Inhibitory Activity of Flavonoid Scaffolds on SARS-CoV-2 3CLpro: Insights from the Computational and Experimental Investigations.

The emergence of the COVID-19 situation has become a global issue due to the lack of effective antiviral drugs for treatment. Flavonoids are a class of plant secondary metabolites that have antiviral activity against SARS-CoV-2 through inhibition of the main protease (3CLpro). In this study, 22 flavonoids obtained from natural sources and semisynthetic approaches were investigated for their inhibitory activity against SARS-CoV-2 3CLpro, along with cytotoxicity on Vero cells. The protein-ligand interactions were examined using molecular dynamics simulation. Moreover, QSAR analysis was conducted to clarify the structural effects on bioactivity. Accordingly, the in vitro investigation demonstrated that four flavonoids, namely, tectochrysin (7), 6″,6″-dimethylchromeno-[2″,3″:7,8]-flavone (9), panduratin A (19), and genistein (20), showed higher protease inhibitory activity compared to the standard flavonoid baicalein. Finally, our finding suggests that genistein (20), an isoflavone discovered in Millettia brandisiana, has potential for further development as a SARS-CoV-2 3CLpro inhibitor.

Microwave-Assisted Extraction of Anticancer Flavonoid, 2',4'-Dihydroxy-6'-methoxy-3',5'-dimethyl Chalcone (DMC), Rich Extract from Syzygium nervosum Fruits.

2',4'-Dihydroxy-6'-methoxy-3',5'-dimethyl chalcone (DMC) is a biological flavonoid that is present in the fruits of Syzygium nervosum (Ma-Kiang in Thai). Microwave-assisted extraction (MAE), which utilizes microwave radiation to heat the extraction solvent quickly and effectively, was used to recover DMC-rich extract from Syzygium nervosum fruit. To determine the DMC content, a highly accurate and precise HPLC technique was developed. The influences of MAE conditions, including the solid-liquid ratio, microwave power, and microwave duration on the content of DMC, were sequentially employed by a single factor investigation and response surface methodology (RSM) exploratory design. The predicted quadratic models were fitted due to their highly significant (p < 0.0001) and excellent determination coefficient (R2 = 0.9944). The optimal conditions for producing DMC-rich extract were a ratio of sample to solvent of 1:35 g/mL, a microwave power of 350 W, and a microwave time of 38 min. Under the optimal MAE setting, the DMC content reached 1409 ± 24 µg/g dry sample, which was greater than that of the conventional heat reflux extraction (HRE) (1337 ± 37 µg/g dry sample) and maceration (1225 ± 81 µg/g dry sample). The DMC-rich extract obtained from MAE showed stronger anticancer activities against A549 (human lung cancer cells) and HepG2 (human liver cancer cells) than the individual DMC substance, which makes MAE an effective method for extracting essential phytochemicals from plants in the nature.

Microwave-Assisted Improved Extraction and Purification of Anticancer Nimbolide from Azadirachta indica (Neem) Leaves.

Nimbolide, a limonoid present in leaves of the neem tree (Azadirachta indica), is an anticancer compound against a panel of human cancer cell lines. The rapid process of extraction and purification of the nimbolide from the leaves of neem tree through microwave-assisted extraction (MAE) coupled with a chromatographic technique was accomplished. The crude with a maximum content of nimbolide could be recovered from neem leaves through MAE. By using three-factors, three-level Box-Behnken design of response surface methodology (RSM), the optimal conditions for nimbolide extraction (R2 = 0.9019) were solid/liquid ratio 1:16 g/mL, microwave power 280 W, and extraction time 22 min. The enriched extract was further purified by a preparative thin-layer chromatography (PTLC), where nimbolide was obtained as 0.0336 g (0.67% yield, purity over 98%) with ethyl acetate/hexane = 4:6 in 3.0 h. Structural elucidation was performed through spectroscopic techniques, including FT-IR, 1H, and 13C-NMR. This method was simple and had a good potential for the purification of bioactive compounds from a natural product.

Antibacterial activity of Dioscorea bulbifera Linn. extract and its active component flavanthrinin against skin-associated bacteria.

BACKGROUND: Dioscorea bulbifera Linn. has been used for wound care in Thailand. However, a comprehensive evaluation of its antibacterial activity is required. This study aimed to investigate the antibacterial efficacy of D. bulbifera extract against skin-associated bacteria and isolate and characterize its active antibacterial agent, flavanthrinin. METHODS: Air-dried bulbils of D. bulbifera were pulverised and extracted with hexane, dichloromethane, ethyl acetate, methanol, ethanol, and distilled water; vacuum filtered; concentrated; freeze-dried; and stored at -20 ºC. Antibacterial activity of the extracts was assessed using microdilution techniques against several skin-associated bacteria. Thin-layer chromatography (TLC) bioautography was used to identify the active compounds in the extract, which were fractionated by column chromatography and purified by preparative TLC. The chemical structures of the purified compounds were analysed using nuclear magnetic resonance (NMR). The cytotoxicity of the extract and its active compounds was evaluated in Vero cells. RESULTS: The ethyl acetate extract exhibited distinct inhibition zones against bacteria compared to other extracts. Therefore, the ethyl acetate extract of D. bulbifera in the ethyl acetate layer was used for subsequent analyses. D. bulbifera extract exhibited antibacterial activity, with minimum inhibitory concentrations (MICs) of 0.78-1.56 mg/mL. An active compound, identified through TLC-bioautography, demonstrated enhanced antibacterial activity, with MICs of 0.02-0.78 mg/mL. NMR analysis identified this bioactive compound as flavanthrinin. Both D. bulbifera extract and flavanthrinin-containing fraction demonstrated potent antibacterial activity against Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), and S. epidermidis. The flavanthrinin containing fraction demonstrated low cytotoxicity against Vero cells, showing CC50 values of 0.41 ± 0.03 mg/mL. These values are lower than the MIC value, indicating that this fraction is safer than the initial ethyl acetate extract. CONCLUSIONS: Dioscorea bulbifera extract and its bioactive component flavanthrinin demonstrated significant antibacterial activity against the skin-associated bacteria Staphylococci, including MRSA. Flavanthrinin has potential as a complementary therapeutic agent for managing skin infections owing to its potent antibacterial effects and low cytotoxicity.

Microwave-assisted commercial copper-catalyzed aerobic oxidative synthesis of AChE quinazolinone inhibitors under solvent free conditions.

A facile and green one-pot synthesis of AChE quinazolinone inhibitors was developed using microwave irradiation under solvent free conditions. Quinazolinones were synthesized from 2-aminobenzamide derivatives and various alcohols such as benzyl alcohol derivatives and butanol using economical commercially available copper as a catalyst in the presence of base, Cs2CO3. The desired products were achieved in moderate to high yields with up to 92% isolated yield. These quinazolinone products were then evaluated for acetylcholinesterase inhibition so that they can be developed as promising anti-acetylcholinesterase agents.

Polyester-releasing sesamin by electrospinning technique for the application of bone tissue engineering.

Sesamin, a significant lignin compound isolated from sesame (Sesamum indicum Linn), is well known for its antioxidant, anti-inflammatory, and tissue growth promotion properties. Bioabsorbable poly(ε-caprolactone) (PCL) is also a well-known polymer applied to various fields of medicine as biomaterials. The main objective of this research was to produce a prototype material from PCL and sesamin by electrospinning technique for bone tissue engineering applications. Dichloromethane and dimethylformamide (7:3) mixture was used as the solvent system for fabrication of PCL nanofiber with different loads of sesamin concentrations (1-6 wt%). The crystallinity levels decreasing and the entrapment efficiency increasing (86.87%-93.97%) were observed while sesamin concentrations were increased. The infrared spectra of electrospun mats confirmed that sesamin corporated into fibrous networks. The sesamin-loaded PCL nanofibrous membranes showed a significant release of sesamin in the range of 1.28-8.16 µg/mL within 10 weeks. The release data were fitted to zero order, first order, Higuchi and Korsmeyer-Peppas models to evaluate sesamin-releasing mechanisms and kinetics. The releasing kinetics of sesamin followed the Fickian diffusion mechanism of Korsmeyer-Peppas (R2 = 0.99). In vitro experiments with an osteosarcoma cell line (MG-63) revealed cell attachment, biocompatibility, and promotion of bone marker expression, the alkaline phosphatase (ALP) activity were studied. The electrospun PCL nanofiber loaded with sesamin had the potential as a scaffold for sesamin delivery to bone cells and applications in biomedicine.