Normal Lung Fibroblast Cell Line Research Articles

Production of a Polyketide Pigment by Fusarium chlamydosporum

The present study was focused to study the production of secondary metabolite by the fungus, F. chlamydosporum on a non-defined medium with less concentration of nitrogen; the organic nitrogen source being peptone and beef extract. In this context, we have been successful in extracting a polyketide pigment from the fungus by using the homogenization technique. The pigments thus extracted were subjected to various purification techniques via thin layer chromatography, column chromatography, UV-visible spectrophotometry, fourier-transform infrared spectroscopy and finally molecular determination by liquid chromatography coupled to tandem quadrupole mass spectrometry (LC-MS-MS). The polyketide red pigment was finally characterized and identified to be fusarubin following which its cytotoxicity was evaluated in vitro by using normal lung fibroblast cell lines (MRC-5). In the verge of researchers identifying novel compounds for various applications, the production of fusarubin by the fungus can be a major breakthrough as fusarubin has been investigated to exhibit many pharmacological activities. Though fusarubin is reported to be produced by other Fusarium species, this is the foremost study on the production of fusarubin by F. chlamydosporum; the composition of the culture medium is also unique. The production of this polyketide probably correlates in the pathogenesis of F. chlamydosporum as studies comment on this fungus as an opportunistic pathogen.

Zinc(II) Complexes with Dimethyl 2,2′-Bipyridine-4,5-dicarboxylate: Structure, Antimicrobial Activity and DNA/BSA Binding Study

Two zinc(II) complexes with dimethyl 2,2′-bipyridine-4,5-dicarboxylate (py-2py) of the general formula [Zn(py-2py)X2], X = Cl− (1) and Br− (2) were synthesized and characterized by NMR, IR and UV-Vis spectroscopy and single-crystal X-ray diffraction analysis. Complexes 1 and 2 are isostructural and adopt a slightly distorted tetrahedral geometry with values of tetrahedral indices τ4 and τ’4 in the range of 0.80–0.85. The complexes were evaluated for their in vitro antimicrobial activity against two bacterial (Pseudomonas aeruginosa and Staphylococcus aureus) and two fungal strains (Candida albicans and Candida parapsilosis), while their cytotoxicity was tested on the normal human lung fibroblast cell line (MRC-5) and the model organism Caenorhabditis elegans. Complex 1 showed moderate activity against both Candida strains. However, this complex was twofold more cytotoxic compared to complex 2. The complexes tested had no effect on the survival rate of C. elegans. Complex 2 showed the ability to inhibit filamentation of C. albicans, while complex 1 was more effective than complex 2 in inhibiting biofilm formation. The interactions of complexes 1 and 2 with calf thymus DNA (ct-DNA) and bovine serum albumin (BSA) were studied to evaluate their binding affinity toward these biomolecules.

Design, synthesis and anticancer activity studies of novel 4-butylaminophenyl hydrazide-hydrazones as apoptotic inducers

In this study, a series of the novel Tetracaine derivatives bearing hydrazide-hydrazone moiety were designed, synthesized, and evaluated for anticancer activity. The structures of these compounds were characterized by spectral (1H–13C NMR, FT-IR, and HR-MS analyses) methods. All synthesized compounds (2a-l) were screened for anticancer activity against human hepatocellular carcinoma (HepG2) and lung carcinoma (A549) cell lines. Against HepG2 and A549 cell lines, among the synthesized compounds, 4-(Butylamino)-N'-[(2,4-dichlorophenyl)methylidene]benzohydrazide (2i) demonstrated the most potent anticancer activity with IC50 values 28 and 7 μM, respectively. Possible cytotoxic effects of compounds (2a-l) on both normal human lung fibroblast (MCR-5) and normal human dermal fibroblast (HDF) cell lines were assessed. Inhibition of anti-apoptotic protein Bax and Bcl-2 was investigated in HepG2 and A549 cells treated with compound 2i using qRT-PCR. Apoptosis was also detected by Annexin V studies. The flow cytometric analysis results showed that compound 2i treatment of HepG2 and A-549 cells significantly increased apoptotic cell populations while decreasing viabilities in these carcinomas in a dose-dependent manner after 72 h of incubation.

Proteomic Analysis Reveals Differential Expression Profiles in Idiopathic Pulmonary Fibrosis Cell Lines.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, irreversible lung disorder of unknown cause. This disease is characterized by profibrotic activation of resident pulmonary fibroblasts resulting in aberrant deposition of extracellular matrix (ECM) proteins. However, although much is known about the pathophysiology of IPF, the cellular and molecular processes that occur and allow aberrant fibroblast activation remain an unmet need. To explore the differentially expressed proteins (DEPs) associated with aberrant activation of these fibroblasts, we used the IPF lung fibroblast cell lines LL97A (IPF-1) and LL29 (IPF-2), compared to the normal lung fibroblast cell line CCD19Lu (NL-1). Protein samples were quantified and identified using a label-free quantitative proteomic analysis approach by liquid chromatography-tandem mass spectrometry (LC-MS/MS). DEPs were identified after pairwise comparison, including all experimental groups. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Protein–Protein Interaction (PPI) network construction were used to interpret the proteomic data. Eighty proteins expressed exclusively in the IPF-1 and IPF-2 clusters were identified. In addition, 19 proteins were identified up-regulated in IPF-1 and 10 in IPF-2; 10 proteins were down-regulated in IPF-1 and 2 in IPF-2 when compared to the NL-1 proteome. Using the search tool for retrieval of interacting genes/proteins (STRING) software, a PPI network was constructed between the DEPs and the 80 proteins expressed exclusively in the IPF-2 and IPF-1 clusters, containing 115 nodes and 136 edges. The 10 hub proteins present in the IPP network were identified using the CytoHubba plugin of the Cytoscape software. GO and KEGG pathway analyses showed that the hub proteins were mainly related to cell adhesion, integrin binding, and hematopoietic cell lineage. Our results provide relevant information on DEPs present in IPF lung fibroblast cell lines when compared to the normal lung fibroblast cell line that could play a key role during IPF pathogenesis.

Structural characterization and antimicrobial evaluation of chromium(III) and cobalt(III) complexes with 2,2-diMe-1,3-pdta: Tuning dimensionality of coordination polymer and the water content by alkyl substitution

Hexadentate 2,2-dimethyl-1,3-propanediamine-N,N,N’,N’-tetraacetate (2,2-diMe-1,3-pdta) ligand, containing two methyl substituents at the central carbon atom of a 1,3-propanediamine, has been prepared and used for the synthesis of Na[Cr(2,2-diMe-1,3-pdta)]·3.75H2O (1) and Na[Co(2,2-diMe-1,3-pdta)]·3.88H2O (2) complexes. These complexes were characterized by IR and electronic absorption spectroscopy, and single-crystal X-ray diffraction analysis. NMR (1H and 13C) spectroscopy was additionally applied for the characterization of complex 2. Crystallographic data indicate that the two investigated crystals are isostructural and contain 2,2-diMe-1,3-pdta ligand coordinated to metal ion through 2N and 4O atoms forming an octahedral complex in which the six-membered 1,3-propanediamine chelate ring adopts a twist-boat conformation. There are four such complex anions in the symmetry independent part of the unit cell. Each complex anion is further connected to the sodium counterion(s) via the bridging carboxylate group(s). Structural changes in 2,2-diMe-1,3-pdta-Cr(III) complex stimulated solely by the presence of alkyl side groups are discussed. The present study shows that in 1,3-pdta-type complexes of Cr(III) and Co(III), the environment at coordination centre can be modified by introducing substitution in one of the carbon atoms of the diamine and the resulting difference in the subunit structure can bring about noticeable change in molecular and crystal structure. The examples illustrate the importance of the steric effect for the fine tuning of the dimensionality of the resulting coordination polymer and the water content. The antimicrobial activity of complexes 1 and 2 was evaluated against different bacterial and Candida spp., while their cytotoxic effects were tested on the normal human lung fibroblast cell line (MRC-5).

Synthesis, Characterization, Biological Activity and Molecular Docking Studies of Novel Organotin(IV) Carboxylates.

Four new carboxylates complexes with general formula R2SnL2 and R3SnL, where R = n-butyl (1, 3), methyl (2, 4) and L = 4-Chlorophenoxyacetate, were synthesized in significant yields. FT-IR analysis revealed a chelating (1 and 2) and a bridging bidentate (3 and 4) coordination modes for the carboxylate ligand in solid state which was further confirmed by the single crystal X-ray analysis of complex 4. The NMR data (1H, 13C and 119Sn) revealed a higher coordination number around the tin center in R2SnL2 (1 and 2) compared to R3SnL (3 and 4). A close matching was observed between the experimental and calculated structures (obtained at B3LYP/6-31G* + LANL2DZ basis set). Quantum chemical analysis indicates that the carboxylate moiety has the major contribution in the formation of filled and unfilled orbitals as well as in ligand to ligand intramolecular charge transfer during the electronic transitions. The cytotoxicity data of the screened compounds evaluated against lung cancer cell line (A549) and normal lung fibroblast cell line (MRC-5) revealed that 1, 3 and 4 have shown dose dependent cytotoxic effects while HL and 2 have shown steady and low cytotoxic activities. The antibacterial activity of complexes 1–4 is higher than that of HL. Molecular docking study showed an intercalation binding mode for complex 3 with DNA (docking score = −3.6005) involving four polar interactions. Complex 3 docking with tubulin (PDB ID 1SA0) with colchicine as a target protein resulted in three polar interactions (docking score −5.2957). Further, the docking analysis of the HL and 1–4 has shown an adequate interactions with the coronavirus SARS-CoV-2 spike protein, nucleocapsid protein and human angiotensin converting enzyme (ACE2).

Structure-Based Discovery and Optimization of Furo[3,2-c]pyridin-4(5H)-one Derivatives as Potent and Second Bromodomain (BD2)-Selective Bromo and Extra Terminal Domain (BET) Inhibitors

Pan-bromodomain and extra terminal (Pan-BET) inhibitors show profound efficacy but exhibit pharmacology-driven toxicities in clinical trials. The development of domain-selective BET inhibitors to separate efficacy and toxicity is urgently needed. Herein, we report a series of furo[3,2-c]pyridin-4(5H)-one derivatives as novel BD2-selective BET inhibitors. The representative compound 8l (XY153) potently bound to BRD4 BD2 with an half-maximum inhibitory concentration (IC50) value of 0.79 nM and displayed 354-fold selectivity over BRD4 BD1. Besides, 8l exhibited 6-fold BRD4 BD2 domain selectivity over other BET BD2 domains. Compound 8l displayed potent antiproliferative activity against multiple tumor cell lines, especially MV4-11 (IC50 = 0.55 nM), while showing weak cytotoxicity against the normal lung fibroblast cell line. It highlights the safety profile of this series of BD2 inhibitors. 8l also demonstrated good metabolic stability in vitro. These data indicate that 8l may serve as a new and valuable lead compound for the development of potential therapeutics against acute myeloid leukemia (AML).

DNA damage induced cellular senescence and it's PTEN-armed exosomes-the warriors against prostate carcinoma cells.

Exosomes are one type of small extracellular vesicles (EVs) having a size range of 30-150nm and secreted by the endosomal compartment of most eukaryotic cells. It has been found that exosomes (EVs) can serve as a communicating vehicle to transfer information among cells and thus can be associated with numerous physiological and pathological functions. In this study, we have isolated exosomes (EVs) from two different human cancer cell lines. Isolated exosomes (EVs) were characterized by scanning electron microscopy, nanoparticle tracking analysis, DLS, and by western blotting. It was observed that exosomes (EVs) isolated from mock-treated human lung epithelial carcinoma (A549) cells or HeLa cells exerted growth arrest to the human prostate carcinoma (PC3) cells, but no growth arrest was observed in case of normal human lung fibroblast cell line (WI-38). Additionally, exosomes (EVs) isolated from PC3 cells have no effect on PC3 cells. This is also true for exosomes (EVs) isolated from H2O2-induced senescent human lung cancer cells (A549). Analysis of exosome (EVs) content by western blotting reveals the presence of PTEN in the exosome (EVs) of lung cancer cells. Functional analysis of PTEN pathways in PC3 cells indicates the inactivation of Akt in exosome (EV)-treated cells. Therefore, from our study we have concluded that exosomes (EVs) secreted from A549 cells which contain functional PTEN may be used for delivery of PTEN to cancer cells without functional PTEN.

Cytotoxic Proteins from King Tuber Oyster Medicinal Mushroom, Pleurotus tuber-regium (Agaricomycetes), Sclerotium against Human MDA-MB-231 Breast Cancer Cells.

Numerous studies have reported the vast medicinal values of proteins from mushrooms. The present study aimed to investigate the potential of protein extracts from the sclerotium of Pleurotus tuber-regium (Fr.) Singer for antitumor activities against a breast cancer cell line. Protein from P. tuber-regium sclerotium was fractionated using ammonium sulphate at concentrations of 30%, 60%, and 90% and designated as PS30, PS60, and PS90, respectively. All protein extracts were assessed for cytotoxicity toward breast cancer cell line MDA-MB-231 and normal lung fibroblast cell line MRC-5 in the MTT assay. The ability of the protein extracts to inhibit cellular migration was evaluated using the antimigration assay. The most promising protein extract against MDA-MB-231 cells was PS60, with a half-maximal inhibitory concentration of 0.75 ± 0.57 μg/mL and a selectivity index of 14.00. Cytotoxicity and antimigration effects on cancer cells were best exhibited by PS60, with absolute migration capability values between 5.4142 ± 0.6916 and 5.6581 ± 0.2015 nm/h. PS60 was shown to exert cytotoxic effects associated with the induction of apoptosis and cell cycle arrest in MDA-MB-231 cells at G1/G0 and S phase. In conclusion, PS60 protein of P. tuber-regium sclerotium has good potential to be developed into a novel antitumor drug against breast cancer.

Design, synthesis, cytotoxic and enzyme inhibitory activities of 1,3,4-oxadiazole and 1,3,4-thiadiazine hybrids against non-small cell lung cancer

Certain new 1,3,4-oxadiazole and 1,3,4-thiadiazine hybrids attached through thioacetamide linker (3a-p) were synthesized and tested in vitro for their cytotoxic activities towards a non-small cell lung cancer (NSCLC) cell line A549. The results indicated that eight compounds (3b, 3d, 3e, 3f, 3 g, 3 h, 3 k and 3n) exhibited more potent activity than the standard drug doxorubicin with IC50 range of 0.44 ± 0.02 – 2.84 ± 0.14 µM. Compound 3 g which displayed the highest cytotoxicity caught our interest for further investigation. It arrested the cell cycle at G2/M phase and showed proapoptotic action as reflected by Annexin-V-FITC assay. The activities of 3 g against certain enzymes which are reported to be overexpressed in NSCLC were studied. Promising inhibitory activities were demonstrated against vascular endothelial growth factor receptor2 (VEGFR2) (IC50 = 0.268 ± 0.006 µM), B-RAF (IC50 = 0.176 ± 0.009 µM), matrix metalloproteinases (MMPs) MMP1 (IC50 = 0.277 ± 0.006 µM) and MMP9 (0.084 ± 0.002 µM) with respect to corresponding reference drugs. Moreover, 3 g inhibited Telomerase activity by 36.55%. Docking study showed that 3 g bound to these target enzyme active sites similar to the co-crystallized ligands. Also, 3 g was more selective to A549 than WI-38 (a normal human fetal lung fibroblast cell line), with selectivity index (SI) = 59.79. In conclusion, 3 g is a promising multitarget anticancer compound which targets key pathways in NSCLC and requires further investigation.

CD44-targeted, indocyanine green-paclitaxel-loaded human serum albumin nanoparticles for potential image-guided drug delivery

The use of multifunctional nanomedicines for image-guided drug delivery is currently being universally evaluated as a means of efficiently managing cancers and other diseases. In this study we evaluated the potential of an indocyanine green (ICG) and paclitaxel (PTX) loaded human serum albumin (HSA) nanoparticles that was conjugated with hyaluronic acid for use in image-guided drug delivery targeted to CD44-positive non-small cell lung cancer (NSCLC). Series of NSCLC cell lines were evaluated for the expression of CD44 using both western blot analysis and qRT-PCR and compared to a normal lung fibroblast cell line (MRC-5). Using Fluorescence microscopy and photoacoustic imaging (PA), we explored the ability of these targeted nanoparticles to selectively accumulate in NSCLC cell lines in comparison to MRC-5 and their potential for biomedical imaging towards their use for theranostic application. Results obtained suggest that these targeted nanoparticles have potential for application in both imaging and treatment of NSCLC.

New mixed ligand copper(II) hydrazone‐based complexes: Synthesis, characterization, crystal structure, DNA/RNA/BSA binding, in vitro anticancer, apoptotic activity, and cell cycle analysis

AbstractFour new mixed ligand copper(II) complexes with the general formula [Cu(L)(B)]n (1)–(4) (H2L = dehydroacetic acid benzoyl hydrazone Schiff base, B = H2O (1), DMSO (2), imidazole (imz) (3), n = 1) or pyridine (py) (4), n = 2) were synthesized. Characterization was performed by elemental analyses, FTIR, 1H NMR, electron paramagnetic resonance (EPR), ESI‐mass, and UV–visible) spectroscopy, together with magnetic susceptibility and molar conductivity measurements. The molecular structure of (2) and (4) complexes was investigated by X‐ray crystallography. The ligand behaves as a di‐basic tridentate ONO donor, coordinating to copper(II) center via deprotonated hydroxyl group, azomethine nitrogen, and deprotonated amide oxygen in a square planar (1)–(3) /square pyramidal geometry (4). The intercalation binding mode of the compounds with calf thymus DNA (ct DNA) and yeast (tRNA) and strong static interaction with bovine serum albumin (BSA) protein have been evaluated by absorption and emission spectroscopy. Further, in vitro cytotoxic activity of the compounds was examined on three human cancer cell lines; breast cancer (MCF7), colon cancer (HCT116), liver cancer (HepG2), and a normal lung fibroblast cell line (WI38) using cisplatin as a standard. Based on IC50 and therapeutic indices (TI), complexes (3) and (4) showed better activity than that of cisplatin. So both complexes were subjected for further studies viz, DNA fragmentation, cell apoptosis (annexin), and cell cycle analysis. They induced DNA fragmentation, and the HCT116 and HePG2 cell death were induced using (3) and (4) complexes, respectively, by apoptosis and cell cycle arrest at the G2/M phase.

Synthesis and characterization of a mononuclear nickel(II) complex with N,O-donor ligand: Its DNA/HSA protein binding properties and tumor suppressive function

A mononuclear Ni(II) complex, ([Ni(HL)2]Cl2•H2O) (Complex 1), where HL = 1-((2-piperidin-1-yl)ethylimino)methyl)naphthalene-2-ol has been synthesized under mild conditions. It has been characterized by elemental analysis, cyclic voltammetry, FT-IR, UV-vis, fluorescence and mass spectral analyses. Square planar geometry of the mononuclear complex has been confirmed by single crystal X-ray diffraction analysis. Complex 1 interacts with CT DNA with a binding constant of 3.6 × 103 M‒1 which suggests electrostatic interaction between the complex and DNA. The metal complex cleaves plasmid DNA efficiently in the presence of H2O2. On the other hand, complex 1 binds with the carrier protein, HSA, quite efficiently with a binding constant value of 7.16 × 105 M‒1. Cytotoxicity of the complex has been checked with lung carcinoma (A549) and normal lung fibroblast cell lines (WI-38). From MTT assay, it has seen that the complex could inhibit the cell proliferation. Western blot analysis suggests that nickel complex is a tumor suppressor.

Design, synthesis and biological evaluation of novel thiohydantoin derivatives as antiproliferative agents: A combined experimental and theoretical assessments

In our endeavors to develop potent anticancer agents, novel two series of 3,5-disubstituted-2-thiohydantoins 5a–c and the S-methyl counterparts 6a–c were designed and synthesized. The structural modifications were based on some reported thiohydantoins derivatives which showed promising antiproliferative activity. The activity of the compounds was verified via the in vitro cytotoxicity assay against prostate (PC-3) and breast (MCF-7) cancer cell lines along with normal lung fibroblast cell line (WI-38) to determine their safety on the normal cells. The tested compounds 5a–c and 6a–c showed promising cytotoxicity with IC50 ranges from 1.980 to 19.089 µM and from 1.619 to 16.976 µM against PC-3 and MCF-7, respectively. Interestingly, compound 6c is the most potent derivative against both PC-3 and MCF-7 (IC50 = 1.980 and 1.619 µM, respectively). Compound 5c is the second most potent derivative against PC-3 and MCF-7(IC50 = 2.385 and 2.084 µM, respectively). The antiproliferative activities on the normal lung fibroblast cell line (WI-38) implied that the most potent cytotoxic derivatives 5c and 6c are safe on the healthy cells. The docking solutions of 5c and 6c into the active site of TOP1 inferred comparable binding interactions to the co-crystallized ligand with docking scores = - 6.859 and 6.939 Kcal/mol, respectively. The computational studies using DFT calculations with B3LYP/6-31+G (d,p) level were performed to study the electronic and geometric properties obtained from the stable structure of the investigated compounds. A good correlation was found between the quantum chemical parameters and experimental observations. Therefore, these promising thiohydantoins can be subjected for further optimization to develop more potent counterparts.

Abstract 1006: Combination therapy with MEK inhibitors and a novel anti-neoplastic drug, imipridone ONC212, demonstrates synergy in pancreatic ductal adenocarcinoma cell lines

Abstract Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with limited therapeutic options. Although KRAS is mutated or amplified in nearly 95% of PDAC, no effective KRAS directed therapy has entered clinical practice. There is emerging data suggesting that inhibition of downstream targets of KRAS, such as MEK or ERK, increases autophagic flux and dependence in KRAS-mutated tumors. Autophagy, a catabolic process used by many tumors to maintain viability, can be targeted with various agents such has hydroxychloroquine and chloroquine, and recent preclinical data has shown that the combination of these agents with inhibitors of MEK/ERK can synergistically induce cell death. Using viability assays, we established sensitivity of multiple KRAS-mutated and KRAS wild-type pancreatic cancer cell lines, BxPC3, PANC1, HPAF-II, AsPC-1 to ONC212, trametinib, chloroquine and combinations of these agents. Results were analyzed after 72 hours of incubation using Compusyn and Combenefit. We also performed Western Blotting on cell lysates after treatment for 48 hours with these agents. We found that trametinib exhibited strong synergy with both chloroquine and ONC212 in all cell lines. The synergistic potential appeared to be most profound in those cell lines treated with ONC212 and trametinib. We used MRC5, a normal lung fibroblast cell line, as a control and found that these combinations did not significantly induce cell death. Using Western Blotting, we assessed markers of autophagy, including Beclin-1 and LC3B, as well as key second messenger pathway activation/suppression with p-AKT/AKT and p-ERK/ERK. At increasing doses, trametinib increased conversion of LC3-I to LC3-II, more notably in KRAS mutant cell lines, suggesting increased autophagic flux. ONC212, however, reduced conversion of LC3-I to LC3-II and increased degradation of Beclin-1 suggesting inhibition of autophagy. The combination, did not appear to have as striking of an effect on this pathway. In addition, p-ERK, as expected, was downregulated with MEK inhibition. ONC212 also appeared to reduce p-ERK levels and, interestingly, increased p-AKT. This suggests possible PI3K pathway upregulation, however, this could be influenced by timing and dose of treatments. Western blot analysis also revealed increased cPARP/PARP with combination therapy across cell lines. In conclusion, the combination of trametinib and ONC212 produces a synergistic cytotoxic effect on tumor cells at doses that are non-toxic to normal cells. In KRAS-mutated pancreatic cancer cell lines, we hypothesize that this may, in part, be due to autophagy suppression by ON212. Similar synergistic effects were noted in a KRAS wild-type pancreatic cancer cell lines, suggesting an autophagy-independent mechanism. Thus, this combination may represent a potential therapeutic modality for PDAC. Citation Format: Alexander G. Raufi, Casssandra Parker, Lanlan Zhou, Varun V. Prabhu, Josh Allen, Wafik S. El-Deiry. Combination therapy with MEK inhibitors and a novel anti-neoplastic drug, imipridone ONC212, demonstrates synergy in pancreatic ductal adenocarcinoma cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1006.

Oridonin improves the therapeutic effect of lentinan on lung cancer

Oridonin, a compound from Rabdosia rubescens, has been shown to exhibit a potent ability to improve the antitumor effects of lentinan (LNT). In the present study, the effects of oridonin, LNT, and the combination of these treatments were assessed on the normal human fetal lung fibroblast cell line MRC-5, as well as the non-small cell lung cancer cell line A549. Next, their effects on metastasis and survival in vivo were assessed in a mouse model of lung cancer. The effects of the treatments on the mRNA and protein expression levels of several regulatory factors in A549 cells and lung tissues were determined using reverse transcription-quantitative PCR and western blotting. The results showed that the viability of MRC-5 and A549 cells were not affected by 0-20 µg/ml oridonin; 0-300 µg/ml LNT did not affect the viability of MRC-5 cells, but 50-400 µg/ml LNT reduced the viability of A549 cells. Thus, 20 µg/ml oridonin and 100 or 300 µg/ml LNT were used in the subsequent experiments. Treatment with oridonin and LNT, alone or combined, had no effect on MRC-5 cell viability. Oridonin treatment had no effect on A549 cell viability; however, LNT suppressed A549 cell viability, and oridonin promoted the suppressive effects of LNT on A549 cells. In vivo analysis showed that oridonin alone had no effect on metastasis and survival, but LNT decreased metastasis and survival in mice. Oridonin augmented the effects of LNT against metastasis and further improved the survival rates of mice. In both A549 cells and lung tissues, LNT increased the mRNA and protein expression levels of caspase-3, caspase-8, caspase-9, Bax, p53, p21 and inhibitor of nuclear factor-κB (NF-κB)-α, and reduced the mRNA and protein expression levels of Bcl-2 and NF-κB. Oridonin augmented all the effects of LNT on expression of these proteins in the cells. Together, the results showed that oridonin enhanced the antitumor effects of LNT, and may thus serve as an adjuvant alongside LNT as a novel anticancer regimen for treatment of lung cancer.

Assessment of anti-hemolytic, cytotoxicity, antioxidant activities and molecular docking study based on thienopyrazole scaffold as pharmacophore

Thienopyrazole compound was used as a key synthon for a one- two- and three- pot component synthesis of new fused heterocyclic moieties which are known to have different biological activities. The investigated compounds were previously synthesized by environmentally benign techniques besides the conventional heating. The current study focused on biological evaluation of cytotoxicity, anti-hemolytic and antioxidant activities of some thienopyrazole compounds. The results showed the strongest anti-hemolytic and antioxidant activities are compounds 3a and 14c, respectively. All compounds were examined against normal human lung fibroblast cell line (WI-38) and four human cancer cell lines as human prostate cancer (PC-3), Hepatocellular carcinoma (HEPG-2), Colorectal carcinoma (HCT-116) and Epitheliod carcinoma (Hela). Molecular docking was performed inside the active site of P53 for compounds 1, 2, 11b and 14c. Moreover, quantum chemical calculations were performed on the selected compounds. Both techniques suggested that compound 1 displays high binding affinity in the P53 active site with respect to compound 2 while compound 14c owns a lower affinity with respect to 11b thereby suggesting that compound 11b may possibly act as P53 inhibitor and thus may participate in anticancer activity.

Synthesis of (4R)‐2‐(3‐hydroxyphenyl)thiazolidine‐4‐carboxylic acid substituted phthalocyanines: Anticancer activity on different cancer cell lines and molecular docking studies

In this study, firstly, (4R)‐2‐(3‐hydroxyphenyl)thiazolidine‐4‐carboxylic acid (1) and (4R)‐2‐(3‐(3,4‐dicyanophenoxy)phenyl)thiazolidine‐4‐carboxylic acid (2) were prepared. Then, the novel type metallophthalocyanines (ZnPc (3), CuPc (4), and CoPc (5)) bearing thiazolidine groups in peripheral positions were synthesized using by compound (2). The synthesized new compounds (1–5) were characterized by the combination of standard spectroscopic methods such as FT‐IR, 1H NMR, 13C NMR, UV–Vis spectral data, and MALDI‐TOF. Aggregation behaviors of peripheral tetra‐substituted metallophthalocyanines were investigated in dimethyl sulfoxide (DMSO) media. Fluorescence properties and fluorescence quantum yield of the new type zinc phthalocyanine (3) were performed in DMSO at room temperature. The anticancer activity of novel type metallophthalocyanines bearing thiazolidine groups in peripheral positions were investigated on rat glioma cancer (C6), human prostate carcinoma (DU‐145), and normal human lung fibroblast (WI‐38) cell lines. Finally, the biological and chemical activities of (4R)‐2‐(3‐(3,4‐dicyanophenoxy)phenyl)thiazolidine‐4‐carboxylic acid (2) and its novel type metallophthalocyanines (ZnPc (3), CuPc (4), and CoPc (5)) have been compared with many parameters obtained using theoretical methods that are the Gaussian software and molecular docking.

The Antimicrobial, Antioxidant, and Anticancer Activity of Greenly Synthesized Selenium and Zinc Composite Nanoparticles Using Ephedra aphylla Extract.

The current work aimed to synthesize selenium and zinc nanoparticles using the aqueous extract of Ephedra aphylla as a valuable medicinal plant. The prepared nanoparticles were characterized by TEM, zeta potential, and changes in the phytochemical constituents. Hence, the phenolic, flavonoid, and tannin contents were reduced in the case of the prepared samples of nanoparticles than the original values in the aqueous extract. The prepared extract of Ephedra aphylla and its selenium and zinc nanoparticles showed high potency as antioxidant agents as a result of the DPPH• assay. The samples were assessed as anticancer agents against six tumor cells and a normal lung fibroblast (WI-38) cell line. The selenium nanoparticles of Ephedra aphylla extract revealed very strong cytotoxicity against HePG-2 cells (inhibitory concentration (IC50) = 7.56 ± 0.6 µg/mL), HCT-116 cells (IC50 = 10.02 ± 0.9 µg/mL), and HeLa cells (IC50 = 9.23 ± 0.8 µg/mL). The samples were evaluated as antimicrobial agents against bacterial and fungal strains. Thus, selenium nanoparticles showed potent activities against Gram-negative strains (Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Escherichia coli), Gram-positive strains (Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, and Staphylococcus epidermidis), and the fungal strain Candida albicans. In conclusion, the preparation of nanoparticles of either selenium or zinc is crucial for improved biological characteristics.

Design, synthesis and molecular modeling of new quinazolin-4(3H)-one based VEGFR-2 kinase inhibitors for potential anticancer evaluation

Globally cancer is the second leading cause of death. So that this work is an attempt to develop new effective anti-cancer agents. In line with pharmacophoric features of VEGFR-2 kinase inhibitors, new nineteen quinazolin-4-one derivatives were designed, synthesized and biologically evaluated for their potential anticancer activity. All target compounds were evaluated in vitro for VEGFR-2 tyrosine kinase inhibition. Then, nine compounds of best results were further investigated by in vitro assay against three human cancer cell lines, namely HepG2, PC3 and MCF. N’-{2-](3-Ethyl-6-nitro-4-oxo-3,4-dihydroquinazoline-2-yl)thio[acetyl}benzohydrazide (36) was found to be the most potent candidate as it showed IC50 = 4.6 ± 0.06 µM against VEGFR-2 kinase. It also exhibited IC50 = 17.23 ± 1.5, 26.10 ± 2.2 and 30.85 ± 2.3 µg/mL against HepG2, PC3 and MCF, respectively. At the same time it showed IC50 = 145.93 ± 1.1 µg/mL against the normal human lung fibroblasts cell line (WI-38), indicating good selectivity index. Further investigation into HepG2 cell cycle showed the ability of compound 36 to induce apoptosis and arrest cell growth at G2/M phase. Moreover, docking studies demonstrated the ability of compound 36 to bind VEGFR-2 in a correct manner making three essential hydrogen bonds with the key residues Glu885, Asp1046 and Cys919. In sum, this work suggests that compound 36 can serve as a lead for development of effective anticancer agents targeting VEGFR-2.