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.
Tags: Posters, Toxicology & Safety
Learn more about our PBMC proliferation assay including:
Tags: Fact Sheets, Toxicology & Safety
As the CNS is a complex organ system, neurotoxicity can be difficult to assess and detect during preclinical development in animal models.
In this poster, we focus on:
Read our poster to learn more about our research!
Tags: Posters, Toxicology & Safety
Cardiotoxicity in response to pharmaceutical drugs, chemicals and environmental toxicants can develop as a result of changes in structural integrity of cardiac tissue or functional changes in cardiac electrophysiology. The development of new approach methodologies (NAMs) is important in predicting these cardiotoxic liabilities.
In this poster, we focus on:
Read our poster to learn more about our research!
Tags: Posters, Toxicology & Safety
Targeting RNA represents a paradigm shift for drug discovery. The ability to seek out and destroy, or change, a faulty RNA template, before the toxic protein has even been made, has only recently begun to be harnessed for the benefit of patients.
As of this blog, only 16 oligonucleotide drugs have been marketed, with an exponential increase in clinical trials and development exploding in this area.
There exist different mechanisms of action for an oligonucleotide drug, all of which are transient and reversible effects and do not include alteration of the DNA, unlike Gene therapy.
Antisense Oligonucleotides harness endogenous systems already existing within a cell to achieve their purpose, with the only limitation being accessibility of the target tissue. Once bound with great specificity to its RNA target, a short synthetic oligonucleotide can cause degradation, upregulation of the translated protein, or alteration of a splicing event leading to correctly folded protein. Longer Oligonucleotides can fold into 3 dimensional shapes called Aptamers with similar target affinities and applications as antibodies, and shorter oligonucleotides can act as miR mimetics or antagonists to alter multiple targets or pathways at the same time with subtle but broader effect.
The precision accuracy of an oligonucleotide and its ability to correct a faulty RNA produced by an error in the genetic code, lends itself to the rare disease therapeutic area and toxic gain of function mutations. The field of oligonucleotide therapeutics is moving to address this as a whole and to innovate a new preclinical and regulatory path that could be adapted for these more unique diseases to make this type of therapy more accessible.
Evotec is a leader in integrated Research and Development (EVOiR&D) and has built substantial drug discovery expertise and technical capabilities that can drive new innovative, diverse modalities into the clinic. In addition, Evotec has built a deep internal knowledge base in key therapeutic areas including neuroscience, pain, immunology, respiratory, women’s health, aging, fibrosis, inflammation, oncology, metabolic and infectious diseases. Leveraging these skills and expertise, Evotec successfully delivers on superior science-driven discovery alliances with pharmaceutical and biotechnology companies.
The global interest in this new modality area has led to high demand in oligonucleotide synthesis and the chemistry surrounding it, such as covalent linkages and complex formulations. Evotec has oligonucleotide manufacturing capabilities as well as ligand and linker chemistry expertise to support discovery projects and is now expanding its capacity to support development stage oligonucleotide projects.
We are extremely proud to share that we have installed the first Cytiva AKTA oligosyntTM in Europe at our Evotec site in Verona.
This new state of the art equipment will allow for the synthesis of complex modified oligonucleotides (ASOs, siRNAs etc) on a scale from 0.5 to 50 g (up to 12 millimoles) to support the initial preclinical development studies.
This is a key milestone for Evotec, and, together with the brand-new AKTA flux 6 and AKTA Pure 150, this new oligonucleotide synthetiser will complete the fully integrated oligo suite Evotec Campus Levi-Montalcini in Verona. Currently both the Verona and Toulouse Evotec sites are equipped to support Drug Discovery programmes in the RNA therapeutics field with the synthesis of oligonucleotides on a research scale and now, Evotec, at the Verona site, has the capability to also support the preclinical and clinical development studies, i.e. analytical and bioanalytical activities.
We look forward to discussing with you – our partners from new or existing collaborations- how we can best help your oligo project succeed.
Tags: Medicinal Chemistry, Blog, Formulation & CMC, IND Enabling Studies/Preclinical Development, Toxicology & Safety
Featured in Nature, this article interviews experts, Paul Walker, Rüdiger Fritsch, and Carla Tameling, and provides the latest insights into transcriptomics and how the technology is transforming toxicology prediction.
It includes:
Tags: Articles & Whitepapers, Toxicology & Safety, Modelling and Simulation
According to a study from 2020, a total of 133 drugs were withdrawn from the market due to safety reasons between 1990 and 2010. Major causes were hepatotoxicity (27.1%), cardiac disorders (18.8%), hypersensitivity (12.8%), and nephrotoxicity (9.8%), accounting for 69.2% of all drugs withdrawn. In most cases, these withdrawals were initiated because of spontaneous reports and/or case reports. Another study looking into drug withdrawals between 1953 and 2013 revealed that 18% of drug withdrawals from the market in this period were due to liver damage.
Add to these withdrawals of marketed drugs the attrition rate of drug candidates in clinical trials: 90 percent of all drug candidates fail in clinical trials, and 30 percent of these failures are due to unmanageable toxicity issues.
These failures occur despite thorough preclinical work and intensive animal studies. It is estimated that only 50% of the compounds that cause liver toxicity in humans are detected by animal studies. Furthermore, some adverse reactions or idiosyncratic toxic effects are typically not detected until the drug in question has gained large exposure in a broad patient population.
Interestingly, a study evaluating the attrition of drug candidates from AstraZeneca, Eli Lilly and Company, GlaxoSmithKline and Pfizer came to the conclusion that there is a strong link between physicochemical properties of compounds and clinical failure due to safety issues. The results also suggest that further control of physicochemical properties is unlikely to have a significant effect on attrition rates and that additional work is required to address safety-related failures.
These failures are not only costly (according to the FDA, drug development takes over 10–15 years with an average cost of over $1–2 billion for each new drug to be approved), but are also putting the health and the life of patients in danger.
Consequently, Cyprotex and its parent company Evotec are very focused on assessing toxicology issues from the very beginning of its drug R&D process and have invested a significant amount of time and resources to expand its technologies for the toxicological evaluation of drug candidates.
“The idea is to make better informed decisions earlier in your discovery campaign when you can select potentially safer compounds, rather than finding a safety liability later on,” says Paul Walker PhD, Vice President, Head of Toxicology at Cyprotex, in Cheshire, UK.
This improved discovery and selection is implemented by Cyprotex by using the unbiased view of transcriptomics and its potential to predict drug-induced toxicity. Transcriptomics involves sequencing thousands of mRNA molecules to identify which processes are active in the cell and allows for a better understanding of the cell’s reaction to known and novel drugs.
This is by no means a purely academic endeavour. As an example, the Cyprotex team demonstrated via transcriptomics it was able to identify problems in liver cells treated with fasiglifam, a promising diabetes drug candidate, which was withdrawn from late-stage clinical trials by its developer, following signs of liver damage in trial participants. This example proves that transcriptomics could have raised a red flag during preclinical development and might have saved hundreds of millions of dollars.
“Our studies have found potential effects on mitochondrial function, which were previously missed in preclinical studies” says Walker.
Therefore, transcriptomics has the potential to supplement or reduce in vivo toxicology studies by effectively identifying safety issues early in drug development, saving time and money — and animal testing.
Sophisticated Human Cell-Based Models
A key advantage of transcriptomics is its use of human cells and Evotec as well as Cyprotex are not just looking at 2D cell cultures, but investigating 3D organoids. These structures formed of thousands of cells that mimic organ-specific tissues are much closer to the real organ and have valuable features: For example, 3D-organoids of the heart exhibit regular contractions, beating like a living heart, and liver organoids secrete typical liver enzymes for days.
“On top of that, a 3D system allows repeat dosing, mimicking dosing regimens in vivo and potentially helps to detect effects due to toxic metabolites,” says Walker.
As they are small, the organoids can be placed in 384-well plates and individually molecular barcoded for simultaneous sequencing. This combination of miniaturization and high-throughput screening is implemented in Evotec’s EVOpanOmics platform and allows a wider adoption of transcriptomics in preclinical toxicology studies allowing for the repeat testing of dozens or even hundreds of compounds at several doses and in multiple organs.
“People have thought about using transcriptomics for toxicology before, but it was always a numbers game,” explains Rüdiger Fritsch PhD, Principal Scientist and Project Lead for EVOpanOmics. “For any compound that’s a real troublemaker, the evidence will show up in the transcriptomics data if you profile it in a relevant model. You just need to test appropriate dosing scenarios with the breadth of genome-wide off-target effects so that you have a chance to find it.”
Complex Analysis of Transcriptomics Data
Evotec, in conjunction with Cyprotex, offers transcriptomics services to drug developers and carries out the entire process in-house, from growing the organoids to sequencing and analysis. This streamlined process allows its researchers to screen hundreds of compounds a day, each delivering tens of thousands of data points on RNA levels. To analyze all of these vast amounts of data, Evotec has developed a software platform called EVOpanHunter that allows among others the analysis of these transcriptomics in an interactive manner.
“We want to democratize data analysis for the biologists who know the biological pathways and processes, without them needing to rely on additional experts from the bioinformatics department for routine tasks,” says Carla Tameling PhD, Head of Sales and Application for EVOpanHunter at Evotec.
On top of the interactive multi-omics analysis platform machine learning is used to trawl through this immense amount of data in order to find specific patterns hinting for toxicological effects and alert the researchers to dig deeper. “The more data we get, the harder it is for a human to dig through it all,” adds Tameling. “Transcriptomics is an unbiased view. You don’t need to define what to look at prior to your studies — you get all the data, and you might see things that you didn’t think would be relevant initially.”
From publically available sources, Cyprotex has compiled a broad and highly valuable transcriptomics reference database for drug-induced liver injuries.. Machine learning is being applied to predict whether a compound is likely to have issues by comparing the observed pattern of gene activity to the activity patterns of known toxic molecules. Furthermore, this is not restricted to hepotoxicity. Cyprotex is already building databases of other organs, such as heart, kidney and brain, using publicly available drug development trial results to select a broad space of reference comounds. “We’re running reference compounds from all kinds of sources where we know there are either late-stage clinical findings or withdrawals from the market,” states Walker.
Given the rapid advancements of the technology, it may be only a matter of time before transcriptomics and other omics technologies become a regulatory standard approach for preclinical toxicity testing.
Tags: Blog, In vitro Biology, Toxicology & Safety, Modelling and Simulation
Toxicokinetic evaluation is a regulatory and scientific requirement in the drug development process. To obtain plasma, blood is generally withdrawn by a conventional venous collection method. Microsampling is a less invasive sampling technique, which allows to reduce the stress correlated to the conventional blood sampling and to decrease the number of rodents for a preclinical study. The implementation of microsampling in particular, in non-human primate can reduce the stress and promote a positive interaction with technical staff which improves the overall well-being of the animal (refinement).
In this poster we summarise the work done to evaluate the possible influence of the blood sampling method on drug plasma concentrations, using LC-MS/MS methods in non-human primate for four drugs selected based on acid-base and volume of distribution properties.
The poster was presented by our expert Rossella Cardin at the 24th International Reid Bioanalytical Forum held in Cambridge, UK, on June 13-16, 2022.
Tags: Posters, ADME/DMPK, IND Enabling Studies/Preclinical Development, Toxicology & Safety
Tags: Fact Sheets, Formulation & CMC, ADME/DMPK, IND Enabling Studies/Preclinical Development, Toxicology & Safety
Learn more about the use of transcriptomics in safety prediction including:
Tags: Fact Sheets, Toxicology & Safety