Mechanistic Toxicity

Evotec and wholly owned subsidiary Cyprotex have combined expertise in toxicology which spans from early discovery through to the clinic so we have a full picture of the entire drug discovery and development continuum and can support you at every stage of your project. The scientific expertise and capabilities can be accessed as fee-for-services or as part of partially or fully integrated programs.

The first step in our understanding of drug- or chemical-induced toxicity is identifying potential mechanisms by evaluating specific adverse outcome perturbations, whether these are molecular, organelle or cellular effects and relating these to organ-specific toxicity.

Cyprotex offers a number of different assays to elucidate specific mechanisms of toxicity.

These include:

• Cell Stress Panel - assesses 36 biomarkers that represent cellular stress signalling pathways, organelle health and cellular cytotoxicity.

• Apoptosis and Necrosis Assay – uses flow cytometry to assess mechanism of cell death through apoptosis and necrosis.

• Phospholipidosis - lysosomal storage disorder characterised by excessive accumulation of intracellular phospholipids in tissues.

• Steatosis - hepatic lipid processing disorder, leading to accumulation of triglycerides within the liver cells.

• Lysosomal Trapping – investigates accumulation of the drug in the lysosome.

• Haemolysis - evaluates hemoglobin release in the plasma (as an indicator of red blood cell lysis).

• Mitochondrial Toxicity (Glu/Gal) – compares cytotoxicity using cells grown in glucose and galactose media to determine mitochondrial impairment.

• Functional Mitochondrial Toxicity  – uses the Seahorse XFe96 extracellular flux analyser to determine oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) to assess cellular bioenergetics and potential mechanisms of mitochondrial toxicity.

• Mitochondrial Respiratory Complex Assay (using Permeabilised Cells)  – uses the Seahorse XFe96 extracellular flux analyser to decipher which complex is involved in the mitochondrial toxicity.

• Reactive Metabolite Assessment – uses glutathione (GSH), potassium cyanide (KCN) or cysteine (Cys) trapping studies with human liver microsomes and LC-MS/MS detection.

• Cell Viability - general cell viability assessment using single endpoints such as LDH (lactate dehydrogenase), neutral red or MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide).

• Transcriptomics – using high throughput RNA-seq combined with a sophisticated bioinformatics platform to determine the drug or chemical-induced effects on the transcriptome.

• Custom Toxicology – flexible service where we can investigate different cell lines, endpoints and time points to address particular customer issues.