The installation of nursing universities has rapidly increased from the late 1990s, there are 263 institutions in Japan as of 2018. Pharmacology and Clinical Pharmacology education in undergraduate and postgraduate nursing universities is highly important, however, the lack of human resources involved in pharmacology education due to the rapid increase of nursing universities cannot be denied. The Ministry of Education, Culture, Sports, Science and Technology of Japan established a " Nursing education model・core・curriculum" for quality assurance of education at nursing universities in 2017. To meet the needs of a new phase of nursing education, re-examination of curriculum of Pharmacology education for nurses is necessary.

Doctors give prescriptions after considering the medical conditions of patients based on the concept of Personal drug (P-Drug). Pharmacists check the prescription, and give information about the effects of the drugs to the patients, including their side effects. Nurses observe patients to detect the effects and side effects of the administered drugs, and then report these to doctors and pharmacists. Each specialist plays a role, allowing medication to be completed. However, medication errors by nurses unfortunately caused because of lack of knowledge of therapeutic drugs. 

In this symposium, I would like to propose a novel concept "integrated drug (iDrug)" and "simple medication guide" based on "Patient-oriented Pharmacology" for nursing Pharmacology education to maintain high level of safety medication.

It is widely recognized that the concept of core curriculum with options is a response to the major problem of content overload. Pharmacology education systems in medical schools are continuously challenged to reform in Japan as well as other countries. The goals of medical workers in patient management are optimization of the pharmaceutics for their patients. To this aim, various new trials have now being conducted. We'll discuss future prospects of pharmacology education keeping designing a next-generation education system in mind.

There are few nationwide reports on pharmacology education to show actual total school hours, how to evaluate students' knowledge, what textbook is recommended to students, or what are performed as pharmacological practice etc. The present study investigated such facts on pharmacology education, based on the results of a nationwide survey asked to the chiefs in the Department of Pharmacology or Clinical Pharmacology in Medical Schools. Around 70 percent of total invited chiefs replied to the survey with answering many kinds of questions. Currently, data have being aggregated. For example, average total school hours are around 90 hours per year. Examination is mainly performed by a combination of description-type and multiple-choice questions. Main textbook is recommended but not specified in a half of the facility. Staffs spend 28 hours for the pharmacological practice per year, etc. Overall, survey results exhibited that the chiefs in the Department of Pharmacology or Clinical Pharmacology in Medical Schools are suffering from performing long-time lecture and pharmacological practice with fewer staffs, but continuously make efforts to perform better pharmacology education with worrying about properly evaluating such educational efforts from the executive in the school. I will discuss about the future plan for pharmacology education with taking into consideration of nationwide cooperation in this session.

To maintain the quality of education in laboratory training with limited human resources, we developed an original method to carry out exercises led by undergraduate students themselves. Our concept is to let all students play individual roles during the seven-day exercises in pharmacology. Laboratory exercises are mainly animal studies using mice and guinea pigs, or a computer simulation of therapeutic drug monitoring. Each student is primarily allocated to a distinct role in the practice of his/her own choice. Faculty members coach the students assigned to respective exercises in advance, so that they learn how to guide their group on the day of practice. With the usage of this "YANEGAWARA" system, laboratory exercises in a large number of students could be operated under the supervision of a few faculty members. Detailed scheme of our trial will be presented at the symposium.

We are using movies as a secondary task of pharmacology practice.　We show several movies suitable as a teaching material of pharmacology or medicine and let students to write a report on drugs and treatment used in the movie. The suitable examples contain Beautiful mind, The awaking, Rain man and Patch Adams and so on. These movies are also useful as education of medical ethics. In this symposium, we will discuss this subject using concrete examples.

Pharmacological practice is important for understanding pharmacological effects from the point of view of active learning in medical education. However, it is very difficult to perform the practices using experimental animals in recent years, because of animal welfare, cost and so on.

We, therefore, developed novel simulation system for animal pharmacological practice using iPad. In our system, each student can access to the Cloud using iPad and simulate the situations for application of various drugs and reagents to atrium, ileum and circulation in whole animal. In addition, students are given various assignments about the pharmacological effects of drugs and reagents and try to solve them. Reports from students about the simulator are prepared by the processes to solve the assignments and their results. Therefore, our system is one of the "problem-based active learning" system.

It is difficult for complete replacement of animal-based pharmacological practice to simulator-based one in present time. However, our simulation system may be useful for reduction of the number of animal use in pharmacological practice.

Endoplasmic reticulum (ER)-associated mitochondrial membranes (MAMs) are highly functionalized domains of interaction between ER and mitochondria in the cells, stabilized by proteic bridges and sequestering numerous proteic assemblies. Through MAMs, ER and mitochondria exchange Ca²⁺, proteins, lipids or small signaling molecules. Ca²⁺ exchanges are driven by IP₃ receptors (IP₃Rs) on ER and VDAC1/MCU complexes, mNCS, UCPs, and Letm1 on mitochondrial membranes. MAM dysfunctions are responsible of several mitochondrial diseases or genetic syndroms, and directly contribute to the neurodegenerative processes in Alzheimer, Parkinson, Huntington diseases and amyotrophic lateral sclerosis.

Targeting MAMs fonctionality by drug candidates is a novel pharmacological area of research and it may offer effective and potentially wide spectrum therapeutic strategies. We are interested since several years in developing drug candidates targeting the sigma-1 chaperone protein (S1R). Highly expressed in MAMs, S1R interacts with IP₃Rs, ER stress sensor proteins (BIP, IRE1), or steroids (pregnenolone) to potentiate focused Ca²⁺ exchanges between the organelles, stabilize mitochondrial physiology (particularly by interacting with complex I) and maintain MAM integrity. We will here detail the physiopathological importance of targeting MAMs and describe new drug development programs with new S1R agonists or positive modulators that already showed very promising efficacy in several neurodegenerative pathologies.

Keywords:

ER-mitochondria communication; sigma-1 chaperone protein; drug development; neurodegenerative diseases.

ASD is a developmental disorder with core symptoms of social impairments and restrictive repetitive behaviors. Its prevalence in a male is 4-times higher than in the female. Dysfunction in the GABAergic system has been implicated in ASD but the exact cause is unclear yet. Thus, no appropriate drugs are currently available for ASD. We recently found that SKF105111 (SKF), a type I 5α-reductase inhibitor, reduced the brain level of allopregnanolone (ALLO), a positive allosteric GABA_A receptor modulator and thereby caused ASD-like behaviors only in male mice, suggesting that SKF treatment provides a novel animal model of ASD. Moreover, to elucidate if Kamishyoyosan (KSS), a Kampo formula used for the treatment of neuropsychiatric symptoms in the menopause female, is available for ASD therapy, we examined the effects of KSS on ASD-like behaviors in the ALLO deficient mouse model. KSS ameliorated sociability deficits via improving the mechanisms mediated by the dopamine D₁ receptor, while it reduced repeated grooming behaviors via improving the mechanisms mediated by dopamine D₁ and D₂ receptors and the GABA_A receptors. Our findings suggested that KSS is beneficial for the treatment of ASD.

Mast cells are crucial effector cells in allergic reactions, where IgE is the best-known mechanism to trigger their degranulation and the release of allergic mediators. Mast cells are also found activated in inflammatory conditions such as autoimmune diseases, chronic inflammatory conditions in upper and lower airways, intestine and skin, neuroinflammation, and cancer, conditions generally not paralleled with an increased antigen-specific IgE expression. However, mast cells can be activated by numerous other stimuli such as complement factors, toll-like receptor ligands, neuropeptides, cytokines, sphingolipids, ATP/ADP/adenosine, histamine and various exogenous compounds. Previously, we have shown that immunoglobulin free light chains (FLC) may also trigger antigen-specific mast cell activation in the absence of IgE. FLC were shown to be increased in various inflammatory disease such as rheumatoid arthritis, non-allergic rhinitis, COPD, inflammatory bowel disease, and cancer and correlated with disease activity. In various preclinical models, we demonstrated that FLC may be of importance to induce mast cell activation and the onset of an inflammatory response. Interestingly, current experiments indicate that FLC may synergize with IgE to stimulate mast cell degranulation. Under conditions of minimal mast cell stimulation via IgE-receptor crosslinking, co-stimulation with FLC strikingly enhanced mast cell activation. These novel findings suggest that this synergy may be relevant under conditions of low local or systemic IgE production.

Keywords: mast cell, IgE, immunoglobulin free light chain, allergy, inflammatory diseases, cancer

The negative regulation of inflammation is very important for the resolution of inflammatory diseases. Although many components and mediators in inflammation have been identified, how inflammation is finely regulated remains lack of evidence. Here we show that Src homology-2 domain containing protein tyrosine phosphatase-2 (SHP2) translocates to mitochondria in the early time of inflammatory stimulation. The RRWFH motif in SH2 domain is essential for the translocation of SHP2. This phosphatase finally localizes in the mitochondrial matrix with the help of Tom20/Tom40 in outer membrane and Tim23 in inner membrane. Then SHP2 interacts with ANT1, a central molecule controlling mitochondrial permeability transition. This mechanism prevents the collapse of mitochondrial membrane potential, mtDNA release and ROS production, and thus inhibits the hyperactivation of NLRP3 inflammasome. Ablation or inhibition of SHP2 in macrophage causes intensified NLRP3 inflammasome activation, production of IL-1β and IL-18, and increased sensitivity to peritonitis. Collectively, our data highlight that SHP2 orchestrates an intrinsic regulatory loop for the mitochondrial homeostasis to limit excessive NLRP3 inflammasome activation, suggesting a possibility to treat inflammatory diseases via maintaining mitochondrial homeostasis.

*This study is supported by National Natural Science Foundation of China (81330079 and 81730100).

Key words: SHP2, resolution of inflammation, mitochondrial homeostasis, NLRP3 inflammasome.

The functions of the lower urinary tract, to store and periodically releaseurine, are organized by complex central and peripheral neural controlmechanisms.  Thus, various disease conditions in the central and peripherallevels can produce lower urinary tract symptoms (LUTS) such as urinary urgency,incontinence or voiding difficulty, urinary retention.  Currently, standardpharmacotherapies for overactive bladder (OAB) include muscarinic receptorantagonists and, more recently, beta₃-adrenoceptor agonists and formale LUTS due to benign prostatic hyperplasia (BPH) utilizealpha₁-adrenoceptor antagonists. Phosphodiesterase type 5 (PDE5)inhibitors and 5alpha-reductase inhibitors have also been prescribed forBPH-associated male LUTS. Furthermore, based on the pathophysiology of LUTS, newpharmacological targets have been identified in bladder urothelium-afferentpathway interactions (e.g., P2X, PGE2 receptors), the spinal cord (e.g.,serotonin, glycine system) and the brain (e.g., adenosine, CRF system). Inaddition, a new chemogenetic approach using "DREADD" may be useful to achievethe subpopulation-specific silencing of sensory pathways using designerreceptors with synthetic ligands for the LUTS treatment without affecting theendogenous system.

Serotonin have been thought to play an important role in regulation of emotional states as well as decision making. Previously we have reported that the activity of the dorsal raphe serotonin neurons was increased by ketamine and olanzapine, promising therapeutic agents for treatment-resistant depression, as well as conventional antidepressants such as selective serotonin reuptake inhibitors and tricyclic antidepressants. It is, however, still unclear whether the activation of the dorsal raphe serotonin neurons is sufficient for eliciting antidepressant-like effects. Furthermore, identification of responsible serotonergic circuits for a variety of behaviors and disorders has been hampered by wide-ranging projection of serotonin neurons. To address these issues, we developed viral vectors capable of optogenetic manipulation of serotonergic neurons specifically. In this symposium, we would like to show our recent findings on the role of serotonin neurons in the regulation of mood and decision making.

Stress is a risk factor for mental illness. Acute stress activates the sympathetic nervous system and HPA axis as a survival mechanism whereas prolonged and excess stress induces cognitive decline and behavioral abnormalities. For instance, repeated social defeat stress, an animal model of depression, induces behavioral changes such as social avoidance.

We previously reported that prostaglandin E₂ which is derived from microglia suppresses mesocortical dopaminergic pathway to induce social avoidance. However, we still do not know whether or how stress activates microglia, or whether microglial activation plays a role in stress-induced behavioral changes.

To identify a molecule which is induced by repeated social defeat stress in the medial prefrontal cortex (mPFC), we performed transcriptome analysis. Repeated social defeat stress upregulated S100A8 and S100A9 in the mPFC. These molecules form a heterodimer to exert their function. Intracellularly, S100A8/A9 migrates phagocytes in a calcium dependent manner. Extracellularly, S100A8/A9 activates Toll-like receptor 4 (TLR4). In this symposium, I will introduce the roles of TLRs in repeated stress-induced behavioral changes and the latest findings on inflammation-like responses.

Fear is an important physiological function for survival. It appears when animals or humans are confronted with an environmental threat. The amygdala has been shown to play a highly important role in emergence of a fear-related behavior such as freezing that only occurs when the environment contains some elements suggestive of danger. Previous studies showed that hypothalamic orexin neurons are activated by fearful stimuli to evoke a 'defense reaction' with an increase in arousal level and sympathetic outflow to deal with the imminent danger. However, how this system contributes to the emergence of fear-related behavior is still unclear. We took cued and contextual fear conditioning experiment using orexin 1 receptor knockout (OX1R^-/-) and orexin 2 receptor knockout (OX2R^-/-) and orexin double receptor knockout (OX1R^-/-; OX2R ^-/-) and prepro-orexin knockout (preproOX^-/-) mice. We found that OX1R^-/- and OX1R^-/-; OX2R ^-/- and preproOX^-/- showed abnormality in both cued and contextual fear conditioning test suggesting the orexin system might be important for fear memory formation and/or consolidation and/or retrieval mainly via OX1R signaling. Orexin neurons in the hypothalamus send excitatory innervations to noradrenergic neurons in the locus coeruleus (NA^LC) which abundantly express OX1R and send projections to the lateral amygdala (LA), which is known as the critical region for fear memory formation. Opto/chemogenetic inhibition of this di-synaptic orexin → NA^LC → LA pathway or pharmacological blockade of OX1R reduces cue-induced fear expression. Inversely, excitatory manipulation of this pathway after fear conditioning induces fear-related behavior. Although, fear memory helps animals respond precisely to a context or cue previously paired with an aversive stimulus, fear-related behavior is sometimes evoked even in a distinct context containing some similar elements, which is known as fear generalization. Our data suggests that the orexin → NA^LC → LA pathway might explain a part of the mechanism of fear generalization. We also discuss about the potential effectiveness of OX1R antagonists for treating excessive fear response or overgeneralization seen in anxiety disorder and post-traumatic stress disorder (PTSD).

A method that promotes the retrieval of lost long-term memories has not been well established. Histamine in the central nervous system is implicated in learning and memory as well as sleep and wakefulness, feeding and drinking, and neuroendocrine regulation, and treatment with antihistamines impairs learning and memory. Since histamine H₃ receptor inverse agonists upregulate histamine release, histamine H₃ receptor inverse agonists may enhance learning and memory. However, whether H₃ receptor inverse agonists promote the retrieval of forgotten long-term memory has not yet been determined. Here, we employed multidisciplinary methods including the mouse behavior, calcium imaging and chemogenetic manipulation to examine whether and how the histamine H₃ receptor inverse agonists, thioperamide and betahistine, promote the retrieval of a forgotten long-term object memory in mice. The treatment of H₃ receptor inverse agonists induced the recall of forgotten memories even 1 week and 1 month after training in mice. In addition, we found a betahistine treatment promote memory retrieval in humans. These results highlight a novel interaction between the central histamine signaling and memory engrams.

Primary sensory cortexes provide massive descending axons to the thalamus to modulate sensory responsiveness of thalamic relay neurons. These top-down controls are pivotal for shifting neuronal firing between burst and tonic modes. The impact of the corticothalamic pathways on the firing mode and sensory gain of thalamic neurons has only been extensively examined in anesthetized animals, but has yet to be established in the awake state. We investigated what change were caused by lesions of the barrel cortex in responses of thalamocortical and thalamic reticular neurons to a single vibrissal deflection during wakefulness. Our results showed that the cortical lesions shifted the response of thalamic neurons towards bursting, elevated the response probability and the gain of thalamocortical neurons, predominantly of recurring responses. In addition, after the lesions, the spontaneous activities of the vibrissa-responsive thalamic neurons were typified by rhythmic spiking with frequent bursting. In single neuron recording/labeling experiments, layer 6 corticothalamic neurons responded to a single vibrissal deflection with short latencies in awake rats, strongly suggesting the existence of an immediate corticothalamic feedback. Because these results showed the importance of corticothalamic neurons in shaping thalamic activities during wakefulness, we next explored what neural circuits in the cortex affected the activities of the corticothalamic circuits. We will further present morphological analyses of corticocortical networks which input to the barrel cortex.

Astrocytes intimately contact synapses with their fine astrocyte processes regulating synaptic functions. Fine astrocytic processes can receive information from synapses and also transmit their information to synapses. Thus, astrocytes may regulate synapses through bidirectional information processing. Recently, we found astrocytes regulate synapse remodelling in primary somatosensory cortex in Ca²⁺ dependent manner, contributing to the induction of aberrant sensation in neuropathic pain model mice. It has been thought that Ca²⁺ dynamics at fine astrocytic processes regulate synaptic functions/neuronal excitability. However, its dynamic interaction between neurons and astrocytes is largely unknown. To this end, we took viral introduction of red fluorescent genetically encoded Ca²⁺ indicators (GECI) and green fluorescent GECI into neurons and astrocytes, respectively. We have succeeded to monitor both spontaneous and evoked Ca²⁺ signals simultaneously from neurons and astrocytes in the hippocampus in situ. Enhancement of astrocytic Ca²⁺ signals by selective manipulation of astrocytic receptors altered dendritic Ca²⁺ excitation. The approach described here may provide useful information on bidirectional information processing at synapses.

Migraine is an episodic neurological condition which prominent symptom is a recurrent, severe, unilateral pulsating headache, which prevalence is 8.4% in Japan. Clinical characteristics of migraine, in addition to severe headache, include photophobia, phonophobia or autonomic nervous symptoms such as nausea and vomiting. Although migraine is not life threatening, it has a significant impact on migraine patient's quality of life. Cortical spreading depression (CSD) is defined as a transient neuronal and glial depolarization and disruption of membrane ionic gradients that propagates slowly across the cerebral cortex. There is a growing evidence from animal experiments, suggesting that CSD is the electrophysiological event underlying migraine aura and activates the trigeminovascular system and upper central nervous system, resulting in the headache. In addition, the functional MRI performed on patients during migraine with aura attacks revealed a causal role of CSD. These basic and clinical findings suggest that CSD plays a pivotal role in the pathophysiology of migraine.

Here, I review the potential correlation between the migraine pathophysiology and CSD for the knowledge necessary to develop the new therapy for migraine.

The inferior alveolar nerve (IAN) is a mandibular branch of the trigeminal nerve and innervates to orofacial region. IAN transection (IANX) induces allodynia in the maxillary nerve-projecting region. However, it is not fully understood whether and how the nociceptive information processing in the cerebral cortex is changed by nerve injury. To address this question, we assessed neural activity during nociception in IANX rats using optical imaging and two-photon calcium imaging techniques. Since the dental pulps principally consist of Aδ and C fibers that transmit nociceptive information, we used dental pulp stimulation to investigate the cortical response to nociception. Optical imaging at macroscopic level revealed that cortical excitation induced by the upper molar pulp stimulation was enhanced in 1 week after IANX. Two-photon calcium imaging revealed that hyperexitability of both excitatory and inhibitory neurons. Whole-cell patch-clamp recording in slice preparation revealed inhibitory postsynaptic inputs to pyramidal neurons were decreased in IANX rats. These results suggest that IANX induces enhancement of neural activity during nociceptive information processing, and plastic changes of the local circuits in the cerebral cortex might contribute to the enhancement.

Relationship between pharmacokinetics (PK) of candidate substances and pharmacodynamic changes caused by candidate substances, i.e., PK/PD is requested to maintain consistency among different R&D stages from in vitro to in vivo or from nonclinical to clinical. It is essential to anticipate clinical efficacy and to verify its outcome in clinical trials. However, there are few data available for modeling and simulation when selecting molecular targets in the early stages of drug discovery and screening candidate substances from various compounds, and it is extremely challenging to quantitatively consider the extrapolation to the clinical. Furthermore, efficacy assessments often take into account species differences in targets and limited suitability with the clinical manifestations of animal models, which often require individualized assessment depending on the target molecule or disease state.

In this topic, we will discuss the improvement of the success rate of drug discovery and development from the viewpoint of efficacy through the introduction of some cases for our type 2 diabetes drugs from the discovery to the post-launch stage.

Predictive biomarkers being used as companion diagnostic agents will help to avoid critical failures of drug treatment due to a lack of efficacy. Therefore, the predictive biomarkers can reduce the number of patients needed to demonstrate the efficacy in clinical trials by enriching the populations with biomarker-positive patients. In addition, this allows avoiding adverse effects by unnecessary treatment. The presence or absence of the target molecule-related gene expression can be such a biomarker. As an example, aberrant activation of anaplastic lymphoma kinase (ALK) after being fused with echinoderm microtubule-associated protein-like 4 (EML4) shows oncogenic potential and the underlying genetic rearrangement is the oncogene in a small subset of non-small cell lung cancer (NSCLC). ALK inhibitors and companion diagnostic agents to detect the fusion gene have been developed simultaneously and approved for ALK fusion gene-positive NSCLC. This represented a landmark in oncology drug development and a significant step toward precision medicine. In this presentation, the current situation of predictive biomarker-driven drug development will be discussed.

Our target diseases with high unmet needs are getting more and more complicated and heterogeneous. Accordingly, to predict clinical efficacy of drugs in the preclinical stages, we should avoid relying too much on phenotypically resemble animal models, but seek more mechanistic, human-predictive models. Thanks to accumulation of publicly-available biomedical information, understanding of pathophysiological mechanisms for both patients and disease animal models can be improved. Systems Biology / Pharmacology are practical Translational Science tools to handle such large-scale biomedical data. For instance, Systems Biology analysis with patient transcriptome data can decipher disease mechanisms, and support creating competitive drug concepts. Quantitative systems pharmacology (QSP) modeling & simulation can provide us with rational clinical efficacy prediction based on the currently-best knowledge. Through the model-based approach, we can continuously and rationally decrease the uncertainties, and identify key research questions to be addressed by both preclinical and clinical researches. I will show some examples for applications of translational science tools to address some research questions in the drug development researches.

The largely unknown etiology and pathophysiology of psychiatric disorders has significantly hampered effective drug discovery and development; to wit, nearly all first-generation antipsychotics and mood stabilizers have been discovered under serendipitous circumstances. However, recent advances in genetics and pathological technologies have provided new avenues into investigating and connecting genetic components in the etiology of psychiatric disorders with pathophysiological changes in the patient's brain. With this newfound synergy, we are now able to commence truly innovative drug discovery based on strong scientific rationale.

Our approach is to pinpoint common biological pathways disturbed in both psychiatric patients and genetically-engineered animal models (targeting both individual genes and CNVs discovered in patients) and use these commonalities to generate testable working hypotheses. We have obtained patient datasets via participation in the Lieber Institute for Brain Development's (LIBD) Pharma RNA-Seq Consortium, BrainSEQ^TM, a precompetitive research collaboration aimed at completing RNA sequencing of up to 800 brain samples from DLPFC and hippocampus. This approach facilitates streamlined, comprehensive drug target selection where candidate compounds are evaluated using behavioral testing and pathophysiological measures, with the eventual goal of reversing the disturbed pathways in gene-manipulated animal models and, ultimately, human patients. Our objective is to achieve precision medicine in psychiatry. In doing this, we aim to tackle emerging problems, including disease- and patient-specific biomarkers used for diagnosis and stratification.

It is well known that there is a high correlation between the relapse rate of mental disorders such as schizophrenia and medication adherence. Therefore, it is possible to prevent acute exacerbation of symptoms and to prevent re-hospitalization by continuing taking medication. On the other hand, it is also well known that it is very difficult for patients to keep taking medication every day for various reasons - patients don't believe drugs work, troublesome side effects or lack of insights on their diseases.

It is very important to understand their medication adherence and their reasons non-adherence, we should provide enough support for them to continue their medication, raise awareness of their disease and educate them on the necessity and importance of taking their medication. However, it is not easy for healthcare professionals, care givers and families to know how good or bad the patient's adherence is because it depends on the patients self-report or checking of the residual medicine which is not always accurate. As a result, when the patient relapses, it is challenging for the healthcare professional to determine whether it comes from poor adherence or whether the medication is effective.

Otsuka and Proteus Digital Health (hereinafter referred to as "Proteus") developed the first digital medicine "Abilify MyCite" that can measure medication adherence. It embeds a very small sensor developed by Proteus into an antipsychotic Abilify tablet marketed by Otsuka. When a patient swallows tablets, a chip emits a signal in the stomach and it is captured by a small patch which sticks to the patient's abdomen and this is recorded. The patch transmits adherence information to the patient's smartphone as well as capturing activity and rest from sensors on the patch. The transmitted information is captured on both the patient's phone as well as shared with healthcare professionals, caregivers and families with the patients' consent. Based on this information, healthcare professionals can measure the condition of the patient and assess the risk if there is a sign of recurrence such as poor adherence or abnormal sleep pattern, then take the appropriate intervention.

In November 2017, Otsuka received approval from the US Food and Drug Administration (FDA) for this world's first digital medicine. We will continue to contribute to patients, families and healthcare professionals through not only medicines as a pharmaceutical company but also to provide important medical information and solutions for better patient health.

Digital Health has emerged as innovative technologies to transform healthcare industry. Many Digital Health solutions are being developed to solve the healthcare problems using artificial intelligence (AI), Internet of things (IoT), wearable devices etc. Pharmaceutical companies are also accelerating the development of Digital Health solutions to optimize their value chain from drug discovery to post marketing, and to improve patients' quality of life. Successful implementation of these solutions requires new competencies such as agile development and digital technological expertise, and open innovation is an essential part of fostering the competencies. At the presentation, we will share our digital activities and ‘G4A' which is an open innovation program for Digital Health.

Recently, "digitization", a kind of buzz word, has been widely used in many industries. In addition, it has been estimated that "disruptive changes" would be induced by such a wave of digitization. The current trend gives me a lot of questions; The media has reported to use of digital technology advances in the health care industry, but what about? The media has reported that information technology (IT) companies have entered the healthcare industry, or what? How Google, Apple, Facebook, and Amazon (GAFA) have entered the healthcare industry? The changes in external environments have made sense for us in the health care sector？

In this symposium, I would like to disclose my understanding of "Implications of digital evolution for the pharmaceutical industry and future prospects", referring to the digital changes in other industries.

In 2017 Mitsubishi Tanabe Pharma established the Future Design Department. The Future Design Department is advancing a transformation to a data-driven company with a view to the use of Big Data, AI, IoT, Digital Medicine and other digital technologies with the objective of fostering "disruptive innovation" in the pharmaceutical business. We are working to create new methods that transcend the pharmaceutical industry's conventional pharmaceutical categories. Mitsubishi Tanabe Pharma is promoting open innovation with some universities and companies of different industries about issues with impact within the drug discovery value chain.

For example, Mitsubishi Tanabe Pharma and Hitachi have initiated collaborative creation for improving the efficiency of clinical trials using Hitachi's advanced digital technology such as AI.

In another example, in 2018, we will start an accelerator program. With the key words of "creating the future of healthcare" we will build a business model that transcends the conventional pharmaceutical business framework and strive to contribute to the happiness and health of patients and those around them.

In order to expand our business "around the pill" and "beyond the pill", we are working on the challenge of creating new value in the fields of medicine and healthcare through cross-industry collaboration with cutting edge technologies and different industries. In this presentation, we would like to introduce the above contents along the case of Future Design Department in Mitsubishi Tanabe Pharma.

In the drug development in pharmaceuticals, development of drugs may be discontinued due to the toxicity and clinical side effect, therefore, safety assessment is one of the important factors in drug development. Recently, pre-clinical studies are screening for compounds by the in vitro screening system using human iPS cell-derived cells which are expected to predictively of human clinical. Human iPS Cell Applied Safety Assessment Consortium (CSAHi). CSAHi focuses on hepato-, cardio-, and neuro-toxicities as important toxicity organs which are attributed to the causes of discontinuation of drug development. In neurotoxicity, seizure is an important finding because of high frequency expression in nonclinical. Multi-electrode array (MEA) systems have recently attracted attention as useful for evaluating seizure risk because they can non-invasively measure the electrophysiological activities of neural networks. We are evaluating the electrophysiological responses to several seizure compounds using MEA in cultured hiPSC-derived neurons. It is important to establish an analytical method to detecting seizure-like activities. We have focused the periodicity of synchronized burst firings as one of the effective analytical parameters for detecting seizure risk. We identify the parameter sets that separate the responses between positive and negative compounds using principal component analysis of 10 parameters. The principal component analysis using parameter set focused on periodicity is useful method to detect the seizure risk.

Functional assays using human induced pluripotent stem cell (hiPSC)-derived sensory neurons are expected to predict the drug safety and side effects in human peripheral nervous system. However, evaluation assays in hiPSC-derived sensory neurons has not been established, and electrophysiological response to pain-related molecules are not known. In this study, we aimed to evaluate the physiological responses to pain-related molecules including anti-cancer drugs in cultured hiPSC-derived sensory neurons using high-throughput multi-electrode array (MEA) system. In capsaicin, menthol, and AITC administration, evoked responses depending on TPRV1, TRPM8, and TRPA1 channel were detected, respectively. We found that hiPSC-derived sensory neurons are a heterogeneous cell population that have different responses to temperature and pain molecules, like living body. Cold hypersensitivity responses were also detected in concentration dependent manner of anti-cancer drug oxaliplatin. These results indicated that this MEA evaluation method using human iPSC-derived sensory neurons is effective as a pain-related toxicity in human peripheral nervous system.

As drug-induced seizures have severe impact on drug development, evaluating seizure induction potential of candidate drugs at the early stages of drug discovery is important. A novel assay system using zebrafish has attracted interest as a high throughput toxicological in vivo assay system, and we have established an experimental method for drug-induced seizure liability on the basis of locomotor activity in zebrafish. Our established experimental method included monitoring locomotor activity at high-speed movement (> 20 mm/sec), extending exposure time or conducting flashlight stimulation (10 Hz) which is a known seizure induction stimulus. The validation study using our methodology was used to assess 52 commercially available drugs, and the prediction rate was approximately 70%. The experimental protocol using zebrafish is considered useful for seizure potential screening during early stages of drug discovery.

At present, owing to the species differences, we have little options for preclinical studies to predict adverse effects (AEs) in the human CNS.  However, as introduced in this symposium, a series of new in vitro experiments have been developed to improve the predictability of CNS AEs. This trend is not limited to Japan inside, but also seen worldwide. Our research project ‘iPSC Non-Clinical Experiments for Nervous System（iNCENS）', aiming at the establishment of the evaluation systems to predict seizure risks of new drugs, is now a committee member of an international think tank, International Life Sciences Institute (ILSI),　Health and Environmental Sciences Institute (HESI), NeuTox micro-electrode array (MEA) sub team. Most megapharmas and regulatory agencies are participating in the pilot study to clarify the efficiency of the electrophysiological recordings of MEA. We are providing some promising data recorded from hiPSC-neurons. In this symposium, I will introduce our on-going activities together with the global trends concerning the development of preclinical in vitro evaluation systems to predict CNS AEs.

We are now facing the third wave of artificial intelligence (AI). AI refers to simulated intelligence in machines that have been programmed to mimic human brain behavior such as learning, reasoning and problem solving. Many computer algorithms had contributed to previous waves, but this latest wave of AI was created by deep learning algorithm which is inspired by how human brain processes information through interaction of neurons. The first simplified mathematical model of neuron, the formal neuron, was proposed by McCulloch and Pitts in 1943. In this model, a neuron receives a set of binary inputs from other neurons, multiplies each input with the weight (i.e. the strength of synapse), and activates if the sum of these weighted inputs is greater than a threshold. More than 70 years have passed since this simple model was proposed and we are now seeing many examples where AI beats humans. In this symposium, we have speakers from two pairs of pharmaceutical and IT companies that have recently announced big partnership for AI technology, and they will present their recent development and application of AI, and their future prospects. I hope we can learn many cases of applications of AI to biology, pharmacology, drug discovery, and pharmaceutical business.

After a lot of initial artificial Intelligence (AI) hype, many useful AI applications have been identified, and now AI is transforming many industrial areas. Pharma companies also realize the application of AI can positively disrupt the traditional business processes in various ways, such as smarter therapy target hunting, quicker drug candidate molecule generation, efficient non-clinical/ clinical development, and more precise drug market penetration. Currently many pharma companies are actively trying to apply AI technologies to their business. However, before AI implementation into their business processes, pharma companies need to make some decisions on how to approach to AI technologies, such as collaborations with AI companies or internal development of AI capabilities. In this session, points needed to be considered upon collaborations with AI companies from pharma perspectives will be discussed. Also, potential applications of AI to pharmaceutical business processes will be discussed including some real use cases.

Deep learning has dramatically improved in recent years and has achieved breakthrough results in image recognition, speech recognition, machine control, anomaly detection, etc. In the field of biotechnology and life science, deep learning is widely used. Deep learning is a promising technology in the field of biotechnology and life sciences for its ability to extract useful features in its process of learning, achieve multi-modal and multi-task learning naturally, and successfully handle data on genomic sequence, low molecular weight compounds, and protein, which are of varying size. In this presentation, we will introduce the latest applications of deep learning and explain how deep learning can be used in the process of drug discovery.

To accelerate the drug-discovery and -development stages, one of solutions is to shorten the term of each task contained in the stages. AI has potential to solve this issue and has been applied to several tasks. Furthermore, some AI systems are in development. I will introduce three examples in the drug-discovery stage and one in the development stage, 1: Prediction of human cardiotoxicity, 2: Phenotype screening derived from cell morphological image features, 3: ADMET prediction for the designed compounds, 4: Efficiency improvement of clinical trials collaborating with Hitachi. Hitachi will talk about the fourth one in detail. The aim of these examples is to reduce time-consuming works, to compensate experimental data with highly accurate predictions, or to possess better observing-eyes instead of human eyes.

Clinical trials are time consuming, expensive, and often burdensome on patients. Poor protocol design can reduce a success rate of clinical trials. In order to improve protocol design, pharmaceutical companies often spend a lot of time searching for information thoroughly regarding protocol design. In the past years, we have dedicated tremendous efforts to explore and implement novel Artificial Intelligence (AI) applications to advance clinical development. During these projects with Mitsubishi Tanabe Pharma,　we have gained experiences with developing AI applications. Especially, we're focusing on AI applications for protocol design. In these applications, we're leveraging NLP & Deep Learning to help pharmaceutical companies optimize their protocol designs. Our system learns from all kinds of sources, like ClinicalTrials.gov, past clinical trials, journal articles and provide recommendations for clinical trial optimization. This presentation will discuss our early findings on the AI in protocol design.

In vivo studies are mandatory to confirm whether a principle generated from in vitro experiments can be reproducible under physiological and pathological conditions, which has become difficult to perform under the principles of 3Rs for animal welfare. Pigs have been expected as an alternative laboratory animal; however, neither regular pigs nor miniature pigs may be suitable for confirming in vivo proof, since they need larger dosage of expensive test articles under developmental phase. Lately, extraordinarily small-sized miniature pigs; namely, microminipigs, weighing approximately 7 kg at 6 months of age when they are young mature, were developed by Fuji Micra Inc. (Shizuoka), which have been provided to several research organizations in Japan as a non-rodent experimental animal optimized for life-science research. We have characterized microminipigs by assessing cardiovascular effects of various drugs and medical devices, and found that most of the cardiovascular responses observed in microminipigs were similar to those in human subjects as well as other non-rodent animals. In this workshop, we will present a brief summary of the development of microminipigs along with recent publications of it in the field of cardiovascular physiology/pharmacology.

Axcelead Drug Discovery Partners, Inc. (Axcelead) was separated from Takeda Pharmaceutical Company Limited and established in July, 2017 to support drug development of academia, pharmaceutical companies, and startups. We provide various services including drug screening, compound synthesis, pharmacological studies, pharmacokinetics examinations, and toxicological studies. We put great efforts into developing large animal models including pigs as pharmacological studies with large animals are desired in the area of regenerative medicine such as cell therapy. An original immunosuppressed pig model has been developed by two methods at Axcelead. The first is to remove thymus and spleen which regulate immune system in pigs. The second is to utilize immunosuppressive agents, which we measure those concentrations in plasma and control dosages to reach the targeted plasma concentration. The targeted concentration in vivo is decided based on in vitro assay using peripheral blood mononuclear cells isolated from pigs and calculated protein binding rate. To reduce stress for animals, the drug is administered via gastrostomy and blood samples are collected from central venous catheter. Furthermore, heart failure and type 1 diabetes pig models have been developed at Axcelead. We will introduce our immunosuppressed models and the two disease models at this workshop.

Pigs are occasionally used in toxicity and pharmacology studies in European countries but rarely in Japan. On the other hand, in pharmacology studies, pigs have been used in skin irritation and absorption tests as one of the alternate animals. However, in recent years, the production of min-pigs maintained under the quality management has been established in Japan and these animals are used actively in each field. The accumulation of the basic data for mini-pigs increases steadily and pigs are used while confirming whether the projects can be correctly conducted only in pigs each time.

Pigs (including training pis) weighing about 10 mg - about 100 kg can be obtained as experimental animals. The body weights of these animals are within the range of human body weights and the conditions capable of coping properly with various variations such as thickness of the blood vessels, size of the visceral organs, load on the joints can be satisfied in pigs. Therefore, pigs are useful animals in preclinical studies. Pigs are remodeling animals showing the bone metabolic turnover similar to that in humans. Since the implantation of bones into the long bones is actively conducted, soft bones such as meniscus and intervertebral disc are actively used in these animals and the sizes of these bones are appropriate for performing the surgical treatment, the use of pigs is more recommendable than that of rabbits, dogs and monkeys and pigs have advantages in evaluating the test substance clearly over the other animals.

In the field of dentistry the use of pigs are also gradually started but since there are still difficulties in obtaining appropriate thickness of the skull and extracting the teeth, the preparation of animal models and the evaluation of the test substance are now in the process of trial and error.

As for the visceral organs, pigs are actively used in the field of transplantation and especially in the circulatory system, kidneys and vascular system. In the extrapolation to humans, many organs of pigs are similar to those of humans in eating habits and anatomical features and are similar to those of humans in the size of organs. Therefore pigs are highly useful in preclinical studies.

On this occasion, we would like to investigate the usability and usefulness of pigs further through the introduction of part of cartilage regeneration test, bone implantation test and validation test for prevention of visceral adhesions which are performed in our Tsukuba Research Center.

Various agents (e.g., prescription drugs, OTC-medications, agricultural chemicals) have a potential to cause neurological disorders including cognitive impairment, movement disorders, convulsions and peripheral neuropathy. Among them, extrapyramidal motor disorders (e.g., parkinsonian symptoms and tremor) are often caused by drugs acting on central nervous system, deteriorating quality of life in many patients. Recent progress in pharmacology/drug safety research revealed detailed mechanisms for the pathogenesis and treatment of drug-induced extrapyramidal side effects (EPS). Although the dopaminergic system dysfunction has long been considered to be the primary cause of parkinsonian symptoms, it is now known that multiple serotonergic (5-HT_1A, 5-HT_2A, 5-HT₃ and 5-HT₆ receptors), adrenergic (α_2A and α_2C receptors) or cholinergic (muscarinic) receptors play crucial roles in modulating the EPS induction. In this presentation, we introduce the mechanisms underlying the pathogenesis and treatment of EPS, with a focus on drug-induced parkinsonian symptoms and tremor, and discuss the prediction and management of EPS at clinical settings (e.g., treatment of schizophrenia, mood disorders, Alzheimer's disease) and chemical intoxication.

Glutamate transporters (GLTs), which regulate glutamatergic transmission are mainly localized on glial cells. Glial disruption results in decreased uptake of glutamate and an elevation in extracellular glutamate levels. Elevated extracellular glutamate may cause cytotoxic damage to neurons and glia. Abnormalities of GLTs cause some neurodevelopmental disorders, such as ADHD and schizophrenia. A recent study found an increased incidence of a rare genetic variant in the human gene encoding glial glutamate and aspartate transporter (GLAST; one of GLT family) in schizophrenia. The loss of GLAST is predicted to cause glutamate excess that provoke glutamate release.  In fact, the deficient of GLAST induces some behavioral abnormalities and morphological changes in mice. During the adolescent period, further, mice that had received the combination of neonatal viral infection and adolescent drug abuse exhibits some psychobehavioral abnormalities and increased expression of GLAST. In this symposium, we will present the recent advances of our researches, regarding functional roles of GLAST in neurodevelopment under the physiological and pathological conditions using the mice with varying expression of GLAST.

The central nervous system is a target organ for environmental toxicants and medicinal drugs. Since the central nervous system is complex and it is difficult to obtain human neuronal cells, establishment of complete neuronal functionality of in vitro cellular models is a great challenge for chemical/drug safety issues. Here we present the framework of iPS cell-based assays regarding neurodevelopmental studies and drug safety assessment toward global standardization. Neurotoxicity can be divided into structural and functional aspects and we have collaborated with OECD developmental neurotoxicity group as well as international consortium HESI (NeuTox subteam) to make new test methods. I would like to show our research activities and discuss future perspectives toward in vitro neurotoxicity method using iPS cell technology.

Human induced pluripotent stem cell-derived neurons are promising for use in toxicity evaluations in nonclinical studies. One of the major adverse events affecting the central nervous system observed during clinical trials is convulsions. Micro-electrode array (MEA) systems have recently attracted attention for use in evaluating the convulsion potential of a drug because they non-invasively measure the electrophysiological activity of neural networks at multiple sites in a high-throughput manner. Over twelve compounds including convulsants were tested at 5 concentrations. Using principal component analysis, clustering analysis, and artificial intelligence (AI), we have succeeded in separating the responses between convulsive drugs and negative control, and in classifying the mechanism of actions of drugs from spontaneous firing data. The MEA assay using hiPSC-derived neurons and our analysis method will be effective for predicting the seizure liability and mechanism of action of new drugs.

Imaging techniques are indispensable methods for recent biological science. Japanese Society of Histochemistry and Cytochemistry is a society focusing on the advance of morphological tchniques and its appplication to life science. In this symposium, three morphology specialists introduce three important imaging techniques that is useful for pharmacological research.

1) Basic technique for immunocytochemistry and the pitfalls

2) Key techniques for multilabeling immunostaing using cultured cells

3) Imaging of physiological functions using fluorescent dye in living tissues

Immunohistochemistry is extremely useful for exploring and visualizing specific substances present in cells, tissues, organs and individuals by utilizing diverse and specific molecular recognition mechanisms of the body called antigen-antibody reaction. 

There are several important points in this step, but the first of which is to obtain and use a "good primary antibody". It is possible to detect substances (antigens) to be searched clearly, easily, and with high reproducibility by using a good primary antibody. 

Secondly, we need to immobilize substances (antigens) present in cells and tissues by fixation. However, the task of fixation bridges between molecules or molecules within a molecule in proteins, nucleic acids, lipid molecules, and forms a state called polymer. It is also an important point of immunohistochemistry to make this contradictory phenomenon successfully compatible.

The third point is to visualize the antigen-antibody reaction. There are several visualization methods. It should be visible not only to look, but also specific, clearly judged.

It is important to make it possible to discriminate reliable immunohistochemical reaction while conscious of these points to prevent incorrect detection results and detection judgment.

　In this presentation, I will explain the principle and basis of immunohistochemistry centering on the above three points.

Immunofluorescence microscopy is a powerful method for analysis of the subcellular localization of the protein of interest. The use of fluorescence is very effective for multiple labeling and for higher magnification observation with a laser confocal microscope as well as a conventional fluorescence microscope. The technique is easy and generally used for researchers; however, it is also true that they sometimes show the results inappropriately and incorrectly. I would like to introduce the basic methods with some helpful suggestions of the immunofluorescent staining in cultured cells, especially focusing on the followings:

1) Cell culture for immunocytochemistry

2) Fixation

3) Treatment before immunostaining

4) Key points for multiple labeling: choices of the secondary antibodies and fluorescence dyes

5) Observation and storage of the samples

I hope this talk could help researchers in the Pharmacological Society to get better and beautiful results in immunofluorescence microscopy.

Histochemistry, which enables us to visualize the precise distribution of target molecules within living organisms, is an essential strategy in life science research. Of various histochemical modalities, confocal microscopy provides not only the existence of target molecules, but also their spatiotemporal dynamics. Because of the high spatial resolution of the confocal images, fluorescence intensities for specific molecules reflect their concentrations: the higher the intensities, the more abundant the molecules present. In addition, rapid scanning confocal imaging provides precise concentration changes in functional molecules, reflecting cellular functions. Here I would like to exemplify fluorescent imaging of intracellular Ca²⁺ dynamics, cellular membrane potentials, and mitochondrial states of the cardiomyocytes in the heart. Photodynamic modulation of cellular functions will also be demonstrated.

Drugs targeting the immune system have been widely exploited in the treatment ofinflammatory, allergic and autoimmune disorders during the second half of the20th century. The recent advances in immunopharmacological research madeavailable new classes of clinically relevant drugs, comprising protein kinaseinhibitors and biologics, such as monoclonal antibodies that selectivelymodulate the immune responses. ImmuPhar is the Immunopharmacology Section ofIUPHAR, providing a unique international expert-lead platform aiming to dissectand promote the growing understanding of immune system as well as to challengethe identification and validation of drug targets and lead candidates for thetreatment of many forms of debilitating immunological diseases. In thissymposium I will outline the current movement on immunopharmacology in Japan aswell as the activity of ImmuPhar, and discuss future perspectives in thistrend.

High mobility group box-1 (HMGB1) was originally identified as an intranuclear chromatin DNA binding protein. HMGB1 has unique feature in its mobilization, translocation and release. Responding to ischemic/hypoxic and traumatic injuries, HMGB1 may be released from different types of cells through cytosolic compartment. In the previous studies, we demonstrated that HMGB1 was released from neurons by ischemic as well as hemorrhagic injuries in the brain, leading to the disruption of integrity of blood-brain barrier (BBB). Intravenous injection of anti-HMGB1 mAb protected BBB and ameliorated brain damage by reducing inflammatory responses including cytokine expression. In the epileptic animal model, we found that therapeutic anti-HMGB1 mAb administered systemically accumulated on vascular endothelial cells while protecting brain inflammation and neural damage. These findings strongly suggested that vascular endothelial cells may be one of major targets of anti-HMGB1 mAb action. In this symposium, I will show our recent data on HMGB1 mobilization in vascular endothelial cells induced by different stimuli, associated with cytokine production and cellular responses. The results imply that HMGB1 released from endothelial cells played fundamental roles in triggering inflammatory responses.

Intestines act as not only a digestive and absorption tissue of food but also asthe largest immunological organ by containing various numbers and kinds ofimmunocompetent cells. Accumulating evidence indicates that that the functionsof intestinal immunity are affected by gut environments such as dietarymaterials and commensal bacteria, which is tightly related to the development ofimmune diseases (e.g., allergy and inflammation) at various tissues. Recenttechnological advancements in the metabolome analysis, metagenomic analysis ofcommensal bacteria, and bio-informatics technology allow us to understandtripartite interaction between diets, commensal bacteria, and host immunity,leading to the establishment of new concept of drug design. In this talk, I willintroduce our studies related to lipids derived from diets and commensalbacteria-mediated immune regulation and the application to the development ofanti-allergic and anti-inflammatory medicines.