Sepsis is defined as "a life-threatening organ dysfunction caused by a dysregulated host response to infection" and is one of the most frequent causes of death worldwide. In 2020, the World Health Organization released their first global report on sepsis: "Global Report on the Epidemiology and Burden of Sepsis." This report recommended inclusion of sepsis in all public health efforts such as national action plans. Bacteria, viruses and fungi can all cause sepsis, but bacteria are the most common cause. Bacterial endotoxin, the major cell wall component of Gram-negative bacteria, triggers massive systemic inflammation and plays a pivotal role in sepsis pathogenesis. Targeting endotoxin has thus long been a major therapeutic strategy for treating sepsis. Sepsis due to Gram-positive bacteria occurs at a similar rate as that for Gram-negative bacteria, but the causative agents of Gram-positive sepsis remain poorly understood. A variety of microbial components trigger distinctive inflammation by acting as pathogen-associated molecular patterns (PAMPs), which are recognized by specific pattern recognition receptors (PRRs) on host immune cells. Due to the often-fast progression of sepsis, rapid and accurate identification of its causative agents is crucial for effective treatment. This review provides an overview of the author's research for overcoming sepsis.
In recent years, therapeutic antibodies with various molecular modifications, such as antibody-drug conjugates, glycoengineered antibodies, and bispecific antibodies, have been developed to address unmet medical needs, resulting in increasingly complex and diverse molecular characteristics. Antibody therapeutics are multifunctional molecules capable of binding not only target antigens but also to Fcγ receptors (FcγRs) and complement. To ensure their proper effector functions, exemplified by antibody-dependent cell-mediated cytotoxicity (ADCC), it is essential to identify and appropriately control their quality attributes that influence the effector functions. To date, we have conducted studies on (1) development of novel analytical methods utilizing antibody-FcγR binding affinity, (2) control of characteristics of antibody molecules through peptide conjugation, and (3) establishment of methods for evaluating the higher-order structure of antibodies using spectroscopic and statistical approaches. The molecular mechanism underlying the effector function of therapeutic antibodies and the advanced quality assessment methods developed in this study are expected to contribute to the promotion of development and quality assurance of next-generation therapeutic antibodies that continue to evolve through diverse molecular modifications.
Dementia is a progressive neurodegenerative condition that severely impairs memory and activities of daily living. In an aging society like Japan, the physical, mental, and economic burdens associated with caregiving and medical expenses are substantial, making it an increasingly serious social issue. This review focuses on Centella asiatica, used in Ayurveda for its memory-enhancing effects, and introduces examples of plant-derived components from traditional medicine applied in drug discovery and preventive medicine. We have fractionated Centella asiatica extracts and identified the active component araliadiol. We have also demonstrated that Centella asiatica extracts and araliadiol exhibit potent cytoprotective effects against oxidative stress and endoplasmic reticulum stress. Furthermore, araliadiol improved drug-induced cognitive impairment. This review summarizes the preventive effects of Centella asiatica extracts and araliadiol against dementia, based on our previous research findings.
Active learning methods, such as team- and problem-based learning, are known to improve students' learning attitudes and teamwork skills in undergraduate education. This study conducted an experiment that incorporated an escape room (ER) in classes with the aim of improving students' motivation to learn and knowledge acquisition through collaborative learning. An ER was designed and administered to third-year undergraduate pharmacy students. Students were randomly assigned to a control group (regular lecture focused on knowledge transfer) or an ER group. Both groups completed pre- and post-tests. Learning outcomes and educational usefulness were evaluated using statistical analyses (t-test and chi-squared test) and text mining. The ER group had significantly higher average post-test scores compared with the control group, particularly in terms of new knowledge acquisition. The results of the questionnaire survey, which included free-form responses, indicated improvements in enjoyment, concentration, and motivation to learn. The ER provided opportunities for collaborative, active learning and demonstrated significant benefits in knowledge acquisition and educational usefulness, including increased motivation. These results could be attributed partly to the fact that ER provides a learning experience combining challenge and a sense of accomplishment, leading to a flow-like psychological state. Therefore, ER demonstrates its potential as an effective method of knowledge education in pharmacy education. Future research can clarify the effectiveness of ER by further examining the conditions that maximize the learning benefits of ER, such as the learning effects of review content and its application to other pharmacy-related subjects.
We have engaged in clinical practice and pharmaceutical research aimed at the appropriate use of medications. In pharmacist-to-pharmacist collaboration using medication guidance information-sharing documents, it was possible to monitor treatment efficacy and adverse effects through sharing clinical data, or making prescription proposals. Our hospital's pharmacist services contributed to expanding the calculation of drug management and guidance fees by pharmacists in clinics with beds. The drug interaction that occurred between enteric-coated omeprazole tablets and Maalox® granules suggested the possibility of an unexpected drug interaction through a mechanism distinct from that reported for fluoroquinolones and Maalox® suspension. It has been suggested that certain components of black vinegar strongly inhibit P-glycoprotein (P-gp) function in the intestines, potentially enhancing the absorption of P-gp substrates. It has been suggested that aojiru strongly binds to cationic drugs with cyclic structures, significantly reducing their absorption. Based on these findings, we will continue to conduct pharmaceutical research in clinical practice and promote the appropriate use of medications.
This study aimed to evaluate the drug cost-saving effects of drug vial optimization (DVO) using an anticancer drug compounding robot in a single institution. Robotic preparations, drug costs, and wastage were evaluated from April to July 2022. Cost-saving simulations were conducted assuming same-day or 7-day DVO. From September to December 2022, DVO was performed for paclitaxel, carboplatin, and etoposide, and actual cost savings were evaluated. During the initial period, 2613 doses (42.0%) were prepared by the robot, with a wastage rate of 2.7%. The simulated cost savings were approximately ¥2.8 million (same-day) and ¥7.49 million (7-days). After the introduction of DVO, 291 vials were saved, equivalent to ¥790000. In conclusion, DVO using an anticancer drug compounding robot can reduce anticancer drug wastage and may contribute to drug cost savings in clinical practice.
Pharmacological and toxicological assessments of a wide array of chemicals typically involve estimating human clearance through allometric extrapolation of in vivo animal data, using empirical compartmental and physiologically based pharmacokinetic (PBPK) models. Parameters for pharmacokinetic absorption, distribution, and metabolic clearance can be calculated and applied to models that replicate observed plasma drug concentration-time profiles. In patients with genetically impaired cytochrome P450 (P450) enzymes who are prescribed only certain drugs, moderate plasma exposure should be noted as a precaution on drug labels, as genetic variants can cause changes in blood concentrations similar to those caused by drug-drug interactions. For new approach methodologies that do not rely on experimental data, human PBPK model input parameters for various compounds have been effectively estimated using in silico-generated chemical descriptors and machine learning tools to assess internal exposure in humans. Many drug oxidations are facilitated by species-dependent and polymorphic P450s and flavin-containing monooxygenases (FMOs). For instance, in rats, the main oxidation product of thalidomide was deactivated 5'-hydroxythalidomide, along with sulfate and glucuronide conjugates. However, the species-specific teratogen thalidomide induces human P450 3A and is activated by P450 3A through the primary human metabolite 5-hydroxythalidomide, leading to its conjugation with nonspecific proteins. The metabolic capacity of polymorphic FMO3 was evaluated based on urine tests for food-derived trimethylamine N-oxide levels. This pharmacokinetic modeling approach, incorporating polymorphic drug-metabolizing enzyme information, could be applied in clinical settings and during computational data-driven evaluations of potential risks associated with a broad spectrum of chemicals.
Supramolecular gels are soft materials formed through the self-assembly of low-molecular-weight gelators via noncovalent interactions. Because their molecular structures can be precisely designed, supramolecular gels are promising functional materials with stimuli-responsive properties. This review summarizes our studies on the design and development of supramolecular gels for pharmaceutical applications, as well as the mechanistic elucidation of supramolecular gelation. First, disaccharide-coupled amphiphilic ureas were designed as low-molecular-weight gelators for small intestine-targeted drug delivery systems. Maltose- and lactose-coupled amphiphilic ureas, Mal-Cn and Lac-Cn, were synthesized in three steps. The resulting supramolecular gels showed enzymatic hydrolysis-responsive phase transitions triggered by disaccharidases. In particular, the supramolecular gel formed from Lac-C8 was degraded by β-galactosidase, and the entrapped model drug, Rhodamine 6G, was released as the gel degraded. These results demonstrate the potential of enzymatic hydrolysis-responsive supramolecular gels as drug carriers. Second, the gelation mechanism of a urea-based low-molecular-weight gelator was investigated using high-speed atomic force microscopy. Direct observation revealed that gel-forming fibers grew through repeated elongation and pause phases. A block-stacking model was proposed to explain the intermittent growth of fibers. Through these observations and additional experimental analyses, we elucidated the mechanism of supramolecular gelation.
The field of epitranscriptomics, an area of genetics concerning the regulation of gene expression via post-transcriptional RNA modification, is currently attracting substantial research attention. In epitranscriptomics, proteins, collectively termed writers, erasers, and readers, enter into complex interactions that contribute to modifying RNA, thereby maintaining biological homeostasis. However, abnormalities in the expression or function of these proteins can lead to the onset and progression of cancers and neuropsychiatric disorders. Using prostate cancer clinical specimens, I cloned a novel gene, prostate cancer antigen-1 (PCA-1), containing a domain similar to the 2-oxoglutarate, iron(II) [Fe(II)]-dependent oxygenase domain of the Escherichia coli AlkB protein and characterized by enzymatic activity associated with the demethylation of methylated RNA. This was accordingly designated AlkB homolog 3 (ALKBH3). I demonstrate that ALKBH3 is highly expressed in tumor cells in prostate, pancreatic, lung, and other cancers, and its activity is correlated with a poor prognosis. In addition, I developed novel compounds that inhibit the RNA demethylase activity of ALKBH3, thereby providing a basis for developing a first-in-class cancer therapeutic. I also succeeded in cloning the ALKBH8 gene. High ALKBH8 expression was also observed in bladder cancer cells. Furthermore, abnormalities in development and behavior were noted in the generated Alkbh8 knockout mice. On the basis of the experience gained from ALKBH3 drug discovery research, I have established a foundation system for supporting academic drug discovery research. In this review, I describe the pathway followed in integrating the findings of basic pharmaceutical and drug discovery research and further developments.
Patients diagnosed with metastatic or recurrent cancer experience uncertainty and distress; however, their consultation needs remain insufficiently quantified. We conducted a web survey in Japan (January 23-29, 2025) among adults (≥18 years) with a history of metastatic or recurrent cancer (n=522). Participants selected concerns from 21 items across four domains, and for each endorsed concern, they indicated whether they wished to consult hospital and/or community pharmacists; consultation intention was calculated among those endorsing each item. Hospital-community differences were evaluated using McNemar's test or Mid-P exact test using a significance threshold of p<0.001. The mean age was 58.9±12.9 years; cancers were colorectal (22.2%), breast (18.2%), lung (11.9%), and gastric (11.1%). 88.7% reported at least one concern. The most common concerns were treatment-related side effects (51.0%), anticancer drug mechanism/efficacy (46.9%), treatment costs (42.1%), mental distress (35.1%), and medications used to alleviate cancer- or treatment-related physical discomfort (34.7%). Consultation intention was higher for hospital than community pharmacists for issues including side effects (43.2 vs. 16.2%), mechanism/efficacy (42.9 vs. 18.0%), and symptom-relief medications (48.6 vs. 24.9%). Although concerns regarding medications other than cancer treatment were uncommon (<10% each), consultation intention exceeded 40% when present. These findings indicate that patients with metastatic or recurrent cancer may perceive different consultation roles for hospital and community pharmacists, particularly according to the type of concern. Because these results are based on self-reported consultation intentions rather than actual consultation behavior, they should be regarded as hypothesis-generating and as a basis for future studies on coordinated pharmacist support.
Managing adverse events is important for optimizing cancer treatment and ensuring high patient satisfaction. Studies have assessed (1) anti-epidermal growth factor receptor (EGFR) monoclonal antibody-induced skin toxicities, (2) development of severe neutropenia by renally excreted anticancer drugs in patients with renal impairment (RI), and (3) pharmaceutical care in the treatment of immune checkpoint inhibitors (ICIs). We identified liver metastasis as a risk factor and preemptive systemic antibiotic administration with anti-inflammatory effect as a preventive factor for grade ≥2 overall skin toxicities in anti-EGFR treatment for metastatic colorectal cancer (mCRC). Additional prophylactic topical steroids to systemic minocycline significantly prevented grade ≥2 rashes, but did not mitigate overall skin toxicities. Patients receiving trifluridine/tipiracil (FTD/TPI)-based chemotherapy for mCRC were assessed, resulting in significantly higher early severe neutropenia development among patients with RI. Additionally, we assessed the impact of RI on severe neutropenia development in carboplatin+pemetrexed-based chemotherapy for thoracic cancer. Consequently, severe neutropenia in the first cycle and all-treatment cycles was significantly more confirmed in patients with RI. We assessed the usefulness of pharmaceutical interventions in ICI treatment, which suggested that pharmaceutical care may improve quality of outpatient ICI treatment, and pharmaceutical intervention during the first three months after initiation of ICI treatment is crucial. Our studies have found clinically important outcomes that support the provision of less onerous chemotherapy.
Antibodies are endogenous proteins that are involved in humoral immunity. Although various therapeutic monoclonal antibodies (mAbs) targeting cancer and immune diseases have been widely used, the development of mAbs for the treatment of infectious diseases has remained limited. During the coronavirus disease 2019 (COVID-19) pandemic, various mAbs targeting the spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were developed, received Emergency Use Authorization (EUA), and were used for treating and preventing COVID-19. As successive SARS-CoV-2 variants emerged and the pandemic spread, mAbs with high target specificity proved less effective against these variants. Consequently, their EUAs were suspended within a short period, highlighting the limitations of antiviral mAbs developed against emerging infectious diseases. Since 2021, guidance documents on mAbs for prevention and treatment of COVID-19 have been issued by the WHO and US FDA, and efforts to prepare a regulatory environment supportive of drug development for future emerging and re-emerging infectious diseases have accelerated. The EMA published a concept paper on development of guidelines for non-clinical and clinical evaluation of mAbs for COVID-19. These trends indicate that the international regulatory landscape for mAbs against infectious diseases is entering a new era. We here provide an overview of the guidance documents issued by regulatory agencies, based on insights obtained from anti-SARS-CoV-2 mAbs during the COVID-19 pandemic. We also discuss the challenges and future prospects for the development of mAbs against infectious diseases, particularly in situations requiring rapid responses during pandemics.
The blood-retinal barrier (BRB) consists of the inner and outer blood-retinal barrier (abbreviated as inner BRB and outer BRB). The inner and outer BRB's together regulate the transport of substances between the blood and neural retina via various membrane transporters. To utilize the oral administration for the pharmacotherapy of retinal diseases, it is necessary and important to understand many drug transport mechanisms in the BRB's. This review discusses the BRB transport research reported over more than a quarter of a century, particularly the development of in vitro and in vivo transport evaluation systems as well as newly discovered BRB-specific cationic drug transport mechanisms.
Structure-based drug design (SBDD) plays a crucial role in modern drug discovery. SBDD is a methodology that utilizes the three-dimensional (3D) structural information of a target protein to computationally search for compounds that bind to it and/or to optimize the binding of known compounds. This article provides an overview of recent advances in SBDD. Furthermore, we introduce 2 case studies conducted by the authors using single-particle analysis via Cryo-electron microscopy (Cryo-EM), a powerful technique for obtaining protein structures. The first example is the sodium ion (Na+)-translocating NADH-quinone oxidoreductase derived from Vibrio cholerae. This enzyme is suggested to undergo large conformational changes during the reaction cycle. By performing classification in single-particle analysis, we successfully captured a partial view of these conformational dynamics. The second example involves the angiotensin-converting enzyme 2 (ACE2) decoy, which binds to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein and inhibits infection. Focusing on ACE2, which is critical for SARS-CoV-2 infection, an ACE2 decoy was developed to achieve higher binding affinity than the wild-type ACE2. We successfully elucidated how this ACE2 decoy binds to the spike protein.
Equivalence of OTC formulations is generally assessed based on pharmaceutical attributes such as active ingredient content, dissolution behavior, and stability. However, for topical liquid formulations that are directly applied to the skin or scalp, the container design and usability may also influence the applied dose and the amount that reaches the target site. Appropriate design of containers or applicator devices is therefore essential for ensuring the effectiveness and safety of OTC topical products. In this study, we investigated the effects of container-tip geometry and hair density on scalp delivery of minoxidil (MXD) using four marketed MXD topical formulations. A scalp model was prepared by covering filter paper with human hair, and we evaluated: (1) the amount of MXD dispensed per actuation from each container, (2) the amount of MXD that reached beneath the hair layer, and (3) the in vitro skin permeation using porcine ear skin. Increased hair density reduced MXD delivery to the scalp for all formulations, especially for containers with larger tip ends. These findings suggest that ensuring the efficacy and reproducibility of MXD-containing OTC topical formulations requires not only optimizing drug permeability and formulation characteristics, but also considering hair density and container-tip geometry in designing appropriate usage conditions.
Amiodarone (AMD) is characterized by complex pharmacokinetics involving high lipophilicity and extensive protein binding. Serum AMD concentrations are influenced by triglyceride (TG) levels. Although the impact of TG on AMD concentrations at steady state has been previously investigated, data regarding the initial treatment period are limited. Given the prolonged half-life of AMD and the need for dose adjustments prior to reaching steady state, elucidating the effect of TG during this early phase is important. This retrospective study analyzed 64 patients who received AMD therapy at Sapporo City General Hospital between January 2016 and December 2024. Serum levels of AMD, its active metabolite desethylamiodarone (DEA), TG, and albumin were measured during the initial period (4-6 months) and steady-state period (7-12 months). Concentration-to-dose (C/D) ratios were calculated to assess pharmacokinetic variability during each period. Significant positive correlations were observed between serum TG levels and C/DAMD during the initial (r=0.424, p<0.05) and steady-state (r=0.492, p<0.01) periods. A modest but significant correlation was observed between TG and C/DDEA during the steady-state period (r=0.304, p<0.05). No significant correlations were found between albumin and C/DAMD or C/DDEA at any period. Serum TG levels affected AMD concentrations from the initial treatment period; however, the pharmacologically active unbound fraction remained stable. Consequently, dose adjustments based solely on fluctuations in TG levels are not required during the initial period. These findings contribute to the development of more precise AMD dosing strategies to maintain antiarrhythmic efficacy from treatment initiation.
ClpP protease is a highly conserved serine protease that plays a crucial role in bacterial protein quality control alongside its partner AAA+ ATPases. ClpP assembles into a barrel-shaped tetradecamer that degrades unfolded or misfolded proteins translocated by ATP-driven unfoldases, such as ClpC, ClpX, or ClpA. Acyldepsipeptide (ADEP) antibiotics bind to the hydrophobic pockets of ClpP, mimicking the natural interaction with ATPases, thus activating ClpP in an ATP-independent manner. ADEP binding induces major conformational changes that open the axial pores, enabling ClpP to degrade large protein substrates such as the cell division protein FtsZ, ultimately causing cell death. Our recent studies revealed that in Bacillus subtilis the ClpP proteolytic system regulates the intracellular levels of nonribosomal peptide synthetases SrfAA, SrfAB, and SrfAC, which are responsible for surfactin biosynthesis. Moreover, ADEP1-activated ClpP directly degraded SrfAA and SrfAB both in cells and in vitro, identifying new physiological substrates of the ADEP1-ClpP complex. High-speed atomic force microscopy (HS-AFM) analysis visualized the stepwise oligomerization of B. subtilis ClpP from monomers to heptamers and then to tetradecamers upon ADEP1 binding, revealing dynamic assembly processes underlying its activation. These findings enhance our understanding of bacterial protein degradation mechanisms and provide a molecular basis for the rational design of ClpP-targeting antibiotics with novel modes of action.
In recent years, cancer chemotherapy has been administered in an outpatient setting; therefore, patient self-management of adverse events is an important issue. Pharmaceutical interventions were performed by pharmacists at the following 3 time points: before the physician consultation, after the physician consultation, and during multidisciplinary conferences at the Ibaraki Prefectural Central Hospital Cancer Chemotherapy Center. In this study, prescription proposals and their effectiveness for adverse events, were investigated. Of the 338 cases in which pharmaceutical interventions were performed, 280 were accepted by physicians, and the acceptance rates were 77% in conference, 89% before physician consultations, and 85% after physician consultations. Pharmaceutical interventions for the management of nausea and vomiting were most frequently accepted (96 cases), with improvement observed in 58 cases (60%). Improvement rates for hypertension and skin disorders were 76% and 56%, respectively, and improvement rates for peripheral neuropathy and dysgeusia were 33% and 22%, respectively. Of all interventions made by pharmacists, 68% were for the 1st 5 courses, with proposals continuing beyond the 6th course, suggesting that there is demand for long-term intervention. Pharmacists collecting patient information and implementing pharmaceutical interventions may contribute to the management of adverse events. As the degree of improvement varies according to the type of adverse event, it will be necessary to establish optimal pharmaceutical interventions in the future.
Targeted protein degradation (TPD) is an emerging approach that selectively eliminates specific proteins using synthetic molecules, such as proteolysis-targeting chimeras (PROTACs). It has attracted increasing attention in medicinal chemistry and chemical biology, with several PROTACs being tested in clinical settings. Unlike traditional small molecules, such as enzyme inhibitors and receptor antagonists, PROTACs exhibit a fundamentally different mechanism. Conventional drugs block enzymatic activities or receptor interactions, whereas PROTACs induce the degradation of target proteins, decreasing their cellular levels and abolishing all associated functions. PROTACs targeting enzymes in protein complexes disrupt both their catalytic activity and involvement in complex formation. In some cases, they also degrade other proteins in complexes, facilitating the elimination of entire assemblies. Our study leverages these unique features of TPD. We are currently developing various PROTACs targeting the enzymes responsible for lysine acetylation or methylation in proteins. Recently, the TPD concept has been extended beyond proteins to include nucleic acids, and ribonuclease-targeting chimeras (RIBOTACs) that selectively degrade RNA have been developed. We are also actively exploring new RNA-targeted degradation strategies. Herein, we highlight our recent work on TPD-inducing small molecules and provide an overview of RIBOTACs, which represent an area of growing research interest.
