Medicine and Pharmacology

Abstract: This study focuses on the development and optimization of topical cream formulations based on biopolymers and plant-derived extracts for potential wound-healing applications. Initial formulations prepared using conventional excipients exhibited insufficient viscosity and structural stability. The incorporation of xanthan gum as a stabilizing agent significantly improved the consistency and homogeneity of the systems. Hydroalcoholic extracts of Hypericum perforatum and Calendula officinalis were subsequently incorporated into the optimized formulations, resulting in satisfactory consistency and improved sensory properties. Formulations containing individual extracts demonstrated better homogeneity compared to combined extract systems. In parallel, an alternative formulation strategy based on biopolymers, namely gellan gum and chitosan, was investigated. The gellan-based formulation exhibited superior structural integrity and stability due to the formation of a three-dimensional polymeric network, whereas the chitosan-based system showed limited stability, likely associated with pH-dependent solubility and interpolymer interactions. Preliminary physicochemical evaluation confirmed the formation of semisolid systems with shear-dependent flow behavior suitable for topical application. These findings highlight the importance of polymer selection and formulation strategy in the development of stable cream systems. Overall, the developed formulations demonstrate promising potential for topical wound-healing applications. Further studies, including detailed pH analysis, rheological characterization, and biological activity evaluation, are required to confirm their therapeutic efficacy.

Abstract: Antibody–drug conjugates (ADCs) are a pivotal technology for precision cancer therapy, harnessing the synergistic effects of antibody targeting and toxin delivery. However, traditional ADCs encounter limitations in efficacy that stem from tumor resistance, heterogeneity, and intense target competition. Dual-payload ADCs (DP-ADCs) represent a promising solution to these challenges, as they leverage dual mechanisms of action that mitigate acquired drug resistance and enhance adaptability to tumor heterogeneity. The complex structure of DP-ADCs presents substantial quality control hurdles. In this manuscript, we review the current payload selection and conjugation strategies of DP-ADCs and examine recent advances in quality control research. Specifically, we analyze the analytical challenges related to the quantification of free toxins, the determination of the total antibody content, and the characterization of the drug-to-antibody ratio and its distribution. Ultimately, the aim of this work is to provide valuable guidance for future DP-ADC quality control analyses to facilitate their clinical translation and application.

Abstract: This study used chemometric models to measure caffeine (CAF), aspirin (ASP), and paracetamol (PAR) in the presence of three hazardous impurities: salicylic acid (SAL), P-nitrophenol (PNP), and P-chloroacetanilide (PCA). A molecular docking study was used to examine how impurities might inhibit cyclooxygenase-2, highlighting the importance of controlling their levels if present in the dosage form. Four sustainable chemometric models: principal component regression, multivariate curve resolution-alternating least squares, artificial neural networks, and partial least squares, were developed. The quantitative analytical performance of all proposed models was evaluated using the per cent recovery, standard error of prediction, and root mean square error of prediction. The developed models covered the concentration ranges (in µg/mL) of PNP (2.00-6.00), PCA (0.50-0.90), SAL (6.00-14.00), ASP (6.00-14.00), CAF (3.00-19.00), and PAR (2.00-10.00). These models effectively addressed collinearity and spectral overlaps. Seven innovative tools, including the Carbon Footprint Reduction Index, Click Analytical Chemistry Index, Multicolour assessment tool, Analytical Green Star Area, spider chart, green solvent selection tool, and Modified Green Analytical Procedure, were used to calculate the sustainability and whiteness of the developed models.

Abstract: Dissolving microneedles (DMN) could be considered as a promising platform for transdermal delivery of naltrexone hydrochloride (NTX), providing a minimally inva-sive alternative to conventional administration routes. In the present study, DMN patches with an advanced design were developed via a two-step micromoulding tech-nique. The systems were composed of drug-free polyvinylpyrrolidone (PVP) and poly-vinyl alcohol (PVA) blend microneedle tips, combined with a drug-loaded backing layer based on PVP and the thermoresponsive polymer Poloxamer 407. The influence of polymer concentration into DMN tips and backing layer composition on morpholo-gy, mechanical properties, drug release and permeation was evaluated. Mechanical studies as well as SEM observation revealed that intermediate polymer concentration (formulation MN-20%/2:1), used for DMN tips preparation, provided optimal mi-croneedle geometry, superior structural integrity and penetration efficiency. Incorpo-ration of NTX into backing layer allowed high and uniform drug loading. In vitro per-meation studies demonstrated significantly enhanced NTX delivery from DMN sys-tems compared to simple matrix patches, with the thermoresponsive backing layer contributing to controlled drug release. These findings highlight the importance of polymer composition in DMN design and demonstrate the potential of the developed systems as an effective platform for transdermal delivery of NTX.

Abstract: Diabetic neuropathy (DN) is a progressive nerve disorder caused by diabetes, characterized by chronic high blood sugar that leads to nerve inflammation and damage, resulting in pain, numbness, and loss of sensation. Current synthetic therapies often produce adverse effects such as hypoglycaemia, weight fluctuations, and organ toxicity, which hinder patient compliance and increase interest in herbal alternatives. This study evaluated a novel polyherbal extract (PHE) against diabetic neuropathy induced by a high-fat diet and streptozotocin (HFD-STZ) in rats. The PHE was prepared in a 2:2:1 composition ratio by decoction method. In the rat model, subjects received different doses of PHE (100, 200, and 400 mg/kg p.o) or gabapentin (300 mg/kg p.o) for 45 days. At the highest dose of 400 mg/kg, PHE significantly alleviated neuropathic symptoms, as shown by reduced mechanical allodynia and thermal hyperalgesia. It also improved glycemic control, normalized lipid profiles, and decreased oxidative stress markers by lowering lipid peroxidation while increasing levels of glutathione, superoxide dismutase, and catalase. Additionally, PHE enhanced gastric emptying time and exhibited neuroprotective effects by preserving the architecture of the sciatic nerve. In MIN6 β-cells, PHE maintained cell viability and stimulated glucose-dependent insulin secretion, highlighting its therapeutic potential for managing diabetic neuropathy.

Abstract: Phosphodiesterase-1 (PDE1) represents an attractive target for the treatment of idiopathic pulmonary fibrosis (IPF). However, the limited chemical diversity of current PDE1 inhibitors has hindered the development of potential anti-IPF drugs, primarily due to an ambiguous understanding of interactions between inhibitors and PDE1. Herein, we report an integrated virtual screening strategy containing pharmacophore modeling, molecular docking, and molecular dynamics simulations, which markedly accelerated the discovery of novel PDE1 inhibitors. Enzymatic assays identified eleven active compounds with moderate inhibition from twenty-six purchased candidates, encompassing nine distinct scaffold types. Notably, 6484-0008 and 6484-0032 exhibited more than 50% inhibition at a concentration of 1 μM. Hydrogen bonding analysis and residue-based energy decompositions revealed key recognition mechanisms involving crucial residues Gln421, His373, and Phe424, as well as the unique Thr271 in the flexible H-loop region, providing insights for the rational design of inhibitors with enhanced potency.

Abstract:

Objective: To compare the efficacy, safety, weight reduction and treatment adherence of oral versus subcutaneous semaglutide in adults with uncontrolled T2DM and obesity. Methods: A multicenter retrospective cohort study was conducted between January 2023 and January 2024. Adult patients (≥18 years) with T2DM (HbA1c ≥ 7%) and obesity (BMI ≥ 30) who received either oral or subcutaneous semaglutide were included. Demographic, clinical, and biochemical variables including body weight, BMI, HbA1c, side effects, and adherence were extracted from electronic medical records. Adverse effects were categorized by severity. Comparative analyses between groups used Chi-square and Mann Whitney U tests, with p<0.05 considered statistically significant. Results: A total of 208 patients were included: 89 on oral semaglutide and 119 on subcutaneous semaglutide. Baseline demographics, including gender, age, and physical activity, were comparable between groups (all p>0.05). The severity of adverse effects predominantly gastrointestinal symptoms such as nausea, vomiting, constipation, and diarrhea did not differ significantly between groups (p=0.994). However, dizziness was significantly more frequent in the subcutaneous group (p = 0.04). Adherence was markedly higher with oral semaglutide (p<0.05), with cost identified as the primary barrier among oral users, while subcutaneous users more frequently cited side effects, forgetfulness, and limited weight loss. Weight reduction was comparable at 3 months (p=0.23), but significantly greater with oral semaglutide at 6, 9, and 12 months (all p<0.01). Conversely, HbA1c reduction favored subcutaneous semaglutide at 3 and 6 months (p=0.03 and 0.02), although baseline glycemic control was similar. Conclusions: This study demonstrates that while subcutaneous semaglutide may provide a faster early HbA1c decline, oral semaglutide offers superior long-term weight reduction and significantly better adherence, likely attributable to easier administration. Both formulations exhibited comparable safety profiles.

Abstract: This study addresses the challenge of transdermal delivery of cyanocobalamin (vitamin B12), a hydrophilic macromolecule with low permeability, by developing biodegradable polymeric microneedle (MN) patches. Conventional methods often suffer from poor bioavailability, but microneedle technology can bypass the stratum corneum barrier, thereby improving drug delivery efficiency. We fabricated MN patches using hydroxypropyl methylcellulose (HPMC K4M), polyvinylpyrrolidone (PVP K30), and polyethylene glycol (PG 4000) through a mold-casting technique, followed by characterization of drug content, release kinetics, and mechanical properties. The optimized formulation (M18) demonstrated high drug content (95.2%) and sustained release (96.4% at 24 hours), while FTIR confirmed no drug-polymer interactions, ensuring stability. Moreover, SEM revealed uniform needle dimensions (867.25 ± 7.35 µm in height), and texture analyzer tests validated robust mechanical integrity. The patches exhibited low moisture content (3.42%) and high folding durability (&gt;200 folds), indicating suitability for storage and application. These results highlight the potential of polymeric MN patches as a non-invasive, efficient alternative for transdermal delivery of hydrophilic macromolecules. The study contributes to the field by providing a scalable, stable, and high-performance delivery system, which could significantly impact treatments for vitamin B12 deficiency and similar therapeutic needs.

Abstract: Background:Advances in oncology have led to the development of novel targeted therapies with demonstrated efficacy in clinical trials; however, their real-world economic impact prior to and after market introduction remains insufficiently characterized [1,2]. Cancer-related healthcare costs vary significantly depending on disease stage, time since diagnosis, tumor type, and therapeutic approach[3–6], making inter-hospital comparisons challenging due to heterogeneity in patient populations and information systems [7]. Therefore, integrating cost analysis with clinically meaningful patient stratification is essential to improve resource allocation and outcome evaluation[8–12]. Methods: A multicentre working group comprising four tertiary hospitals in Madrid (Spain) was established to develop and validate a novel classification system for adult oncohematological patients. A standardized methodology was designed to stratify patients into homogeneous groups (PATONCO categories) based on tumor location, therapeutic objective, and clinically relevant biomarkers. A cost indicator was defined as the average cost per patient per month for each PATONCO category. Data were extracted from pharmacy dispensing systems and analyzed using descriptive and inferential statistics, including Kruskal–Wallis and post hoc Dunn tests. Results: A total of 3,659 patients were included (3,168 oncology; 491 hematology), distributed across 62 programmes (54 oncology; 8 hematology). The PATONCOS tool enabled the identification and validation of a cost indicator (average cost/patient/month per category), allowing inter-hospital comparison. Significant differences in costs were observed across most high-prevalence categories, reflecting variability in therapeutic strategies and adoption of innovative treatments. The model demonstrated its capacity to detect intra-group homogeneity and inter-group variability, improving the identification of high-cost patient subgroups and supporting benchmarking across centres. Conclusions: The PATONCOS tool provides a novel, clinically oriented stratification methodology that integrates pharmacotherapy, biomarkers, and disease stage with economic evaluation. This approach enables more accurate comparisons of oncology treatment costs between institutions and supports data-driven decision-making in resource allocation. Its implementation may contribute to more sustainable healthcare systems by aligning clinical practice with economic outcomes.

Abstract: Background: The domain of microencapsulation technology is considered to be at the level of an advanced scientific discipline that includes the fields of materials science, pharmaceutical engineering, and food technology in the formulation of very specific matrices of polymeric or lipid nature. Method: In this review, a comprehensive analysis of sixteen different techniques of microparticles preparation has been presented: Solvent Evaporation, Solvent Extraction, Coacervation, Spray Drying, Spray Congealing, Ionic Gelation, Interfacial Polymerization, Air Suspension, Pan Coating, In-situ Polymerization, Supercritical Fluid Technology, Electrospraying, Microfluidics, Sol-Gel Process, Hot Melt Encapsulation, and Salting Out. Each technique has been explained by describing the basic physical and chemical phenomena that govern the process of microparticles formation. Results: The review has been presented with a critical analysis of the operating parameters, along with the core and shell material, as well as the applications of the technique, which are of interest in the field of pharmaceuticals, cosmetics, food, and medicine. Conclusion: The types of drugs that are best suited for the particular technique, as per their physical and chemical properties, i.e., solubility in water, lipid solubility, acid–base properties, as well as their thermoreactive properties, have been discussed in the review. The possibility of scaling up the technique from the laboratory scale to the industrial scale has been evaluated by searching the patent database, as well as the grant status of the patents, presented in the review. The prospective industrial applications of the technique, as well as the current limitations that restrict the scaling up of the laboratory-scale protocol, have been discussed in the review.

Abstract: Background/Objectives: Although pharmaceutical formulations on the market are more varied than in previous decades, developing tablets remains a continuing interest due to increased patient compliance. Simultaneously, the development of multifaceted excipients has remained a requirement of the pharmaceutical industry. This study aimed to develop a granular co-processed excipient for tablets and to evaluate it using the SeDeM expert system. Methods: Six granule formulations (obtained via wet layering granulation) were developed using different binder concentrations, fillers, and core types. The binders’ concentration (AquaPolish® STA) varied on three levels: 10%, 15%, and 20%. Three formulations used microcrystalline cellulose as the filler, while the remaining three replaced it with lactose (six formulations coded E1-E6). The granules obtained were evaluated using the SeDeM expert system for all 12 characteristic parameters, encompassing six incidence factors. The unloaded granules were compressed to yield uncoated tablets, which were verified for dimensional parameters, mechanical properties, and disintegration ability in accordance with the in-force European Pharmacopoeia requirements. Results: The binder concentration influenced particle size, with a 20% AquaPolish® STA concentration yielding large granules. It has been observed that the type of core used to prepare the granules played an important role in establishing mechanical strength; thus, the formulation in which Cellets® was used exhibited lower resistance than those in which sugar was used. During the SeDeM evaluation, it was observed that two formulations (E4 and E5) exhibited good results in terms of the parameter index (PI), parameter profile index (PPI), and Good Compressibility Index (GCI). The recorded disintegration times were less than 15 minutes for all the tablets obtained from the formulated granules. Conclusions: For granule development, binder concentration had the greatest influence on particle size, mechanical strength, and lubricity; also, the type of core used played an important role in tablet mechanical strength. With the help of the SeDeM expert system, the excipients most suitable for developing uncoated tab-lets were highlighted.

Abstract: Lipid-based drug delivery systems have emerged as a cornerstone in modern pharmaceutical research, offering innovative solutions to overcome limitations associated with conventional drug therapies. The article provides comprehensive information about liposomes and lipid nanoparticles by showing their structural foundations and various ways to categorize them and their different formulation techniques which follow Quality by Design principles. The development of advanced nanocarriers which include tocosomes and nanoliposomes and programmable lipid nanoparticles has transformed drug delivery systems from their original liposome design through enhancements in product stability and product targeting capabilities and product delivery control mechanisms. The study compares liposomal systems with surfactant-based systems to show that carrier selection needs to match both therapeutic needs and formulation capabilities of the drug. The article describes how lipid nanoparticles function in targeted cancer therapies and neurological disease treatments and systemic disease management while demonstrating their capacity to enhance drug absorption and decrease toxic effects throughout the body. Emerging delivery platforms such as oral, dermal, and rapidly dissolving systems highlight advancements in patient-centric drug administration. The combination of smart programmable nanocarriers and artificial intelligence technology has enabled accurate drug delivery control which supports the creation of customized medical treatments. The existing progress needs to address three major challenges which include problems with producing products at an industrial scale and obtaining regulatory authorization and maintaining long-term safety. Future perspectives focus on developing multifunctional nanocarriers which respond to stimuli and provide solutions for diverse medical treatment requirements. The current medical field has achieved a revolutionary change through lipid-based drug delivery systems which connect traditional drug delivery methods with cutting-edge medical treatment methods.

Abstract: Recent studies have shown that the solubilisation of poorly water-soluble drugs can be enhanced by using infant formula as a lipid-based formulation. In those studies, digestion of the triglycerides in infant formula to produce more polar lipids, namely fatty acids and monoglycerides, produced a high-capacity solubilisation environment for weakly basic drugs such as clofazimine, driven mainly by ion pairing of the fatty acid with the drug. However, digestion of lipid-based formulations is not expected to provide the same effect for nonionised or acidic drugs and in fact may present a reduced solubilisation capacity for weakly acidic drugs. In this study, a weakly acidic drug, tolfenamic acid, was dispersed in reconstituted infant formula and the infant formula was digested under in vitro simulated intestinal conditions. The quantity of tolfenamic acid that was solubilised in the infant formula during digestion was determined by high-performance liquid chromatography and small-angle X-ray scattering. Unexpectedly, digestion of the infant formula increased the solubilisation capacity for tolfenamic acid. Reconstituting infant formula at a higher fat content also increased the rate and extent of solubilisation of tolfenamic acid during digestion. The quantity of tolfenamic acid that was solubilised during digestion correlated approximately linearly with the quantity of free fatty acids produced during digestion. These results show that a weakly acidic drug can also exhibit digestion-driven solubilisation in a lipid-based formulation in the absence of ion-pairing and highlights the need to better understand drug response to digestion of lipid-based foods and formulations, and their versatility as a formulation option even for poorly water-soluble acidic drugs.

Abstract: Background/Objectives: The development of dry powder formulations for pulmonary delivery of therapeutic antibodies requires careful stabilization strategies to preserve protein integrity during spray-drying and long-term storage. This study investigates the impact of various sugar-derivatives, a polyol (D-mannitol), a disaccharide (D-sucrose) and a polysaccharide (dextran 10kDa) used individually or in combination, on the physical stability of bovine polyclonal immunoglobulin G (pAb) in dry powders for inhalation (DPIs). Methods: A design of experiments (DoE) approach was employed to evaluate the effects of these excipients on residual moisture (RM), low-order aggre-gates (LOA) and high-order aggregates (HOA), immediately after spray-drying (T0) and after 10 months of storage at room temperature (T10) in a desiccator. Results: All DPIs exhibited a high amorphous content and a favorable glass transition temperature, with RM decreasing over time. A combination of D-mannitol and dextran 10kDA (DPI-MD) demonstrated the best stabilization, minimizing LOA and HOA formation, both at T0 and T10. A ternary mixture, including also D-sucrose (DPI-MSD), showed enhanced short-term stability, but was less stable over time. The aerodynamic perfor-mance of these carrier-free DPIs, assessed via laser diffraction and a Next Generation Impactor, confirmed that DPI-MD and DPI-MSD formulations produced aerosol with suitable size distribution and fine particle fractions (FPFn of 70 ± 5% for DPI-MSD), for deep pulmonary deposition. Conclusions: These findings highlight the importance of combining excipients with complementary physical properties to achieve robust pro-tein stabilization. The DPI-MD emerged as the most promising candidate for pAb lung delivery, balancing protein integrity, powder stability, and aerodynamic efficiency.

Abstract: Breast cancer remains one of the leading causes of cancer-related morbidity and mortality worldwide, with tumour recurrence and therapeutic resistance largely driven by the immunosuppressive tumour microenvironment (TME). Conventional systemic chemotherapy and immunotherapy often suffer from poor tumour selectivity, systemic toxicity, and limited immune activation within the acidic and hypoxic TME. In this context, 4D-printed pH-responsive nanofiber implants have emerged as a next-generation platform capable of delivering spatiotemporally controlled therapy tailored to dynamic tumour conditions. Unlike static 3D constructs, 4D systems incorporate stimuli-responsive polymers that undergo programmed structural or functional transformations in response to environmental triggers such as acidic pH, enabling site-specific drug release. This review critically examines the design principles of pH-responsive nanofiber implants, including polymer selection, fabrication strategies, cytokine nano-assembly, and controlled release kinetics. Special emphasis is placed on TME modulation, highlighting how localised delivery of immune-stimulatory agents such as interleukin-15 and interleukin-2 can enhance natural killer cell activation, promote artificial immune synapse formation, and induce tumour apoptosis while minimising systemic toxicity. Furthermore, we analyse the translational challenges associated with manufacturing scalability, sterilisation, regulatory classification, and long-term implant safety. By integrating smart biomaterials engineering with immunotherapeutic strategies, 4D-printed nanofiber implants represent a transformative approach for localised breast cancer treatment. However, successful clinical translation will require interdisciplinary optimisation across materials science, pharmaceutical engineering, and regulatory frameworks. This review outlines future directions toward personalised, microenvironment-responsive cancer immunotherapy platforms.

Abstract: Background/Objectives: Human and canine leishmaniasis are neglected diseases with limited therapeutic options. The nitrochalcone NAT22, a high-affinity inhibitor of the essential parasite enzyme tryparedoxin peroxidase (cTXNPx), has emerged as a promising antileishmanial candidate. Interestingly, NAT22 demonstrated superior efficacy when administered orally rather than intralesionally, suggesting metabolism-driven enhancement of activity. Since in vivo studies with chalcones have been conducted exclusively in mice, this study aimed to evaluate whether mice are suitable models for oral chalcone therapies for human and canine leishmaniasis and to identify metabolites with potential antileishmanial activity. Methods: NAT22 hepatic metabolism was investigated using in silico prediction and in vitro liver microsomal assays from rats, mice, humans, and dogs. Metabolites were identified by LC-MS/MS and NMR, and docking studies were performed against cTXNPx. Results: In silico analysis predicted metabolism mainly by CYP1A2, CYP2A6, CYP2C8, and CYP3A4. Seven metabolites (M1–M7) were identified by LC-MS/MS and NMR in all species except mice, whose microsomes did not generate M6. Structural analyses indicated preservation of the α,β-enone system and nitro-substituted B ring in all metabolites. Docking studies showed that metabolites M2 and M4 displayed stronger predicted binding energies than NAT22. Conclusions: NAT22 undergoes hepatic phase I metabolism generating two metabolites with enhanced predicted interaction with cTXNPx. The similarity between human and canine metabolic profiles supports the translational relevance of oral NAT22 therapy in leishmaniasis, while metabolites M2 and M4 emerge as candidates for validation in local treatment of cutaneous leishmaniasis.

Abstract: The optimization of membrane permeability is a decisive strategy for mitigating late-stage failures in peptide drug development. By leveraging linker chemical diver-sity, stapled peptides utilize linker engineering to precisely modulate key physico-chemical parameters—such as lipophilicity and conformational constraints—to over-come the desolvation energy penalty. This review systematically evaluates link-er-based strategies for enhancing the permeability of stapled peptides, categorized into two primary dimensions: (1) High-throughput screening (HTS) compatibility, focusing on the integration of functionalized linkers into mRNA display, phage display, and DNA-encoded libraries (DELs) to identify lead scaffolds with inherent permeability potential during early discovery ; and (2) Post-screening structural refinement, cover-ing rational design strategies including intramolecular hydrogen bond (IMHB) shield-ing, "chameleonic" adaptations, and stimuli-responsive reversible stapling . Further-more, we analyze the paradigm shift in assessment methodologies from qualitative imaging to quantitative cytosolic delivery assays, which have deepened our under-standing of mechanisms such as the charge/lipophilicity threshold balance and meta-bolic-driven trapping. Overall, linker engineering provides a robust technical roadmap for developing the next generation of cell-permeable stapled peptide therapeutics.

Abstract: Background: Medications for Opioid Use Disorder (MOUD) have historically been the cornerstone of evidence-based treatment for opioid use disorder (OUD). However, the widespread proliferation of illicit fentanyl has introduced unprecedented challenges that threaten the effectiveness of traditional MOUD approaches. Objective: This paper examines the multifaceted barriers to MOUD implementation and effectiveness in the context of widespread fentanyl use, organized across patient, provider, and system levels. Methods: This paper synthesizes current literature on fentanyl-related challenges to MOUD delivery, focusing on precipitated withdrawal, pharmacological complications, patient perceptions, and healthcare system barriers. Key insights: Fentanyl's unique pharmacokinetic properties- including high lipophilicity, prolonged tissue accumulation, and variable elimination rates - fundamentally disrupt standard buprenorphine induction protocols. These pharmacological challenges cascade into patient-level barriers including fear of precipitated withdrawal, treatment avoidance, and premature dropout. Provider-level barriers include clinical uncertainty, inadequate training on fentanyl-specific protocols, and challenges with individualized dosing. System-level barriers encompass regulatory constraints, access limitations, and lack of integration between emergency, inpatient, and outpatient care settings. Conclusion: The fentanyl era has exposed critical gaps in MOUD delivery that span multiple levels of the healthcare system. Understanding these interconnected barriers is essential for developing targeted interventions and policy reforms. Future research should focus on fentanyl pharmacokinetics, optimized induction protocols, and MOUD efficacy in poly-substance use contexts.

Abstract: Background/Objectives: Pharmacogenomics (PGx) enables personalized therapy by identifying genetic variants that influence drug response. Despite the advantages of next-generation sequencing (NGS), few clinically validated, guideline-aligned panels comprehensively detect common, rare, and structurally complex pharmacogenetic variants. Methods: We developed and analytically validated Action PharmaKitDx, a targeted NGS panel covering 335 pharmacogenes, including all priority genes recommended by CPIC, DPWG, and CPNDS. Performance was assessed using Coriell HapMap and GeT-RM reference materials across multiple library preparation workflows and Illumina platforms. Clinical feasibility was evaluated in 41 patient samples from diverse specialties. Results were compared with established reference methods, including PCR-based assays, STR analysis, Sanger sequencing, and whole-exome sequencing. Results: More than 99% of target bases achieved ≥30× coverage. Analytical accuracy, sensitivity, specificity, and positive predictive value exceeded 99.3%, with repeatability and reproducibility &gt;99.7%. Concordance with GeT-RM haplotypes reached 98% after star-allele harmonization. The panel accurately detected complex variants, including CYP2D6 copy-number changes and hybrid alleles. Full concordance with prior genotyping was observed in clinical samples. Beyond the initial testing indication, each sample harbored a mean of six actionable variants (range 2–10). Thirty-six rare (minor allele frequency &lt;1%) potentially actionable variants were additionally identified. Conclusions: Action PharmaKitDx demonstrates high analytical performance and broad clinical applicability, supporting its implementation as a scalable solution for comprehensive pharmacogenetic testing and precision prescribing.

Abstract: Pharmaceutical regulation and healthcare governmental agencies are central to protecting public health by governing clinical trials, market authorization, and post-market safety monitoring of medicinal products worldwide. Although substantial literature describes major established systems, particularly the United States Food and Drug Administration (FDA), Japan's Pharmaceuticals and Medical Devices Agency (PMDA), and the European Union regulatory network coordinated by the European Medicines Agency (EMA) together with national competent authorities, comparative analysis that integrate both established and emerging regulatory authorities remain limited. This review examines the core functions of regulatory affairs across the product life cycle and compares key features of global regulatory frameworks, including approval pathways, evidentiary expectations, data transparency, and pharmacovigilance approaches. It emphasizes the need for systems thinking to balance innovation with safety, efficacy, and quality, while anticipating unintended consequences of new therapies. This evaluation also highlights how region‑specific constraints and enabling infrastructures, such as national drug‑utilization registries in parts of Europe, can shape regulatory decision‑making and post‑market evaluation. Finally, this paper discusses opportunities for stronger international alliances and greater harmonization to improve efficiency, support timely patient access to essential therapies, strengthen risk management, and reinforce global health security in an increasingly interconnected healthcare environment.