Assay and drug development technologies最新文献

Freeze-drying is popular for producing pharmaceutical formulations with structurally complicated active components and drug delivery system carriers. It is the process of eliminating water from ice crystals through the sublimation mechanism. Some formulations may require drug-specific excipients such as stabilizers, buffers, and bulking agents to maintain the appearance and assure the long-term stability of the drug product. This approach is utilized for therapeutic compounds that are moisture sensitive, thermolabile, and degrade in the atmosphere. Freezing and primary and secondary drying are critical processes in the lyophilization process because they directly impact the end result. This approach is effective for producing a variety of dosage forms, including oral, inhalation, and parenteral. As a result, lyophilization may be an important method for improving the therapeutic efficacy and delivery of various dosage forms delivered via different routes. Additionally, lyophilization is used in pharmacological research to preserve biological samples, stabilize reference/standards, and increase the solubility and bioavailability of poorly soluble drugs. Thus, lyophilization is critical for maintaining the stability, efficacy, and safety of pharmaceutical products throughout their development and lifecycles. This article includes a broad overview of the lyophilization process, principle, excipients for lyophilized medicine compositions, and new lyophilization technologies as well as their applications in a variety of fields.

Buzhong Yiqi decoction (BZYQD) is a traditional Chinese medicine prescription for treating neurogenic bladder (NB). However, the underlying pharmacological mechanism remains unclear. This study aims to clarify the related molecular mechanism. Molecular structure information and targets of core components of BZYQD were obtained from Traditional Chinese Medicines Systems Pharmacology Platform (TCMSP) and SwissTargetPrediction databases. Genes involved in NB were obtained from Comparative Toxicogenomics Database, DisGeNet, GeneCards, and Online Mendelian Inheritance in Man databases. The hub targets of BZYQD in NB treatment were identified by protein-protein interaction (PPI) network analysis with STRING platform and analyzed by gene ontology analysis and the Kyoto Encyclopedia of Genes and Genomics pathway enrichment analysis. Molecular docking was used to verify the binding affinity between the hub targets and the bioactive components of BZYQD. Subsequently, the neuroprotective and anti-inflammatory effects of main bioactive components of BZYQD were investigated with in vitro assays. A total of 131 candidate compounds and 925 predicted target genes were screened. PPI network analysis suggested that ESR1, EGFR, HSP90AA1, MAPK3, AKT1, and CASP3 were the hub targets. BZYQD treatment was associated with hypoxia inducible factor-1 (HIF-1) signaling pathway. Dehydroglyasperin C (DGC), N-cis-feruloyltyramine, shinpterocarpin (SHI), gancaonin M, and glyasperin B, as the main bioactive components of BZYQD, had good binding affinity with hub target proteins. DGC and SHI treatment could significantly inhibit the injury of neurons and inflammatory response of microglia stimulated by oxidized low-density lipoprotein (ox-LDL), respectively. In summary, BZYQD and its main bioactive components DGC and SHI show good potential to ameliorate the symptoms of NB.

Glycerosomes signify a groundbreaking advancement in drug delivery technology. Comprising glycerol, phospholipids, and water, glycerosomes offer superior drug stability, penetration, entrapment efficiency, fluidity, and viscosity compared with conventional liposomes. Their formation process eliminates the need for specific transition temperatures, streamlining production. Glycerol's plasticizing properties enhance vesicle elasticity and flexibility, enabling enhanced skin penetration. These vesicles demonstrate immense promise across a range of drug delivery pathways. In dermal and transdermal applications, glycerosomes augment drug permeation by moisturizing the stratum corneum and improving membrane fluidity. For oral delivery, they shield drugs from the harsh gastrointestinal environment and boost intestinal absorption. Pulmonary delivery benefits from glycerosomes' capacity to stabilize and disperse aerosolized vesicles, facilitating deep penetration into lung tissues. Ophthalmic applications profit from increased corneal penetration and extended retention. Intranasal use of glycerosomes enhances mucosal penetration and enables direct drug delivery to the central nervous system by circumventing the blood-brain barrier. Ongoing advancements in glycerosome technology concentrate on integrating diverse functional ingredients like essential oils, β-sitosterol, sodium hyaluronate, and trimethyl chitosan to develop specialized formulations. These variants include STO-glycerosomes, S-glycerosomes, PO-S-glycerosomes, HY-glycerosomes, TMC-glycerosomes, glycethosomes, and glycerospanlastics, all offering enhanced stability, permeability, and therapeutic efficacy. This review delves into the mechanisms of drug transport within glycerosomes, their applications in various delivery routes, and the latest technological developments, highlighting their substantial potential as versatile carriers in contemporary drug delivery systems.

Cytotoxicity assays are essential in the field of research as they enable the examination of cellular responses to stimuli and shed light on complex mechanisms involved in multiple diseases and drug development. This review covers a range of cytotoxicity assays, including trypan blue and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assays, to more advanced techniques like caspase activity assays, Lactate dehydrogenase release assays, comet assays, and micronucleus assays for DNA damage assessment. Apart from these, other relevant assays like Alamar Blue, Bromodeoxyuridine incorporation, and clonogenic cell survival are also discussed. In this study, significance of these assays in drug development, toxicology studies, and biomedical research is discussed in detail, highlighting their role in ensuring safety and unraveling disease mechanisms. Furthermore, we explore emerging technologies such as chip-based assays, organ-on-a-chip systems, and high-throughput screening, which enhance precision and efficiency in research. Despite these advancements, challenges remain that necessitate standardization efforts and the development of more refined models. In conclusion, this review reflects on the evolving landscape of cytotoxicity assays, finding a balance between traditional methodologies and cutting-edge technologies.

To enhance brain delivery of efavirenz (EFV), optimized nanostructured lipid carriers (NLCs) were developed using a melt-emulsification technique and probe sonication. Box-Behnken design was chosen to systematically analyze the effects of variables on formulation outcomes, enabling efficient optimization with fewer experimental trials. This selection helped to improve the formulation by allowing us to refine key characteristics such as particle size, entrapment efficiency, and polydispersity index (PDI). The optimized EFV-NLCs had a mean particle size of 91.41 ± 7.90 nm, a PDI of 0.28 ± 0.04, a zeta potential of -17 mV, an entrapment efficiency of 85 ± 7%, and a drug loading of 14 ± 1%. Transmission electron microscopy confirmed that the EFV-NLCs were spherical with uniform size distribution. In vitro release tests showed prolonged drug release, with release rates ranging from 63.09 ± 2.76% to 84.43 ± 4.24% at pH 1.2 and 87.66 ± 6.31% to 92.56 ± 1.48% at pH 6.8. This was significantly better than the EFV suspension, which showed moderate and unsustainable release rates over 8 h. Furthermore, dissolution studies in both fasted and fed state simulated-intestinal-fluids (FaSSIF and FeSSIF) over 6 h revealed that % cumulative drug release was significantly higher in FeSSIF (94.06 ± 1.62%) compared with FaSSIF (65.21 ± 3.95%), indicating enhanced absorption in the presence of food. In vitro gut permeation studies revealed that EFV-NLCs had a 2.05-fold higher drug permeability than the suspension. These findings suggest that EFV-NLCs are promising for targeted brain delivery, are safe for oral administration, and could be instrumental in managing neuro-acquired immunodeficiency syndrome.

The present study highlighted enhancing the therapeutic effectiveness of curcumin (CUR) co-loaded lawsone (LS) through a solid lipid nanoparticles (SLNs)-based delivery system. The cetyl palmitate (CP), polyethylene glycol 400 (PEG), and probe sonication time (PS) were considered as independent variables whereas particle size and % entrapment efficiency (EE) were selected as dependent variables. The CUR-LS-SLN was developed by hot emulsification followed by probe sonication. A 2³ factorial design was utilized in formulation development using JMP software version 17. Notably, the particle size and %EE of all the formulations were about 500 nm and greater than 75%, respectively. The zeta potential value was found to be -46.8 mV. From leverage plots significant and sensitive factors on particle size and %EE were identified. Contour plots led to the identification of an optimized formula whereby maintaining CP at 100 mg, PEG 400 at 6 mL, and PS at 10 min the desired particle size and %EE was achieved. TEM studies indicated the spherical shape of the particles. MTT assays of Michigan Cancer Foundation-7 (MCF-7) cells showed enhanced efficacy and greater cell inhibition of CUR-LS-SLN and combining both drugs using nanocarriers gave superior inhibition as compared with using either of the drugs evident from IC₅₀ values of 3.7, 9.4, and 2.5 μM, respectively, for CUR, LS, and CUR-LS-SLN. The cells in the combination mostly had irregular cell walls and cell shrinkage was noted and greater cell reduction was also seen. It was found that the enhanced cytotoxicity effect of MCF-7 cells on the developed formulation was attributed to the drug's synergistic actions, more efficient nanocarrier internalizations, and sustained drug release from the formulation. Stability studies indicated that the optimized SLN was stable for 6 months.

Methotrexate (MTX) is an effective anticancer agent with limited water solubility, resulting in lower absorption in the gastrointestinal tract when administered orally. The present aim of the study is to construct sustained-release formulation of MTX-loaded microsponges with enhanced intestinal absorption and bioavailability using a quasi-emulsion solvent diffusion method. The Box-Behnken design (BBD) was adopted for this purpose. Particle size, encapsulation efficiency (EE), Q 2 h % (% drug release in 2 h), and Q 24 h % (% drug release in 24 h) were used as dependent factors, and polyvinyl alcohol, solvent, and stirring speed were used as independent factors. The prepared microsponges were characterized to assess their particle size and encapsulation efficacy (%). Attenuated total reflectance-Fourier transform infrared spectroscopy and differential scanning calorimetry were used to verify the compatibility study. Moreover, the cytotoxicity study was conducted on the HT-29 cell line. The optimized formulation exhibited a % encapsulation efficacy of 87.191% and a particle size of 2.176 µm. Furthermore, the optimized formulation demonstrated sustained drug release (85.71%) in Simulated Gastric Fluid (SGF) fluid at different pHs 1.2, 6.8, and 7.4. The stability study of the optimized formulation revealed good stability in terms of drug release, % encapsulation efficacy, and particle size. The results of the optimized formulation demonstrated that the viability of HT-29 colon cancer (CC) cells was dose-dependently decreased by MTX-loaded microsponges. BBD was successfully employed for the development and optimization of MTX microsponges filled in Eudragit S-100-coated hard gelatin capsule, depicting their potential release of MTX from microsponges capsule only at the colonic region and found to be potential carrier system for CC.

This study aimed to elucidate the hepatoprotective mechanisms of microalgal fatty acids (MFA) from Schizochytrium against alcoholic liver disease (ALD) through network pharmacology and in vivo analysis. Network pharmacology and molecular docking methodologies were employed to predict the potential mechanisms of MFA against ALD. To substantiate these predictions, an acute alcoholic liver injury mouse model was utilized to assess the impact of MFA on serum levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), total protein (TP), and albumin (ALB). Additionally, liver histopathology and the expression levels of phosphatidylinositol 3 kinase (PI3K) and protein kinase B (AKT) protein were evaluated. Seven active ingredients and 53 potential targets (including 7 core targets) for ALD treatment were identified in MFA. Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated that these seven core targets are implicated in various biological pathways, notably those associated with cancer, viral infections, and the PI3K/AKT signaling pathway. Furthermore, molecular docking studies demonstrated that docosahexaenoic acid and docosapentaenoic acid in MFA exhibited strong binding affinity for these seven crucial targets. Animal experiments demonstrated that administration of MFA significantly decreased the levels of AST, ALT, and ALP, while increasing the levels of ALB and TP in mice with acute alcoholic liver injury. Moreover, MFA ameliorated liver tissue pathology and markedly down-regulated the expression of PI3K and AKT proteins in the liver. These results suggest that MFA may possess therapeutic potential for ALD by targeting multiple pathways, with its mechanisms likely involving the inhibition of the PI3K/AKT signaling pathway.

Viral diseases remain a significant challenge for global health with rising fatalities each year. In vitro assays are crucial techniques that have been utilized by researchers in the quest to develop antiviral therapies. These assays mimic the internal conditions of a living system and make it possible to study how antiviral compounds interact with such systems in a laboratory setting. Thus, the importance of in vitro assays cannot be overemphasized, as they provide an accurate means for assessing the efficacy of potential antiviral compounds. This review offers an overview of in vitro antiviral assays, the different types of cell lines used, and emerging techniques and applications that have been developed in recent times. The current review also assesses challenges that are encountered in antiviral drug research, as well as emerging technologies like microfluidics and three-dimensional cell cultures. The integration of computational models and multiparametric assays into antiviral research was noted to significantly improve antiviral drug development process.

The skin is a dynamic tissue that consists of different layers such as stratum corneum, the site for keratinocyte development and maturation for the natural changeover of skin. In psoriasis, this natural development of keratinocytes gets disturbed and aggregation of nucleated keratinocytes takes place in the epidermis of the skin, leading to the presence of scaly skin, which makes the patient physically, socially, and psychologically ill. Various natural, semisynthetic, and synthetic treatments are available. Still, semisynthetic or synthetic are mainly used to treat psoriasis with side effects on different parts of the body, which is life threatening. Various molecular target sites are getting upregulated such as Janus kinase/Signal transducer and activator of transcription (JAK/STATs), phosphodiesterase 4 (PDE4), mitogen-activated protein kinase (MAPK), platelet selectin (Pan Selectin), Tumor Necrosis Factor Alpha (TNF-α), Interleukin-23 (IL-23), Interleukin-17 (IL-17), and Tyrosine Kinase 2 (Tyk2) in psoriasis. Plants and their bioactive compounds of flavonoids, alkaloids, resins, tannins, glycosides, and terpenoids category are used in the treatment of psoriasis as topical, oral, and biological forms. Using a computational approach, the inhibition of these molecular targets can be studied and potential molecules can be identified. This research article aims to find out the potential molecules that can inhibit the molecular sites and are effective than synthetic ones.