Anticancer Screening Research Articles

Design, synthesis, and biological evaluation of dinaciclib and CAN508 hybrids as CDK inhibitors.

The scaffolds of two known CDK inhibitors (CAN508 and dinaciclib) were the starting point for synthesizing two series of pyarazolo[1,5-a]pyrimidines to obtain potent inhibitors with proper selectivity. The study presented four promising compounds; 10d, 10e, 16a, and 16c based on cytotoxic studies. Compound 16a revealed superior activity in the preliminary anticancer screening with GI % = 79.02-99.13 against 15 cancer cell lines at 10 μM from NCI full panel 60 cancer cell lines and was then selected for further investigation. Furthermore, the four compounds revealed good safety profile toward the normal cell lines WI-38. These four compounds were subjected to CDK inhibitory activity against four different isoforms. All of them showed potent inhibition against CDK5/P25 and CDK9/CYCLINT. Compound 10d revealed the best activity against CDK5/P25 (IC50 = 0.063 µM) with proper selectivity index against CDK1 and CDK2. Compound 16c exhibited the highest inhibitory activity against CDK9/CYCLINT (IC50 = 0.074 µM) with good selectivity index against other isoforms. Finally, docking simulations were performed for compounds 10e and 16c accompanied by molecular dynamic simulations to understand their behavior in the active site of the two CDKs with respect to both CAN508 and dinaciclib.

Improving the power of drug toxicity measurements by quantitative nuclei imaging

Imaging-based anticancer drug screens are becoming more prevalent due to development of automated fluorescent microscopes and imaging stations, as well as rapid advancements in image processing software. Automated cell imaging provides many benefits such as their ability to provide high-content data, modularity, dynamics recording and the fact that imaging is the most direct way to access cell viability and cell proliferation. However, currently most publicly available large-scale anticancer drugs screens, such as GDSC, CTRP and NCI-60, provide cell viability data measured by assays based on colorimetric or luminometric measurements of NADH or ATP levels. Although such datasets provide valuable data, it is unclear how well drug toxicity measurements can be integrated with imaging data. Here we explored the relations between drug toxicity data obtained by XTT assay, two quantitative nuclei imaging methods and trypan blue dye exclusion assay using a set of four cancer cell lines with different morphologies and 30 drugs with different mechanisms of action. We show that imaging-based approaches provide high accuracy and the differences between results obtained by different methods highly depend on drug mechanism of action. Selecting AUC metrics over IC50 or comparing data where significantly drugs reduced cell numbers noticeably improves consistency between methods. Using automated cell segmentation protocols we analyzed mitochondria activity in more than 11 thousand drug-treated cells and showed that XTT assay produces unreliable data for CDK4/6, Aurora A, VEGFR and PARP inhibitors due induced cell size growth and increase in individual mitochondria activity. We also explored several benefits of image-based analysis such as ability to monitor cell number dynamics, dissect changes in total and individual mitochondria activity from cell proliferation, and ability to identify chromatin remodeling drugs. Finally, we provide a web tool that allows comparing results obtained by different methods.

Exploration of isoxazole analogs: Synthesis, COX inhibition, anticancer screening, 3D multicellular tumor spheroids, and molecular modeling

In this study, a new series of Isoxazole-carboxamide derivatives were synthesized and characterized via HRMS, 1H-, 13CAPT-NMR, and MicroED. The findings revealed that nearly all of the synthesized derivatives exhibited potent inhibitory activities against both COX enzymes, with IC50 values ranging from 4.1 nM to 3.87 μM. Specifically, MYM1 demonstrated the highest efficacy among the compounds tested against the COX-1, displaying an IC50 value of 4.1 nM. The results showed that 5 compounds possess high COX-2 isozyme inhibitory effects with IC50 value in range 0.24–1.30 μM with COX-2 selectivity indexes (2.51–6.13), among these compounds MYM4 has the lowest IC50 value against COX-2, with selectivity index around 4. Intriguingly, this compound displayed significant antiproliferative effects against CaCo-2, Hep3B, and HeLa cancer cell lines, with IC50 values of 10.22, 4.84, and 1.57 μM, respectively, which was nearly comparable to that of doxorubicin. Compound MYM4 showed low cytotoxic activities on normal cell lines LX-2 and Hek293t with IC50 values 20.01 and 216.97 μM respectively, with safer values than doxorubicin. Furthermore, compound MYM4 was able to induce the apoptosis, suppress the colonization of both HeLa and HepG2 cells. Additionally, the induction of Reactive oxygen species (ROS) production could be the mechanism underlying the apoptotic effect and the cytotoxic activity of the compound. In the 3D multicellular tumor spheroid model, results revealed that MYM4 compound hampered the spheroid formation capacity of Hep3B and HeLa cancer cells. Moreover, the molecular docking of MYM4 compound revealed a high affinity for the COX2 enzyme, with energy scores (S) −7.45 kcal/mol, which were comparable to celecoxib (S) −8.40 kcal/mol. Collectively, these findings position MYM4 as a promising pharmacological candidate as COX inhibitor and anticancer agent.

Abstract LT07: Signalling Inhibition by Ponatinib Disrupts Productive Alternative Lengthening of Telomeres (ALT)

Abstract Enabling replicative immortality is one prominent feature of cancer cells. To that end, cancer cells activate means to elongate their telomeres. While 85% of cancers re-express telomerase, around 15% of all cancers rely on the ALT (Alternative Lengthening of Telomeres) mechanism. Despite its frequency, no clinically approved ALT-targeted therapy exists to date. By performing anti-cancer compound library screen on isogenic cell lines and using extrachromosomal telomeric C-circles, as a bona fide marker of ALT activity, we identified a receptor tyrosine kinase inhibitor, ponatinib, that deregulates ALT mechanisms, induces telomeric dysfunction, reduced ALT- associated telomere synthesis, and targets, in vivo, ALT-positive cells. By scrutinizing the mode of action of ponatinib on ALT, we identified an ABL1-JNK-JUN signalling circuit to be inhibited by ponatinib and to have a role in suppressing extrachromosomal telomeric C-circle formation. Furthermore, transcriptome and interactome analyses of JUN suggested a role of JUN in DNA damage repair pathways, independently of its capacity as a transcription factor. These results were corroborated by new synergistic drug interactions between ponatinib and either DNA synthesis or repair inhibitors such as triciribine and KU-60019, respectively. Overall, we identified a novel signalling pathway impacting ALT which can be targeted by a clinically approved kinase inhibitor. Citation Format: Frances Kusuma, Aishvaryaa Prabhu, Galen Tieo, Syed Moiz Ahmad, Pushkar Dakle, Wai Khang Yong, Elina Pathak, Vikas Madan, Yan Yi Jiang, Wai Leong Tam, Dennis Kappei, Peter Dröge, H. Phillip Koeffler, Maya Jeitany. Signalling Inhibition by Ponatinib Disrupts Productive Alternative Lengthening of Telomeres (ALT) [abstract]. In: Proceedings of Frontiers in Cancer Science; 2023 Nov 6-8; Singapore. Philadelphia (PA): AACR; Cancer Res 2024;84(8_Suppl):Abstract nr LT07.

Protease hydrolysed brewers’ spent grain bioactive extracts for applications in human health

About 39 million tons of Brewer’s spent grain (BSG), the main by-product of beer manufacturing, are produced annually and is used for low value applications. To valorise this rich bioresource, the present work entails bioprocessing of BSG with various proteases (Novozymes) at two different concentrations (4% and 9%) to solubilise protein, carbohydrate and polyphenols from the grain and evaluate its effect on gut and brain health. The results show the highest (p < 0.05) FRAP (Ferric Reducing Antioxidant Power) based antioxidant activity was obtained for Pro 5 (4%: 108.10 ± 4.17 µmole Trolox Equivalence (TE)/g protein, 18.06 ± 0.70 µmole TE/g d.w., which was 3.6 times higher than untreated control extracts. The highest DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activity was obtained for the same extract (Pro 5 at 4%: 0.118 ± 0.006 µg AAE/mg d.w., 9%: 0.110 ± 0.006 µg AAE/mg d.w). Proximate composition showed this extract to contain the highest concentration of proteins at 21.66% ± 2.71, and color analysis showed the same Pro 5 (4%) extract to be the darkest (L*53.73) indicating the possible presence of dark polyphenols. Anticancer screening showed Control and Pro 1 to possess cytotoxic effect against colon cancer cells with IC50 of 3.2 and 13.91 mg/mL, respectively. No significant activity was noted against the brain cancer cell line. Thus anti-colon cancer activity of BSG extracts highlights its potential in gut health. The observed bioactivity resulted from a combination of peptides, carbohydrates and polyphenolic compounds in the extract and warrants further characterisation for targeted nutraceutical applications.Graphical abstract

Abstract 6769: Applications of a novel fibrous extracellular matrix for advanced in vitro cancer models

Abstract Compared to traditional in vitro two-dimensional (2D) planar cell culture system, three-dimensional (3D) mammalian cell culture can better recapitulate physiological environments and the complexity of natural tissues, hence they are crucial in modern biomedical research. Extracellular matrices (ECM) play a key role in in vitro mammalian cell culture systems. The role of the ECM on in vitro 3D cell culture are: 1) to simulate an in vivo-like physiological environment, 2) support cell-cell and cell-ECM interactions, 3) promote cell adhesion, migration, and communication, 4) aid in tissue development, morphogenesis, differentiation, and functioning. Although basal membrane extracts (BME) are the most popular type of ECM supporting 3D modeling in vitro, they often have significant limitations including limited long-term stability, significant batch to batch heterogeneity, restricted cell movement, and relatively high cost for culturing many cancer cell lines. In this investigation, we employ a novel micro-fibrous scaffold that closely mimics natural ECM, providing porous and interconnected networks for 3D culturing of hepatic cells, lung cancer, breast cancer, pancreatic cancer. As revealed by optical and scanning electron microscopy, the scaffold promotes significant cell-to-cell and cell-to-matrix interactions. Moreover, we observed improved cell growth in 3D culture over longer periods than 2D culture systems for all cell types examined. We also evaluated the anticancer efficiency of different drugs including Lapatinib, Cisplatin, Docetaxel, Doxorubicin and observed that 2D cultured cells were more sensitive to drugs treatment in comparison to 3D. Furthermore, ECM was observed to support 3D growth and structure formation with cell pools of primary cells achieved from disassociated human tumors. Tumor organoids were able to form within seven days and ready for anticancer screening within 14 days. We achieved an 80% success rate in developing breast cancer organoids from patient tissues. Additionally, tissue granules were also able to be cultured under the same condition, which allowed us to investigate the invasion of cancer cells into tissue. The primary cells from human tumors formed cell clusters resembling the original tumor tissue's structure. In conclusion, the novel micro-fibrous scaffold, closely mimicking natural extracellular matrixes, promotes significant cell-to-cell and cell-to-matrix interactions, while also facilitating cell growth over extended periods providing a more physiologically relevant platform for evaluating drug responses for various cancer cell types, further showing the importance of 3D culture systems in cancer research. Citation Format: Navneet Kaur, Dazhi Yang, Jamel Ali. Applications of a novel fibrous extracellular matrix for advanced in vitro cancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6769.

Abstract 4701: A novel microtubule disruptor exerts broad anticancer efficacy with a tolerable safety profile

Abstract Initial evaluation of the prenylated hydroxy-stilbene isolated from bee propolis, AUS_001, in the NCI 60 human cell line anticancer screen showed significant growth inhibitory effects with the most robust being observed in leukemia, central nervous system and breast cancer cell lines. The aim of the current study was to further assess the potency of AUS_001 in vitro and investigate its safety profile and mechanism of action. A commercially available cell-based profiling screen (OncoPanel®) identified significant AUS_001-induced growth inhibition in 273 out of 280 cancer cell lines with a concentration causing 50% cell growth inhibition in the range of 0.021-0.94 µM, while non-neoplastic cells required an average of 20x higher concentration to generate similar efficacy. These findings indicate that AUS_001 exerts a high degree of potency across 30 cancer types with a selectivity for cancer versus normal cells. Maximum tolerated dose studies in mice illustrated that orally administered AUS_001 is well tolerated at doses 4 times greater than those that conferred antitumor efficacy, with no myelosuppression or other overt toxicities observed. Predictive toxicology and safety screening indicated that AUS_001 exerts a strong safety profile as evidenced by absence of mutagenic potential in the Ames test using up to 100 µM drug doses and low hERG-blocking liability with a half-maximal inhibitory concentration of 65 µM. Notably, AUS_001 is a poor substrate of the drug efflux transporter, P-glycoprotein 1 (Pgp), as it does not inhibit drug-stimulated Pgp ATPase activity and retains efficacy in high Pgp-expressing models in vitro. Subsequently, the effects of AUS_001 were explored in the ToxTracker® assay, consisting of a panel of mammalian stem cell lines that contain fluorescent reporters for the exploration of DNA damage, oxidative stress, and protein damage using flow cytometry. Robust activation of the Rtkn genotoxicity reporter under low cytotoxicity conditions, in combination with accumulation of cells in G2/M phases of the cell cycle within 4 h and an increase in aneuploid cells after 24 h of drug treatment, suggested an aneugenic mode of action, typically caused by tubulin-targeting agents or mitotic kinase inhibitors. Biochemical tubulin polymerization assays then revealed that AUS_001 acts as a direct tubulin destabilizing agent and monitoring microtubule dynamics in GFP-tubulin reporter cells provided evidence that AUS_001 blocks microtubule assembly. Our studies have elaborated the mechanism of AUS_001 as an inhibitor of tubulin polymerization. The favorable safety profile of AUS_001, along with its ability to circumvent Pgp-mediated multidrug resistance, provides potential for efficacy against multiple cancers where microtubule destabilization is proven to be an effective target. Citation Format: Herman Lelie, Inger Brandsma, Giel Hendriks, Lee R. Cavedine, Brogan A. Epkins, Steven M. Garner, Andrew J. Cook, Muthukrishnan Renganathan, Yong Zhao, Alastair J. King, Peter Y.W. Chan, April Risinger, Marina Koutsioumpa. A novel microtubule disruptor exerts broad anticancer efficacy with a tolerable safety profile [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4701.

Synthesis, characterization and anticancer screening of novel phenylbenzylidene thiosemicarbazone derivatives

Ten derivatives of phenylbenzylidene thiosemicarbazones (1–10) were synthesized by multicomponent reaction between hydrazine hydrate, isothiocyanate and substituted aldehydes in the presence of the phase transfer reagent Triton-B and subsequently heating the mixture. The synthesized compounds 1, 2, 5, 8 demonstrated to act as highly potent anticancer agents. They showed anticancer activity by mobilizing cellular 59Fe, inhibiting cellular uptake of 59Fe from 59Fe2-Tf, and mediating the ascorbate oxidation. Further, the structure activity relationship revealed that the donor atom and its stabilization resulted in better anticancer activity. The softness of the chelating donor atoms N and S yielded redox active iron complexes, which easily mediated ascorbate oxidation. This study identified the selective and potential chelators, which may act as anticancer agents and require further in vivo screening in future.

New Benzimidazole-Triazole Derivatives as Topoisomerase I Inhibitors: Design, Synthesis, Anticancer Screening, and Molecular Modeling Studies.

In this study, we designed, synthesized, and evaluated a series of 1,2,4-triazole benzimidazoles for their cytotoxic effects against the A549, C6, and NIH3T3 cell lines. Additionally, these compounds were assessed for their inhibitory activity against DNA topoisomerase I, aiming to develop novel anticancer agents. The synthesized final compounds 4a-h were characterized using 1H NMR, 13C NMR, and HRMS. Among them, compounds 4b and 4h emerged as the most potent agents against the A549 cell line, exhibiting an IC50 value of 7.34 ± 0.21 μM and 4.56 ± 0.18 μM, respectively. These results were compared to standard drugs, doxorubicin (IC50 = 12.420 ± 0.5 μM) and Hoechst 33342 (IC50 = 0.422 ± 0.02 μM). Notably, all tested compounds displayed higher cytotoxicity toward A549 cells than C6 cells. Compounds 4b and 4h demonstrated significant inhibitory activity against topoisomerase I, highlighting their potential as lead compounds in anticancer therapy. Subsequent in silico molecular docking studies were conducted to elucidate the potential binding interactions of compounds 4b and 4h with the target enzyme topoisomerase I. Molecular dynamics studies also assessed and validated the binding affinity and stability. These studies confirmed the promising binding affinity of these compounds, reinforcing their status as lead candidates. According to DFT, compound 4b having the lower energy gap value (ΔE = 3.598 eV) is more chemically reactive than the others, which is consistent with significant inhibitory activity against topoisomerase I. Furthermore, in silico ADME profiles for compounds 4b and 4h were evaluated using SwissADME, providing insights into their pharmacokinetic properties.

Click chemistry-based synthesis of new 1,2,3-triazolo-benzoquinoline-3-carbonitriles: anticancer screening and DFT studies

This study emphasizes the synthesis of a novel series of 1,2,3-triazolo-benzoquinoline-3-carbonitriles by using Cu(i)-catalyzed azide–alkyne cycloaddition known as the click reaction.

Green synthesis, in silico modeling, and biological evaluation of N-substituted (Z)-5-arylidene imidazolidine/thiazolidine-2,4-dione/4-thione derivatives catalyzed by Bu SO3H core-shell nanostructures.

In this effort, the immobilization of guanidine-sulfonate on the surface of Fe3O4 MNPs (magnetic nanoparticles) as a novel acid nanocatalyst has been successfully reported for the synthesis of N-substituted (Z)-5-arylidene thiazolidine-2,4-dione and related cyclic derivatives, including rhodanine (RHD) and hydantoin (HYD) via a one-pot multiple-component reaction under green conditions. The products were characterized by SEM, TEM, TGA, EDS, BET techniques, VSM, and FTIR. The novel compounds synthesized using this methodology, designated as series La (1-9), Lb (1-8), and Lc (1-8), were subjected to anticancer screening against A549 and MCF7cell lines via MTT assays. Notably, several compounds (L1a, L2a, L3a, L1b, L2b, L3b, and L4b) exhibited potent antiproliferative activities, with observed IC50 values as low as 1.23 μM and 1.02 μM against MCF-7 cells, thereby outperforming the established anticancer drugs doxorubicin and cisplatin. Molecular docking and dynamics simulations revealed that ligands L1a, L2a, and L3a strongly interact with the protein target 3CD8, with L1a displaying significant hydrophobic and hydrogen bonding interactions and L2a engaging in unique pi-pi stacking with key residues. For protein 2WGJ, ligand L4b exhibited exceptional binding affinity, characterized by robust hydrogen bonding, hydrophobic interactions, and additional stabilizing mechanisms such as water bridges and pi interactions. Hence, N-substituted (Z)-5-arylidene thiazolidine-2,4-dione and its cyclic derivatives may serve as promising candidates for further exploration in the development of new multi-target cancer chemotherapy agents. These findings introduce promising anticancer agents and establish Fe3O4 MNPs as a versatile and environmentally sustainable catalytic platform in drug discovery.

In Vivo Anti-Hepatocellular Carcinoma Effects of the Chloroform Root Extract of Clausena excavata Burm.

Liver cancer is the most common cancer among males in Africa. The disease has a poor prognosis and its treatment is associated with toxicity and resistance. For this reason, numerous herbal combinations are being subjected to anticancer screening to circumvent the shortcomings of the conventional anticancer drugs. In the current study, the in vivo anti-cancer effects of the chloroform root extract of the herb, Clausena excavata Burm were investigated. Liver cancer was induced in mice by a single intraperitoneal injection of diethylnitrosamine (DEN) followed by oral administration of the promoter of carcinogenesis, 2-aminoacetyl fluorine that was mixed with the mice feed. The cytotoxicity of the root extract of C. excavata on liver cancer cells was investigated using liver enzyme, histology, DNA fragmentation and caspases assays. Real time qPCR was conducted to evaluate the effect of the extract on apoptotic genes. The findings revealed that the extract of C. excavata significantly decreased the progression of hepatocarcinogenesis and the toxicity-induced production of the liver enzymes, alanine and aspartate aminotransferases. The histological analyses of the liver tissues revealed evidence of apoptotic cell death. The extract also provoked significant (p < .05) expressions of caspase 9 protein and gene as well as other apoptotic genes (P53, P27, Apaf-1, cytochrome C, bax and bid). Therefore, we postulate that the chloroform root extract of C. excavata induces apoptosis of liver cancer in mice.

Nickel oxide nanoparticles from Sargassumwightii: Synthesis,Characterization, and biomedical applications

Antimicrobial resistance and cancer claim millions of lives each year, making it important to find effective medications. The current study focuses on the rapid and sustainable manufacture of nickel oxide nanoparticles using Sargassum wightii aqueous extract (SW-NiO NPs) as a novel antibacterial and anticancer agent. The presence of phytochemicals in the Sargassumwightii in all extract was qualitatively screened and water extract, which is found to contain abundant phytochemical was used for the biosynthesis. Thus synthsized SW-NiO nanoparticles was characterised by UV–Visible, FT-IR, XRD, SEM, and TEM spectral studies, and then screend for its invitro anti microbial activity against bacterial strains (E. coli, K. pneumoniae, S. aureus, and B. subtilis) and fungal activity with the fungus strains (C. albicans, C. tropicalis),invitro anticancer and larvicidal activity. The SW-NiO NPs possessed cubic and hexagonal structures with an average crytallite size of 15–20 nm, according to the results of the analytical characterization. SEM and TEM analysis confirms the spherical like morphology and their respective elemental composition were identified by EDAX studies. The antimicrobial activity was carried by well diffusion method.Compared to gram-negative organisms, gram-positive species were more vulnerable to the SW-NiO NPs with the zone of inhibition 17 mm. The fungal activity results showed the zone of inhibition 16 and 14 mm. It was also found that SW-NiO NPs inhibit the growth of tested fungus at higher doses. The cytotoxicity studies with liver cancer cell line through MTT assay revealed the inhibition value at 52.25 ± 0.05 µM, indicating their sensitivity to the HepG2 cell line. SW-NiO NPs made with water extract showed the highest larvicidal activity and 88.33 % death of Aedes agypti(AE). The LC50 and LC90 values for SW-NiO NPs against Aedes agypti(AE) larvae are found to be 31.420 and 134.261 ppm, respectively. The findings of the antimicrobial, larvicidal and invitro anticancer screening of this work provide compelling evidence that SW-NiO NPs made from an aqueous extract of Sargassum wightii have considerable biological applications which can be explored in detail on identifying the potential phyto constituents which are responsible for the activity.

Anticancer, Immunomodulatory, and Phytochemical Screening of Carthamus oxyacantha M.Bieb Growing in the North of Iraq.

Carthamus oxyacantha M.Bieb is a promising repository of active phytochemicals. These bioactive compounds work synergistically to promote the plant's antioxidant, anticancer, and immunomodulatory capabilities. The present study aimed to discover the potential immunomodulatory and cytotoxicity of different extracts of Carthamus oxycantha roots. Aqueous ethanol (70%), aqueous methanol (90%), ethyl acetate, and n-hexane extracts were tested against five cell lines (T47D, MDA-MB231, Caco-2, EMT6/P, and Vero). Among these extracts, ethyl acetate and n-hexane extracts showed significant activity in inhibiting the proliferation of cancerous cells because of the presence of several phytochemical compounds, including flavonoids, phenolics, and alkaloids. The n-hexane extract was the most potent extract against T47D and Caco-2 cell lines and had IC50 values of 0.067 mg/mL and 0.067 mg/mL, respectively. In comparison, ethyl acetate extract was active against T47D and MDAMB231, and IC50 values were 0.0179 mg/mL and 0.03 mg/mL, respectively. Both n-hexane and ethyl acetate extracts reduced tumor size (by 49.981% and 51.028%, respectively). Remarkably, Carthamus oxyacantha extracts decreased the average weight of the tumor cells in the in vivo model. The plant induced significant apoptotic activity by the activation of caspase-3, immunomodulation of macrophages, and triggering of pinocytosis. The implications of these intriguing findings demand additional research to broaden the scope of the understanding of this field, opening the doors to the possibilities of using Carthamus oxyacantha M.Bieb as an effective cancer treatment adjuvant in the future.

Microtubule destabilising activity of selected 7-methoxy-2-phenylbenzo[b]furan derivative against primary and metastatic melanoma cells

Herein, we report the results of anticancer screening of two 2-phenylbenzo[b]furan derivatives functionalised at the 3-position with 4-hydroxy-3,5-dimethoxybenzoyl (BF2) or 3,4,5-trimethoxybenzoyl (BF3) against 60 different cancer cell lines. The results confirmed the anticancer potential of the tested compounds against different cancer cell types, especially colon cancer, brain cancer and melanoma. BF3 was defined as the most potent (also as a tubulin polymerisation inhibitor). Its anticancer activity against melanoma cell lines that originated from different stages, i.e., primary skin-derived A375 and metastatic WM9/MDA-MB-435S, was evaluated (as the clinical success of melanoma therapy strictly depends on the disease stage). Moreover, to determine the BF3 mode of action and its effect on cell proliferation, intracellular microtubule networks, cell cycle phase distribution and apoptosis were evaluated. Our study revealed that BF3 inhibited cell proliferation in a dose-dependent manner, with IC50 yielding 0.09 ± 0.01 μM, 0.11 ± 0.01 μM and 0.18 ± 0.05 μM for A375, MDA-MB435S and WM9, respectively. The strong antiproliferative activity of compound BF3 correlated well with its inhibitory effect on tubulin polymerisation. Molecular docking proved that BF3 belongs to the colchicine binding site inhibitors (CBSIs), and experimental studies revealed that it disturbs cell cycle progression leading to G2/M arrest and apoptosis.

Caspase-9 inhibition triggers Hsp90-based chemotherapy-mediated tumor intrinsic innate sensing and enhances antitumor immunity

BackgroundAntineoplastic chemotherapies are dramatically efficient when they provoke immunogenic cell death (ICD), thus inducing an antitumor immune response and even tumor elimination. However, activated caspases, the hallmark of most cancer...

A Novel Human Cell-Based Spheroid Model to Screen for Oncolytic Viruses and Oncolytic Viral Proteins

Colorectal cancer is one of the major causes of cancer-related deaths worldwide and surgery has been the ideal treatment method. Oncolytic virus therapy uses a modified virus to specifically kill only cancer cells without harming the neighboring non-cancerous cells. A best example is Newcastle Disease Virus (NDV), an avian paramyxovirus which is seen as a promising oncolytic virus shown to combat many different types of cancers by in vitro and few in vivo animal studies. Traditionally, anticancer screening is done through in vitro cell line based assays in the lab and promising candidates are further tested in animal tumor models which is more expensive, time consuming and may not always reflect the truth due to species variations. Further results from the in vitro cell culture system may not always predict the outcome in animal models. 3D cell based models, i.e. spheroids are better than the cell lines as they can mimic the microenvironment of a tumor and can be used as an alternative to animal testing. In this study we demonstrated a novel human colorectal cancer spheroid model, which mimics the tumor microenvironment and better emulates the pathophysiology of human colorectal cancer, as a tool for identification of oncolytic agents. Through cytotoxic and colony forming assays we show the potential application of the spheroid model as a high throughput enabled anticancer screen.

Synthesis, crystal structure, DNA binding, and anticancer activity of the cobalt(II), nickel(II), and copper(II) complexes of 9-benzothiazolanthrahydrazone

In the present study, we synthesized and extensively characterized a novel ligand, 9-THA, based on 9-benzothiazolanthrahydrazone, along with its three corresponding transition metal complexes: 9-THA-Co (1), 9-THA-Ni (2), and 9-THA-Cu (3). The crystallographic analysis revealed distinct coordination geometries - six-coordinate for 9-THA-Co and 9-THA-Ni, binding three and two 9-THA ligands, respectively, and four-coordinate for 9-THA-Cu, binding one 9-THA ligand. In vitro anticancer screening demonstrated that 9-THA-Cu exhibited significant growth inhibition in HeLa-229, NCI-H460, and SK-OV-3 cancer cells with IC50 values of 12.10 ± 0.54, 12.12 ± 0.58, and 17.65 ± 0.66 μM, respectively. Moreover, it displayed comparable cytotoxicity to cisplatin on HL-7702 normal liver cells. In contrast, 9-THA and 9-THA-Co/9-THA-Ni did not exhibit significant growth inhibition in the tested cancer cells, suggesting that Cu(II) played a pivotal role in its anticancer mechanism. Further investigation revealed the intercalative DNA binding mode of 9-THA-Cu by UV–Vis and fluorescent spectral analyses. It also exhibited competitive binding with two typical DNA intercalators, Gel-red (GR) and ethidium bromide (EB). Cell cycle and cell apoptosis experiments indicated that 9-THA-Cu at a concentration of 22 μM could induce cell cycle arrest in the S phase in 40 % of HeLa-229 cancer cells. Simultaneously, it induced 33.2 % apoptosis in HeLa-229 cells, suggesting that 9-THA-Cu may employ multiple mechanisms to exert its anticancer effects.

In Vitro anticancer screening of herbal syrup formulation of podophyllum (May Apple) extract

The in vitro anticancer screening of the medicated herbal formulation of podophyllum as reported earlier. The formulation of herbal syrup was prepared and optimized as per industrial practice guidelines and was subjected to in vitro anticancer assay against a A549-Human lung cancer cell line and MCF7-Huam Breast cancer cell line. The LC50, TGI and GI50 are reported here against standard Adriamycin used in the test. The result showed comparable activity of the herbal syrup of podophyllum extract.

In Vitro anticancer screening of herbal syrup formulation of allium cepa extract

The formulation of herbal syrup of extract of Allium Cepa was reported in the previous paper. The herbal syrup of extract was evaluated here for anticancer activity. The anticancer activity was evaluated against 2 cell lines namely human lung cancer cell line A-549 and human breast cancer cell line MCF7. The results indicate that there is no significant anticancer activity of the herbal syrup formulation of the extract as compared with the standard Adriamycin.