Current Pharmaceutical Design - Online First

Sepsis-induced acute lung injury (S-ALI) is one of the diseases with a very high fatality rate. However, the traditional Chinese medicine compound Buzhong Yiqi Decoction (BZYQD) has an excellent effect in the treatment of S-ALI. Nevertheless, its mechanism of action is still unclear. In this study, we explored the molecular mechanisms of S-ALI injury treated with buzhong yiqi decoction through network pharmacology, in combination with in vivo experimental validation.

Traditional Chinese medicine system pharmacology (TCMSP) database was used to screen thechemical composition of BZYQD and its action targets; Multiple databases were used to collect target genesfor-S-ALI, including OMIM, TTD, GeneCards, and DrugBank; The STRING database was used for the protein-protein interaction (PPI) analysis of the common targets of the BZYQD and the S-ALI; The DAVID databasewas used for GO and KEGG analysis; molecular docking was used to detect the binding capacity of corecomponents and targets. HE staining was used to visualize the pathology of lung tissue in each group; ELISA wasused to detect the levels of inflammatory factors (IL-1β, IL-6, IL-8, NF-κB and TNF-α) and oxidative stressrelatedfactors (LDH, CK-MB, SOD, GSH-Px); The qPCR and Western blot were used to examine the mRNAand protein expression of IL-1β, IL-6, TNF-α NF-κB, p-NF-κB, PI3K, p-PI3K, AKT, and IKKα.

113 chemical components and 226 targets were screened from BZYQD; 9059 S-ALI-related geneswere screened out, with a total of 228 intersecting targets between BZYQD and S-ALI. Stigmasterol, quercetin, and isorhamnetin are the core components of BZYQD, PPI analysis shows that AKT1, IL6, TNF, andIL1B are the core targets of BZYQD for treating S-ALI, and molecular docking results show that the corecomponents have high binding activity with the target; Enrichment analysis shows that these core targets arerelated to the TNF signaling pathway. In vivo experimental studies have found that BZYQD can improve thedegree of inflammatory infiltration and edema in lung tissue of S-ALI model mice, reduce the expression ofIL-6, IL-1β, IL-8, TNF-α, LDH, CK-MB, and NF-κB in serum (P0.05), as well as the mRNA and proteinexpression of IL-6, IL-1β, TNF-α, NF-κB, p-NF-κB, PI3K, p-PI3K, AKT, and IKKα in lung tissue (P0.05),and levels of SOD and GSH-Px were increased (P0.05).

The action targets of the main chemical components of BZYQD are TNF, AKT, and IL6. Thesetargets can promote the activation of PI3K and TNF pathways and mediate the occurrence of inflammationand oxidative stress, which provides inspiration for the treatment of S-ALI. However, the results of this study still need to be verified in combination with in vitro approaches.

This study suggests that the mechanism of BZYQD in treating S-ALI may be achieved by inhibiting the TNF and PI3K signaling pathway and reducing inflammation and oxidative stress levels.

Pulmonary fibrosis (PF) is a chronic pulmonary disorder with unknown etiology and an irreversible course. Traditional Chinese medicine (TCM) possesses promising clinical benefits for PF treatment through a multi-component and multi-target approach. This study evaluates the efficacy of Yangyin Yifei Tongluo Wan (YF), a traditional formulation, in the treatment of PF, and further explores the underlying mechanism.

A bleomycin (BLM)-induced PF mouse model was established. Mice were administered with low-, medium-, and high-dose YF (1.5, 3, and 6 g/kg/d, respectively). The fibrosis degree of mouse lung tissues was evaluated by morphometric measurements and hydroxyproline (HYP) analysis. Network pharmacology-based bioinformatics were employed for constructing a network involving components, targets, and disease, and YF's potential mechanism and molecular targets for PF therapy were explored. This was further validated by TUNEL staining, Western blot, RT-qPCR, and ELISA in BLM-treated mice.

YF could relieve PF in BLM-treated mice in a dose-dependent manner, evidenced by a notable decrease in collagen deposition, and collagen I and III, HYP, fibronectin, vimentin, and α-SMA expressions. Network pharmacology revealed that JAK2/STAT3 signaling pathway-mediated alveolar epithelial cell apoptosis may be a potential therapeutic target for YF in treating PF. In vivo assays confirmed that YF's anti-fibrosis effect on BLM-induced PF was ascribed to the suppression of alveolar epithelial cell apoptosis and disruption of the JAK2/STAT3 signaling pathway.

YF can block alveolar epithelial cell apoptosis through inactivation of the JAK2/STAT3 signaling, subsequently enhancing the resolution of PF.

YF may be a promising therapeutic candidate for PF treatment.

Polycystic ovarian syndrome (PCOS) is a hormonal condition that affects women of reproductive age. The purpose of this study was to identify the undiscovered molecular mechanisms by which Stachys lavandulifolia treats PCOS. Although Stachys lavandulifolia has been used to treat PCOS, its exact biological mechanism of action remains unknown.

We used a multifaceted strategy that included network pharmacology, molecular docking, and molecular dynamics simulations.

Network pharmacology discovered 68 gene targets shared by Stachys lavandulifolia bioactive chemicals and PCOS-associated genes. Subsequent KEGG and Reactome analysis identified 18 enhanced pathways, including steroid hormone production, glucose homeostasis, and insulin resistance. Key genes involved in ovarian steroidogenesis and the hypothalamic-pituitary-ovarian axis (CYP19A1, Kiss1, human androgen receptor, oestrogen receptor alpha, and HSD17B1) were chosen for molecular docking.

Molecular docking indicated that bioactive substances Myrsen, Agnol, Alpha Pyogenin, and Gamma Morolen have high binding affinities for the identified target proteins. Notably, the CYP19A1-Myrsen complex has the highest binding affinity at -9.0 kcal/mol. Additional molecular dynamics simulations indicated that the CYP19A1-Myrsen complex had increased flexibility and mobility, indicating a stable and effective association.

Our findings identify potential gene pathways and interactions through which Stachys lavandulifolia bioactive chemicals exert their therapeutic benefits in PCOS. This study establishes a solid platform for future research into Stachys lavandulifolia as a potential PCOS therapy.

Recombinant protein vaccines against infectious diseases, based on immunogenic antigen identification and employing polymeric nanoparticles as a delivery system, can provoke immune responses comparable to or better than traditional vaccines. The production of a safe and immunogenic vaccine against diphtheria was achieved by preparing sodium alginate nanoparticles containing recombinant diphtheria toxoid (CRM197).

Alginate nanoparticles loaded with CRM197 were prepared using the ionic-gelation method and thoroughly characterized. Safety and immunogenicity studies were conducted in an animal model for comparison with commercial vaccines. Antibody responses were evaluated using both qualitative and quantitative measurements, as determined by the toxin neutralization test (TNT) and indirect ELISA, respectively. IgG subclasses in the sera of immunized mice and possible pathological lesions in vital tissues of all immunized mouse groups were investigated.

Nanoparticles with or without CRM197 were synthesized by the ionic gelation method. LE and LC measurements showed ˃80% and ˃20%, respectively, indicating stable and persistent release without a bursting pattern. In vivo studies showed safety and enhanced immunogenicity in mice immunized with the CRM197-loaded sodium alginate nanoparticles, with higher levels of total anti-CRM197 IgG and subclasses than those induced by conventional vaccines.

Reducing antigen usage in vaccine production while increasing immunogenicity and safety compared with traditional vaccines are the goals of new vaccine development, which were achieved in the current study.

Engineered alginate nanoparticles loaded with recombinant diphtheria antigen (CRM197) demonstrated in vitro controlled and slow release, as well as safety and immunogenicity profiles against diphtheriain vivo. Nanoparticles containing CRM197 antigens equivalent to adult and children doses showed high levels of IgG1 and IgG2a, confirming the combined responses of the humoral and cellular immune systems.

Recent studies indicate that niclosamide demonstrates considerable promise as both an anthelmintic agent and a possible anticancer medication. Given the increasing interest in nano-sized drug delivery methods for cancer therapy, lipid nanocapsules (LNCs) have emerged as a viable approach to enhance the bioavailability of poorly soluble pharmaceuticals due to their beneficial properties. This research intends to develop niclosamide-loaded lipid nanocapsules (NIC-LNCs) using the phase inversion technique, followed by the optimization of these formulations via the Box-Behnken experimental design.

A reverse-phase high-performance liquid chromatography (RP-HPLC) method was devised and validated for quantifying niclosamide in the LNC formulations. Optimal chromatographic separation was attained utilizing an Agilent Eclipse XDB-C18 column (150×4.6 mm, 5 µm i.d.) with a mobile phase of a 50:50 (v/v) mixture of acetonitrile and 0.1% H3PO4 phosphate buffer, at a flow rate of 1.2 mL/min. The detection wavelength was set at 335 nm, and the analysis was performed at 35°C. The developed analytical methodology was validated through a comprehensive evaluation of accuracy, linearity, precision, limit of detection, limit of quantitation, specificity, and stability.

The optimization of the NIC-LNC formulation through the Box-Behnken design resulted in an optimal formulation labeled LNC5, consisting of 4% niclosamide, 20% lipid, and 20% surfactant. The proven RP-HPLC method enables accurate quantification of NIC in the LNC formulations. The refined NIC-LNC formulation exhibited developed attributes as assessed by the design.

NIC-LNCs were successfully prepared with particle sizes below 100 nm, narrow size distributions (PDI<0.2), and negative zeta potential values in accordance with the literature. All formulations exhibited high encapsulation efficiency and sustained drug release profiles. The optimum formulation revealed a particle size of 43.29 ± 0.32 nm, encapsulation efficiency of 99.99 ± 0.02%, and drug release at one week of 68.85 ± 1.76%. The formulation maintained stability throughout the short-term study period.

The findings indicate that LNC systems are a promising method for drug administration, especially for anticancer drugs with limited solubility in water.

Pharmacological studies in vitro demonstrate the preventive and therapeutic potential of green tea and its constituent epigallocatechin-3-gallate (EGCG) in the fight against coronavirus disease 2019 (COVID-19). Previously reported correlations between per capita green tea consumption and COVID-19 morbidity/mortality suggest similar effects in vivo. Considering that some recent SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) sub-variants are less influenced by EGCG, this study aimed to determine whether this affects the aforementioned correlations, focusing on comparisons between the periods before (2021) and after (2022-2024) the emergence of the Omicron variant.

Correlations between per capita green tea consumption and COVID-19 morbidity/mortality were calculated using multiple regression models accounting for several confounding factors in a subset (n=84) of countries/territories worldwide with Human Development Index (HDI) above 0.55.

Higher per capita green tea consumption was associated with lower COVID-19 morbidity and mortality. Statistically significant correlations were observed in 2021-2024. Compared with 2021, the strength of both correlations decreased; the relative decrease in the strength of the correlation between per capita green tea consumption and COVID-19 mortality was notably less pronounced.

This differential decrease at the epidemiological level supports the idea that green tea consumption may have not only preventive but also therapeutic value regarding COVID-19. This aligns with in vitro pharmacological evidence indicating that green tea constituents target distinct molecular pathways responsible for the entry of the virus and its replication.

While promising, these findings require further assessment in observational and interventional studies focused on potential therapeutic benefits.

Ovarian cancer (OC) is a malignancy of the female reproductive system for which cisplatin chemotherapy is one of the first-line treatments. Despite the initial response to chemotherapy, such patients eventually develop resistance, which poses a major obstacle to treatment, along with potential side effects. Phytochemicals function as chemosensitizers, offering novel therapies in OC patients by targeting drug resistance, and are perceived to be less toxic. Plumbagin has emerged as an anticancer compound, with some findings suggesting its anti-ovarian cancer activity. However, there is no study on the potential of plumbagin to target cisplatin resistance in non-high-grade OC. The current study aimed to determine the antitumor activity of plumbagin for cisplatin resistance in OC cells in vitro, and to identify its potential molecular target for therapeutic benefit using in silico studies.

Plumbagin was used for in vitro cytotoxic effects on cisplatin-resistant (A2780-CR) and sensitive (A2780-CS) isogenic cell lines using a crystal violet cell viability assay. The binding of plumbagin to the nine selected molecular targets was estimated by molecular docking and their binding energies were compared. The stabilities of the selected docked complexes were confirmed by molecular dynamics simulation (MDS) and molecular mechanics generalized born surface area (MM-GBSA) calculations, and conclusions were drawn to predict the inhibition potential of plumbagin to its best targets.

Plumbagin demonstrated the potential to kill A2780-CR cells, and, expectedly, the cell death effect on A2780-CS ovarian cancer cells demonstrated its anti-tumor activity in vitro. It was found to be non-effective in killing normal non-tumorigenic RPE cells, even at higher doses. Docking analysis suggested that it potentially inhibits through various pharmacological targets with high affinity for binding to Chk1 (PDB ID=1ia8) and Aurora Kinase (PDB ID=5ORL). Molecular dynamic simulation data revealed strong and stable protein-ligand complex formation, which was measured in terms of root mean square deviation (RMSD), root mean square fluctuation (RMSF), and radius of gyration (Rg). On the other hand, the MM-GBSA study revealed that the binding free energy of the CT1019-1ia8 complex (-84.26 ± 2.99 Kcal/mol) and CT1019-5ORL (-67.04 ± 2.63 Kcal/mol) was better when compared to other complexes.

Plumbagin showed anti-ovarian cancer benefits in cisplatin-resistant ovarian cells, and the potential pharmacological targets identified were Chk1 and Aurora kinase.

Our study offers promising insights into plumbagin, particularly in combating cisplatin-resistance OC. However, further in vivo and mechanistic studies are required to validate plumbagin's potential as a therapeutic candidate for OC treatment.

Ibrutinib is a selective tyrosine kinase inhibitor used to treat chronic lymphocytic leukemia (CLL). However, it has low oral bioavailability (2.9%), which is attributed to low solubility (0.002 mg/mL) and a first-pass effect. Ibrutinib-loaded nanostructured lipid carriers (IBR-NLCs) were prepared and investigated in this study to overcome the solubility and presystemic metabolism issues. The goal of the current study was to formulate IBR-NLCs for enhanced bioavailability. IBR-NLCs were optimized using a 3² factorial design and evaluated using various in vitro and in vivo parameters.

IBR interaction with solid lipid (Glyceryl monostearate) and liquid lipid (oleic acid) was studied using molecular docking. The hot-melt ultrasonication method was used to formulate IBR-NLCs, and a 3² factorial design was used for optimization. Particle size, PDI, zeta potential, entrapment efficiency, DSC, XRD, FTIR, SEM, and in vitro study were used to evaluate the NLCs. HepG2 cell lines were used to study the in vitro cytotoxicity of IBR-NLCs and IBR suspension. IBR-NLCs were administered to male Wistar rats in the presence and absence of cycloheximide (CXI) to compare pharmacokinetic parameters.

Molecular docking confirmed good interaction between IBR-GMS and IBR-oleic acid. The optimized IBR-NLCs exhibited particle sizes, PDI, zeta potentials, and %EE of 154.5 ± 0.7 nm, 0.2 ± 0.0, -25.8 ± 1.1 mV, and 84.0 ± 1.2%, respectively. Differential Scanning Calorimetry (DSC) reveals the development of molecular dispersion of IBR in the melted lipid matrix, and X-Ray Diffraction (XRD) studies show a decline in the crystalline drug peaks in the formulation's diffractogram. SEM images showed uniformity distributed spherical-shaped particles. According to an in vitro investigation, IBR-NLCs exhibited a sustained release pattern of 98.0 ± 0.5% with a Korsmeyer-Peppas model mechanism (R² = 0.9615). The IC50 values of IBR suspension and IBR-NLCs were 4.155 µg/mL and 3.03 µg/mL. The AUC0-24 of IBR-NLCs administered in the absence of CXI was 1.60 times higher than the AUC0-24 values in the presence of CXI, indicating lymphatic transport.

IBR-NLCs appear to be promising as a novel innovative nanocarrier for the management of CLL.

Ovarian cancer (OC) is a common malignant tumor of the female reproductive system and is usually found at an advanced stage. However, the treatment of OC with conventional the efficacy of surgery and chemotherapy is limited. Brusatol (BRU) is a unique nuclear factor erythroid 2-related factor 2 (Nrf2) pathway inhibitor with significant anti-cancer effects. At the same time, the Nrf2 system also plays a vital role in ferroptosis, which can be used as a new way to treat tumors. This study investigated the mechanism of action of BRU as a novel ferroptosis inducer to inhibit OC cells.

Using bioinformatics to screen for key targets and pathways that act on OC in BRU, and then the effects of BRU on OC cells were examined by cell viability assay, clone formation assay, wound healing assay, and apoptosis assay. The intracellular levels of ROS (Reactive Oxygen Species), Fe²⁺, glutathione (GSH), and malondialdehyde (MDA) were also quantified. Western blotting analysis was then performed to verify ferroptosis marker proteins and pathways. In addition, the combination of Ferrostatin-1 (Fer-1) and BRU was further tested for ferroptosis-related markers.

By obtaining BRU and OC targets, 171 potential BRU-OC action targets were screened to the core target NQO1. KEGG enrichment analysis showed that the anticancer effects of IBC were mediated through multiple pathways, including the PI3K-AKT and Ras signaling pathways. In vitro results showed that IBC inhibited the proliferation, invasion, and migration of OC cells and induced ferroptosis in OC cells.

We demonstrated that BRU increased intracellular ROS, Fe^2+, and MDA levels. It also significantly reduced intracellular GSH level and the expression of two marker proteins for ferroptosis, GPX4 and SLC7A11. Meanwhile, BRU could inhibit the Nrf2/HO-1/NQO1 and AKT/mTOR dual signaling pathways in OC cells. Furthermore, the combination of Ferrostatin-1 (Fer-1) and BRU reversed BRU-induced ferroptosis in OC cells.

In this study, we demonstrated for the first time through bioinformatics, molecular docking technology, and experimental validation that BRU acts as a novel inducer of ferroptosis in ovarian cancer cells by targeting the Nrf2/HO-1/NQO1 and AKT/mTOR dual signaling pathways, and may have great potential in the treatment of ovarian cancer cells.

Crizotinib, an inhibitor of the epidermal growth factor receptor (EGFR) tyrosine kinase, holds significant potential for the treatment of lung cancer. However, its toxicities present a major challenge to its clinical use. To enhance the targeted delivery of Crizotinib to lung tumors, polymeric-based nanoparticles were developed.

Crizotinib-loaded polymeric nanoparticles were prepared using a nano-precipitation method, incorporating stearic acid as the lipid, polyethylene glycol as the polymer, and Tween 80 as the surfactant. Key formulation parameters were optimized to achieve high-quality nanoparticles.

The optimized formulation exhibited a mean particle size of 142 nm, a zeta potential of -31.9 mV, an entrapment efficiency of 82.35%, and an in vitro drug release of 60.69%. These nanoparticles were then tested on lung cancer cell lines to assess their cytotoxicity, apoptosis induction, and anti-proliferative effects on the cell cycle. In vitro studies confirmed that the Crizotinib-loaded nanoparticles exerted targeted effects on non-small cell lung carcinoma (NSCLC) cell lines, showing maximum inhibitory effects. One year of storage at 4°C, stability testing demonstrated that the lyophilized nanoparticles maintained their effectiveness.

crizotinib nano-formulations were assessed for a variety of physicochemical and in vitro characterization. Five different formulations were designed and optimized on the basis of Particle size, Zeta potential, %EE, and in vitro drug release. Optimum formulation also showed maximum inhibitory effect on the cancer cell line.

This nanotechnology approach offers a promising targeted drug delivery system for Crizotinib, characterized by small particle size, high encapsulation efficiency (EE), and optimal in vitro drug release.

Cholesterol is considered a major factor contributing to cardiovascular diseases. Statins, the most commonly prescribed cholesterol-lowering drugs, are known to have various limitations. Inhibition of Adenosine Triphosphate-Citrate Lyase (ACLY) has been proposed as an alternative therapeutic strategy for managing hypercholesterolemia by lowering cholesterol levels. This has led to the discovery of a cell-permeable small molecule ACLY inhibitor.

ACLY enzyme activity was assessed using an ACLY Assay Kit with the ADP-Glo Kinase Assay Kit. HepG2 cells were treated with test compounds to demonstrate cholesterol and fatty acid synthesis. Pharmacokinetic studies were performed on CD-1 mice following a single oral dose of the compounds. Hypercholesterolemia was induced in mice through a High-Fat and High Cholesterol Diet (HFHCD), and drugs were administered orally for six weeks. Serum and hepatic lipid profiles were subsequently analyzed.

To increase the pharmacochemical properties, four analogues of BMS-303141, ID0018, ID0023, ID0085, and ID0106, were designed and synthesized. These compounds showed superior ACLY inhibitory activity and dose-dependent suppression of cholesterol and fatty acid synthesis in HepG2 cells. Among the analogues, ID0085 exhibited the most potent ACLY inhibition (IC50: 45 nM, 10-fold lower than BMS-303141) and achieved near-complete suppression in cholesterol and fatty acid synthesis at the highest concentration. Pharmacokinetic studies revealed improved half-lives and systemic exposures for all analogues. 
In hypercholesterolemic mouse models, test compounds significantly reduced serum total cholesterol 
(32.0-57.3%) and low-density lipoprotein cholesterol (67.5-80.2%) levels compared to the vehicle group. Notably, ID0085 also increased high-density lipoprotein cholesterol levels.

Among the synthesized analogues, ID0085 exhibited the most potent ACLY inhibition, superior pharmacokinetic properties, and significant improvements in both serum and hepatic cholesterol profiles compared to BMS-303141.

Based on the results, ID0085 appears to be the most promising therapeutic candidate for the treatment of hypercholesterolemia.

Genetic factors play a significant role in the development of leukemia. The overexpression of MDM2 is associated with the progression of certain leukemias. This meta-analysis investigates the relationship between the MDM2 SNP 309T>G and various forms of leukemia across global populations.

A comprehensive literature search was conducted to retrieve genotyping data from twenty case-control studies related to MDM2 SNP 309T>G polymorphism and leukemia. A random-effects model was used to calculate the pooled odds ratio (OR) and 95% confidence interval (95% CI) for the association analysis. MetaGenyo software was utilized to conduct statistical analyses in this meta-analysis.

The findings indicate a significant association between MDM2 309 SNPT>G polymorphism and leukemia in Asian and Caucasian populations. Additionally, this polymorphism is associated with an increased risk of Acute Myeloid Leukemia (AML) and Chronic Myeloid Leukemia (CML), implying that MDM2 may play a role in the pathogenesis of these specific forms of leukemia.

This meta-analysis suggests that MDM2 may represent a susceptibility gene for leukemia risk.