Cell Proliferation, Migration and TNF-α-Induced VEGF Secretion in Human Immortalized Keratinocytes.

Water steam distillation of rose flowers separates the essential oil from the polyphenol-containing rose oil distillation wastewater. Recently, a strategy was developed to separate rose oil distillation wastewater into a polyphenol depleted water fraction and a polyphenol-enriched fraction [RF20-(SP-207)]. The objective of the present study was to investigate RF20-(SP-207) and fraction F(IV), augmented in quercetin and ellagic acid, for possible antiproliferative effects in immortalized human keratinocytes (HaCaT) since rose petals are known to contain compounds with potential antiproliferative activity.RF20-(SP-207) revealed dose-dependent antiproliferative activity (IC50 of 9.78 µg/mL). In a nontoxic concentration of 10 µg/mL, this effect was stronger than that of the two positive controls LY294002 (10 µM, PI3 K-inhibitor, 30 % inhibition) and NVP-BEZ235 (100 nM, dual PI3 K/mTOR inhibitor, 30 % inhibition) and clearly exceeded the antiproliferative action of quercetin (50 µM, 25 % inhibition) and ellagic acid (1 µM, 15 % inhibition). Time-lapse microscopy detected a significant impairment of cell migration of RF20-(SP-207) and F(IV). At concentrations of 10 µg/mL of both, extract and fraction, cell migration was strongly suppressed (51 % and 28 % gap closure, respectively, compared to 95 % gap closure 24 hours after control treatment). The suppression of cell migration was comparable to the positive controls LY294002, NVP-BEZ235, and quercetin. Furthermore, basal and TNF-α-stimulated VEGF-secretion was significantly reduced by RF20-(SP-207) and F(IV) at 10 µg/mL (44 % vs. untreated control).In conclusion, RF20-(SP-207) showed promising antiproliferative and antimigratory effects and could be developed as a supportive, therapy against hyperproliferation-involved skin diseases.

Abstract B45: Piptandenia africana and Nauclea latifolia protect against N-Nitrosodiethylamine induced hepatic tumors in Wistar rats

Abstract In vitro studies on crude extracts of Piptandenia africana (PA) and Nauclea latifolia (NL) showed promising anti proliferative effects on Hela, MCF7 and ICMST-6 cell lines. Furtherance to these finding, we examined the anti-tumorigenic effect of NL and PA on N-nitrosodiethylamine (DEN) induced hepatic tumors in Wistar rats as potential phytotherapeutic candidates for cancer prevention and treatment. Five groups of rats (n=5) were co-treated as follows for 60 days: Control (1), DEN only (25μg/g body wt.) (2), DEN+PA (3), DEN+ NL (4) and DEN+PA+NL (5). The serum was assessed for markers of toxicity: alanine and aspartate transaminase (ALT and AST) and alkaline phosphatase (ALP). Total bilirubin (TB) conjugated bilirubin (CB) and creatinine were also examined. Numbers, volumes and sizes of DEN-induced tumors were estimated in harvested livers using a DelcastTM caliper. Sections of liver were examined for histopathological changes and hepatic antioxidants profiles: glutathione peroxide (GPx), superoxide dismutase (SOD), glutathione -s- transferase (GST), catalase and reduced glutathione (GSH). Decreases (p<0.05) in food consumption and body weight were observed in grp 2 rats and had mean tumor of one (T1), in the largest lobe (LL) of their liver. In grp 3, two rats had (T0) but three other rats had (T1) each on the LL of their livers. Grp 4, one rat had (T1) on the LL, while two other rats had (T3) and (T1) on the liver’s smallest lobe (SL). Two rats had (T0), one rat had (T1) on the LL of grp 5 rats, one rat had (T3) on LL and (T2) and (T1) on the intermediate lobe (IL) while the last rat had (T0) on LL and (T1) on SL. Mean blood glucose (±S.D) level in grp2 rats was highest (87.5±5.1) compared to all other treated rats. Hepatic GPx, SOD, GST, catalase and GSH levels suggested that grp2 treated rats had lower levels of antioxidant activities compared to grp 1 and 3. Grp 4 rats exhibited higher antioxidant activity relative to animals in grp 3 and grp 2. In addition markers (AST, ALT and ALP, TB and CB) of hepatotoxicity were highest in grp 4 relative to all other groups. Histopathology of liver sections showed severe portal cellular infiltration by mononuclear cells, congestion and multiple foci of severe hepatic necrosis in grp 2, compared to the grp 1. In contrast grp 3 exhibited mild portal cellular infiltration with moderate Kupffer cell hyperplasia. Grp 4 animals presented with severe but diffuse vacuolar degeneration and necrosis of hepatocytes. Mild portal congestion, occasioned by cellular infiltration by mononuclear cells was observed in grp 5 rats. PA and NL co-treatment mitigated DEN-induced hepatic tumorigenesis in rats. PA and NL exhibited phytotherapuetic properties supporting earlier in vitro findings. This may be relevant in preventing hepatic tumor formation and may serve as potential chemoprevention agents. Elucidating the phytochemistry and mechanistic mode of actions of PA and NL are required. Citation Format: Queen-Esther E. Akinade, Sarah O. Nwozo, Olubunmi A. Akinwumi, Solomon E. Owumi. Piptandenia africana and Nauclea latifolia protect against N-Nitrosodiethylamine induced hepatic tumors in Wistar rats. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr B45.

A Rare Class of New Dimeric Naphthoquinones from Diospyros lotus have Multidrug Reversal and Antiproliferative Effects.

Three new dimeric naphthoquinones, 5,4′-dihydroxy-1′-methoxy-6,6′-dimethyl-7,3′-binaphthyl-1,4,5′,8′-tetraone (1), 5′,8′-dihydroxy-5-methoxy-6,6′-dimethyl-7,3′-binaphthyl-1,4,1′,4′-tetraone (2) and 8,5′,8′-trihydroxy-6,6′-dimethyl-7,3′-binaphthyl-1,4,1′,4′-tetraone (3), were isolated from the roots of Diospyros lotus. Their structures were elucidated by spectroscopic techniques, including 1D and 2D NMR, such as HSQC, HMBS, NOESY, and J-resolved. Compounds 1–3 were evaluated for their effects on the reversion of multidrug resistance (MDR) mediated by P-glycoprotein through use of the rhodamine-123 exclusion screening test on human ABCB1 gene transfected L5178Y mouse T-cell lymphoma. Compounds 1–3 were also assessed for their antiproliferative and cytotoxic effects on L5178 and L5178Y mouse T-cell lymphoma lines. Both 1 and 2 exhibited promising antiproliferative and MDR-reversing effects in a dose-dependent manner. The effects of the tested compounds on the activity of doxorubicin were observed to vary from slight antagonism to antagonism.

Deprotometalation–iodolysis and computed CH acidity of 1,2,3- and 1,2,4-triazoles. Application to the synthesis of resveratrol analogues

1-Aryl- and 2-aryl-1,2,3-triazoles were synthesized by N-arylation of the corresponding azoles using aryl iodides. The deprotometalations of 1-phenyl-1,2,3-triazole and -1,2,4-triazole were performed using a 2,2,6,6-tetramethylpiperidino-based mixed lithium–zinc combination and occurred at the most acidic site, affording by iodolysis the 5-substituted derivatives. Dideprotonation was noted from 1-(2-thienyl)-1,2,4-triazole by increasing the amount of base. From 2-phenyl-1,2,3-triazoles, and in particular from 2-(4-trifluoromethoxy)phenyl-1,2,3-triazole, reactions at the 4 position of the triazolyl, but also ortho to the triazolyl on the phenyl group, were observed. The results were analyzed with the help of the CH acidities of the substrates, determined in THF solution using the DFT B3LYP method. 4-Iodo-2-phenyl-1,2,3-triazole and 4-iodo-2-(2-iodophenyl)-1,2,3-triazole were next involved in Suzuki coupling reactions to furnish the corresponding 4-arylated and 4,2′-diarylated derivatives. When evaluated for biological activities, the latter (which are resveratrol analogues) showed moderate antibacterial activity and promising antiproliferative effect against MDA-MB-231 cell line.

Retinoic acid amide inhibits JAK/STAT pathway in lung cancer which leads to apoptosis.

Small cell lung cancer (SCLC) accounts for 12 to 16% of lung neoplasms and has a high rate of metastasis. The present study demonstrates the antiproliferative effect of retinoic acid amide in vitro and in vivo against human lung cancer cells. The results from MTT assay showed a significant growth inhibition of six tested lung cancer cell lines and inhibition of clonogenic growth at 30 μM. Retinoic acid amide also leads to G2/M-phase cell cycle arrest and apoptosis of lung cancer cells. It caused inhibition of JAK2, STAT3, and STAT5, increased the level of p21WAF1, and decreased cyclin A, cyclin B1, and Bcl-XL expression. Retinoic acid amide exhibited a synergistic effect on antiproliferative effects of methotrexate in lung cancer cells. In lung tumor xenografts, the tumor volume was decreased by 82.4% compared to controls. The retinoic acid amide-treated tumors showed inhibition of JAK2/STAT3 activation and Bcl-XL expression. There was also increase in expression of caspase-3 and caspase-9 in tumors on treatment with retinoic acid amide. Thus, retinoic acid amide exhibits promising antiproliferative effects against human lung cancer cells in vitro and in vivo and enhances the antiproliferative effect of methotrexate.