Pharmaceutical Research最新文献

Diseases of the central nervous system represent a significant challenge in the field of medical research and clinical practice. The utilization of gene therapy technology represents a promising therapeutic strategy aimed at ameliorating diseases at the genetic level. Nonetheless, the presence of the blood-brain barrier (BBB) and challenges pertaining to delivery efficiency, expression site and level, along with the potential risks linked to overexpression and prolonged expression, raise considerable safety concerns. The design, production, delivery, and expression of adeno-associated virus (AAV) vectors significantly influence both clinical efficacy and safety considerations. This article examines the factors contributing to and potential strategies for addressing the low transduction efficiency associated with the blood-brain barrier, focusing on the optimization of vector design, delivery methods, specific expression, and distribution. The study examines the optimization of promoter synthesis, the application of machine learning algorithms for the enhancement of AAV capsid evolution, and the evaluation of various administration routes. Additionally, it explores innovative delivery methods, including mannitol intra-arterial delivery and focused ultrasound strategies, aimed at improving efficacy and safety. These initiatives offer valuable guidance and insights pertaining to gene therapies aimed at addressing neurodegenerative diseases and various disorders of the central nervous system.

Introduction: Small molecules are biologically active organic compounds with molecular weight below 1 kDa. Their small size enables efficient transport across cell membranes and modulation of intracellular signaling, making them promising for drug development. Chelidonic acid (ChA) is a small molecule (184 Da) with a wide range of biological effects, but its transport mechanisms and molecular targets remain unknown.

Purpose: The aim of this study is to identify a possibility of ChA uptake by human cells and to search for transporter proteins that may be involved in the intracellular trafficking of ChA using a combination of in silico and in vitro approaches.

Methods: Co-culturing of human MCF-7 cells with ChA was conducted in vitro for 4 h and residual (not absorbed by cells) ChA concentration in solution was measured using HPLC. Candidate transporters were screened from databases. Molecular docking was performed with Autodock Vina, and molecular dynamics simulations were run for 50 ns using GROMACS to assess protein-ligand interactions. Statistical analysis used the R language with Newey-West estimator and Welch's t-test. HOLE and VMD were used for 3D-reconstruction and visualization of transport channels.

Results: MCF-7 cancer cells uptake ChA through one or several of the common cell transport proteins. Initial screening identified six transmembrane proteins, with further analysis pinpointing three candidates (GLUT3, SVCT1, URAT1) demonstrating structural and functional compatibility for ChA transport.

Conclusion: The study contributes to the understanding of the pharmacokinetics and pharmacodynamics of ChA and provides the basis for the rational design of pharmaceutical substances based on it.

Objective: Hydroquinone (HQ) is globally regarded as the gold-standard topical agent for melasma treatment. However, its clinical utility is limited by several challenges: susceptibility to oxidation induced by light, air, and metal ions, which generates harmful byproducts and triggers adverse effects (e.g., skin irritation, pruritus, and dermatitis). Moreover, excessively high concentration in the skin of hydroquinone due to the burst release of HQ may cause melanocyte cytotoxicity, while systemic penetration raises additional safety concerns. To overcome these limitations and enhance the therapeutic applicability of HQ, this study aims to develop strategies that mitigate its instability and optimize its delivery profile.

Methods: In this study, an oil-in-water cream containing hydroquinone cyclodextrin inclusion complex (HQ-HPCD) was designed with PO as the oil phase matrix to improve the treatment of melasma.

Results: As expected, the in vitro release profile of HQ-HPCD cream exhibited near-zero-order kinetics, effectively mitigating the irritation associated with burst release-induced peak concentrations. The HQ-HPCD inclusion complex demonstrated enhanced photostability and oxidative resistance compared to free HQ. Then it was confirmed by B16F10 cells that the HQ-HPCD increased the IC₅₀ of HQ by 4.6 times. Moreover, skin condition, tyrosinase activity, SOD (Superoxide Dismutase), MDA (Malondialdehyde) and melanin content in the mice were detected and the results were in line with the in vitro assays, which showed that PO-HQ-HPCD cream has higher resistance to melanin deposition and lower irritation.

Conclusions: This study successfully developed a topical formulation that enhances the stability of hydroquinone (HQ) while achieving its sustained release in the epidermal layer, thereby improving its clinical applicability of melasma.

Purpose: Establishing bioequivalence (BE) for narrow therapeutic index (NTI) drugs presents unique challenges due to the delicate balance between therapeutic efficacy and toxicity. Recognizing the critical nature of NTI drugs, regulatory agencies such as the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) have adopted stricter, though differing, BE criteria for NTI drugs. This manuscript aims to align with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) M13's goal of harmonizing BE standards globally.

Methods: We propose alternative FDA NTI BE criteria by capping the minimum BE limits, applying alpha adjustment, and applying a point estimate constraint. We compare the operating characteristics-Type I error rate, statistical power, and sample size requirements-of the current and proposed FDA criteria with those of the current EMA criteria and the alternative approach proposed by Paixão et al. Strengths and limitations of each criterion are analyzed, and areas for potential alignment across regulatory frameworks are discussed.

Results: The proposed FDA alternative criteria offer improved performance over the current FDA criteria, particularly its low power for drugs with low within-reference standard deviation (e.g., σ < 0.1), maintenance of Type I error control, and close alignment with Paixao's proposed EMA alternative.

Conclusions: Our proposed alternative FDA criteria provide both theoretical and empirical support, which will help ICH M13 Expert Working Group in developing the ICH M13C guideline with the goal of harmonizing NTI BE study design and criteria across different regulatory agencies.

Purpose: First-generation PARP inhibitors, including olaparib, niraparib, and rucaparib, act as non-selective inhibitors of the PARP family, and are associated with hematological side effects. DM5167 is a selective PARP1 inhibitor, and its selectivity could reduce its side effects; however, its pharmacokinetic properties, tissue distribution, and excretion profiles remain unclear. This study aimed to characterize the pharmacokinetic properties of DM5167 to support the prediction of its pharmacokinetics in humans.

Method: The pharmacokinetics of DM5167 were assessed in mice, rats, and dogs over a dose range of 3-30 mg/kg. Parameters included oral bioavailability, tissue distribution, plasma protein binding, enzyme inhibition, half-life in microsomes and hepatocytes across species, and in vitro-in vivo extrapolation to predict human clearance. Metabolite profiling was performed to identify whether any the human-specific metabolites.

Results: DM5167 demonstrated linear pharmacokinetics across the studied dose ranges in mice and rats. Oral bioavailability was moderate to high, and tissue distribution showed broad organ coverage. DM5167 exhibited high plasma protein binding across species, including humans, and did not inhibit cytochrome P450 enzymes. The half-life of the compound varied among species, with a notably longer half-life in humans. In vitro-in vivo extrapolation results indicated stable predicted clearance values in humans, and metabolite profiling did not reveal any human-specific metabolites.

Conclusion: DM5167 exhibited a favorable pharmacokinetic profile in vivo and in vitro. These findings support its potential for further clinical development and provide valuable insights to guide future preclinical and clinical studies of DM5167 and other PARP inhibitors.

Introduction: Inhibition of Phospholipase D1 (PLD1) is a promising therapeutic strategy for Alzheimer's disease (AD) and related dementia (ADRD), yet clinical progress has been stalled by the inability of potent inhibitors to effectively cross the blood-brain barrier (BBB). While next-generation PLD1 inhibitors have failed in preclinical mammalian models due to poor CNS penetration, we revisit our functionally proven inhibitor VU0155069 (or VU01), demonstrating a definitive solution to this delivery challenge.

Methodology: We engineered an exosome/extracellular vesicle (EVs)-based nanocarrier (Exo-VU01) to encapsulate VU0155069, optimizing drug payload via electroporation and confirming vesicle integrity through Nanoparticle Tracking Analysis. The therapeutic potential of this formulation was tested in a head-to-head intravenous pharmacokinetic study against free VU0155069 in mice.

Results: Relative to free VU0155069, which exhibited rapid systemic clearance and negligible brain retention, Exo-VU01 achieved approximately 20-fold higher brain area under the curve (AUC), underscoring its potential to optimize drug retention and regional biodistribution within the CNS.

Conclusion: Our study validates Exo-VU01 as a viable platform for CNS drug delivery. It provides increased efficacy for VU0155069 as a therapeutic candidate for AD/ADRD and establishes a clear translational pathway for targeted delivery.

Purpose: Cannabidiol (CBD) exhibits antiepileptic, anticonvulsant, and anticancer effects. However, its clinical value is limited by poor oral absorption, low water solubility, and poor stability. Therefore, this study aimed to prepare a novel nanomicelles (NMs) to improve the above aspects of CBD.

Methods: The amphiphilic polymer was synthesized by selecting γ -polyglutamic acid (γ-PGA) and cholesterol (CHOL) as hydrophilic and hydrophobic materials respectively. The optimal preparation process for CBD/(γ-PGA-g-CHOL) NMs was obtained through single-factor and orthogonal tests. The particle size, potential, stability, morphology, apparent solubility and in vitro drug release behavior of the drug-loaded NMs were characterized. Cytotoxicity, uptake and transport experiments on Caco-2 cells and in vivo pharmacokinetics studies in rats were performed for in vitro and in vivo oral absorption of the drugs.

Results: The optimal parameters were a dosage of 2 mg, blank NM concentration of 5 mg/mL, an organic/aqueous ratio of 1:5, and a stirring time of 6 h. The NMs showed pH-sensitive release behavior, with particle size 163.1 ± 2.3 nm, zeta potential -16.5 ± 1.7 mV, encapsulation rate 84.46% ± 0.35%, and drug loading 8.78% ± 0.28%. They were spherical by SEM, stable at 25 ℃ and 37 ℃, and their apparent solubility increased 424-fold. The NMs were biocompatible and improved CBD absorption in uptake/transport tests. Orally administered NMs showed lower apparent clearance value (CL/F), and higher C_max and AUC than those in the free CBD group.

Conclusions: These findings reveal potential of the delivery system, γ-PGA-g-CHOL NMs, to improve the stability of CBD and enhance its apparent solubility and systemic exposure.

The oral bioavailability of drugs is often limited by metabolic barriers, including enzymatic degradation and active efflux processes in the gastrointestinal tract. Piperine, a pungent alkaloid found in black pepper (Piper nigrum), has garnered significant interest as a natural bioenhancer due to its multifaceted ability to inhibit cytochrome P450 enzymes, particularly CYP3A4, and efflux transporters such as P-glycoprotein (P-gp). These actions result in enhanced intestinal absorption and prolonged systemic retention of various therapeutic agents. Additionally, Piperine modulates intestinal permeability and alters the pharmacokinetics of drugs by interfering with first-pass metabolism. Recent developments in nanotechnology have led to innovative formulation strategies, such as nanoemulsions, liposomes, and self-emulsifying drug delivery systems, which further enhance Piperine's solubility, stability, and efficacy. However, despite its promising bioenhancing effects, Piperine exhibits limitations such as poor water solubility, dose-dependent toxicity, reproductive and hepatic concerns, and the potential for significant drug-drug interactions. This review critically examines the mechanistic pathways, formulation advances, pharmacological roles, safety issues, and clinical prospects of Piperine. Furthermore, it emphasizes the need for rigorous clinical trials and regulatory evaluation to validate Piperine's use in pharmaceutical applications. Overall, Piperine represents a potent, yet cautiously applicable, tool in modern drug delivery strategies.

Ocular inflammation is a major contributor to vision-threatening disorders, with phosphodiesterase 4 (PDE4), a key regulator of cAMP playing a central role in pro-inflammatory signaling. Although investigational PDE4 inhibitors like Rolipram (RP) show therapeutic promise, their systemic toxicity limits clinical application, underscoring the need for safer, targeted alternatives. Urolithin A (UA), a gut-derived metabolite of ellagic acid with emerging anti-inflammatory properties, was evaluated as a novel PDE4 inhibitor. Molecular docking revealed that UA binds with high affinity to the A-chain of PDE4A (-8.79 kcal/mol), forming unique π-π stacking and multiple hydrogen bonds. In contrast, RP binds preferentially to the B-chain with slightly lower affinity (-8.42 kcal/mol) and fewer stabilizing interactions. While both ligands engage similar catalytic residues, UA exhibited a more extensive binding profile, suggesting enhanced stability and specificity. In lipopolysaccharide (LPS)-stimulated human retinal pigment epithelial cells (ARPE-19), UA significantly inhibited PDE4A activity, elevated intracellular cAMP, and reduced key inflammatory mediators (NF-κB, IL-6, TNF-α), as demonstrated by immunofluorescence, ELISA, and gene expression analysis. These findings support UA's function as an anti-inflammatory agent by inhibiting PDE4A, highlighting its potential as a safer systemic or localized therapy for ocular inflammatory diseases.

Purpose: This study investigated the potential of a deep eutectic solvent (DES) to enhance the dissolution of two poorly water-soluble drugs, ibuprofen (IBU) and empagliflozin (EMPA). The DES was synthesized from tetrabutylphosphonium bromide (TBPB) and diethylene glycol (DEG).

Methods: The apparent solubility of IBU and EMPA was measured in aqueous solutions containing eleven different mass fractions of the DES at temperatures ranging from 20 to 40°C. Dissolution kinetics were monitored over 24 h to differentiate between true equilibrium solubility and supersaturated states. The collected experimental data were then analyzed and correlated using three thermodynamic models: Wilson, NRTL, and UNIQUAC.

Results: The findings indicated that ibuprofen achieved higher dissolution than empagliflozin in the DES-water system. For both drugs, the dissolution process was endothermic, with solubility increasing as both temperature and DES concentration increased. Among the thermodynamic models tested, the UNIQUAC model provided the most accurate correlation with the experimental dissolution data.