Anticancer potential of fused heterocycles: structural insights and mechanistic advances.

The principal objective of the conducted study is to synthesize enantiomerically pure a class of sugar-based (thio)ureas (9-12) and to investigate their antiproliferative activities against the A549 (lung cancer), MCF-7 (breast cancer), and PANC1 (human pancreatic cancer) cell lines. The synthesis of (thio)urea sugars was performed by two stage procedure. First, the amino sugars (4 and 8) were obtained in three steps (tosylation, substitution and reduction). And secondly, the reaction of 3,5-bis(trifluoromethyl)phenyliso(thio)cyanate with the corresponding amines gave chiral (thio)urea derivatives (9-12). Cell viability was determined in human A549, MCF-7, PANC-1 and noncancer human embryonic kidney (HEK-293) cell lines. Four chiral sugar (thio)ureas (9-12) were synthesized and screened against the A549, MCF-7, and PANC1 cell lines. Of the four chiral sugar derivatives, the compound 9 not only showed the best anticancer activity in the A549 cell line but also provided the highest normal cell (HEK-293) viability. From chiral sugar-derived (thio)ureas obtained, the compound 9 was found to be shown the highest activity against A549 cancer cell line. The compound 9, therefore, gave more promising results for future researches as anti-cancer agents. X-ray studies revealed that amide type hydrogens are playing important roles in anti-cancer activities.

Doxorubicin (Dox) is a highly potent chemotherapeutic agent, approved by FDA since 1974, for treating a broad spectrum of cancers. However, development of severe side effects and drug resistance are main issues that limit the clinical use of Dox. Herein, we aimed to investigate the sensitivity of Dox in a variety of human cancer cell lines. Eleven cell lines were tested in this study including 2 hepatocellular carcinoma (HCC) cell lines (HepG2 and Huh7), 4 bladder cancer (BlCa) cell lines (UMUC-3, VMCUB-1, TCCSUP and BFTC-905), a lung cancer cell line (A549), a cervical carcinoma cell line (HeLa), a breast cancer cell line (MCF-7), a skin melanoma cell line (M21) and a noncancer human kidney cell line (HK-2). The MTT assay was employed for the determination of cytotoxicity. Cells were treated with various concentrations of Dox for 24 h. The half-maximal inhibitory concentration (IC50) of Dox was calculated to compare the Dox sensitivity among tested cell lines. The result showed that IC50 of Dox in HepG2, Huh7, UMUC-3, VMCUB-1, TCCSUP, BFTC-905, A549, HeLa, MCF-7, M21 and HK-2 cells were 12.2, > 20, 5.1, > 20, 12.6, 2.3, > 20, 2.9, 2.5, 2.8 and > 20 mM, respectively. BFTC-905 had the lowest IC50 value, therefore, it was the most sensitive to Dox. In contrast, Huh7, VMCUB-1 and A549 cells were resistant to Dox with IC50 values > 20 mM. Conclusions, we demonstrated that different human malignant cell lines had different sensitivity to Dox. BFTC-905 BlCa cell line had the highest sensitivity to Dox. Huh7, VMCUB-1 and A549 cell lines were resistant to Dox. Perhaps, the different Dox sensitivity in different cell lines was due to the different acquisition of Dox resistance mechanisms. Huh7, VMCUB-1 and A549 cell lines could be suitable cell models to investigate the molecular mechanism of Dox resistance in different cancers. HIGHLIGHTS Different types of cancer cell lines exhibited different sensitivity to doxorubicin. BFTC-905, MCF-7 and M21 cell lines were sensitive to doxorubicin. HepG2, UMUC-3, TCCSUP and HeLa cells were moderately sensitive to doxorubicin. Huh7, VMCUB-1, A549 and HK-2 cells were resistant to doxorubicin. Huh7, VMCUB-1 and A549 cell lines could be suitable cell models for investigating the molecular pathways of Dox resistance in different types of cancers. GRAPHICAL ABSTRACT

Cytotoxic lignans from fruits of Cleistanthus tonkinensis

Thienopyrimidine derivatives exert their anticancer efficacy via apoptosis induction, oxidative stress and mitotic catastrophe

Objective To detect the expressions of interleukin-11 (IL-11) and interleukin-11 receptor α (IL-11Rα) in non-small cell lung cancer (NSCLC) cell lines, and explore their clinical significances. Methods The expressions of IL-11 and IL-11Rα in NSCLC cell lines A549, H2228, healthy lung small airway epithelial cell (SAEC) line cytoplasm, cell membrane and nucleus were detected by Western blot. Results The expressions of IL-11 and IL-11Rα were low in the cell membrance and nucleus (cell membrane: IL-11 0.04±0.03, IL-11Rα 0.05±0.03; nuclear: IL-11 0.45±0.19, IL-11Rα 0.07±0.02; P < 0.01); The expressions of IL-11 and IL-11Rα in A549 and H2228 cell lines were significantly increased compared with those of SAEC cell lines in the cell membrance and cytoplasm (P < 0.01); Among the A549 cell lines, the expressions of IL-11 and IL-11Rα in cell nucleus were much higher than those of the cell membrance and cytoplasm (P < 0.01). Among the H2228 cell lines, the expression of IL-11 in cytoplasm was the highest and the expression of IL-11Rα was the highest in the cell nucleus (P < 0.01). Conclusion The expressions of IL-11 and IL-11Rα are high in NSCLC cell lines, and it is good for the screening and early diagnosis of lung cancer by detecting the expressions of IL-11 and IL-11Rα. Key words: Carcinoma, non-small-cell lung; Interleukin-11; Interleukin-11 receptor alpha subunit; Diagnosis

To evaluate the effects of SGLT2 inhibitors on the proliferation, tumorigenesis, migration, colony formation, apoptosis, selected gene expression pattern, and combination with known chemotherapeutic drugs in different human cancer cell lines. The antiproliferative and combined effects of SGLT2 inhibitors were evaluated by MTT assay. Cell migration was assessed using wound-healing and colony formation assays. Apoptosis assay was conducted using annexin V-FITC/ propidium iodide staining. SGLT2 gene expression was determined using real-time PCR. Canagliflozin, dapagliflozin, and ipragliflozin significantly inhibited the growth of different cancer cell lines in a dose and time-dependent manner. IC50 values after 48 hours of treatment with canagliflozin, ipragliflozin, and dapagliflozin ranged from 41.97 µM to 69.49 µM, 63.67 µM to 255.80 µM, and 167.7 µM to 435.70 µM in the examined cancer cell lines, respectively. The combined treatment of SGLT2 with doxorubicin and raloxifene separately resulted in a synergistic effect in Caco-2 and A-549 cell lines. On the other hand, the combination of SGLT2 inhibitors with cisplatin resulted in an antagonistic effect in A-549, Du-145, and Panc-1 cell lines. Canagliflozin and ipragliflozin inhibited cell migration and colony formation ability at IC50 and Sub-IC50 in the examined cancer cell lines. Canagliflozin and ipragliflozin significantly induced apoptosis at IC50 and Double-IC50 in the Du-145 cell line compared to the control. Real-time PCR showed that the treatment with 0.1 IC50 and 0.2 IC50 of both canagliflozin and ipragliflozin resulted in diminished RNA expression of SGLT2, VEGF, and Bcl-2 genes in the Du-145 cell line. SGLT2 inhibitors have antiproliferation, anti-tumorigenesis, and anti-migration effects and may induce apoptosis in cancer cells. In addition, treatment with SGLT2 inhibitors resulted in the downregulation of selected genes in the Du-145 cell line.

The pyrrolo[2,1-c][1,4]benzodiazepines (PBDs) have been extensively investigated as DNA-interactive agents since their discovery in 1963. The significant cytotoxicity of the naturally derived PBDs has led to the search for synthetic analogues that retain the cytotoxicity of the parent structure while providing an enhanced therapeutic index. There is growing evidence that PBD monomers inhibit the binding of transcription factors to gene promoter regions, blocking their effects on gene expression. For example, GWL-78, a C8-linked PBD-Py-Py (Py = pyrrole) conjugate, has been shown to block interaction of the transcription factor NF-Y, and KMR-28-39, a C8-linked PBD-MPB conjugate inhibits the activity of the transcription factor NF-κB. As up-regulation of NF-κB has long been implicated in the development and progression of several cancer types, this may account for the femtomolar potency observed for PBD monomers such as KMR-28-39. The initial synthesis of a small library of C8-PBD conjugates with benzofused C8-substituents provided notable activity against a panel of leukaemia cell lines. In addition to exhibiting low picomolar IC50 values in a number of cell lines, these compounds also showed selectivity for binding to NF-κB consensus sequences. This small library was used as the starting point for the synthesis of three analogous libraries of PBD C8-conjugates designed to explore the SARs of such structures, with a view to selecting a candidate for pre-clinical development. The design rationale for the novel conjugates involved modification of the shape of the C8-linked substituent with the introduction of specific functional groups in an effort to induce sequence-selective DNA interactivity. The sequence selectivity of the novel benzofused PBD conjugates was investigated using a FRET-based assay in which ligand binding to several different transcription factor binding sequences implicated in cancer was explored (i.e., AP-1, NF-κB1, NF-κB2, STAT3 and EGR). Differential DNA stabilisation compared to control was observed for all analogues, with ΔTm values between 23.4 and 3.4oC at 1 μM ligand concentration (Ligand:DNA = 1:5) for these sequences, indicating the molecules had preferential binding to certain transcription factor binding sequences. Stabilisation was especially pronounced for the AP-1 consensus sequence with two lead compounds (DC-3-28-33 and DC-1-113) providing a high degree of stabilisation (23.9oC and 26oC, respectively). Preliminary cytotoxicity screening studies were carried out in Primary CLL and a Myeloma cell line (JJN3), and low nanomolar IC50 values were observed in the CLL cells for several of the synthesised compounds. These included DC-1-92 (9.7nM), DC-1-115 (14nM), DC-1-36 (16nM), DC-1-170 (17nM), and DC-1-113 (18nM). Control screening in normal lymphocytes is currently underway in order to establish the selective toxicity profiles of these compounds. Citation Format: David B. Corcoran, Paul J. M. Jackson, Ambereen Ajaz, Thomas Lewis, Chris Pepper, David E. Thurston, Khondaker M. Rahman. C8-linked pyrrolobenzodiazepine (PBD)-benzofused hybrids as transcription factor inhibitors. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4555. doi:10.1158/1538-7445.AM2015-4555

1,3,5-triazine derivatives as potential anticancer agents against lung and breast cancer cell lines: Synthesis, biological evaluation, and structure-based drug design studies

Despite the development of novel therapeutic modalities, lung cancer persists as the leading cause of cancer-related mortality. Platinum-based treatments represent the most prominent treatment option, with cisplatin being the most frequently utilized chemotherapeutic agent. However, cisplatin has several serious side effects. A substantial body of evidence has emerged in recent years indicating that boron compounds exhibit anti-cancer properties when administered as monotherapy or in combination with chemotherapy agents. The objective of this study is to examine the anti-cancer effects of Cisplatin (Cis) and sodium pentaborate pentahydrate (NaB), both individually and in combination, on non-small cell lung cancer (NSCLC) cell line A-549 cells and small cell lung cancer (SCLC) cell line DMS-114 cells under in vitro conditions. The effects of cisplatin and NAB on cell survival, apoptosis, the cell cycle, and the expression levels of apoptotic, anti-apoptotic, and tumor suppressor genes were determined by an MTS assay, an Annexin-V assay, a cell cycle analysis, and real-time PCR (qPCR). It was found that the IC-50 value of cisplatin, which was 10 µM when used alone, decreased to 2.5 µM when combined with a non-toxic dose of NaB on the A-549 cell line. BAX and TP53 gene expression levels were elevated by the Nab-Cisplatin combination in the A-549 cell line. The combination was observed to result in an approximately 19-fold increase in CDK2 gene expression in the A-549 cell line and an approximately 6-fold increase in the DMS-114 line, which resulted in S phase and/or G2 phase arrest on the cell cycle. Gene expression levels of Survivin and Ki-67 were decreased by the combination on both cell lines when compared with cisplatin alone. The findings demonstrate that NaB exerts an anti-cancer effect on the A-549 and DMS-114 cell lines. Moreover, when combined with cisplatin, it produces a synergistic anti-cancer effect on the A-549 cell line, whereby apoptosis is activated and cell proliferation is inhibited. The combination of NaB and cisplatin represents a novel approach to the treatment of NSCLC. This is due to the fact that it reduces the IC-50 value of cisplatin and also results in a greater inhibition of cell division and a stronger induction of cell death when used in the context of a combined treatment. Further insight into the effects of the NaB-cisplatin combination will be gained from in vivo experiments and clinical studies.

To explore the effects of icotinib on the proliferation and apoptosis of various lung cancer cell lines. Human lung cancer cell lines HCC827, H1650, H1975, A549 and human epidermal cancer cell line A431 were treated in vitro with icotinib or gefitinib at a concentration gradient of 0 - 40 µmol/L. Their proliferation effects were analyzed by the thiazolyl blue (MTT) assay and the apoptotic effects detected by flow cytometer. The downstream signaling proteins were detected by Western blot. The median inhibitory concentrations (IC(50)) of icotinib for A431 and HCC827 cell lines were (0.04 ± 0.02) and (0.15 ± 0.06) µmol/L respectively. No significant differences existed between the inhibitions of gefitinib and icotinib on A431, HCC827, H1650, H1975 and A549 cell lines (all P > 0.05). Compared with H1650, H1975 and A549 cell lines, icotinib significantly inhibited A431 (P = 0.009, 0.005 and 0.000) and HCC827 (P = 0.001, 0.001 and 0.000) cell lines. And it lowered the expressions of p-AKT, p-ERK and survivin protein expression through the inhibited activity of p-EGFR protein. Icotinib can arrest the proliferation of lung adenocarcinoma cells with EGFR mutation or over-expression by inhibiting the signal pathways of AKT-ERK and survivin.

This study aimed to design and synthesize new ferrocene derivatives for the development of potent anticancer drugs. Cancer is a major cause of death globally. Some small-molecule anticancer drugs have been used in clinics for the treatment of cancer, and several candidates are in different phases of clinical trials. However, cancer chemotherapy is still highly inadequate due to the side effects of the clinical drugs. Thus, developing novel anticancer drugs is essential. Firstly, we synthesized the R-substituted benzaldoxime intermediates (2a-2s) using R-substituted benzaldehyde (1a-1s) and hydroxylamine hydrochloride. Then, the target compounds (3a-3s) were synthesized using ferrocene carboxylic acid and R-substituted benzaldoxime intermediates (2a-2s) as starting materials, and using DCC and DMAP as catalysts. The purity of the target compounds was determined by HPLC, and their structures were characterized using NMR, SC-XRD, and HR-ESIMS. Subsequently, the preliminary in vitro cytotoxicity against HeLa, A549, and A2780 cell lines was evaluated using MTT assay. The results showed that compound 3a exhibited cytotoxicity against both HeLa and A549 cancer cell lines with IC50 values of 0.691 and 0.876 mM, respectively. Compound 3k showed potent cytotoxicity against HeLa cell lines with an IC50 value of 0.097 mM, compounds 3n and 3o exhibited potent cytotoxicity against three cancer cell lines, compound 3q showed potent cytotoxicity against HeLa cell lines with an IC50 value of 0.175 mM, while compound 3s exhibited potent cytotoxicity against HeLa and A549 cell lines with IC50 values of 0.470 and 0.298 mM, respectively. In this work, 19 new ferrocene derivatives containing R-substituted benzaldoxime moieties (3a-3s) were synthesized and their structures were confirmed. Their cytotoxicity against HeLa, A549, and A2780 cell lines was tested, and the results showed that several compounds exhibited potent cytotoxicity against the tested cancer cell lines. This work developed a variety of ferrocene compounds, providing lead compounds based on ferrocene pharmacophore for the development of anticancer drugs.

The purpose of this study was to examine the cytotoxic effects of albendazole, known as anthelmintic in human cancer (A-549, AGS, MKN-74, SNU 601, HCT-116, MDA-MB-231, MCF-7 and U87-MG) and normal (MRC-5, DSC, USMSC and HEK-293) cell lines derived from various tissues. Following 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the cell growth was highly inhibited below 1 μM albendazole treatment, and the 50% inhibitory concentration (IC50) values were especially lower in the HCT-116, A549 and AGS cancer cell lines, compared with others cell lines. A high incidence of cells with activity of senescence-associated ß-galactosidase was also observed in the albendazole-treated cells. Thus, the albendazole treatment was effectively induced the inhibition of cell growth in both cancer and normal cell lines. Moreover, the cell doubling time was significantly lower in the HCT-116, A549 and AGS cancer cell lines with lower IC50 values, compared with others cell lines. Based on present results, albendazole could serve as a potent chemical/drug for cancer therapy, as its treatment was more available in cell lines with a high cell growth capacity.

Introduction: IL-6 has both diagnostic and prognostic significance in pancreatic cancer. We investigated the signaling pathways activated in response to IL-6 in pancreatic cell lines, with a focus on STAT-3 and Pim-1 kinase. Methods: IL-6 receptor (IL-6R) expression was evaluated by Western blot analysis across a panel of human pancreatic cell lines. Panc-1 and MiaPaCa2 cell lines were serum starved for 24 hrs and then treated with 100 ng/mL of IL-6 for various time courses from 0 to 6 hr. STAT-3 activation and Pim-1 kinase protein levels were assessed in the presence and absence of IL-6 by Western blot analysis. The ability of cucurbitacin B to block both STAT-3 and Pim-1 activation, and inhibit cell proliferation was investigated. Stably transfected Pim-1 kinase over-expressing cell lines were generated and characterized for their response to IL-6 in vitro, and their growth rate in vivo as flank tumors in scid mice. Results: IL-6R is broadly expressed across multiple cancer cell lines, with the highest expression observed in the Panc-1 cell line. IL-6 stimulation resulted in increased STAT-3 phosphorylation in Panc-1, and MiaPaCa2 tumor cell lines as well as the HP-DEV transformed pancreatic ductal cell line. Treatment with IL-6 resulted in increased Pim-1 protein expression in the Panc-1 but not the MiaPaCa2 parent cell lines. Induction of Pim-1 protein was also observed in both Panc-1 and MiaPaCa2 stably transfected over-expressing cell lines. Pre-treatment with the STAT-3 inhibitor cucurbitacin B resulted in inhibition of STAT-3 phosphorylation and decreased cell proliferation, but was not associated with suppression of Pim-1 protein expression in the Panc-1 cell lines. Over-expression of Pim-1 in the Panc-1 cell line was associated with no significant changes in basal apoptosis rate, cell growth rate in vitro or in flank xenografts, or IL-6 induced expression of VEGF. Conclusions: IL-6 stimulation activates both STAT-3 and Pim-1 kinase expression in pancreatic cell line models. Induction of Pim-1 kinase was cell line specific, suggesting that the molecular context is essential for this pathway to be activated. Suppression of STAT-3 with a pharmacological inhibitor resulted in exaggerated Pim-1 expression in over-expressing cell lines implying Pim-1 up-regulation may be one of the compensatory and/or stress pathways activated when STAT-3 is inhibited. Stable over-expressing Pim-1 kinase Panc-1 and MiaPaCa2 cells did not exhibit significant changes in growth or apoptosis, unlike previously reported studies in the literature with other pancreatic cancer cell line models. This may be due to the multiple pathways that are dysregulated in these cell lines. Both STAT-3 and Pim-1 kinase have previously been identified as drug targets in multiple malignancies, including leukemia, lymphoma, prostate, and pancreatic cancer. Our study suggests that both pathways may be activated in patients with elevated IL-6 levels. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2930. doi:10.1158/1538-7445.AM2011-2930

BackgroundTo develop a novel therapeutic strategy for human pancreatic cancer using a midkine promoter-based conditionally replicating adenovirus.MethodsWe examined midkine mRNA expression and midkine protein expression by seven human pancreatic cancer cell lines (AsPC-1, BxPC-3, CFPAC-1, HPAC, MIAPaCa-2, PANC-1, and Suit-2), as well as by non-cancerous pancreatic tissue and pancreatic cancers. Midkine promoter activity was measured in cancer cell lines by the dual luciferase reporter assay. Adenoviral transduction efficiency was assessed by fluorescent staining of cancer cell lines using adenovirus type 5 containing the green fluorescent protein gene (Ad5GFP). Replication of adenovirus type 5 containing the 0.6 kb midkne promoter (Ad5MK) was assessed by the detection of E1 protein in cancer cell lines. The cytotoxicity of Ad5MK for cancer cells was evaluated from the extent of growth inhibition after viral infection. Infection and replication were also assessed in nude mice with subcutaneous Suit-2 tumors by intratumoral injection of Ad5MK, Ad5GFP, or vehicle. E1a mRNA expression in the treated tumors and expression of the replication-specific adenoviral hexon protein were evaluated. Finally, the anti-tumor activity of Ad5MK against intraperitoneal xenografts of Suit-2 pancreatic cancer cells was examined after intraperitoneal injection of the virus.ResultsBoth midkine mRNA expression and midkine protein expression were strong in AsPC-1 and CFPAC-1 cell liens, moderate in BxPC-3, HPAC, and Suit-2 cell lines, and weak in PANC-1 and MIAPaCa-2 cell lines. Expression of midkine mRNA was significantly stronger in pancreatic cancers than in non-cancerous pancreatic tissues. The relative luciferase activity mediated by the 0.6 kb midkne fragment in AsPC-1, PANC-1, and Suit-2 cell lines was approximately 6 to 20 times greater than that in midkne-negative MIAPaCa-2 cell lines. Pancreatic cancer cell lines exhibited a heterogeneous adenoviral transduction profile. E1A expression was higher in cell lines with strong midkine expression than in cell lines with weak midkine expression. Ad5MK showed much greater cytotoxicity for midkine-expressing Suit-2 and PANC-1 cell lines than for midkine-negative MIAPaCa-2 cell lines. In the Suit-2 subcutaneous xenograft model, expression of E1A was detected in Ad5MK-treated tumors, but not in untreated and Ad5GFP-treated tumors. In the Suit-2 intraperitoneal xenograft model, the Ad5MK group survived for significantly longer than the Ad5GFP, PBS, and untreated groups.ConclusionAd5MK has an anti-tumor effect against human pancreatic cancer cell lines that express midkine mRNA. Midkine promoter-based conditionally replicative adenovirus might be a promising new gene therapy for pancreatic cancer.

Two novel steroidal saponins, trilliumosides K (1) and L (2), were isolated from the rhizomes of Trillium govanianum led by bioactivity-guided phytochemical investigation along with seven known compounds: govanoside D (3), protodioscin (4), borassoside E (5), 20-hydroxyecdysone (6), 5,20-hydroxyecdysone (7), govanic acid (8), and diosgenin (9). The structure of novel compounds 1-2 was established using analysis of spectroscopic data including 1D and 2D nuclear magnetic resonance (NMR) and high resolution mass spectrometry (HR-ESI-MS) data. All isolated compounds were evaluated for in vitro cytotoxic activity against a panel of human cancer cell lines. Compound 1 showed significant cytotoxic activity against the A-549 (Lung) and SW-620 (Colon) cancer cell lines with IC50 values of 1.83 and 1.85 µM, respectively whereas the IC50 value of Compound 2 against the A-549 cell line was found to be 1.79 µM. Among the previously known compounds 3, 5, and 9, the cytotoxic IC50 values were found to be in the range of 5–10 µM. Comprehensive anti-cancer investigation revealed that Compound 2 inhibited in vitro migration and colony-forming capability in the A-549 cell line. Additionally, the mechanistic analysis of Compound 2 on the A-549 cell line indicated distinctive alterations in nuclear morphology, increased reactive oxygen species (ROS) production, and decreased levels of mitochondrial membrane potential (MMP). By upregulating the pro-apoptotic protein BAX and downregulating the anti-apoptotic protein BCL-2, the aforementioned actions eventually cause apoptosis, a crucial hallmark in cancer research, which activates Caspase-3. To the best of our knowledge, this study reports the first mechanistic anti-cancer evaluation of the compounds isolated from the rhizomes of T. govanianum with remarkable cytotoxic activity in the desired micromolar range.