Current pharmaceutical biotechnology最新文献

Background: Immunotoxins (ITs) represent a novel class of therapeutics with bifunctional structures that facilitate their penetration through cell membranes to induce target cell destruction. Programmed cell death ligand-1 (PD-L1), a human cell surface protein, is overexpressed in various cancers. This study aimed to construct a novel IT by genetically fusing an anti-PD-L1 Nanobody (Nb) to a truncated diphtheria toxin (DT).

Methods: The IT construct comprised a 127-amino acid anti-PD-L1 Nb fused to a 380-amino acid fragment of DT, with an N-terminal 6x-His tag. Molecular cloning techniques were employed, followed by transformation and verification through colony-PCR, enzyme digestion, and sequencing. The anti-PD-L1 Nb was expressed in WK6 E. coli cells induced by Isopropyl β-D-1- Thiogalactopyranoside (IPTG) and purified from periplasmic extracts using immobilized Metal Ion Affinity hromatography (IMAC). The IT was similarly expressed, purified, and validated via SDS-PAGE and Western blot analysis.

Results: ELISA confirmed the binding activity of both Nb and IT to immobilized PD-L1 antigen, whereas truncated DT exhibited no binding. MTT assays demonstrated significant cytotoxicity of IT on A-431 cell lines compared to Nb and truncated DT controls. Statistical analyses underscored the significance of these findings.

Conclusion: This study provides a thorough characterization of the constructed IT, highlighting its potential as a therapeutic agent targeting PD-L1-expressing cancer cells. The results support the potential of this IT in cancer immunotherapy, emphasizing the need for further investigation into its efficacy and safety profiles.

The human microbiota represents the community and diverse population of microbes within the human body, which comprises approximately 100 trillion micro-organisms. They exist in the human gastrointestinal tract and various other organs and are now considered virtual body organs. It is mainly represented by bacteria but also includes viruses, fungi, and protozoa. Although there is a heritable component to the gut microbiota, environmental factors related to diet, drugs, and anthropometry determine the composition of the microbiota. Besides the gastrointestinal tract, the human body also harbours microbial communities in the skin, oral and nasal cavities, and reproductive tract. The current review demonstrates the role of gut microbiota and its involvement in processing food, drugs, and immune responses. The discussion focuses on the implications of human microbiota in developing several diseases, such as gastrointestinal infections, metabolic disorders, malignancies, etc., through symbiotic relationships. The microbial population may vary depending on the pathophysiological condition of an individual and thus may be exploited as a therapeutic and clinical player. Further, we need a more thorough investigation to establish the correlation between microbes and pathophysiology in humans and propose them as potential therapeutic targets.

Background: Guettarda viburnoides Cham. & Schltdl., "veludinho do campo", is used in the Brazilian Amazon for its effects on the central nervous system (CNS) as a "brain tonic"; however, scientific evidence is needed to elucidate its ethnobotanical uses.

Objective: This study evaluated the neurobehavioural effects of an ethanolic extract of G. viburnoides (EEGV). Molecular docking, microchemical and morphoanatomical features of the species were investigated.

Methods: EEGV was investigated by UHPLC‒MS/MS and dereplication and molecular network were investigated using platforms available for natural product chemistry. For the in vivo assay, EEGV was administered to mice orally (3, 30 or 100 mg/kg). The effect of EEGV on spatial memory was measured using the Morris water maze test in mice with scopolamine-induced amnesia. The depression- and anxiety-like effects were assessed by forced swimming, tail suspension, marble burying and elevated plus maze tests. The AChE inhibition was evaluated in the brains of treated mice and molecular docking simulations were carried out with the main constituents. The leaves and stems of G. viburnoides were analysed via optical microscopy, scanning electron microscopy and energy dispersive X-ray spectroscopy.

Results: Secoxyloganin, grandifloroside, hyperin/or isoquercitrin, uncaric acid and ursolic acid were identified by UHPLC‒MS/MS. Molecular networking by three flavonoids, three triterpenes, two coumarins, two iridoids, and one phenolic acid. EEGV reversed these scopolamineinduced effects. In the forced swim and tail suspension test, EEGV (30 and 100 mg/kg) significantly reduced the immobility time. EEGV significantly reduced the number of buried marbles, while in the elevated plus maze test, no changes were observed compared to the Sco group. AChE activity was altered in the hippocampus. Studies of the molecular coupling of iridoid glycosides (grandifloroside and secoxyloganin) and flavonoid hyperin with AChE revealed significant interactions, corroborating the activity indicated by the inhibition assay.

Conclusions: These results might be in accordance with medicinal use for neuroprotetor effects and important microchemical and micromorphological data that support the identification and quality control of G. viburnoides.

Background: Organophosphorus insecticides, widely used in farming and agriculture, have been associated with various health issues. Curcumin, a natural antioxidant, has shown potential in mitigating the adverse effects induced by these insecticides.

Aim: This study aimed to evaluate the nephroprotective effects of Curcumin (CUR) against Chlorpyrifos (CPF)-induced renal damage.

Method: Forty male Wistar albino rats were randomly assigned to five groups, each containing eight rats: control (0.5 mL of olive oil, the solvent for chlorpyrifos), CPF (10 mg/kg of chlorpyrifos), CPF + CUR 25 mg/kg/day, CPF + CUR 50 mg/kg/day, and CPF + CUR 100 mg/k/day. All groups were treated for 90 days. Finally, kidney tissue was assessed for oxidative stress, inflammatory markers, and histopathological changes.

Result: A considerable elevation in urea and Creatinine (Cr) concentrations was observed in the CPF group compared to the control rats (p < 0.01). CUR decreased creatinine and urea levels in the CPF-exposed group compared to the non-CUR-treated animals (p < 0.05). Additionally, the concentrations of NO, MDA, IL-6, IL-1β, and TNFα were significantly increased in the kidneys of the CPF-induced rats compared to the controls (p < 0.001). However, CUR (100 mg/kg) administration significantly reduced the abovementioned parameters in rat kidneys (p < 0.01). CUR (100 mg/kg) also increased superoxide dismutase activity and glutathione concentration in the kidneys of CPF-exposed animals compared to non-CUR-treated rats (p < 0.05). Histopathological analysis revealed severe congestion in the kidney tissue after CPF exposure. However, coadministration of CUR restored the normal structure of the kidney in the experimental rats.

Conclusion: Our findings suggest that curcumin, a potent antioxidant, can help alleviate chlorpyrifos-induced nephrotoxicity.

Cervical cancer has become a major worldwide health concern that demands attention to women's health and often needs more effective and specialized treatment options. Cervical cancer claims the lives of over 300,000 women globally, ranking as the fourth most prevalent cancer among women. The tumor microenvironment (TME) shapes a distinctive landscape for tumor survival, characterized by factors like immunosuppression, hypoxia, acidity, and nutrient scarcity. Comprising tumor cells, immune cells, mesenchymal cells, cancer-associated fibroblasts, and extracellular matrix, the TME reprograms key aspects of tumor development, uncontrolled proliferation, invasion, metastasis, and response to treatments. Recognizing the TME's pivotal role in tumor progression and treatment responsiveness, targeting the TME has emerged as a potential strategy in cancer therapy. This publication delves into recent TME research, offering a comprehensive overview of the specific functions of each TME component in cancer development and progression. Based on the reviewed literature, it appears that women with cervical cancer may benefit from more effective therapy, fewer side effects, and a higher quality of life in the future. By addressing pressing problems and unmet needs in the field, this review has the potential to significantly alter the course of cervical cancer treatment in the future. Furthermore, it outlines the primary therapeutic targets identified by researchers, which may prove valuable in treating tumors.

Neurological disorders are devastating conditions affecting both cognitive and motorrelated functions in aged people. Yet there is no proper medication to treat these illnesses, and the currently available medications can only provide symptomatic relief to the patients. All neurological disorders share the same etiology, such as oxidative stress, mitochondrial dysfunction, neurochemical deficiency, neuronal loss, apoptosis, endoplasmic reticulum stress, neuroinflammation, and disease-related protein aggregation. Nowadays, researchers use antioxidant-based strategies to prevent or halt the disease progression. Nerolidol, a strong antioxidant, possesses various biological activities and properties that treat cardiotoxicity, nephrotoxicity, neurotoxicity, and many other diseases. Many recent publications and research studies highlight the beneficial effect of nerolidol on brain disorders. In Alzheimer's disease, nerolidol shows neuroprotection by decreasing amyloid plaque formation, lipid peroxidation, cholinergic neuronal loss, locomotor dysfunction, neuroinflammation, and hippocampal damage via enhancing antioxidant expression. Also, it shows neuroprotection against rotenone-induced neurotoxicity by inhibiting microglial activation. Another study reported that nerolidol shows antiepileptic effects in animal models by suppressing kindling-induced memory impairment by decreasing oxidative stress. It has been found that NRL administration increases the antioxidant levels, decreasing the proinflammatory cytokine release as well as decreasing the apoptotic protein and cerebral infarct size. In conclusion, nerolidol tends to reverse the harmful effects of disease-related factors, including OS, neuroinflammation, protein aggregation, and apoptosis, making nerolidol a choiceable drug for the management of neurological disorders. The purpose of this review is to discuss the mechanism of nerolidol in treating various neurological disorders.

Objective: Atherosclerosis (AS) is an inflammatory disease of arterial intima driven by lipids. Liver X receptor alpha (LXRα) and peroxisome proliferator-activated receptor alpha (PPARα) agonists are limited in the treatment of AS due to their off-target effects and serious side effects. Therefore, this study was designed to construct a novel nanoparticle (NP) and evaluate its mechanism of action on inflammation inhibition and lipid reduction in AS.

Methods: We synthesized cRGD-platelet@MnO/MSN@PPARα/LXRα NPs (cRGD-platelet- NPs) and confirmed their size, safety, and targeting ability through various tests, including dynamic light scattering and immunofluorescence. In vivo and in vitro experiments assessed cell proliferation, apoptosis, inflammation, and plaque formation. Finally, the NF-κB signaling pathway expression in rat aorta was determined using a western blot.

Results: The synthesis of cRGD-platelet-NPs was successful; the particle size was approximately 150 nm, and the PDI was below 0.3. They could be successfully absorbed by cells, exhibiting high safety in vivo and in vitro. The cRGD-platelet-NPs successfully reduced plaque formation, improved lipid profiles by lowering LDL-cholesterol, total cholesterol, and triglycerides, and raised HDL-cholesterol levels. Additionally, they decreased inflammatory markers in the serum and aortic tissue, suggesting reduced inflammation. Immunohistochemistry and western blot analyses indicated that these NPs could not only promote M2 macrophage polarization but also suppress the NF-κB signaling pathway.

Conclusion: The newly developed cRGD-platelet-NPs with high safety are a promising approach to AS treatment, which can regulate ABCA1, reduce the formation of AS plaques, and enhance cholesterol efflux. The mechanism may involve the suppression of the NF-κB signaling pathway.

Objective: The objective of this study is to search for hydroxysafflor yellow A (HSYA) and Idiopathic sudden sensorineural hearing loss (ISSNHL)-related target genes and to study the treatment effects of HSYA on lipopolysaccharide (LPS)-induced endothelial cell injury.

Methods: We used network pharmacology to screen molecules related to HSYA and ISSNHL, then analyzed these molecules and their enriched biological processes and signaling pathways via Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO). We selected inflammation-related hub genes for molecular docking determination by protein-protein interaction (PPI) analysis, and further verified them with in vitro experiments.

Results: Thirty-four HSYA-ISSNHL-related differential genes were obtained using drug-disease differential gene screening using online tools. Three key proteins, NF-κB, CASP3, and MAPK1, were selected according to Degree > 20. Among them, NF-κB is closely related to inflammation and ISSNHL. In in vitro experiments, HSYA reduced inflammatory (IL-6, TNF- α) and oxidative stress (ROS, SOD and MDA) indicators after LPS intervention, and the expression of NF-κB-related signaling pathway genes.

Conclusion: HSYA may reduce inflammation and oxidative stress by inhibiting the expression of the TLR4 / NF-κB-related signaling pathway, therefore protecting endothelial cells, which might be a potential mechanism of HSYA in ISSNHL treatment.

Aim: This study aimed to determine the phytoconstituents of Crateva religiosa bark (CRB) and evaluate the hypolipidemic effect of bioactive CRB extract by preventing adipocyte differentiation and lipogenesis.

Background: After performing the preliminary phytochemicals screening, the antioxidant activity of CRB extracts was determined through a DPPH (2, 2-diphenyl-1-picrylhydrazyl) assay. Ethyl acetate extract (CREAE) and ethanol extract (CRETE) of CRB were selected for chromatographic evaluation.

Method: The antihyperlipidemic potential was analyzed by molecular docking through the PKCMS software platform. Further, a 3T3-L1 cell line study via In vitro sulforhodamine B assay and western blotting was performed to confirm the prevention of adipocyte differentiation and lipogenesis Results: The total phenolic contents in CREAE and CRETE were estimated as 29.47 and 81.19 μg/mg equivalent to gallic acid, respectively. The total flavonoid content was found to be 8.78 and 49.08 μg/mg, equivalent to quercetin in CREAE and CRETE, respectively. CRETE exhibited greater scavenging activity with the IC50 value of 61.05 μg/ mL. GC-MS analysis confirmed the presence of three bioactive molecules, stigmasterol, gamma sitosterol, and lupeol, in CRETE. Molecular docking studies predicted that the bioactive molecules interact with HMG-CoA reductase, PPARγ, and CCAAT/EBP, which are responsible for lipid metabolism. In vitro, Sulforhodamine B assays revealed that CRETE dose-dependently reduced cell differentiation and viability. Cellular staining using 'Oil Red O' revealed a decreased lipid content in the CRETE-treated cell lines. CRETE significantly inhibited the induction of PPARγ and CCAAT/EBP expression, as determined through protein expression via western blotting.

Conclusion: The influence of CRETE on lipid metabolism in 3T3-L1 cells is potentially suggesting a new approach to managing hyperlipidemia.