HepG2 Cancer Cell Lines Research Articles

Preparation, spectroscopic and anticancer investigations of metal-drug complexes associated between flumequine antibiotic drug with lanthanum(III), samarium(III) and terbium(III) chloride

Flumequine ligand (FLQ) metal complexes of the [M(FLQ)2(Cl)(H2O)].nH2O type, where M are (La(III), Sm(III), and Tb(III)) and FLQ is flumequine have been synthesized. The FLQ metal complexes could be prepared using MCl3 : flumequine in stoichiometry of 1:2 in situ bidentate chelation. The characterization of FLQ complexes obtained have been done using elemental analyses (%C, %H and %N), molar conductivity (Ʌm), infrared spectroscopy (FTIR), electronic spectra (UV-Vis), thermal analysis (TGA), and physicochemical techniques such as X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) measurements at room temperature. Accordingly, these complexes are indicative of an octahedral coordination structure. Flumequine ligand has a bidentate fashion through the oxygen atoms of carbonyl (quinolone) and carboxylic groups. The findings demonstrated the anti-cancer efficacy of these compounds against HepG-2 and MCF-7 cancer cell lines at extremely low doses of up to 58.2 mg/mL. The La(III) complex was shown to have the highest selectivity against cancer cells in comparable with both samarium and terbium(III) complexes.
 KEY WORDS: Flumequine, Lanthanide metal ions, FTIR, EDX, Complexation, Anticancer activity
 Bull. Chem. Soc. Ethiop. 2024, 38(3), 671-684. 
 DOI: https://dx.doi.org/10.4314/bcse.v38i3.10

Dual drug-loaded cubosome nanoparticles for hepatocellular carcinoma: a design of experiment approach for optimization and in vitro evaluation

BackgroundLiver cancer, a formidable and complex disease, poses a significant global health threat, stemming from various causes, including chronic infections like hepatitis B and C, cirrhosis, and lifestyle factors. In liver cancer treatment, targeted delivery revolutionizes precision therapy, minimizing side effects by directing drugs specifically to cancer cells. This study aims to develop and statistically optimize cubosomal formulations containing piperine and quercetin with the goal of augmenting their activity against hepatocellular carcinoma.ResultsEmploying a central-composite design, we utilized Design-Expert® software to guide the experiment. The key formulation variables were the concentration of glyceryl monooleate (GMO) and Poloxamer-407, while the dependent responses were particle size (PS) and entrapment efficiency (EE%). The optimized cubosomal formulation was validated through the utilization of high-resolution transmission electron microscopy (HR-TEM), in vitro release studies, and an in vitro cell proliferation assay conducted on the HepG2 cell line. High-performance liquid chromatography was employed for the determination of piperine and quercetin in the optimized cubosomal nanoparticle. The optimized formulation had a composition of 2.5 (w/w%) GMO and 0.5 (w/w%) Poloxamer 407. The predicted values for PS and EE% were 102.34 and 75.11%, respectively. The cytotoxicity of the optimized cubosomal formulation exhibited enhanced efficacy on the HepG2 cancer cell line, even at lower concentrations, when compared to the standard. Notably, it demonstrated a superior cytotoxic effect on the liver cancer cell line.ConclusionThe findings of the study indicated that cubosomes exhibit promise as an effective carrier for delivering piperine and quercetin, addressing hepatocellular carcinoma effectively.Graphical abstract

Zinc oxide‐cadmium(II) sulfide heterostructure as a potential photocatalyst for preparing substituted chromenes and its anti‐liver cancer activity

There are ongoing studies on the potential use of chromene derivatives in liver cancer therapy. They have shown promising results in preclinical studies for liver cancer, including inhibiting tumor growth and inducing apoptosis. Herein, the nanosized hybrid material zinc oxide (ZnO)–cadmium sulfide (CdS) is prepared using a green and ecofriendly procedure. The nanomaterial was characterized by FE‐SEM, transmission electron microscopy, energy dispersive X‐ray, DRS, X‐ray diffraction, and Fourier transform infrared spectroscopy. The photocatalytic proficiency of ZnO–CdS was then scrutinized in the fabrication of some 4H‐chromenes in a mild condition. The outcomes of this study revealed that the nanophotocatalyst has high photocatalytic reactivity and acceptable reusability in the desired condensation reaction. Furthermore, a preliminary in vitro cellular toxicity assay was performed on ZnO–CdS nanoparticles using HepG2 cancer cell line through MTT assay.

Design and In Silico and In Vitro Evaluations of a Novel Nicotinamide Derivative as a VEGFR-2 Inhibitor

A new nicotinamide derivative, (E)-N-(4-(1-(2-(4-benzamidobenzoyl)hydrazone)ethyl)phenyl)nicotinamide, was designed as a VEGFR-2 inhibitor. Utilizing the density functional theory (DFT) calculations, the three-dimensional structure of the designed compound was determined, shedding light on its stability and reactivity. Molecular docking revealed its capability to inhibit VEGFR-2, which was further supported by molecular dynamics (MD) simulations confirming its binding to the target protein. In addition, molecular mechanics-generalized born surface area (MM-GBSA), protein-ligand interactions profiler (PLIP), and essential dynamics studies provided further validation of the compound’s precise binding with optimal energy. Then, the “compound 10” was synthesized and subjected to in vitro assays. Compound 10 inhibited VEGFR-2 with an IC50 value of 105.4 ± 0.896 nM, comparing sorafenib’s IC50 value of 61.65 ± 0.934 nM. Besides, it exhibited cytotoxicity against HepG2 and MCF-7 cancer cell lines, with IC50 values of 35.78 ± 0.863 μM and 57.62 μM ± 0.871, comparing sorafenib’s IC50 values of 5.95 ± 0.917 μM and 8.45 ± 0.912 μM. Furthermore, compound 10 demonstrated a lower level of toxicity towards Vero cell lines, with an IC50 value of 127.3 μM. Likewise, compound 10 induced apoptosis in HepG2 cell lines through a flow cytometric analysis in addition to an increase in the levels of caspase-3 and caspase-9. Moreover, compound 10 hindered the migration and healing abilities of HepG2 cells. In conclusion, our study positions compound 10 as a promising candidate for further chemical modifications and biological evaluations.

Development of 1,5-diarylpyrazoles as EGFR/JNK-2 dual inhibitors: design, synthesis, moleecular docking, and bioactivity evaluation

The eradication of multifactorial diseases, such as cancer, requires the design of drug candidates that attack multiple targets that contribute to the progression and proliferation of such diseases. Here, 1,5-diarylpyrazole derivatives bearing vanillin or sulfanilamide are developed as potential dual inhibitors of epidermal growth factor receptor (EGFR)/c-Jun N-terminal kinase 2 (JNK-2) for possible anticancer activity. These derivatives inhibited the growths of DLD-1, HeLa, K-562, SUIT-2 and HepG2 cancer cell lines, with minimum concentration required to inhibit half of the cellular growth (IC50) values of 2.7–63 μM. The tests confirmed that 5b and 5d were potent JNK-2 inhibitors, with IC50 of 2.0 and 0.9 μM, respectively, whereas 6 h selectively inhibited EGFR protein kinase (EGFR-PK) (IC50 = 1.7 μM). Notably, 6c inhibited both kinases, with IC50 values of 2.7 and 3.0 μM against EGFR-PK and JNK-2, respectively, offering a reference for designing mutual inhibitors of EGFR/JNK-2. The docking studies revealed the ability of the pyrazole ring to bind to the hinge region of the ATP binding site, thereby supporting the experimental inhibitory results. Furthermore, the developed compounds could induce apoptosis and induce cell cycle arrest at different cell phases.

Investigation of Anticancer Therapy Using pH-Sensitive Carbon Dots-Functionalized Doxorubicin in Cubosomes.

Cancer remains a highly lethal disease due to its elusive early detection, rapid spread, and significant side effects. Nanomedicine has emerged as a promising platform for drug delivery, diagnosis, and treatment monitoring. In particular, carbon dots (CDs), a type of fluorescent nanomaterial, offer excellent fluorescence properties and the ability to carry multiple drugs simultaneously through covalent bonding. In this work, CDs with carbonyl groups on the surface were prepared by aldol condensation and reacted with amine groups in the structure of doxorubicin (DOX) through Schiff base reaction to generate pH-responsive CDs-DOX. On the other hand, cubosomes with three-dimensional lattice structures formed by lipid bilayers have advantageous capabilities of encapsulating various hydrophilic, amphiphilic, and hydrophobic substances. The pH-responsive CDs-DOX are subsequently loaded into cubosomes to form an anticancer therapeutic nanosystem, CDs-DOX@cubosome. Leveraging the unique properties of CDs-DOX and cubosomes, our CDs-DOX@cubosome can enter tumor tissue through the enhanced permeation and retention effect first and conduct membrane fusion with tumor cells to intracellularly release CDs-DOX. Then, the imine bond in CDs-DOX breaks under acidic conditions within human cancer cell lines (HeLa and HepG-2 cells), releasing DOX and achieving enhanced treatment of tumors. Additionally, fluorescent CDs can synchronously achieve real-time in situ diagnosis of tumor tissue. We demonstrate that our CDs-DOX@cubosome works as an excellent drug delivery system with therapeutic efficiency enhancement to the tumor and reduced side effects.

Iron oxide nanoparticles coated with Glucose and conjugated with Safranal (Fe3O4@Glu-Safranal NPs) inducing apoptosis in liver cancer cell line (HepG2)

Magnetic nanoparticles can be considered a reliable tool for targeted drug delivery to cancer tissues. Based on this, in this study, the anticancer effect of iron oxide nanoparticles coated with glucose and conjugated with Safranal (Fe3O4@Glu-Safranal NPs) on a liver cancer cell line (HepG2) was investigated. Physicochemical properties of nanoparticles were characterized using FT-IR, XRD, VSM, EDS-mapping, SEM and TEM imaging, zeta potential, and DLS analyses. MTT test was used to investigate the inhibitory effect of nanoparticles on cancer and normal cell lines. Also, the reactive oxygen species (ROS) level, the population of apoptotic cells, and cell cycle analysis were evaluated in control and nanoparticle-treated cells. The synthesized particles were spherical, in a size range of 17–49 nm, without impurities, with a surface charge of − 13 mV and hydrodynamic size of 129 nm, and with magnetic saturation of 22.5 emu/g. The 50% inhibitory concentration (IC50) of Safranal, Fe3O4, Fe3O4@Glu-Safranal and Cisplatin drug on liver cancer cells were 474, 1546, 305 and 135 µg/mL, respectively. While, the IC50 of Fe3O4@Glu-Safranal for normal cell line was 680 µg/mL. Treating liver cancer cells with nanoparticles significantly increased the population of apoptotic cells from 2.5% to 34.7%. Furthermore, the population of the cells arrested at the G2/M phase increased in nanoparticle-treated cells. Due to the biocompatibility of the constituent compounds of these nanoparticles, their magnetic properties, and their inhibitory effects on cancer cells, Fe3O4@Glu-Safranal NPs can be further considered as a promising anticancer compound.

Synthesis, characterization, molecular modeling studies, and biological evaluation of metal piroxicam complexes (M = Ni(II), Pt(IV), Pd(II), Ag(I)) as antibacterial and anticancer agents.

Four piroxicam metal complexes; NiL2 , PtL2 , PdL2 , and AgL were synthesized and characterized by different techniques with enhanced antibacterial and anticancer activity. Regarding in vitro antimicrobial activity, complex NiL2 displayed potent antibacterial effect against Escherichia coli and Pseudomonas aeruginosa that was 1.9-folds higher than piroxicam (minimum inhibitory concentration [MIC] = 31.85, 65.32 µM), respectively. In case of G+ve bacteria, complex PtL2 had potent activity on Staphylococcus aureus which was 2.1-folds higher than piroxicam (MIC = 43.12 µM), while activity of complex AgL against Enterococcus faecalis was threefolds higher than piroxicam (MIC = 74.57 µM. Complexes PtL2 and PdL2 exhibited higher inhibition of DNA gyrase than piroxicam (IC50 = 6.21 µM) in the range of 1.9-1.7-folds. The in vitro antiproliferative activity depicted that all investigated complexes showed better cytotoxic effect than piroxicam, specifically Pt and Pd complexes which had lower IC50 values than piroxicam on human liver cancer cell line HepG2 by 1.8 and 1.7-folds, respectively. While Pd and Ag complexes showed 2 and 1.6-folds better effect on human colon cancer cell line HT-29 compared with piroxicam. Molecular modeling studies including docking on Stranded DNA Duplex (1juu) and DNA gyrase enzyme (1kzn) that gave good insight about interaction of complexes with target molecules, calculation of electrostatic potential map and global reactivity descriptors were performed.

Novel biologically active paddlewheel-like palladium thiocyanate complex catalyzed cyanation of alkyl bromide utilizing Q-tube system: A green perspective

A novel paddlewheel-like complex of [Pd(SCN)2(3-Acpy)2], (1) was produced at ambient temperature by adding an aqueous solution of palladium chloride to an aqueous acetonitrile solution of 3-acetylpyridine (3-Acpy) and potassium thiocyanate. Elemental analysis, Fourier-transform infrared (FT-IR), ultraviolet–visible (UV–vis), and nuclear magnetic resonance (NMR) spectroscopic methods were used to characterize complex 1 comprehensively. Single crystal X-ray diffraction was used to analyze the crystal structure of complex 1. The palladium atom is bonded to two 3-Acpy ligands and two thiocyanates groups in trans mode giving square planer geometry in complex 1. The discrete units of paddlewheel-like complex 1 through hydrogen bonding extend to form a 3D supramolecular network structure. The synthesized and characterized new palladium complex 1 was utilized as a precatalyst for the cyanation of alkyl bromide using potassium ferrocyanide as a benign cyanide source reagent. The suggested catalytic test reaction under high-pressure conditions via a Q-tube system in green solvent cyclopentyl methyl ether introduced high yields and a short reaction time. Moreover, the cytotoxic activity of complex 1 was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against HePG-2 of liver cancer and MCF7 of breast cancer cell lines. According to the findings, complex 1 has significant strong activity against the cancer cell lines HePG-2 and MCF-7. Additionally, the luminescence spectra of complex 1 were investigated.

Synthesis and molecular docking simulations of novel azepines based on quinazolinone moiety as prospective antimicrobial and antitumor hedgehog signaling inhibitors

A series of novel azepine derivatives based on quinazolinone moiety was synthesized through the reaction of quinazolinone chalcones (2a–d) either with 2-amino aniline in acidic medium to give diazepines (3a–d) or with 2-aminophenol to offer oxazepine (4a–d). The structure of the synthesized compounds was confirmed via melting points, elemental analyses, and different spectroscopic techniques. Moreover, these newly compounds mode of action was investigated in-silico using molecular docking against the outer membrane protein A (OMPA), exo-1,3-beta-glucanase for their antimicrobial activity, and against Smoothened (SMO), transcription factor glioma-associated homology (SUFU/GLI-1), the main proteins of Hedgehog signaling pathway to inspect their anticancer potential. Our results showed that, diazepine (3a) and oxazepine (4a) offered the highest binding energy against the target OMPA/ exo-1,3-beta-glucanase proteins and exhibited the potent antimicrobial activities against E. coli, P. aeruginosa, S. aureus, B. subtilis, C. Albicans and A. flavus. As well, diazepine (3a) and oxazepine (4a) achieved the best results among the other compounds, in their binding energy against the target SMO, SUFU/GLI-1 proteins. The in-vitro cytotoxic study was done for them on panel of cancer cell lines HCT-116, HepG2, and MCF-7 and normal cell line WI-38. Conclusively, it was revealed that molecular docking in-silico simulations and the in-vitro experiments were agreed. As a result, our findings elucidated that diazepine (3a) and oxazepine (4a), have the potential to be used as antimicrobial agents and as possible cancer treatment medications.

Green-Synthesized Silver and Selenium Nanoparticles Using Berberine: A Comparative Assessment of In Vitro Anticancer Potential on Human Hepatocellular Carcinoma Cell Line (HepG2).

A well-known natural ingredient found in several medicinal plants, berberine (Ber), has been shown to have anticancer properties against a range of malignancies. The limited solubility and bioavailability of berberine can be addressed using Ber-loaded nanoparticles. In this study, we compared the in vitro cytotoxic effects of both Ber-loaded silver nanoparticles (Ber-AgNPs) and Ber-loaded selenium nanoparticles (Ber-SeNPs) in the human liver cancer cell line (HepG2) and mouse normal liver cells (BNL). The IC50 values in HepG2 for berberine, Ber-AgNPs, Ber-SeNPs, and cisplatin were 26.69, 1.16, 0.04, and 0.33 µg/mL, respectively. Our results show that Ber and its Ag and Se nanoparticles exerted a good antitumor effect against HepG2 cells by inducing apoptosis via upregulating p53, Bax, cytosolic cytochrome C levels, and caspase-3 activity, and the down-regulation of Bcl-2 levels. Similarly, incubation with Ber and both Ber-NPs (Ag and Se) led to a significant dose-dependent elevation in inflammatory markers' (TNF-α, NF-κB, and COX-2) levels compared to the control group. In addition, it led to the arrest of the G1 cell cycle by depleting the expression of cyclin D1 and CDK-2 mRNA. Furthermore, Ber and both Ber-NPs (Ag and Se) caused a significant dose-dependent increase in LDH activity in HepG2 cells. Furthermore, our findings offer evidence that Ber and its nanoparticles intensified oxidative stress in HepG2 cells. Furthermore, the migration rate of cells subjected to berberine and its nanoforms was notably decreased compared to that of control cells. It can be inferred that Ber nanoparticles exhibited superior anticancer efficacy against HepG2 compared to unprocessed Ber, perhaps due to their improved solubility and bioavailability. Furthermore, Ber-SeNPs exhibited greater efficacy than Ber-AgNPs, possibly as a result of the inherent anticancer characteristics of selenium.

Synthesis, Characterization and Biosafety Evaluation of Hollow Gold Nanospheres.

In order to assess the biosafety of HAuNS using zebrafish models and the cancer cell lines HepG2, HEK293, and A549, this study prepared HAuNS in a variety of sizes and alterations. By oxidizing cobalt nanoparticles encased in gold shells, HAuNS were created. In the meantime, PEG- and PEI-coated HAuNS were created. The diameters of the HAuNS that were produced were 30~40 nm, 50~60 nm, and 70~80 nm. MTT assay was used to assess the toxicity of HAuNS on HepG2, HEK293, and A549 cells. For the investigation of their toxicities, HAuNS (50~60 nm) of various concentrations were incubated with zebrafish embryos. Then, cell death was determined using acridine orange staining. In a cell line model, it was demonstrated that purified HAuNS exhibit lower toxicity than unpurified HAuNS. Meanwhile, it was discovered that surface-modified HAuNS was less hazardous than unmodified HAuNS. Unpurified HAuNS (50.60 nm) exposure to embryos caused deformity and increased mortality. Moreover, embryos exposed to HAuNS displayed an increase in cell death, showing that HAuNS can put zebrafish under physiological stress. The possible toxicity of HAuNS is now more understood thanks to this investigation. The details could improve our comprehension of the nanotoxicity of medication delivery systems. Comparing HAuNS (50~60 nm) to the other two particle sizes, its toxicity was quite low. Compared to unpurified HAuNS, purified HAuNS displayed less toxicity. Comparing PEI-HAuNS and HAuNS to PEG-HAuNS, cytotoxicity was found to be lower. Our data support the use of pure HAuNS, HAuNS-PEG, and HAuNS (50~60 nm) as possible photothermal conductors when seen as a whole.

Biosynthesis of Gold Nanoparticles Using Aqueous Extract of Paramignya trimera Roots and Its Anticancer Activity

Developing environment-friendly processes for metal nanoparticle synthesis without using toxic chemical is very necessary to meet the demand for biological and medical applications. In this work, biosynthesis of gold nanoparticles (AuNPs) using aqueous extract of Paramignya trimera roots as the reducing and stability agents was carried out. The formation of gold nanoparticles was confirmed by the presence of an absorption peak at 550 nm using UV–Visible spectrophotometry. The size and morphology of gold nanoparticles was monitored by Field emission scanning electron microscopy (FE-SEM) and High resolution transmission electron microscopy (HR-TEM). Analysis of these particles showed the synthesized AuNPs were spherical in shape, with the particle size range of 10 nm–15 nm. The element gold and the crystalline nature of AuNPs were finalized using Energy dispersive X-ray (EDX) and X-ray diffraction (XRD) spectrometer, respectively. Fourier transform infrared (FTIR) spectroscopy revealed possible involvement of reductive groups on the surfaces of nanoparticles. The synthesized AuNPs are evaluated for its activity against MCF-7, HepG2, KB, LU-1 and MKN7 cancer cell lines with inhibition concentration 50% (IC50) of 2.01, 7.60, 3.89, 3.99 and 3.53 ppm respectively after 72 hours of treatment.

Tet1 peptide and zinc (II)-adenine multifunctional module functionalized polycations as efficient siRNA carriers for Parkinson's disease

A bioreducible Zn (II)-adenine multifunctional module (BS) and Tet1 peptide were used to modify low-molecular-weight PEI3.5k (polyethyleneimine with molecular weight of 3.5 kDa)into a siRNA vector Zn-PB-T with high transfection efficiency in neurons. A GSH-responsive breakable disulfide spacer was introduced into BS to realize the controlled release of siRNA from the polyplexes in cytoplasm. Zn-PB showed >90% transfection rates in multiple cell lines (3 T3, HK-2, HepG2, 293 T, HeLa, PANC-1)，and 1.8-folds higher EGFP knockdown rates than commercial Lipo2k in normal cell line 293 T and cancer cell line HepG2. And Zn-PB-T1 showed 4.7–4.9- and 8.0–8.1-folds higher transfection efficiency comparing to commercial Lipo2k and PEI25k (polyethyleneimine with molecular weight of 25 kDa) in PC12 cells respectively, 2.1-fold EGFP gene silencing efficiency (96.6% EGFP knockdown rates) superior to commercial Lipo2k in neurons. In Parkinson's model, Zn-PB-T1/SNCA-siRNA can effectively protect neurons against MPP+-induced cell death and apoptosis, increasing the cell survival rate to 84.6% and reducing the cell apoptosis rate to 10.8%. This work demonstrated the promising application prospects of the resulting efficient siRNA carriers in siRNA-mediated gene therapy of Parkinson's disease.

Cytotoxic activity of Combretum quadrangulare leaf extracts on HepG2 cancer cell line

Studies on anticancer using plant extracts/compounds are promising. However, information regarding in vitro cytotoxicity is incompleted. Herein, this study aims to evaluate the cytotoxicity potentials of hexan:ethyl acetate (50:50, v:v) and ethanol extracts of Combretum quadrangulare leaves on HepG2 cancer cell line. Using cell viablity MTT assay, hexan:ethyl acetate and ethanol extracts were defined as potential cytotoxicity with IC50 at 38 and 47 µg/mL respectively. In addition, by microscopy observation, we found that morphology of the cells apparently change in a dose and time dependent manner. The data showed in this study shed new light for further investigation of the anticancer effect of C. quadrangulare extracts and its compounds.

Screening of Multitarget Compounds against Acetaminophen Hepatic Toxicity Using In Silico, In Vitro, and In Vivo Approaches.

Combination therapy and multitarget drugs have recently attracted much attention as promising tools to fight against many challenging diseases and, thus, represent a new research focus area. The aim of the current project was to screen multitarget compounds and to study their individual and combined effects on acetaminophen-induced liver injury. In this study, 2 of the best hepatoprotective multitargeting compounds were selected from a pool of 40 major compounds present in Curcuma longa and Cinnamomum zeylanicum by using molecular docking, ADMET profiling, and Pfizer's rule of five. The two selected compounds, quercetin and curcumin, showed a high binding affinity for the CYP2E1 enzyme, MAPK, and TLR4 receptors that contribute to liver injury. The candidates caused the decreased viability of cancer cell lines (HepG2 and Huh7) but showed no effect on a normal cell line (Vero). Examination of biochemical parameters (ALT, AST, ALP, and bilirubin) showed the hepatoprotective effect of the candidate drugs in comparison with the control group, which was confirmed by histological findings. Taken together, quercetin and curcumin not only satisfied the drug-like assessment criterion and proved to be multitargeting by preventing liver damage but also showed anticancer activities.

Production of natural flavor compounds using Bacillus subtilis-fermented soybean meal extract and their biological potential: a comprehensive in vitro study.

This study aims to investigate the production of natural flavor compounds through the utilization of Bacillus subtilis-fermented soybean meal extract and evaluate their biological potential. The experiment involved a comprehensive in vitro investigation to assess the capabilities and effects of the produced flavor compounds. The resulting flavor compounds were subjected to various in vitro tests to assess their properties, including cytotoxicity, antioxidant activity, anticancer potential, antiviral activity, and antimicrobial activity. To enhance the fermentation process, soybean meal extract was fortified with a combination of L-Lysine and L-Threonine. Gas chromatography-mass spectrometry (GC/MS) analysis was conducted on the fermented soybean meal using two strains of Bacillus subtilis, namely NRCH123 and NRCZ144. This analysis revealed the presence of various volatile compounds in all extracts, including Butylated hydroxytoluene. The fermented soybean extract with bacillus subtilis NRCZ144 (B2) fortified with a combination of 2.5% (w/w) L-Lysine and 2.5% w/w L-threonine (SLT2) exhibited a rich profile of flavor compounds, with Eucalyptol being identified as the predominant compound. The antioxidant activity of the SLT2 extract was found to be 72.04% at a concentration of 100 μg/mL, indicating significant antioxidant potential. Furthermore, when tested against the human liver cancer cell line HepG2, the extract demonstrated anticancer activity with an IC50 value of 2.26 μg/mL. The extract exhibited potent cytotoxicity, with an IC50 value of 1.02 μg/mL. Importantly, the SLT2 extract displayed strong antibacterial and antifungal activity, even at very low concentrations. The extract's antimicrobial properties indicate its potential for inhibiting the growth of bacteria and fungi.

Evaluation of Stability and In vitro Anti-Cancer Activity of Dihydroquercetin Nanoemulsion

Background: Dihydroquercetin (DHQ), also known as taxifolin, is a flavonoid commonly found in many plants. Dihydroquercetin has been documented to have powerful antioxidant activity and many beneficial properties for human health, especially its ability to inhibit certain types of cancer cells. However, its low solubility and bioavailability are major obstacles to biomedical applications. Moreover, DHQ is chemically unstable and quickly degrades when exposed to alkaline conditions. background: Dihydroquercetin (DHQ), also known as taxifolin, is a flavonoid and commonly found in many plants. Dihydroquercetin has been documented to have powerful antioxidant activity and many beneficial properties for human health, especially its ability to inhibit certain types of cancer cells. However, its low solubility and bioavailability are major obstacles to biomedical applications. Moreover, DHQ is chemically unstable and quickly degrades when exposed to alkaline conditions. Objective: In the present study, a DHQ nanoemulsion formulation was prepared by Self Nano- Emulsifying Drug Delivery System (SNEDDS) technique to overcome the above disadvantages. Methods: The obtained nanoemulsion system was evaluated for its micro-properties, stability, and in vitro cytotoxic activity against some cancer cells using tetrazolium dyes (MTS assay). Results: Measurement results showed that the DHQ nanoemulsion was successfully synthesized with typical mean droplet sizes from 9 to 11 nm, and revealed excellent stability over time. Dihydroquercetin in nanoemulsion form is more stable than the non-encapsulated form, as evidenced by the maintenance of droplet size in the nanometer range when dispersed in aqueous solution for up to 48 hours. This stability is particularly pronounced in both acidic and neutral environments. In vitro experiments on cytotoxic activities against A549, Hela, and HepG2 cancer cell lines indicated that the prepared DHQ nanoemulsion effectively inhibited the growth of all these cell lines with IC50 values (μg/mL) of 8.0, 20.4, and 29.5 respectively. Conclusion: From the detailed results above, it is evident that the solubility and bioavailability of DHQ can be improved by creating its nanostructure in the form of nanoemulsions. Furthermore, the nano form of DHQ carried within stable nanoemulsions exhibited better performance in inhibiting cancer cells compared to free DHQ. Therefore, further research is required to explore the development of cancer therapeutics utilizing nano DHQ emulsions.

Comparative Analysis of Phytochemical Profiles and Selected Biological Activities of Various Morphological Parts of Ligustrum vulgare.

Ligustrum vulgare (LV), widely cultivated in Europe and often used in hedges, has been historically recognized in folk medicine for its potential health benefits. This study focused on exploring the untargeted identification of secondary metabolites in ethanol extracts (70% v/v) from different morphological parts (young shoots, leaves, flowers and fruits) of LV at various stages of plant development, using ultra-high-performance liquid chromatography with high-resolution mass spectrometry (UHPLC-HRMS). Additionally, the selected biological activities (antioxidant activity, cyclooxygenase-2 inhibition (COX-2), α-amylase inhibition and cytotoxicity) of the tested extracts were determined. Untargeted metabolomics showed that LV extracts were a rich source of phenylethanoid compounds, flavonoids, iridoids and their derivatives. The flowers of LV had the highest content of oleuropein (33.43 ± 2.48 mg/g d.w.). The lowest antioxidant activity was obtained for ripe and post-seasonal fruits, while in the case of other samples, the activity was at a similar level. All tested extracts showed α-amylase and COX-2 inhibitory activity. In addition, LV extracts showed strong antiproliferative properties in colorectal (HT29) and liver (HepG2) cancer cell lines. The obtained results show the difference in the content of bioactive compounds in various morphological parts of Ligustrum vulgare. These differences may influence the multifaceted medicinal potential of this plant.

Five undescribed aryltetralin lignans with cytotoxic activities from the fruits of Cleistanthus eberhardtii

Five undescribed lignans, cleiseberharnins A–D (1–4), cleiseberharside A (5) were isolated from the fruits of Cleistanthus eberhartii (Phyllanthaceae), together with six known aryltetralin lignans, cleistantoxin (6), picroburseranin (7), neocleistantoxin (8), 7-hydroxypicropolygamain (9), cleisindoside D (10), and cleisindoside A (11). Their structures and relative configurations were established by analysis of HRESIMS and NMR data, and quantum chemical calculations of JH,H coupling constants. The absolute configurations of 1–5 were determined by analysis of their experimental CD spectra and comparison with calculated electronic circular dichroism (ECD) spectra. All compounds (1−11) were evaluated for their cytotoxicity against KB, MCF-7, HepG-2, and Lu-1 human cancer cell lines. Among the tested compounds, compounds 6 and 7 showed strong activity against KB, MCF7, HepG2 and Lu-1 cell lines with IC50 values in the range of 0.02–0.62 μM. Compound 1 showed activity against three cancer cell lines KB, HepG2, and Lu-1 with IC50 values of 6.98, 7.61 and 11.75 μM, respectively. Compound 2 exhibited a selective inhibition with moderate cytotoxicity against Lu-1 with IC50 value of 15.30 μM. Compounds 4, 5 and 9 showed moderate activity against the three cancer cell lines with IC50 values in the range of 8.73–19.70 μM.