Science Pool

While simple combinations of dosage forms expedite access to First-in-human (FIH) studies, the demand for robust formulations and processes challenges quick development, particularly for tablet production. Early clinical phases face hurdles like dose uncertainty and limited active pharmaceutical ingredient availability. The study addresses these challenges by proposing a compressibility assessment method using small-scale experiments. The aim is to establish a pre-formulation screening approach based on compressibility data for active ingredients, utilizing round punches of varying diameters. By employing the smallest tooling, the study achieves a remarkable 75% reduction in the amount of active ingredient required for compressibility analysis.

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• The genotoxic potential is one of the major risks associated to potential pitfalls in drug development.
• True positive results in the genotoxicity screening observed during the Lead Optimization phase not always represent a showstopper, but need to be carefully characterized.
• In this scenario, the toxicologist could provide a strategy supporting the next IND enabling phase by adopting the most effective risk management to allow the highest success rate possible.
• The process proposed can be adapted to any scenario, and provides data supporting the risk assessment ensuring the lowest attrition and the ethical use of animals.

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Read our latest poster on Differentiated transcriptomic signatures detectable in primary human hepatocytes transduced by AAV-based vectors: A potential enabler for in vitro safety profiling.

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Hepatic transporter inhibition, mitochondrial dysfunction, and reactive metabolite formation are some of the most common mechanisms associated with intrinsic DILI. The cytochrome P450 (CYP450) superfamily of enzymes play an important role in phase 1 metabolism within the liver. For certain chemical entities, reactive metabolites may form with increased toxicity compared to the parent. These reactive metabolites may result in hepatotoxicity through the formation of reactive oxygen species, DNA damage, mitochondrial dysfunction, and endoplasmic reticulum (ER) stress. In our research the pan-specific CYP450 inhibitor, 1-aminobenzotriazole (1-ABT), was used in combination with high-content imaging to evaluate the effects of potential reactive metabolites on cell health parameters in hepatocytes. The endpoint assessed included nuclear features, glutathione (GSH) content, mitochondrial dysfunction, and reactive oxygen species (ROS) formation, as well as cellular ATP content. A calculated fold-shift in cell health features between the plus and minus 1-ABT dosing conditions was used to determine reactive metabolite formation. A panel of known DILI reference compounds associated with the formation of reactive metabolites were assessed through this HCI bioactivation assay within metabolically competent HepaRG cells, primary human hepatocytes (PHH) and primary mouse hepatocyte (PMH).

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Peripheral neuropathy can be induced by many chemotherapeutics. Symptoms include numbness, tingling or abnormal sensations which can impact on the long-term quality of a patient’s life. Animal models used to evaluate these side effects may be difficult to interpret and are labor-intensive. In this study, we further optimized a previously developed DRG assay by comparing the neurite outgrowth responses of a group of chemotherapeutics from different classes at 24 h and 72 as an in vitro cell-based model for peripheral neuropathy. Cytotoxicity was assessed alongside neurite outgrowth. Taxanes (paclitaxel, docetaxel), microtubule interfering agents (vincristine, vinblastine, colchicine, nocodazole) and epothilones (ixabepilone) were assessed.

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The generation of anti-drug antibodies (ADA) towards a therapeutic agent can have severe implications with respect to drug safety and efficacy. Therefore, identification of this risk during the initial phase of development is imperative to improve both patient outcomes and downstream attrition rates. As preclinical species often fail to mimic the complexity of the human immune system, the immunogenic potency of therapeutic agents and their potential to elicit a human-specific proinflammatory response can be grossly underestimated during in vivo safety assessment. In our research, a high-throughput in vitro approach to immunogenicity screening was developed using two distinct models: i) peripheral blood mononuclear cells (PBMC) and ii) monocyte-derived dendritic cells (MoDC) isolated from the blood of healthy human donors. The effect of serum-free culture conditions was also evaluated. The immunogenicity of monoclonal antibodies, small molecules associated with delayed-onset hypersensitivity reactions and oligonucleotides were assessed in each model through lymphocyte proliferation and cytokine release.

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The periodic activation and inactivation of various cardiac ion channels enables the regular and synchronous contraction of the heart. Therefore, any disruption of the cardiac ion channel signaling network can result in a change in membrane potential and in the shape of cardiac action potential (AP). As a result, the cardiac excitation-contraction coupling can be modified which may lead to arrhythmias and potential patient death. Local Extracellular Action Potential (LEAP) is a technique which enables non-invasive, label-free monitoring of cardiac action potential in a high-throughput real-time format. In our research, we used the Axion LEAP MEA assay to analyse the effect of various selective and non-selective ion channel inhibitors on cardiac AP by quantification of action potential morphology, repolarization irregularities, and arrhythmic risk factors such as triangulation.

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Drug-induced cardiotoxicity may result from a functional change in cardiac electrophysiology (acute alteration of the mechanical function of the myocardium) and/or from a structural change, resulting in damage to the cardiac tissue. In our research, the effects of 42 reference compounds in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were investigated in a combined risk assessment strategy. Functional cardiotoxicity was evaluated through kinetic monitoring of calcium transients (CaT), while structural morphology changes and gross cytotoxicity were assessed using high-content imaging (HCI) and cellular ATP measurements. In addition, whole genome high-throughput RNA-sequencing (ScreenSeq) was performed in matched-sister plates. Data were analysed to determine differentially expressed genes (DEGs) and any associated perturbed pathways.

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Mitochondrial dysfunction has been implicated in numerous drug induced adverse events, such as liver failure and cardiac toxicity. Read about how Cyprotex uses a combination of in vitro approaches to detect mitochondrial toxicity and analyse the potential mechanism of action in our poster.

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