Human Embryonic Kidney Cell Line Research Articles

Microwave-assisted synthesis, spectral characterization, and biological activities of complexes of a Schiff base derived from S-benzyl dithiocarbazate

Co(II), Ni(II), Zn(II), and Cd(II) complexes were synthesized from a Schiff base ligand, benzyl-2-(1-(thiophen-2-yl)propylidene)hydrazine-1-carbodithioate (BTPHC), by conventional as well as microwave irradiation methods. The ligand and its metal complexes were characterized by spectroscopic techniques (FT-IR, UV-vis, mass spectrometry, and 1H NMR) and powder X-ray diffraction. Based on magnetic measurements and spectral data, octahedral environments for Co(II) and Ni(II) complexes and tetrahedral environments for Zn(II) and Cd(II) complexes are proposed. The metal(II) complexes were active toward bacterial and fungal strains. Antidiabetic studies showed that the Cd(II) complex possesses greater potential as an antidiabetic agent than the other metal complexes and the ligand. Antioxidant studies indicate that the Zn(II) and Cd(II) complexes possess high radical scavenging activities. The synthesized ligand and metal(II) complexes were screened for their anticancer properties against two human cancer cell lines MCF-7 and HeLa using an MTT cell viability assay. The Ni(II) complex excelled in halting the proliferation of HeLa and MCF-7 cancer cell lines with IC50 values of 46.63 µM and 80.90 µM, respectively, whereas it displayed insignificant cytotoxicity against the human normal embryonic kidney cell line (HEK293) with IC50 = 300.41 µM.

Design, synthesis, and biological evaluation of novel quinoxaline aryl ethers as anticancer agents.

We designed and synthesized thirty novel quinoxaline aryl ethers as anticancer agents, and the structures of final compounds were confirmed with various analytical techniques like Mass, 1 H NMR, 13 C NMR, FTIR, and elemental analyses. The compounds were tested against three cancer cell lines: colon cancer (HCT-116), breast cancer (MDA-MB-231), prostate cancer (DU-145), and one normal cell line: human embryonic kidney cell line (HEK-293). The obtained results indicate that two compounds, FQ and MQ, with IC50 values < 16 μM, were the most active compounds. Molecular docking studies revealed the binding of FQ and MQ molecules in the active site of the c-Met kinase (PDB ID: 3F66, 1.40 Å). Furthermore, QikProp ADME prediction and the MDS analysis preserved those critical docking data of both compounds, FQ and MQ. Western blotting was used to confirm the impact of the compounds FQ and MQ on the inhibition of the c-Met kinase receptor. The apoptosis assays were performed to investigate the mechanism of cell death for the most active compounds, FQ and MQ. The Annexin V/7-AAD assay indicated apoptosis in MDA-MB-231 cells treated with FQ and MQ, with FQ (21.4%) showing a higher efficacy in killing MDA-MB-231 cells than MQ (14.25%). The Caspase 3/7 7-AAD assay further supported these findings, revealing higher percentages of apoptotic cells for FQ-treated MDA-MB-231 cells (41.8%). The results obtained from the apoptosis assay conclude that FQ exhibits better anticancer activity against MDA-MB-231 cells than MQ.

Functional analyses of TRAF6 gene in Argopecten scallops

The tumor necrosis factor (TNF) receptor-associated factor (TRAF) family has been reported to be involved in many immune pathways. In a previous study, we identified 5 TRAF genes, including TRAF2, 3, 4, 6, and 7, in the bay scallop (Argopecten irradians, Air) and the Peruvian scallop (Argopecten purpuratus, Apu). Since TRAF6 is a key molecular link in the TNF superfamily, we conducted a series of studies targeting the TRAF6 gene in the Air and Apu scallops as well as their hybrid progeny, Aip (Air ♀ × Apu ♂) and Api (Apu ♀ × Air ♂). Subcellular localization assay showed that the Air-, Aip-, and Api-TRAF6 were widely distributed in the cytoplasm of the human embryonic kidney cell line (HEK293T). Additionally, dual-luciferase reporter assay revealed that among TRAF3, TRAF4, and TRAF6, only the overexpression of TRAF6 significantly activated NF-κB activity in the HEK293T cells in a dose-dependent manner. These results suggest a crucial role of TRAF6 in the immune response in Argopecten scallops. To investigate the specific immune mechanism of TRAF6 in Argopecten scallops, we conducted TRAF6 knockdown using RNA interference. Transcriptomic analyses of the TRAF6 RNAi and control groups identified 1194, 2403, and 1099 differentially expressed genes (DEGs) in the Air, Aip, and Api scallops, respectively. KEGG enrichment analyses revealed that these DEGs were primarily enriched in transport and catabolism, amino acid metabolism, peroxisome, lysosome, and phagosome pathways. Expression profiles of 28 key DEGs were confirmed by qRT-PCR assays. The results of this study may provide insights into the immune mechanisms of TRAF in Argopecten scallops and ultimately benefit scallop breeding.

Precise control of microfluidic flow conditions is critical for harnessing the in vitro transfection capability of pDNA-loaded lipid-Eudragit nanoparticles.

Microfluidics is widely regarded as a leading technology for industrial-scale manufacture of multicomponent, gene-based nanomedicines in a reproducible manner. Yet, very few investigations detail the impact of flow conditions on the biological performance of the product, particularly biocompatibility and therapeutic efficiency. Herein, this study investigated the engineering of a novel lipid-Eudragit hybrid nanoparticle in a bifurcating microfluidics micromixer for plasmid DNA (pDNA) delivery. Nanoparticles of ~150 nm in size, with uniform polydispersity index (PDI = 0.2) and ξ-potential of 5-11 mV were formed across flow rate ratios (FRR, aqueous to organic phase) of 3:1 and 5:1, respectively. The hybrid nanoparticles maintained colloidal stability and structural integrity of loaded pDNA following recovery by ultracentrifugation. Importantly, in vitro testing in human embryonic kidney cell line (HEK293T) revealed significant differences in biocompatibility and transfection efficiency (TE). Lipid-Eudragit nanoparticles produced at FRR 3:1 displayed high cellular toxicity (0-30% viability), compared with nanoparticles prepared at FRR 5:1 (50-100% viability). Red fluorescent protein (RFP) expression was sustained for 24-72 h following exposure of cells to nanoparticles, indicating controlled release of pDNA and trafficking to the nucleus. Nanoparticles produced at FRR 5:1 resulted in markedly higher TE (12%) compared with those prepared at FRR 3:1 (2%). Notably, nanoparticles produced using the bench-scale nanoprecipitation method resulted in lower biocompatibility (30-90%) but higher RFP expression (25-38%). These findings emphasize the need for in-depth analysis of the effect of formulation and flow conditions on the physicochemical and biological performance of gene nanomedicines when transitioning from bench to clinic.

Antibacterial Evaluation of Gallic Acid and its Derivatives against a Panel of Multi-drug Resistant Bacteria.

Infectious diseases are the second leading cause of deaths worldwide. Pathogenic bacteria have been developing tremendous resistance against antibiotics which has placed an additional burden on healthcare systems. Gallic acid belongs to a naturally occurring phenolic class of compounds and is known to possess a wide spectrum of antimicrobial activities. In this study, we synthesized thirteen derivatives of gallic acid and evaluated their antibacterial potential against seven multi-drug resistant bacteria, as well as cytotoxic effects against human embryonic kidney cell line in vitro. Methods: 13 compounds were successfully synthesized with moderate to good yield and evaluated. Synthesized derivatives were characterized by using nuclear magnetic resonance spectroscopy, mass spectrometry, and Fourier transformation infrared spectroscopy. Antibacterial activity was determined using microdilution while cytotoxicyt was assessed using MTT assay. The results of antibacterial assay showed that seven out of thirteen compounds exhibited antibacterial effects with compound 6 and 13 being most potent against Staphylococcus aureus (MIC 56 μg/mL) and Salmonella enterica (MIC 475 μg/mL) respectively. On the other hand, most of these compounds showed lower cytotoxicity against human embryonic kidney cells (HEK 293), with IC50 values ranging from over 700 μg/mL. Notably, compound 13 was found to be non-toxic at concentrations as high as 5000 μg/mL. These findings suggest that the present synthetic derivatives of gallic acid hold potential for further studies in the development of potent antibacterial agents.

Cytotoxicity of vanadium dioxide nanoparticles to human embryonic kidney cell line: Compared with vanadium(IV/V) ions

Vanadium dioxide (VO2) is a class of thermochromic material with potential applications in various fields. Massive production and wide application of VO2 raise the concern of its potential toxicity to human, which has not been fully understood. Herein, a commercial VO2 nanomaterial (S-VO2) was studied for its potential toxicity to human embryonic kidney cell line HEK293, and two most common vanadium ions, V(IV) and V(V), were used for comparison to reveal the related mechanism. Our results indicate that S-VO2 induces dose-dependent cellular viability loss mainly through the dissolved V ions of S-VO2 outside the cell rather than S-VO2 particles inside the cell. The dissolved V ions of S-VO2 overproduce reactive oxygen species to trigger apoptosis and proliferation inhibition via several signaling pathways of cell physiology, such as MAPK and PI3K-Akt, among others. All bioassays indicate that the differences in toxicity between S-VO2, V(IV), and V(V) in HEK293 cells are very small, supporting that the toxicity is mainly due to the dissolved V ions, in the form of V(V) and/or V(IV), but the V(V)’s behavior is more similar to S-VO2 according to the gene expression analysis. This study reveals the toxicity mechanism of nanosized VO2 at the molecular level and the role of dissolution of VO2, providing valuable information for safe applications of vanadium oxides.

Design, synthesis and biological evaluation of novel pyrido-[2,3-d]-pyrimidin-2-amine analogues as antimycobacterial agents

Thirty-eight novel Pyrido-[2,3-d]-pyrimidin-2-amine derivatives (P1-P38) were designed, synthesized, and their structures confirmed using analytical techniques viz.,1H NMR, 13C NMR, and mass spectrometry. Using the Microplate Alamar Blue Assay (MABA), the final derivatives, P1-P38, were tested for their anti-mycobacterial activity against the Mycobacterium tuberculosis (Mtb) H37Rv strain. Amongst all the assessed compounds, P35 displayed significant inhibition with MIC 7.18 μg/mL. The remaining compounds exhibited MIC from 18.11 to >100 μg/mL. The potent compounds with MIC 〈 40.96 μg/mL, were evaluated for toxicity on HEK293T a normal human embryonic kidney cell line. The most active compound P35 exhibited a selectivity index 〉 20, proving its selective behavior towards Mtb H37Rv strain than human cells. To further evaluate the binding pattern in the active site of Thymidylate Kinase (TMPK) from Mtb (PDB-1G3U), a molecular docking analysis of compound P35 was performed using the glide module of Schrodinger software. Finally, molecular dynamics simulations were performed for 100 ns in order to elucidate the stability, confirmation, and intermolecular interactions of the co-crystal ligand and significantly active compound P35 on the selected target protein.

Crocin-loaded Niosomal Nanoparticles Reversing Cytotoxicity and Oxidative Stress in HEK293 Cell Line Exposed to Paraquat: An In vitro Study.

Paraquat (PQ) is an effective herbicide which is widely used around the world to remove weeds in agriculture. As a water-soluble carotenoid, crocin is a pharmacologically active constituent of C. sativus L. (saffron). In the present study, we investigated the effects of crocin-loaded niosomes (Cro-NIO) compared to free crocin on PQ-induced toxicity in the eukaryotic human embryonic kidney (HEK293) cell line. The Cro-NIO was synthesized and characterized. Cell viability was determined using the MTT assay in PQ-exposed HEK293 cell lines. The activities of biochemical markers were quantitatively determined to reveal the potential mechanism of PQ-induced oxidative stress in HEK293 cell line. The particle size, zeta potential, polydispersity index (PDI), DL, and EE of Cro-NIO were 145.4 ± 19.5 nm, -22.3 ± 3.11 mV, 0.3 ± 0.03, 1.74 ± 0.01%, and 55.3 ± 7.1%, respectively. PQtreated HEK293 cell lines decreased cell viability. The results of oxidative status showed that PQ significantly could increase ROS accumulation, accompanied by a decreasing antioxidant defense system. However, treatment with Cro-NIO, compared to crocin, not only did dose-dependently improve the cell viability but also significantly attenuated the ROS accumulation and increased antioxidant markers. According to these results, Cro-NIO, compared to crocin, was superior to ameliorating PQ-induced cytotoxicity and oxidative damage in HEK293 cells.

Tuning plasmid DNA amounts for cost-effective transfections of mammalian cells: when less is more

Transient gene expression (TGE) in mammalian cells is a well-known approach to the fast expression of recombinant proteins. The human cell line HEK (human embryonic kidney) 293F is widely used in this field, due to its adaptability to grow in suspension to high cell densities in serum-free media, amenability to transfection, and production of recombinant proteins in satisfactory quantities for functional and structural analysis. Amounts of plasmid DNA (pDNA) required in transfections for TGE remain high (usually 1 µg pDNA/mL, or even higher), representing a noticeable proportion of the overall cost. Thus, there is an economic need to reduce amounts of coding pDNA in TGE processes. In this work, amounts of both pDNA and transfecting agent used for TGE in HEK 293F cells have been explored in order to reduce them without compromising (or even improving) the productivity of the process in terms of protein yield. In our hands, minimal polyethyleneimine (PEI) cytotoxicity and optimum protein yields were obtained when transfecting at 0.5 µg pDNA/mL (equal to 0.5 µg pDNA/million cells) and a DNA-to-PEI ratio of 1:3, a trend confirmed for several unrelated recombinant proteins. Thus, carefully tuning pDNA and transfecting agent amounts not only reduces the economic costs but also results in higher recombinant protein yields. These results surely have a direct application and interest for the biopharmaceutical industry, always concerned in increasing productivity while decreasing economic costs.Key points• Mammalian cells are widely used to produce recombinant proteins in short times.• Tuning DNA and transfecting agent are of great interest to optimize economic costs.• Reducing DNA and transfecting agent amounts result in higher protein yields.Graphical

7SK small nuclear RNA (Rn7SK) induces apoptosis through intrinsic and extrinsic pathways in human embryonic kidney cell line.

Rn7SK, a highly conserved small nuclear non-coding RNA, controls Polymerase II transcription machinery by activating of the Positive Transcriptional Elongation Factor b (P-TEFb). Apart from its role in transcriptional regulation, the potential functions of Rn7SK in cell apoptosis are poorly understood. In a previous study, we demonstrated that overexpression of 7SK induces apoptosis in HEK cells. However, it remains unclear whether 7SK-mediated apoptosis induction is exerted through the intrinsic or extrinsic pathways. Rn7SK was overexpressed in HEK 293T cell line using Lipofectamine 2000 reagent to investigate its potential apoptotic functions. The overexpression of Rn7SK resulted in reduced cell viability through the induction of apoptosis, as evidenced by MTT assay and Annexin V/PI staining. Concurrently, alterations in the expression levels of key apoptosis-related genes were observed, as determined by quantitative RT-PCR. Furthermore, Rn7SK overexpression led to a decrease in cell proliferation, as assessed by colony formation assay and growth curve analysis. This reduction was associated with downregulated expression of key proliferative-related genes. Additionally, the migration and invasion capabilities of cells were significantly inhibited upon upregulation of Rn7SK, as demonstrated by transwell assays. This study suggests the apoptotic role of 7SK through both intrinsic and extrinsic pathways, necessitating further investigation into its underlying mechanisms.

Anticancer Study on IrIII and RhIII Half-Sandwich Complexes with the Bipyridylsulfonamide Ligand.

Organometallic half-sandwich complexes [(η5-Cp)IrCl(L)]PF6 (1) and [(η5-Cp)RhCl(L)]PF6 (2) were prepared using pentamethylcyclopentadienyl chloride dimers of iridium(III) or rhodium(III) with the 4-amino-N-(2,2'-bipyridin-5-yl)benzenesulfonamide ligand (L) and ammonium hexafluorophosphate. The crystal structures of L, 1, and 2 were analyzed in detail. The coordination reactions of the ligand with the central ions were confirmed using various spectroscopic techniques. Additionally, the interactions between sulfaligand, Ir(III), and Rh(III) complexes with carbonic anhydrase (CA), human serum albumin (HSA), and CT-DNA were investigated. The iridium(III) complex (1) did not show any antiproliferative properties against four different cancer cell lines, i.e., nonsmall cell lung cancer A549, colon cancer HCT-116, breast cancer MCF7, lymphoblastic leukemia Nalm-6, and a nonmalignant human embryonic kidney cell line HEK293, due to high binding affinity to GSH. The sulfonamide ligand (L) and rhodium(III) complex (2) were further studied. L showed competitive inhibition toward CA, while complexes 1 and 2, uncompetitive. All compounds interacted with HSA, causing a conformational change in the protein's α-helical structure, suggesting the induction of a more open conformation in HSA, reducing its biological activity. Both L and 2 were found to induce cell death through a caspase-dependent pathway. These findings position L and 2 as potential starting compounds for pharmaceutical, therapeutic, or medicinal research.

Cortisol controls endoplasmic reticulum stress and hypoxia dependent regulation of insulin receptor and related genes expression in HEK293 cells.

Objective. Glucocorticoids are important stress-responsive regulators of insulin-dependent metabolic processes realized through specific changes in genome function. The aim of this study was to investigate the impact of cortisol on insulin receptor and related genes expression in HEK293 cells upon induction the endoplasmic reticulum (ER) stress by tunicamycin and hypoxia. Methods. The human embryonic kidney cell line HEK293 was used. Cells were exposed to cortisol (10 µM) as well as inducers of hypoxia (dimethyloxalylglycine, DMOG; 0.5 mM) and ER stress (tunicamycin; 0.2 µg/ml) for 4 h. The RNA from these cells was extracted and reverse transcribed. The expression level of INSR, IRS2, and INSIG2 and some ER stress responsive genes encoding XBP1n, non-spliced variant, XBP1s, alternatively spliced variant of XBP1, and DNAJB9 proteins, was measured by quantitative polymerase chain reaction and normalized to ACTB. Results. We showed that exposure of HEK293 cells to cortisol elicited up-regulation in the expression of INSR and DNAJB9 genes and down-regulation of XBP1s, XBP1n, IRS2, and INSIG2 mRNA levels. At the same time, induction of hypoxia by DMOG led to an up-regulation of the expression level of most studied mRNAs: XBP1s and XBP1n, IRS2 and INSIG2, but did not change significantly INSR and DNAJB9 gene expression. We also showed that combined impact of cortisol and hypoxia introduced the up-regulation of INSR and suppressed XBP1n mRNA expression levels. Furthermore, the exposure of HEK293 cells to tunicamycin affected the expression of IRS2 gene and increased the level of XBP1n mRNA. At the same time, the combined treatment of these cells with cortisol and inductor of ER stress had much stronger impact on the expression of all the tested genes: strongly increased the mRNA level of ER stress dependent factors XBP1s and DNAJB9 as well as INSR and INSIG2, but down-regulated IRS2 and XBP1n. Conclusion. Taken together, the present study indicates that cortisol may interact with ER stress and hypoxia in the regulation of ER stress dependent XBP1 and DNAJB9 mRNA expression as well as INSR and its signaling and that this corticosteroid hormone modified the impact of hypoxia and especially tunicamycin on the expression of most studied genes in HEK293 cells. These data demonstrate molecular mechanisms of glucocorticoids interaction with ER stress and insulin signaling at the cellular level.

Quaternary nanorods: promising versatile agents for cancer therapy, antimicrobial strategies and free radical neutralization

Cu2XSnS4 (CXTS; X: Zn, Mn or Co) nanorods (NRs) were prepared by a simple hot-injection method and their cytotoxic activities on human breast adenocarcinoma (MCF-7), metastatic melanoma (451Lu) and embryonic kidney (HEK-293) cell lines were investigated for the first time. The antibacterial activities of these nanoparticles on both gram-positive (S. aureus) and gram-negative (E. coli) pathogenic bacteria as well as their DPPH (2,2-diphenyl-1- picrylhydrazyl) scavenging activities were evaluated. The structural characterizations show that the obtained nanoparticles have the same crystal structure and are shaped as nanorods with an average 10–40 nm edge length. The data obtained in cytotoxicity studies revealed that the synthesized CXTS NRs have a significant inhibition effect, especially on cancer cell lines (MCF-7 and 451Lu), in a dose-dependent manner over a period of 24 h, demonstrating their potential for use as anticancer agents. In addition, the findings indicate that the synthesized nanorods possess strong antibacterial properties against the pathogenic microorganisms tested and display a marked antioxidant effect through efficient elimination of DPPH activity. This comprehensive study sheds light on the creation and development of CXTS NRs as promising versatile agents in various biomedical and environmental applications such as cancer therapy, antimicrobial strategies and the neutralization of free radicals.

Effects of 2.4 GHz radiofrequency electromagnetic field (RF-EMF) on glioblastoma cells (U -118 MG).

Mobile phones and Wi-Fi are the most commonly used forms of telecommunications. Initiated with the first generation, the mobile telephony is currently in its fifth generation without being screened extensively for any biological effects that it may have on humans or on animals. Some studies indicate that high frequency electromagnetic radiation emitted by mobile phone and Wi-Fi connection can have a negative effect upon human health, and can cause cancer, including brain tumour. The aim of the study was to investigate the influence of 2.4 GHz radiofrequency electromagnetic field (RF-EMF) on the proliferation and morphology of normal (human embryonic kidney cell line Hek-293) and cancer cells (glioblastoma cell line U-118 MG). The cell cultures were incubated in RF-EMF at the frequency of 2.4 GHz, with or without dielectric screen, for 24, 48 and 72h. In order to analyse the influence of the electromagnetic field on cell lines, Cytotoxicity test Cell Counting Kit-8 was performed. To protect cells against emission of the electromagnetic field, a dielectric screen was used. It was found that 2.4 GHz RF electromagnetic field exposure caused a significant decrease in viability of U-118 MG and Hek-293 cells. The impact of the electromagnetic field was strongest in the case of cancer cells, and the decrease in their survival was much greater compared to the healthy (normal) cells of the Hek-293 line. Results of the study indicate that using a radio frequency electromagnetic field (2.4 GHz) has a clearly negative effect on the metabolic activity of glioblastoma cells. RF-EMF has much less impact on reducing the viability of normal cells (Hek -293) than cancer cells.

MYCT-1 Gene Expression in Patients with Gastric Cancer: an Ex Vivo Study.

Ploidy, p53, bcl-2, and c-myc genes are associated with gastric cancer. Myc target 1 (MYCT1) gene is an oncogenic gene and is associated with cancer progression through different signal transduction pathways identifying the corresponding genes The objective of the study was to evaluate the association between MYCT1 gene expression and gastric cancer. Real-time polymerase chain reaction (RT-PCR), western blot analysis, cell growth study, and TUNEL assay were performed for the human gastric cancer cell lines and human embryonic kidney cell line. β-Actin gene preferred as a control in RT-PCR. The ratio of MYCT1 gene expression to β-actin gene expression less than 0.5 was considered as downregulation. Using SDS-PAGE MYCT1 gene expression was measured in western blot analysis. Cells with and without the MYCT1 gene were incubated in 35 mm plates with 10% fetal bovine serum in the cell growth study. TUNEL assay was performed to detect the effect of the MYCT1 gene on the apoptosis of cells. The ratio of MYCT1 gene expression to β-actin gene expression was 0.47 ± 0.01 and 0.76 ± 0.01 for human gastric cancer cell lines and human embryonic kidney cell lines, respectively. MYCT1 gene expression was downregulated in the human gastric cancer cell lines than human embryonic kidney cell line (p < 0.001). MYCT1 gene decreased cell growth (p = 0.041) during 6 days of incubation study of cells. TUNEL assay showed only the fluorescence of PI in BGC823 cells without the MYCT1 gene. MYCT1 gene expression was downregulated in the human gastric cancer cell lines, and MYCT1 gene accelerates the apoptotic process.

Structural, luminescent and in vitro studies of europium‐doped soda lime phosphate glasses

AbstractIn this article, we have reported the effect of varying concentration of europium (Eu) in (50 − x)% P2O5–25% Na2O–24% CaO–% Eu2O3, where x = 1, 3, 5. The glass samples were synthesised via conventional melt‐quench method. The impact of europium ion (Eu3+) on the structural, optical and luminescent properties of phosphate soda lime glasses has been studied using X‐ray diffraction (XRD), Fourier‐transformed infrared (FTIR) spectroscopy, ultraviolet‐visible spectroscopy, scanning electron microscopy coupled with energy‐dispersive spectroscopy and photoluminescent techniques. The amorphous nature of glass samples was confirmed by XRD patterns. FTIR confirmed the presence of various functional groups. The emission spectra of synthesised samples exhibited intense emission peaks corresponding to Eu3+ under excitation at 393 nm. Among all the peaks, the maximum intensity was observed for 5D0 → 7F2 transition. Judd–Ofelt (J–O) parameters (Ω2, Ω4) and other radiative parameters such as band width, radiative transition probabilities, stimulated emission cross‐sections and branching ratio were determined from emission spectra. The other photometric parameters such as CIE coordinates and colour purity were also determined. Furthermore, cytotoxic studies were carried out on normal cell line human embryonic kidney cells (HEK‐293) using MTT assay. Results showed that the prepared samples significantly enhanced growth in glass sample‐treated cells as compared to control cells. These findings suggest that synthesised glass samples are biocompatible in nature and have potential for applications in display devices and biomedical research area.

Phytochemical analysis and high‐performance thin‐layer chromatography guided quantification of Boeravinone B from Boerhaavia diffusa root extracts: Assessment of in vitro safety analysis

AbstractAn Indian traditional medicinal plant, Boerhaavia diffusa (Punarnava), is reported for its diuretic activity and helps in impaired kidney function. The present study indicates that alkaloids, carbohydrates, saponins, phytosterols, flavonoids, and glycosides are absent in the petroleum ether extract and present chiefly in the methanolic extract of roots. The phytosterols were found to be present in the ethyl acetate extract, while other phytochemical constituents, that is, alkaloids, carbohydrates, saponins, flavonoids, and glycosides, were absent. From high‐performance thin‐layer chromatography, the results indicated that out of all the three extracts and two marketed formulations, ethyl acetate root extract (LEA) has the highest amount of Boeravinone B content. Further, different root extracts of B. diffusa and two marketed formulations were evaluated on the human embryonic kidney cell line to understand pharmacological safety. From the results, it was observed that LEA and petroleum ether root extracts showed a higher percentage of cell survival and a better safety profile up to the dose of 1 mg/mL. The overall results indicated that further preclinical evaluation of B. diffusa is needed to understand the mechanism behind nephroprotective potential which can further expedite the drug discovery process of nephroprotective molecules.

Evaluation of Cytotoxicity of 1H-1,2,4-Triazole-3-carboxamide Derivatives in Various Cancer Cell Lines using MTT Assay: Synthesis, Characterization and Molecular Docking Studies

The pursuit of discovering efficacious novel anticancer agents is an ongoing and perpetual endeavour. The ongoing research endeavour involves the discovery of a new set of various substituted 1,2,4-triazole carboxamide derivatives. The synthetic process involves multiple steps: converting thiooxamic acid ethyl ester to ethyl b-N-boc-oxalamidrazone (2), preparing ethyl esters of 5-substituted 1,2,4-triazole-3-carboxylic acid (3) and synthesizing 5-substituted 1,2,4-triazole-3-carboxylic acid amides (4a-n) using potassium tert.-butoxide. The resulting compounds are characterized using 1H and 13C NMR spectroscopy and HRMS. Molecula docking experiments were performed using the synthesized compounds against two cancer targets, viz. EGFR (6LUD) and CDK-4 (7SJ3). The compounds were assessed for their anticancer potential against four different cancer cell lines, namely non-small cell lung cancer (A-549), pancreatic cancer (PANC-1), colorectal cancer (HCT-116) and cervical cancer (HaLa) cell lines. Additionally, a normal human embryonic kidney cell line (HEK-293) was used for comparison. The results obtained from molecular docking analysis demonstrated that the 1,2,4-triazole carboxamides (4c, 4e, 4h, 4m and 4n) exhibited more favourable binding interactions compared to the ligands that were co-crystallized. All the compounds exhibited a satisfactory cytotoxicity profile in comparison to the reference drug doxorubicin. Among the compounds that tested, it was observed that compounds 4e and 4m displayed the most pronounced inhibitory activity across all the cell lines that were tested.

Abstract B146: Design and preclinical evaluation of CPL976-MMAE - novel, potent AXL-PD-L1 bispecific antibody conjugated with MMAE in targeted anticancer therapy

Abstract Background: Antibody-drug conjugates (ADCs) combine the specificity of antibodies with the potency of highly cytotoxic agents, reducing the off-target effects and improve effectiveness with more precising delivery of the payload to the tumor site. Strong correlation has been established between AXL and PD-L1 expression levels in many types of cancer. Therefore, targeting both proteins simultaneously ensure high cancer specificity in delivering the toxic payload. We have developed a bispecific antibody, that strongly induces receptor internalization. Our bispecific ADC combines anti-AXL and anti-PD-L1 construct with toxic MMAE (Monomethyl auristatin E, a potent tubulin inhibitor toxin), for better targeting to solid tumor cells and greatly reducing off-target effects. Materials and Methods: CPL976-MMAE was prepared with site-specific conjugation technology using Fc glycan remodeling and click-chemistry. Briefly, after enzymatic trimming of the N-linked glycans in the Fc pocket of antibody, the terminal GlcNAc was extended with 6-N3-GalNAc using GalNAc-transferase, allowing metal-free click conjugation of the MMAE payload. Bispecific antibodies chosen for conjugation were first characterized in vitro for the interaction with the extracellular domain of human AXL and PD-L1 using surface plasmon resonance and flow cytometry. After the conjugation with MMAE, ADCs cytotoxicity, and selectivity were evaluated in the human breast cancer model (MDA-MB-231) with high expression of both targets, AXL and PD-L1, using SytoX and Hoechst. The human embryonic kidney cell line (HEK-293) with minimal expression of targets, was used as a negative control. The efficacy of conjugates were evaluated in xenograft mouse model. Results: Analytical characterization by RP-HPLC confirmed efficient conjugation and homogeneity of the conjugates. Conjugates showed improved cellular uptake, selectivity, and anticancer activity, compared to unconjugated MMAE in the AXL/PD-L1 overexpressing cancer cell lines. Additionally, HEK-293 cells were not affected by conjugates in concentrations applied in the assay with use of the breast cancer cell lines. This implies enhanced safety by reducing off-target side effects compared to free auristatin. The efficacy of the compound was confirmed in xenograft mouse model. Conclusions: We have designed a new, potentially first-in-class anti-AXL/PD-L1 bispecific antibody conjugated with MMAE, which showed robust and selective antitumor effects in human cancer models. Citation Format: Delfina Popiel, Krzysztof Lacek, Anna Jabłońska, Aleksandra Sowińska, Agnieszka Bojko - Matuszek, Damian Kołakowski, Filip Mituła, Sebastian Kwiatkowski, Magdalena Bojko, Tomasz Kornatowski, Beata Kliszcz, Tomasz Banach, Michał Górka, Jerzy Pieczykolan, Maciej Wieczorek, Olga Abramczyk. Design and preclinical evaluation of CPL976-MMAE - novel, potent AXL-PD-L1 bispecific antibody conjugated with MMAE in targeted anticancer therapy [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr B146.

Quinoline-based thiazolyl-hydrazones target cancer cells through autophagy inhibition.

Heterocyclic pharmacophores such as thiazole and quinoline rings have a significant role in medicinal chemistry. They are considered privileged structures since they constitute several Food and Drug Administration (FDA)-approved drugs for cancer treatment. Herein, we report the synthesis, in silico evaluation of the ADMET profiles, and in vitro investigation of the anticancer activity of a series of novel thiazolyl-hydrazones based on the 8-quinoline (1a-c), 2-quinoline (2a-c), and 8-hydroxy-2-quinolyl moiety (3a-c). The panel of several human cancer cell lines and the nontumorigenic human embryonic kidney cell line HEK-293 were used to evaluate the compound-mediated in vitro anticancer activities, leading to [2-(2-(quinolyl-8-ol-2-ylmethylene)hydrazinyl)]-4-(4-methoxyphenyl)-1,3-thiazole (3c) as the most promising compound. The study revealed that 3c blocks the cell-cycle progression of a human colon cancer cell line (HCT-116) in the S phase and induces DNA double-strand breaks. Also, our findings demonstrate that 3c accumulates in lysosomes, ultimately leading to the cell death of the hepatocellular carcinoma cell line (Hep-G2) and HCT-116 cells, by the mechanism of autophagy inhibition.