Secondary pharmacology has become an established component of modern drug discovery and nonclinical safety assessment, supporting the early identification of off-target activities that may contribute to unintended pharmacological clinical effects. Although current regulatory guidance does not prescribe specific panel compositions, secondary pharmacology data are commonly expected to support first-in-human submissions and non-clinical safety assessments. However, practices remain heterogeneous with respect to panel design, assay format, testing strategy, and data interpretation.This perspective provides a contract research organization (CRO)-based overview of current secondary pharmacology practices, based on anonymized operational data from more than 550 sponsor organizations and contextual insights from voice-of-customer activities and routine scientific interactions.Standard catalog panels accounted for more than 90% of panel use and were favored across sponsor categories, whereas customized panels were used less frequently and mainly to address specific scientific or safety questions. Small panels (≤50 targets) were the most commonly used format. Binding assays remained the predominant screening modality, particularly for early compound triage, while functional assays were mainly applied for follow-up characterization or tiered testing strategies. Panel selection was influenced by regulatory considerations, historical experience, target coverage, operational constraints, and project-specific needs.Our analysis highlights ongoing challenges related to dataset sufficiency, regulatory expectations, and the biological interpretation of secondary pharmacology data. Greater harmonization of testing strategies, interpretation frameworks, and regulatory guidance will be important to maximize the value of secondary pharmacology in drug discovery while supporting implementation of the 3Rs principles.
Integrating safety pharmacology (SP) endpoints into repeated-dose toxicology studies enables the simultaneous assessment of organ toxicity and determination of functional safety risks within a single study, a practice endorsed by regulatory guidelines. Initiated by the Chinese Safety Pharmacology Society, this questionnaire-based survey assessed the implementation of such integrated studies across 19 Chinese institutions compliant with good laboratory practice and qualified to support submissions to the National Medical Products Administration (NMPA, China), FDA, and EMA. The survey compared integration practices for new chemical entities (NCEs) versus biopharmaceuticals, including animal models, monitoring technologies, statistical design, regulatory acceptance, and technical challenges. Furthermore, the survey solicited recommendations regarding revisions to the current ICH S7A guideline. For biopharmaceuticals, integration was common, with 16 of 19 institutions applying it in more than 75% of projects, primarily using non-human primates. Endpoint coverage in these studies included the cardiovascular system in all 19 institutions, the respiratory system in 16 of 19, and the central nervous system in 11 of 19, alongside an average 18% reduction in animal use. Respondents reported that data from these studies were fully accepted by the NMPA, FDA, and EMA in regulatory submissions. Conversely, integration was rarely used for NCEs, with 18 of 19 institutions reporting limited implementation. The primary barriers cited were interference with toxicokinetic sampling and lower statistical power compared to standalone crossover designs. As the first systematic survey of SP-integration practices across Chinese GLP institutions, these findings provide valuable real-world evidence to optimize non-clinical safety assessment strategies and support future revisions of the ICH S7A guideline.
The blood-brain barrier (BBB), which tightly regulates the exchange of substances between the blood vessel and the brain, maintains central nervous system (CNS). Therefore, the development of BBB in vitro model is expected to significantly contribute to the CNS-targeted drug development. Organ-on-a-chip systems using microfluidic devices under flow conditions have recently attracted attention for enabling cell culture environment that closely resemble in vivo. In particular, organ-on-a-chip systems utilizing closed, dual-channel microfluidic devices with a porous membrane are valuable tools for analyzing inter-organ interactions, such as those at the BBB. In this study, we aimed to investigate the influence of shear stress, induced by perfusion, on the gene expression in human induced pluripotent stem cell-derived brain microvascular endothelial cells (iBMECs) cultured in such a device for the development of a BBB-on-a-chip.iBMECs were seeded into the upper channel of a closed, two-channel microfluidic device and cultured under static conditions for 12 h. Following this, the culture medium was perfused for 48 h, and the cells were then for gene expression analysis. As a control, iBMECs cultured under static conditions using conventional cell culture inserts were similarly analyzed.We first optimized the perfusion parameters to prevent cell detachment. As a result, a trend toward increased expression of genes, including those encoding drug efflux transporters, was observed in the perfused group compared to the static culture. Notably, gene expression of Breast Cancer Resistance Protein markedly increased to levels comparable to those observed in vivo. In addition, gene expression of a glycosyltransferase C1GALT1, which has been reported to contribute to the barrier function of BBB, also showed a tendency to increase in the perfused group.In this study, we successfully established perfusion culture condition for iBMECs within a microfluidic device and demonstrated that flow-induced mechanical stimuli can significantly enhance the expression of key BBB-related genes. These findings suggest that the perfusion culture of iBMECs is a valuable approach for developing a functional BBB-on-a-chip model.
Cardiovascular diseases, with high mortality and incidence rates, drive development of drugs and treatments. Off-target effects resulting from insufficient models are responsible for 45% of drug withdrawals. Engineered heart tissues (EHT) from hiPSC cardiomyocytes are gaining traction in New Approach Methodologies (NAM) as they better recapitulate human physiology and functions, however, lack of standardization and throughput are impeding their industrial adoption. Objectives: Using the Ethica M platform, with the MChip to:1.Study tissue formation and contractile maturation of EHTs2.Validate function with force frequency relationship (FFR) and compound addition3.Validate the ability of an automated high-throughput system to generate robust, reproducible EHTs for drug screening applicationsCell/gel matrix was dispensed and tissues formed in 2 days. Spontaneous beating was tracked and videos recorded daily during the study. Continuous electrical pacing started on day 9 up to endpoint assays of FFR and compound exposure. After initial tissue formation (85%), irregular spontaneous beating began at d2, stabilizing at 0.6 Hz (day 5). Maturation was assessed by recording contraction amplitude, beat rate and pulse width. Within 14 days, spontaneous contraction amplitudes of previously stimulated tissues increased by 64%, while pacing the same tissues increased amplitudes by 165%. Contraction increases were accompanied by a beat rate reduction of 38% (0.91–0.56 Hz) and a 15% pulse width decrease, indicating a shift from fetal to adult phenotype as maturation processes in sarcomere structures, electrophysiology and calcium handling result in stronger, shorter contractions and a lower beat rate. Tissues were exposed to inotropic compounds (d31). Isoprenaline showed increased contraction strength (positive inotrope), EC50 = 9 ± 2 nM, while Nifedipine showed negative inotropy with an amplitude reduction, IC50 = 41 ± 3 nM. In both cases, tissues showed dose-dependent responses mimicking the response of mature cardiomyocytes. Functional maturity was confirmed by FFR. Electrical stimulation ranged from 0.8 to 4 Hz showing positive FFR correlation for frequencies up to 2.2 Hz, recapitulating the response of human myocardium. We demonstrate the ability of Ethica M to produce mature EHTs suitable for screening workflows that may recapitulate expected drug compound and FFR responses of the human myocardium.
