Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan最新文献

Deficiency of adenosine deaminase 2 (DADA2) is an autosomal recessive genetic disorder caused by loss-of-function mutations in the gene encoding adenosine deaminase (ADA) 2. This enzyme catalyzes the deamination reaction of adenosine/2'-deoxyadenosine to inosine/2'-deoxyinosine. DADA2 exhibits a complex clinical presentation, with systemic vasculitis with stroke, bone marrow failure, and immunodeficiency as the major pathologies. Since its discovery in 2014, more than 400 cases have been reported, and the phenotype has expanded significantly. DADA2 generally presents in childhood, although diagnosis in adulthood has also been reported, indicating the need to raise awareness of this disease beyond pediatrics. ADA2 is believed to be relevant in the regulation of the human immune system. Currently, knowledge is accumulating on the association between macrophage polarization into an inflammatory phenotype and systemic vasculitis, upregulation of the type I/II interferon pathway, and neutrophil function. The biochemical characteristics of ADA2 that differ from those of its isozyme ADA1 are a subject of significant research. One of these characteristics, N-glycosylation, plays a vital role in controlling the formation of the functional three-dimensional structure of ADA2. Moreover, with the accumulation of knowledge regarding the dysregulation of innate and adaptive immunity in DADA2 and the biochemical properties of ADA2, effective treatments and diagnostic methods are being established. This review provides an overview of ADA2 properties and DADA2.

Antigen-binding fragments (Fab) are a type of antibody fragment that contains an antigen-binding site in a therapeutic antibody format. To further improve their utility as therapeutic antibodies, this study aimed to enhance Fab functionality through protein engineering. A Fab expression system using the yeast Pichia pastoris was constructed, and recombinant Fabs were efficiently prepared. Second, a Fab mutant suitable for conjugation with polyethylene glycol (PEG) was generated to increase the serum half-life of the Fab. The interchain disulfide bond normally formed at the C-terminus (H: Cys224-L: Cys214) was shifted to a novel position (H: Cys177-L: Cys160), allowing a free cysteine residue at the C-terminus to be used for site-directed PEGylation without conformational destabilization of the Fab. The prepared PEGylated Fab displayed an increased serum half-life. Several additional sites for the introduction of interchain disulfide bonds, which contribute to conformational stability, have been identified in the Fab constant region, and a Fab with an N-glycosylation site introduced at position 178 of its heavy chain (H: L178N) was expressed in P. pastoris. The high-mannose type N-glycan attached to Fab showed the inhibited Fab aggregation under pH shift-induced stress, and the immunogenicity of the glycosylated Fab was lower than that of the wild-type Fab. These protein engineering results are expected to contribute to the design of Fab molecules with increased functional value and greater safety.

As the role of pharmacists expands from product-focused to patient-focused care, the development of pharmacist scientists with a balanced clinical and research mindset is expected in Pharmaceutical education. The 2022 revision of the Model Core Curriculum for Pharmacy Education emphasizes scientific inquiry as a fundamental competency for pharmacists. Although graduation research is crucial in promoting this exploration, the current approach to the 6-year pharmacy education program that began in 2006 needs to be re-evaluated. We surveyed undergraduate pharmacy students and practicing pharmacists to investigate their perspectives on graduation research. Those who value the cognitive and methodological skills gained through research have a positive view, whereas those who judge it based on its direct applicability to practice are more critical. The 2022 revision of the Model Core Curriculum for Pharmacy Education aims to develop the ability to identify and solve unknown problems through scientific thinking and appropriate research activities from a pharmaceutical perspective. Graduation research is the only opportunity in undergraduate education for students to experience the full research process-formulating clinical questions into research questions, planning studies, collecting and analyzing data, examining results, and presenting findings-thereby cultivating logical thinking skills and an understanding of research ethics. Universities are responsible for training pharmacists in the research capabilities needed to contribute to future healthcare. Hence, this issue requires collective consideration with clinical pharmacists. It is expected that all stakeholders will share a common understanding that the goal of graduation research is to cultivate a research mindset that pharmacists should naturally possess.

Bio-related nanoparticles are key players in intercellular communication and represent the ultimate goal of nanotechnology as material-transport systems. Biological membranes support the functionality of bio-related nanoparticles; therefore, understanding the correlation between membrane molecular characteristics and the functions of bioparticles is essential. In this study, we developed foundational technologies for measuring the biological membranes of bioparticles by integrating fields such as separation science, materials chemistry, and omics. Furthermore, we developed a new methodology for incorporating the functions of natural bioparticles into artificial nanomaterials.

LM1010, a high-performance liquid chromatography system for therapeutic drug monitoring (TDM) of various agents in clinical practice, is currently approved in Japan. Unlike conventional systems, LM1010 can rapidly measure clinical samples, and data analysis is based on the absolute calibration curve method and fully automated. However, the analytical performance of the LM1010 for the measurement of serum concentrations of linezolid (LZD) has not yet been evaluated. In this study, we aimed to evaluate the analytical performance of LM1010 and confirm the long-term stability of LZD at -30°C. One hundred forty-eight serum samples were collected from 25 patients (17 men and 8 women) who received LZD. Serum concentrations of LZD were measured using LM1010 and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Three LM1010 instruments were used to confirm the differences in the measured values. A Passing-Bablok regression analysis demonstrated a strong correlation between LM1010 and LC-MS/MS (r=0.991), and comparable results were observed among the three LM1010 instruments (r=0.982-0.998). Bland-Altman analyses revealed that the average difference between LM1010 and LC-MS/MS methods was 1.7%, and similar results were observed among the three LM1010 instruments. The long-term stability of serum LZD in samples stored at -30°C for 48 weeks was also confirmed. These results suggest that LM1010 can be used for TDM of LZD in clinical practice.

Synaptic plasticity is a key mechanism underlying long-term memory and learning, with proposed associations with altered actin cytoskeleton and gene expression in neurons. Myocardin-related transcription factor (MRTF) family members, abundantly expressed in the brain, have actin-binding motifs at the N-terminus and act as cofactors of serum response factor (SRF). Rho signaling may promote MRTF's release from G-actin and subsequent translocation into the nucleus, and increases SRF-dependent gene expression. MRTF is therefore thought to act as a link between morphological change and gene expression in neurons. In this review, we focus on the neurotrophin, brain-derived neurotrophic factor (BDNF). BDNF plays crucial roles in neuronal survival, gene expression, and dendritic growth, which are essential for neuronal plasticity. As previous studies suggest that BDNF triggers the activation of MRTF/SRF-mediated signal transduction, we have studied how this supposed regulatory ligand is involved in the functional MRTF changes in neurons. We review the BDNF-mediated roles of MRTF in neuronal morphology and gene expression and briefly discuss the possible involvement of MRTF in neurodevelopmental disorders, such as autism spectrum disorder and intellectual disability.

Nitriles are important organic compounds that are frequently found in pharmaceuticals, natural products, agrichemicals, and functional materials. In addition, nitriles serve as versatile building blocks in synthetic chemistry, as the nitrile moiety can be readily transformed into a variety of valuable functional groups, such as carbonyl derivatives, amines, and heterocycles. However, conventional methods for nitrile synthesis often rely on highly toxic metal cyanides, such as copper(I) cyanide (CuCN) and potassium cyanide (KCN). In this review, I introduce our recent works on a cyanide- and transition metal-free approach for the synthesis of (hetero)aromatic and aliphatic nitriles from N-acyl (2-nitrophenyl)sulfonamides via a base-promoted desulfonylative Smiles rearrangement. The reactions proceeded under operationally simple conditions using potassium acetate as a base and 1,3-dimethyl-2-imidazolidinone as a solvent. This method enables the efficient preparation of a wide range of nitriles, including those bearing primary, secondary, and tertiary carbon centers, with broad functional group compatibility and in good to excellent yields.