Development of an Icariin-Loaded Bilosome-Melittin Formulation with Improved Anticancer Activity against Cancerous Pancreatic Cells

Pancreatic cancer currently represents a severe issue for the entire world. Therefore, much effort has been made to develop an effective treatment against it. Emerging evidence has shown that icariin, a flavonoid glycoside, is an effective anti-pancreatic cancer drug. Melittin, as a natural active biomolecule, has also shown to possess anticancer activities. In the present study, with the aim to increase its effectiveness against cancerous cells, icariin-loaded bilosome-melittin (ICA-BM) was developed. For the selection of an optimized ICA-BM, an experimental design was implemented, which provided an optimized formulation with a particle size equal to 158.4 nm. After estimation of the release pattern, the anti-pancreatic cancer efficacy of this new formulation was evaluated. The MTT assay was employed for the determination of half maximal inhibitory concentration (IC50), providing smaller IC50 for ICA-BM (2.79 ± 0.2 µM) compared to blank-BM and ICA-Raw (free drug) against PNAC1, a human pancreatic cancer cell line isolated from a pancreatic carcinoma of ductal cell origin. Additionally, cell cycle analysis for ICA-BM demonstrated cell arrest at the S-phase and pre-G1 phase, which indicated a pro-apoptotic behavior of the new developed formulation. The pro-apoptotic and anti-proliferative activity of the optimized ICA-BM against PNAC1 cells was also demonstrated through annexin V staining as well as estimation of caspase-3 and p53 protein levels. It can be concluded that the optimized ICA-BM formulation significantly improved the efficacy of icariin against cancerous pancreatic cells.

Early Optimization Stages of Agave lechuguilla Bagasse Processing toward Biorefinement: Drying Procedure and Enzymatic Hydrolysis for Flavonoid Extraction

Agave lechuguilla agro-waste is a promising renewable material for biorefining purposes. The procurement of added-value co-products, such as bioactive phytochemicals, is required to improve bioprocesses and promote the bio-based economy of the productive areas of Mexico. In this study, we aimed to evaluate the effect of post-harvest management and enzymatic pretreatment as the first stages of the A. lechuguilla valorization process. Four drying methods were compared, and enzymatic hydrolysis was optimized to obtain a flavonoid-enriched extract applying ultrasound-assisted extraction. In both experiments, the total phenolic (TPC) and flavonoid (TFC) contents, HPLC-UV flavonoid profiles, and radical scavenging capacity (DPPH) were considered as response variables. The results demonstrated that light exposure during the drying process particularly affected the flavonoid content, whereas oven-dehydration at 40 °C in the dark preserved the flavonoid diversity and antioxidant functionality of the extracts. Flavonoid glycoside recovery, particularly anthocyanidins, was 1.5–1.4-fold enhanced by enzymatic hydrolysis using the commercial mix Ultraflo© under optimized conditions (pH 4, 40 °C, 180 rpm, and 2.5 h) compared to the unpretreated biomass. The extraction of flavonoids from A. lechuguilla bagasse can be carried out using a scalable drying method and enzymatic pretreatment. This study confirmed the potential of this agro-waste as a source of marketable natural products.

Effects of Cynaroside on Cell Proliferation, Apoptosis, Migration and Invasion though the MET/AKT/mTOR Axis in Gastric Cancer

The Chinese medicine monomer cynaroside (Cy) is a flavonoid glycoside compound that widely exists in plants and has a variety of pharmacological effects, such as its important role in the respiratory system, cardiovascular system and central nervous system. Studies have reported that Cy has varying degrees of anticancer activity in non-small cell lung cancer, cervical cancer, liver cancer, esophageal cancer and other cancers. However, there are no relevant reports about its role in gastric cancer. The MET/AKT/mTOR signaling pathway plays important roles in regulating various biological processes, including cell proliferation, apoptosis, autophagy, invasion and tumorigenesis. In this study, we confirmed that Cy can inhibit the cell growth, migration and invasion and tumorigenesis in gastric cancer. Our finding shows that Cy can block the MET/AKT/mTOR axis by decreasing the phosphorylation level of AKT, mTOR and P70S6K. Therefore, the MET/AKT/mTOR axis may be an important target for Cy. In summary, Cy has anti-cancer properties and is expected to be a potential drug for the treatment of gastric cancer.

Scutellarin Reduces Cerebral Ischemia Reperfusion Injury Involving in Vascular Endothelium Protection and PKG Signal

Flavonoid glycoside scutellarin (SCU) has been widely applied in the treatment of cerebral ischemic diseases in China. In this article, we conducted research on the working mechanisms of SCU in hypoxia reoxygenation (HR) injury of isolated cerebral basilar artery (BA) and erebral ischemia reperfusion (CIR) injury in rat models. In isolated rat BA rings, HR causes endothelial dysfunction (ED) and acetylcholine (ACh) induces endothelium-dependent vasodilation. The myography result showed that SCU (100 µM) was able to significantly improve the endothelium-dependent vasodilation induced by Ach. However, SCU did not affect the ACh-induced relaxation in normal BA. Further studies suggested that SCU (10–1000 µM) dose-dependently induced relaxation in isolated BA rings which were significantly blocked by the cGMP dependent protein kinase (PKG) inhibitor Rp-8-Br-cGMPs (PKGI-rp, 4 µM). Pre-incubation with SCU (500 µM) reversed the impairment of endothelium-dependent vasodilation induced by HR, but the reversing effect was blocked if PKGI-rp (4 µM) was added. The brain slice staining test in rats’ model of middle cerebral artery occlusion (MCAO) induced CIR proved that the administration of SCU (45, 90 mg/kg, iv) significantly reduced the area of cerebral infarction. The Western blot assay result showed that SCU (45 mg/kg, iv) increased brain PKG activity and PKG protein level after CIR surgery. In conclusion, our findings suggested that SCU possesses the ability of protecting brain cells against CIR injury through vascular endothelium protection and PKG signal. Graphic Abstract

Rutin (Bioflavonoid) as Cell Signaling Pathway Modulator: Prospects in Treatment and Chemoprevention

Cancer is a complex ailment orchestrated by numerous intrinsic and extrinsic pathways. Recent research has displayed a deep interest in developing plant-based cancer therapeutics for better management of the disease and limited side effects. A wide range of plant-derived compounds have been reported for their anticancer potential in the quest of finding an effective therapeutic approach. Rutin (vitamin P) is a low-molecular weight flavonoid glycoside (polyphenolic compound), abundantly present in various vegetables, fruits (especially berries and citrus fruits), and medicinal herbs. Numerous studies have delineated several pharmacological properties of rutin such as its antiprotozoal, antibacterial, anti-inflammatory, antitumor, antiviral, antiallergic, vasoactive, cytoprotective, antispasmodic, hypolipidemic, antihypertensive, and antiplatelet properties. Specifically, rutin-mediated anticancerous activities have been reported in several cancerous cell lines, but the most common scientific evidence, encompassing several molecular processes and interactions, including apoptosis pathway regulation, aberrant cell signaling pathways, and oncogenic genes, has not been thoroughly studied. In this direction, we attempted to project rutin-mediated oncogenic pathway regulation in various carcinomas. Additionally, we also incorporated advanced research that has uncovered the notable potential of rutin in the modulation of several key cellular functions via interaction with mRNAs, with major emphasis on elucidating direct miRNA targets of rutin as well as the process needed to transform these approaches for developing novel therapeutic interventions for the treatment of several cancers.

Rutin Ameliorates Malaria Pathogenesis by Modulating Inflammatory Mechanism: An In-Vitro and In-Vivo Study

Rutin (3,3’,4’ 5,7-pentahydroxyflavone-3-rhamnoglucoside) is a flavonoid glycoside, found in many edible plants such as buckwheat and berries. Rutin as a food supplement is recommended for the treatment of various diseases, which directed us to investigate its valuable effects in malaria induced pathogenesis. In the present study, Rutin was tested for its anti-plasmodial activity against chloroquine sensitive and resistant strains (NF-54 and K1) of Plasmodium falciparum and studied for its anti-oxidative and anti-inflammatory potential against LPS stimulated macrophage cells. In vitro observations were further validated using an in-vivo physiological rodent model of Plasmodium berghei-induced malaria pathogenesis. Rutin was also tested for its effect in combination with chloroquine.Rutin was found to have potent anti-plasmodial activity against both chloroquine sensitive and resistant strains of P. falciparum (NF-54 and K1). It was able to reduce the oxidative stress induced by LPS in macrophage cells with decreased production of pro-inflammatory cytokines (IL-6, TNF-α and IL-1β). Rutin was found to significantly suppress the parasitaemia, increase the mean survival time and restored the haemoglobin and glucose level in in vivo assays. This was corroborated by reduced production of malondialdehyde (MDA) and pro-inflammatory mediators in rutin treated mice in P.berghei-induced malaria pathogenesis. Interestingly, the combination of rutin with chloroquine had shown synergy in both in vitro and in vivo experiments. The findings of the present study thus highlighted the suitability of rutin for further study in the management of drug resistant malaria, alone or in combination with other compounds.

Phenolic compounds as antiviral agents: An In-Silico investigation against essential proteins of SARS-CoV-2

Plant biodiversity is endowed with a huge composition and variability of active molecules known for their therapeutic effects against several diseases. In this current work, several phenolic compounds are subject of in silico evaluation of their interactions with six severe acute respiratory syndrome coronavirus (SARS-CoV) enzymes to evaluate the binding mode and mechanism of phenolic compounds interactions with SARS-CoV-2 enzymes. Results of molecular docking and data analysis revealed that the importance of interactions was dependent to the phenolic class of tested ligand; tannin, biflavone and flavonoid glycoside were the most interactive classes. Among the top three ranked molecules recording lower binding energy against each virus protein target, In conclusion, it was found that Amentoflavone, Dieckol, Bilobetin, Punicalagin, Tellimagrandin-I, Tannic acid, Sciadopitysin, Ginkgetin and Chebulagic acid could be a promising antiviral drug since they present more important binding energy than conventional ones. Their interactions were justified by the Wenn diagram and Ramachandran plot. However, these phenolic compounds recorded an important bioavailability score and found fulfilling most of the drug-likeness criteria such as Lipinski's rule. Clearly, all observations point to further required works aiming to examine more deeply the possibility of using these molecules that could be probably a subject of pre-clinical studies.

Antimalarial Flavonoid-Glycoside from Acacia pennata with Inhibitory Potential Against PfDHFR-TS: An In-silico Study

Drug resistance, toxicity, and adverse effects of current antimalarial drugs have mandated the need to search for newer antimalarial agents. The present study aims to identify promising flavonoid-glycosides (FGs) from Acacia pennata as possible antimalarial agents effective against PfDHFR-TS (PDB ID: 3DGA) by in-silico studies. The co-crystal inhibitor (RJ1) of PfDHFR-TS was used as the reference standard compound. A compound library of 17 FGs reported to be isolated from A. pennata was prepared and subjected to molecular docking simulation studies. PyRx 0.8 and AutoDock Vina revealed Pinocembrin-7-O-β-D-glucopyranoside (FG17) as the best PfDHFR-TS inhibitor with a binding affinity of -10.4 kcal/mol and -10.8 kcal/mol, respectively. In both methods, FG17 has a better binding affinity than the co-crystal inhibitor, RJ1 (-7.9 kcal/mol). The docking protocols were validated by RMSD calculation with Discovery Studio Visualizer software (2020). FG17 interacted with amino acids ALA16, LEU40, SER167, GLY41, GLY44, MET55, PHE58, ILE112, LEU119, GLY166, and TYR170 at the active binding site of PfDHFR-TS. Further, FG17 was computed as a non-toxic, bioavailable, synthetically accessible compound and a better enzyme inhibitor than RJ1. Hence, we conclude that FG17 may be used as a lead scaffold to design antimalarial agents against PfDHFR-TS in the future.

Based on Activation of p62-Keap1-Nrf2 Pathway, Hesperidin Protects Arsenic-Trioxide-Induced Cardiotoxicity in Mice

Background: Hesperidin (HES) is a flavonoid glycoside found in the tangerine peel and has antioxidant properties. Arsenic trioxide (ATO) is an anti-tumour drug; however, its serious cardiotoxicity limits its clinical application. In addition, the protection of HES against ATO-induced cardiotoxicity has not been explored.Objective: The study aims to investigate and identify the underlying effect and mechanism of HES on ATO-induced cardiotoxicity.Methods: Fifty mice were randomly assigned to five groups. Mice were orally given HES:100 or 300 mg/kg/day concurrently and given ATO intraperitoneal injections: 7.5 mg/kg/day for 1 week. Blood and heart tissues were collected for examination. Evaluated in serum was the levels of creatine kinase (CK), lactate dehydrogenase (LDH) and cardiac troponin I (cTnI). In addition, evaluated in heart tissues were the levels of reactive oxygen species (ROS), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), catalase (CAT), tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), Caspase-3, cleaved-Caspase-3, p62, Kelch-like ECH-associated protein 1 (Keap1), and nuclear factor erythroid 2-related factor 2 (Nrf2). The heart tissues were also examined for histopathology and mitochondrial ultrastructure.Results: Compared with the ATO group, the HES treatment groups reduced the levels of CK, LDH, cTnI, ROS, MDA, TNF-α, IL-6, Bax, Caspase-3, cleaved-Caspase-3 and Keap1 and enhanced the levels of SOD, GSH, CAT, Bcl-2, p62 and Nrf2.Conclusions: The results demonstrate that HES protects against ATO-induced cardiotoxicity, through inhibiting oxidative stress, and subsequent inflammation and apoptosis. The underlying results are closely related to the regulation of the p62-Keap1-Nrf2 signalling pathway.

Tilianin Promotes the Proliferation and Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells

Osteoporosis is a systemic bone disease characterized by a decrease in bone mineral density and mass. To examine the mechanism(s) underlying the pathogenesis of osteoporosis, we have used an in vitro model of osteoporosis induced by exposure to high glucose. Tilianin is a flavonoid glycoside isolated from Dracocephalum moldavica L. that has been reported to exhibit a variety of pharmacologic activities. However, the utility of tilianin in the treatment of osteoporosis remains unexplored. To this end, we have examined the effect of tilianin on bone marrow mesenchymal stem cells exposed to high glucose. Our data revealed that tilianin suppressed apoptosis, promoted osteogenic differentiation, and survival of the bone marrow mesenchymal stem cells in the presence of high glucose. Our data therefore confirmed that tilianin could serve as a promising drug for the treatment of osteoporosis.