Pharmaceutical Research最新文献

Introduction: The unbound brain-to-plasma partition coefficient (K_p,uu,BBB) is an essential parameter for predicting central nervous system (CNS) drug disposition using physiologically-based pharmacokinetic (PBPK) modeling. K_p,uu,BBB values for specific compounds are however often unavailable, and are moreover time consuming to obtain experimentally. The aim of this study was to develop a quantitative structure-property relationship (QSPR) model to predict the K_p,uu,BBB and to demonstrate how QSPR-model predictions can be integrated into a physiologically-based pharmacokinetic model for the CNS.

Methods: Rat K_p,uu,BBB values were obtained for 98 compounds from literature or in house historical data. For all compounds, 2D and 3D physico-chemical and structural properties were derived using the Molecular Operating Environment (MOE) software. Multiple machine learning (ML) regression models were compared for prediction of the K_p,uu,BBB, including random forest, support vector machines, K-nearest neighbors, and (sparse-) partial least squares. Finally, we demonstrate how the developed QSPR model predictions can be integrated into a CNS PBPK modeling workflow.

Results: Among all ML algorithms, a random forest showed the best predictive performance for K_p,uu,BBB on test data with R² value of 0.61 and 61% of all predictions were within twofold error. The obtained K_p,uu,BBB were successfully integrated into the LeiCNS-PK3.0 CNS PBPK model.

Conclusions: The developed random forest QSPR model for K_p,uu,BBB prediction was found to have adequate performance, and can support drug discovery and development of novel investigational drugs targeting the CNS in conjunction with CNS PBPK modeling.

Purpose: To improve the quality of aerosol delivery to infants, the iDP-ADS was advanced to include dual-prong nose-to-lung aerosol administration with a bifurcating interface, consistently monitor lung pressures and control ventilatory parameters with a pressure monitoring and control (PMC) unit, and implement flexible nasal prongs for use across a range of subject sizes.

Methods: Four bifurcating flow pathways were integrated into the iDP-ADS and tested in vitro with a full-term infant nose-throat (NT) model for comparison to the performance of a single-prong interface. After selecting the best-performing flow pathway, flexible prong designs were evaluated in the same model and chosen for additional testing. Realistic pulmonary mechanics (PM) and age-appropriate tidal volumes were used to simulate ventilation with the PMC unit and aerosol delivery in full-term and 34-week gestational age preterm NT models.

Results: Three of the four bifurcating flow pathways matched the performance of the single-prong design (tracheal filter delivery of ~55%), and the FP4 design with co-flow was selected. A flexible prong version of FP4 produced similar performance to the rigid version. Measurements from the PMC unit demonstrated that consistent air volumes under safe operating pressures could be delivered with a PEEP between 4-6 cmH₂O. Considering aerosol delivery, PM conditions resulted in ~4% decrease in filter deposition but high lung delivery efficiencies of ~45% and ~34% for the full-term and preterm models, respectively.

Conclusions: The best-performing interface with flexible prongs matched the lung delivery efficiency of a high-transmission single-prong interface and delivered high aerosol doses through late-preterm to full-term NT models.

Purpose: The transition from high Global Warming Potential (GWP) propellants such as HFA134a to low-GWP alternatives such as HFA152a and HFO1234ze(E) in pressurised metered dose inhalers (pMDIs) poses a number of challenges for inhaled pharmaceutical product development. Changes in chemicophysical properties will alter product performance, impacting in-vitro bioequivalence metrics. This study investigates those differences using equivalent pMDI hardware and formulations.

Methods: Aerodynamic particle size distribution (APSD) measurements, laser diffraction and high-speed imaging were used to compare the performance of HFA134a, HFA152a and HFO1234ze(E) solution formulations of beclomethasone dipropionate. Propellant-only placebos, cosolvent-free solutions, and ethanol solutions at 8% and 15% w/w were investigated.

Results: HFA152a formulations had increased drug deposition on the actuator and throat while HFO1234ze(E) produced comparable APSD performance to HFA134a formulations. Plumes from HFA152a formulations spread more rapidly and were less stable and repeatable than those from HFA134a. HFO1234ze(E) plumes spread more slowly than HFA134a, but converged with HFA134a ex-mouthpiece. Differences between propellants were moderated by the addition of ethanol.

Conclusion: Plume stability is a driver of differences between formulations in the near-orifice region. Shot-to-shot repeatability differences are more pronounced ex-mouthpiece, where mixing with ambient air is dominant. Modifications to low-GWP pMDI product actuator orifice and mouthpiece geometries may provide a route to improved in-vitro product bioequivalence relative to current pMDIs. Differences between formulations are modest and may be managed through a combination of formulation, orifice and mouthpiece geometry changes. These generic formulations provide a database of benchmark data against which the performance of low-GWP products may be compared.

Purpose: Therapeutic monoclonal antibodies (mAbs) are prone to degradation via aggregation and fragmentation. In this study, forced degradation of trastuzumab (TmAb) was explored in saline and in-vitro models having H₂O₂ and exposed to UV light (case study 1)_, both bleomycin (BML) formulation and ferrous ions (Fe²⁺) (case study 2)_, and sodium hypochlorite (NaOCl) (case study 3).

Methods: Size exclusion chromatography, dynamic light scattering, spectroscopic analysis, and fluorescence microscope image processing was carried out for characterizing TmAb degradation.

Results: Saline samples containing TmAb and 0.1% H₂O₂ incubated at 40ºC for 1 h in the presence of UV light showed increased monomer loss by more than 40% compared to TmAb sample without H₂O₂ exposed to UV light. Saline containing TmAb having both 0.1-unit BML and 0.25 mM Fe²⁺ showed increased monomer loss by more than 50% compared to TmAb in saline having only Fe²⁺ or BML. A higher TmAb degradation was also observed in saline containing 0.01% NaOCl compared to saline without NaOCl. Samples containing aggregates of mAb showed altered protein structure. Degradation of TmAb in saline increased with time, temperature, and concentrations of H₂O₂, Fe²⁺_, and NaOCl. At different analysis time points, TmAb monomer loss was higher in saline compared to human serum filtrate, an in-vitro model. Aggregate particles (> 2 µm size) of TmAb were also observed in serum containing both Fe²⁺ and BML.

Conclusion: It can be concluded that rapid TmAb degradation significantly enhanced due to various stress factors, and the aggregates could result in enhanced immunogenic risk to the patients.

As of now, more than 15 oligonucleotide drugs, primarily small interfering RNAs and antisense oligonucleotide classes, have been approved by the US FDA for therapeutic use, and many more are under clinical trials. However, safe and effective delivery of the oligonucleotide-based drugs to the target tissue still remains a major challenge. For enhanced plasma half-life, effective endosomal release, and other multiple functionalities, various carrier molecules have been used over the years. The successful therapeutic application of antibody-drug conjugates has made antibodies a popular choice for the delivery of oligonucleotide payloads into the target tissues. Single-chain variable domains of heavy chain antibodies (nanobodies) have proven a promising alternative to antibodies in recent years due to their small size, high affinity for the target, cell-penetrating potency, simple and easy production. The present review highlights the oligonucleotide drug types and their conjugation with nanobodies called NucleoBodies for effective targeted delivery, detection and diagnostics.

Objective: Biologic drug molecules such as antibodies are exposed to the physiological stress conditions of pH 7.4 and 37°C during their long circulation lifetime in vivo. The stress on biologic molecules in vivo is more severe compared to that under typical storage conditions of low pH formulation and cold temperature. Chemical degradation of critical residues such as asparagine may occur in vivo, leading to potential loss of biological activity. This study describes a physiologically relevant and convenient in vitro PBS stress condition of pH 7.4 and 40°C for pre-clinical stability screening of biologic molecules.

Methods: As benchmarks, multiple commercial antibodies (alirocumab, evolocumab, golimumab, ramucirumab, and trastuzumab) were tested in parallel for formulation stability at storage and accelerated temperature conditions and for physiological stability at pH 7.4 and 40°C stress both for 3-4 weeks. The stressed antibodies were monitored for chemical modification and target binding, without requiring affinity purification.

Results: The major CDR chemical modifications observed in PBS-stressed commercial antibodies were deamidations of asparagine residues. Although slight decreases in target binding were observed for two antibodies, the affinities overall remained strong after PBS stress.

Conclusions: This benchmarking study of commercial antibodies would be useful as a guide to screen discovery-stage biologic molecules both for drug product stability at formulation pH under storage and accelerated temperature conditions and for physiological stability under in vivo-mimicking pH and temperature stress condition.

Background: Osteoarthritis is the prevailing form of inflammatory condition in joints of adults and the aging population, leading to long-term disability and chronic pain. Current therapeutic options have variable therapeutic efficacy and/or several side effects.

Methods: A randomized, placebo-controlled, double-blind clinical trial was conducted in 62 participants using a nutraceutical [standardized Boswellia serrata Roxb. gum resin (300 mg) and Apium graveolens L. seed extract (250 mg)], to determine its safety and efficacy for supporting cartilage health and reduction in knee osteoarthritis symptoms. All participants were assessed for physical function and pain with the help of WOMAC, VAS, Physicians' Global Assessment for the six-minute walk test/pain. Knee X-ray, KOOS questionnaire score, and FACIT-F score were assessed. Additionally, inflammatory, cartilage degeneration and regeneration biomarkers in serum and urine were evaluated at baseline and after 90 days of treatment.

Results: Oral administration of the nutraceutical resulted in prolonged symptomatic relief with reduced pain, stiffness, and swelling. Inflammatory (serum IL-7, IL-1, IL-6, hs-CRP, TNF-α, ESR) and cartilage degeneration biomarkers (serum CTX-II, COMP, MMP-3 and urinary CTX-II) were decreased in the nutraceutical group compared to baseline and placebo. Furthermore, serum N-propeptide of collagen IIA (PIIANP) and procollagen-type-C propeptide (PIICP) levels were increased in the nutraceutical group, suggesting collagen synthesis contributing to cartilage regeneration. At given doses for 90 days, there were no adverse effects based on the clinical examination, biochemical, hematological, and ECG analysis.

Conclusions: Taken together, the combination of Boswellia and celery could be a safe and promising herbal nutraceutical option for managing osteoarthritis and cartilage health effectively.

Purpose: Tylvalosin Tartrate (TAT), a new-generation macrolide antibiotic, undergoes significant degradation in the stomach and in vivo rapid elimination upon oral administration, resulting in poor bioavailability. This study developed TAT enteric amorphous pellets by liquid layering (TAT/EAP-LL) with pH-sensitive and burst release characteristics, to enhance drug stability in the stomach and concentration enrichment in the duodenum.

Methods: The drug loading layer, isolation layer and enteric layer were formed on the surface of the blank core pellets. Investigation into the characteristics of TAT/EAP-LL revealed that stable amorphous solid dispersions in the drug loading layer were formed by liquid layering. Then, DSC analysis confirmed that triethyl citrate significantly improved the film-forming properties of Methacrylic-ethyl acrylate copolymer. Additionally, TAT/EAP-LL was confirmed to exist in the amorphous state by DSC、PXRD and PLM.

Results: In vitro, TAT/EAP-LL demonstrated a similar 4.07% release within 2 h at pH 1.0 as TAT enteric pellets (TAT/EP-LL) and a much faster burst release at pH 6.8, with complete release within 15 min. In vivo, the oral bioavailability of TAT/EAP-LL was improved to 1.71 times compared to commercial formulations and 1.47 times compared to TAT/EP-LL.

Conclusion: This study offers a novel platform for the enhanced oral delivery of TAT and proposes effective formulation strategies for pulsatile drug delivery.

Background: Iron overload is implicated in many neurodegenerative diseases, where there is also blood-brain barrier (BBB) dysfunction. As there is a growing interest in the role of iron in modulating key BBB proteins, this study assessed the effect of iron on the expression and function of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) and claudin-5 in primary mouse brain endothelial cells (MBECs) and their abundance in mouse brain microvessel-enriched membrane fractions (MVEFs).

Methods: Following a 48 h treatment with ferric ammonium citrate (FAC, 250 µM), MBEC protein abundance (P-gp, BCRP and claudin-5) and mRNA (abcb1a, abcg2, and cldn5) were assessed by western blotting and RT-qPCR, respectively. Protein function was evaluated by assessing transport of substrates ³H-digoxin (P-gp), ³H-prazosin (BCRP) and ¹⁴C-sucrose (paracellular permeability). C57BL/6 mice received iron dextran (100 mg/kg, intraperitoneally) over 4 weeks, and MVEF protein abundance and iron levels (in MVEFs and plasma) were quantified via western blotting and inductively coupled plasma-mass spectrometry (ICP-MS), respectively.

Results: FAC treatment reduced P-gp protein by 50% and abcb1a mRNA by 43%, without affecting ³H-digoxin transport. FAC did not alter BCRP protein or function, but decreased abcg2 mRNA by 59%. FAC reduced claudin-5 protein and cldn5 mRNA by 65% and 70%, respectively, resulting in a 200% increase in ¹⁴C-sucrose permeability. In vivo, iron dextran treatment significantly elevated plasma iron levels (2.2-fold) but did not affect brain MVEF iron content or alter P-gp, BCRP or claudin-5 protein abundance.

Conclusions: Iron overload modulates BBB transporters and junction proteins in vitro, highlighting potential implications for CNS drug delivery in neurodegenerative diseases.

The U.S. Food and Drug Administration (FDA) and the Center for Research on Complex Generics (CRCG) hosted a public workshop on May 2-3, 2024, titled "Considerations and Potential Regulatory Applications for a Model Master File". The workshop aimed to discuss the application of the Model Master File (MMF) concept in regulatory submissions that contain model integrated evidence (MIE), improving model sharing, model standardization, regulatory consistency, and regulatory efficiency. On Day 1, there was a session dedicated to MMF applications for long-acting injectables (LAIs). This perspective summarizes presentations, panel discussion, and small group discussion for the potential applications of MMFs in LAI product development, including case studies and potential situations in which MMFs can support regulatory submissions. The scientific presentations discussed the application of MMFs in mechanistic physiologically based pharmacokinetic (PBPK), multiphysics simulation, and population pharmacokinetics (popPK) models, as well as the potential utility of a model-integrated bioequivalence (MI-BE) framework. Additionally, challenges and considerations of implementing MMFs for LAIs were discussed in the panel and small groups. The anticipated benefits of MMFs are recognized among model developers, industries, and regulators.