Outbreaks of hand, foot, and mouth disease (HFMD) that occur worldwide are mainly caused by the Coxsackievirus-A16 (CV-A16) and Enterovirus-A71 (EV-A71). Unfortunately, neither an anti-HFMD drug nor a vaccine is currently available. Rupintrivir in phase II clinical trial candidate for rhinovirus showed highly potent antiviral activities against enteroviruses as an inhibitor for 3C protease (3Cpro). In the present study, we focused on designing 50 novel rupintrivir analogs against CV-A16 and EV-A71 3Cpro using computational tools. From their predicted binding affinities, the five compounds with functional group modifications at P1′, P2, P3, and P4 sites, namely P1′-1, P2-m3, P3-4, P4-5, and P4-19, could bind with both CV-A16 and EV-A71 3Cpro better than rupintrivir. Subsequently, these five analogs were studied by 500 ns molecular dynamics simulations. Among them, P2-m3, the derivative with meta-aminomethyl-benzyl group at the P2 site, showed the greatest potential to interact with the 3Cpro target by delivering the highest number of intermolecular hydrogen bonds and contact atoms. It formed the hydrogen bonds with L127 and K130 residues at the P2 site stronger than rupintrivir, supported by significantly lower MM/PB(GB)SA binding free energies. Elucidation of designed rupintrivir analogs in our study provides the basis for developing compounds that can be candidate compounds for further HFMD treatment.

Janus kinase (JAK) deregulation of the JAK/signal transducers and activators of transcription pathway leads to myelofibrosis that can be treated by JAK inhibitors including Ruxolitinib and Tofacitinib. Even though both inhibitors are effective against myelofibrosis, each of them has a different mode of action in the cells. Ruxolitinib is an inhibitor for selective JAK1/2, and Tofacitinib is an inhibitor for JAK3. This study evaluated the chemical fingerprints of TF-1 cells after JAK inhibitor treatments by the synchrotron Fourier transform infrared microspectroscopy (S-FTIR) spectrum. Tofacitinib and Ruxolitinib treatments in TF-1 cells were applied with a chemical fingerprint approach in S-FTIR spectroscopy and in vitro cytotoxicity in a cell-based assay. Principal component analysis or PCA was utilized to classify three cell treatments with three biochemical alteration absorbances of lipid vibration by the C-H stretching, protein amide I that appeared from the C=O stretching, and a P=O phosphodiester bond from nucleic acids. The results showed that the inhibition effect of Ruxolitinib on the TF-1 cell lines was two-fold higher than Tofacitinib. PCA distinguishes untreated and drug-treated cells by detecting cellular biochemical alteration. The loading plots identify that proteins and nucleic acids were the different main components in disparate cell treatments. Tofacitinib was distinct from the others in lipid and nucleic acid. The second derivative spectra of the three molecular components had decreased lipid production and accumulation, changes in secondary structures in proteins, and a high level of RNA overexpression in cell treatment. The JAK inhibitors caused different spectroscopic biomarkers of the modifications of secondary protein conformation, stimulated cell lipid accumulation, and phosphorylation from untreated cells. The alteration of cellular biochemical components suggests that FTIR is a potential tool to analyze specific patterns of drug cellular responses at the molecular level.

Context: Inhibiting α-glucosidase activity has been proven to regulate hyperglycemia for treating diabetes, particularly type 2 diabetes mellitus (T2DM). Plant secondary metabolites consist of phenolic acids and flavonoids, which have been identified as α-glucosidase inhibitory agents. Aims: To assess the ability of the tested phenolic acids and flavonoids as α-glucosidase inhibitors and to predict their binding interactions with α-glucosidase, as well as their pharmacokinetic properties. Methods: The tested phenolic acids and flavonoids were evaluated for α-glucosidase inhibition by in vitro study, and their binding interactions were observed by an in silico molecular docking study utilizing AutoDock 4.2 software, while their pharmacokinetic properties were predicted using SwissADME and pkCSM online software. Results: The results displayed that quercetin (QE; IC50 = 0.85 μg/mL) and gallic acid (GA; IC50 = 26.19 μg/mL) significantly inhibited α-glucosidase activity than acarbose (IC50 = 139.4 μg/mL). Molecular docking analysis revealed that QE exhibited a higher binding affinity (ΔG; -7.53 kcal/mol) compared to acarbose (ΔG; -7.00 kcal/mol). This suggested that QE formed more favorable interactions with the enzyme, as evidenced by its binding to catalytic residues D215 and D352. In contrast, acarbose interacted with only one catalytic amino acid, i.e., D215. Furthermore, GA displayed a lower binding affinity (ΔG; -4.63 kcal/mol) than acarbose and bound to a distinct site on the α-glucosidase involving amino acids K156, S241, and D242. The pharmacokinetic properties predictions revealed that the potential compounds could bind and inhibit the α-glucosidase and were not toxic. Conclusions: Therefore, this study is useful for modifying the structures of phenolic acids and flavonoids to improve their ability to inhibit the α-glucosidase activity.