AAPS Journal最新文献

The rising prevalence of metabolic-associated steatotic liver disease emphasizes the need to understand its lipid metabolism. Dapagliflozin may improve hepatic steatosis but could also increase the risk of ketoacidosis by elevating β-hydroxybutyrate (KB) levels. This study investigates dapagliflozin's effects on hepatic lipid metabolism and quantifies KB levels in vivo. Male Sprague-Dawley rats were fed either a normal diet or a high-fat diet (HFD) for 12 weeks. The HFD rats were then divided into four subgroups to receive vehicle, 0.5 mg/kg, 1 mg/kg, and 3 mg/kg of dapagliflozin for four weeks. Free fatty acids (FFA) and KB levels were monitored, while protein and gene expression were analyzed. And a dynamic model of KB was developed for humans based on preclinical data. Dapagliflozin decreased body weight and visceral fat in HFD rats, increasing KB by upregulating CPT1a, HMGCS2, and HMGCL, and downregulating ACC. These changes correlated with reduced liver/fat index, liver pathology score, and oil-red staining area. A pharmacokinetic/pharmacodynamic (PK/PD) model was created from preclinical data to quantify KB levels in rats and validated in humans. Dapagliflozin reduces hepatic steatosis by enhancing fatty acid β-oxidation and ketogenesis and inhibiting fat synthesis. A dynamic model accurately predicts ketone body levels in treated individuals.

Lung cancer is a major public health problem across the globe, since it is the second most frequent cancer and the leading cause of cancer fatalities. This necessitates careful assessment of current therapies for lung cancer and discovery of novel drug candidates. 1,2,3 triazole compounds have emerged as an important class of prospective chemotherapeutic drugs for the treatment of lung cancer, with promising anti-lung cancer activity shown via a variety of pathways. They may interact with a various enzymes and receptors in cancer cells, causing cell cycle arrest and the activation of apoptosis. The present study aims to investigate the cytotoxic potential of institutional molecule based on 1,2,3 triazole [IIIM(S)-RS98] on multiple cancer cell lines. The compound was found to be most active on A549 cells and displayed the selectivity index as 8.16 in normal cells (e.g. HEK293). The in vitro findings revealed that IIIM(S)-RS98 induced apoptosis, loss of mitochondrial membrane potential, enhanced ROS and nitric oxide levels, and arrest cells in the G1 phase of the cell cycle. It inhibits the cell migration and clonogenic potential of A549 cells. Additionally, the downregulation of PI3K and p-Akt pathway leads to the activation of pro-apoptotic proteins Bax, downregulation of bcl2, activation of caspase 9, cleaved caspase 3, and cleaved parp1 expression and finally contribute towards apoptosis. Furthermore, molecular docking analysis indicated the interactions of IIIM(S)-RS98 with the apoptotic target proteins. The results demonstrated the potential of IIIM(S)-RS98 in the therapy of lung cancer.

Protein aggregates and particles in biopharmaceuticals can induce adverse immune responses in patients. Thus, suppression of the formation of protein aggregates and particles is important for the successful development of therapeutic proteins. Mechanical stresses, including agitation, are widely recognized as stress factors that generate protein aggregates and particles. However, although refrigerators and storage chambers generate weak vibration, there have been no studies of the impact of such weak vibration on aggregate and particle formation during storage. In this study, monomer loss and aggregate formation of a CTLA4-Ig were evaluated during storage in a refrigerator (having a vibration acceleration less than 0.006 G) with or without three vibration isolators. The vibration isolators reduced the vibration acceleration, thereby decreasing the rate of monomer loss and nanometer-sized aggregate formation. The increase in the aggregation rate due to the weak vibration was not mitigated by adding poloxamer 188 or eliminating the air-liquid interface, which are processes known to be effective in preventing protein aggregation due to mechanical stresses. Thus, reducing vibration should be an effective way to mitigate the risk of aggregate formation.

Protein-based therapeutics may elicit undesired immune responses in a subset of patients, leading to the production of anti-drug antibodies (ADA). In some cases, ADAs have been reported to affect the pharmacokinetics, efficacy and/or safety of the drug. Accurate prediction of the ADA response can help drug developers identify the immunogenicity risk of the drug candidates, thereby allowing them to make the necessary modifications to mitigate the immunogenicity. In this study, we leveraged the rich clinical study data collected by Roche/Genentech to identify factors that impact drug immunogenicity. We focused on conventional monoclonal antibodies, but have included a variety of additional drug modalities in the analysis. We show that the clinical ADA incidences are associated with the mechanism of action of the drugs, the mechanism of action of comedications, the routes of drug administration and the diseases of the patient cohort. By combining these clinical factors with the in silico epitope prediction, we improved the prediction accuracy of drug immunogenicity in clinical trials (AUC of cross validation improved from 0.72 to 0.93).

Minimizing harm is a cornerstone of ethical research practices. A drug that has undergone extensive clinical pharmacological testing in healthy participants (HPs) and a diverse selection of patients can be described with a sufficiently predictive population pharmacokinetic (PopPK) model. In impaired clearance trials, recruitment is minimized and underpowered for all but major exposure differences. Virtual HP arms have been reported to support similar conclusions to conventional impaired clearance studies, and further minimize potential harm of drug exposure without medical benefit by eliminating an arm of the study. However, the extent to which the conventional analysis of impairment studies compare to the simulation approach is unknown. Here we assess the operating characteristics of the virtual cohort approach along with the conventional approach through controlled simulations. These simulations included a simple, widely accessible PopPK model and several internal models that have been used in a previous meta-analysis of the virtual cohort approach. In the pairwise comparisons assessed, the virtual cohort simulation approach had greater power per sample size than the conventional approach and the same power under the null hypothesis. Given key methodological differences, it is recommended that the simulation and conventional approaches be treated as having approximately the same power under equivalent conditions. These results provide a strong justification for the use of the virtual cohort approach when an adequate PopPK model is available, minimizing unnecessary exposure to study drugs that will not benefit healthy study participants.

Bile salts are biosurfactants released into the intestinal lumen which play an important role in the solubilisation of fats and certain drugs. Their concentrations vary along the gastrointestinal tract (GIT). This is significant for implementation in physiologically based pharmacokinetic (PBPK) modelling to mechanistically capture drug absorption. The aims of this meta-analysis were to collate all appropriate data on intestinal bile salt concentrations in healthy adults across all GIT segments in fasted and fed states for the purpose of PBPK modelling. Terms relating to bile composition were searched in PubMed and Google Scholar from inception to May 2024. Selected studies included aspirated intestinal fluid collected via gastric tubes or colonoscopy. Results showed high variability across studies and a time-dependency for the fed state. Data were rich for the duodenum, which showed a two-fold increase for the fed state versus the fasted state within multiple studies. Peaks and troughs in bile salt concentrations along the GIT were observed for both fasted and fed states, likely due to segmental water absorption differences. The highest between subject variability was observed for the duodenum in the fasted and fed state and the fed proximal jejunum, distal ileum, and colon. The findings from this meta-analysis can be used for the purpose of PBPK modelling to capture segmental drug solubilisation and absorption in fasted and fed states. However, data are lacking under different fed conditions, especially following low-fat meals, so the impact of different fat content associated with different meals on bile salt concentrations cannot be discerned.

In addition to the known therapeutic indications for cannabidiol, its administration by inhalation appears to be of great interest. Indeed, there is evidence of cannabidiol's efficacy in several physiological pathways, suggesting its potential for a wide range of applications for both local and systemic pulmonary administration like cancers. Significant advances in pulmonary drug delivery have led to innovative strategies to address the challenges of increasing the respirable fraction of drugs and standardizing inhalable products. Among different devices, dry powder inhalers offer significant advantages including high stability and ease of use. Particle engineering using techniques such as spray drying is now the focus of research and is expected to improve upon, rather than completely replace, traditional carrier-based formulations. The development of carrier-free powders (without lactose-carrier) is mainly used for medicines with low active ingredient doses, which limits the technology. Previously, we demonstrated the benefits of using a cyclodextrin to obtain deflated spherical-shaped powders by spray drying. In this study the potential of this excipient with a very poorly water-soluble active molecule was investigated. Inhalable cannabidiol powders were developed by spray drying, using the solubility enhancers hydroxypropyl-beta-cyclodextrin and ethanol to optimize cannabidiol water-solubility. Electron microscopy images revealed consistent deflated spherical shapes, while particle size analysis showed low polydispersity and suitable sizes for deep lung deposition (2 µm). The selected engineered powders (without ethanol) had very high fine particle fractions (> 60%) due to their deflated surface. Finally, the powder was instantly solubilized leading to drug dissolution, which is important for therapeutic efficacy. In conclusion, this study successfully develops a cannabidiol inhalation powder by particle engineering having suitable aerosolization behavior. Due to the speed of the process and the performance of the finished product, this work opens the door for future studies. It has been shown that active molecules that are only slightly soluble in water can be formulated effectively as a powder for inhalation. Other molecules could be tested and subsequent in vivo studies conducted to demonstrate correlation with these in vitro results.

Freeze drying is one of the common methods to extend the long-term stability of biologicals. Biological products in solid form have the advantages of convenient transportation and stable long-term storage. However, long reconstitution time and extensive visible bubbles are frequently generated during the reconstitution process for many freeze-dried protein formulations, which can potentially affect the management efficiency of staff, patient compliance, and product quality. The reconstitution time has been extensively studied, but the influence of the formulations on the formation of visible bubbles is often overlooked. This paper investigated the effect of freeze-drying formulation factors (i.e., protein concentrations, surfactant concentrations, and sucrose/mannitol compositions) on product stability and visible bubbles generated during reconstitution of freeze-dried etanercept formulations. The generating and breakup mechanisms of visible bubbles were detected via internal microstructure of cake, surface tension, and viscosity measurement. Under the same protein concentration, the formulation of mannitol mixed with sucrose in a weight ratio of 4:1 produces fewer visible bubbles during the reconstitution process compared to the formulation of sucrose with the same total mass. This has been proven to be due to the large number of smaller radius pores distributed in the pores of the freeze-dried cake of the former, while the average internal structure pores of the latter are much larger than those of the former. As an amorphous stabilizer, sucrose can ensure the long-term stability of protein and greatly reduce the generation and maintenance of foams in the reconstitution process, making it a more robust excipient for freeze-dried protein formulations.

While the branched DNA (bDNA) assay is an established bioanalytical method for measurement of lipid nanoparticle (LNP)-encapsulated messenger RNA (mRNA) pharmacokinetic parameters, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) has been considered as an alternative platform. RT-qPCR and bDNA platforms were compared for sensitivity, specificity, correlation, and overall assay performance using serum and tissue samples from 2 nonclinical mouse studies of a therapeutic mRNA candidate, LNP-PAH-mRNA, which encodes for human phenylalanine hydroxylase enzyme. Pharmacokinetic parameter noncompartmental analysis was completed using Phoenix WinNonlin. The assays were compared using simple linear regression and Bland-Altman analyses. Sensitivity ranged from 0.05 to 6.40 ng/mL for the bDNA assays, from 0.00000761 to 7.61 ng/mL in serum, and from 0.000179 to 179 ng/g in tissue for the RT-qPCR assay. Inter-assay accuracy was within ± 10%, inter-assay precision was ≤ 10%, and the total error for both assays was ≤ 20%. RT-qPCR serum mRNA concentrations were 2- to fourfold lower compared with the bDNA assay, whereas tissue samples were comparable between assays. A linear relationship with - 0.37 to - 0.02 systematic bias demonstrated acceptable concordance. Bland-Altman plots demonstrated close equivalence, with a negative bias of < 0.5, and ≥ 95% of the data points were within the 95% limits of agreement. The comparison of the RT-qPCR with bDNA assay platforms for quantification of pharmacokinetic properties of an mRNA-LNP therapeutic has demonstrated acceptable concordance. This comparison reinforces the use of the RT-qPCR, a widely accessible strategy, as an alternative platform for the quantification of subsequent mRNA-LNP therapeutics.