Revolutionizing Drug Testing: The Advancement of Non-Animal Methodologies – a New Era of Drug Testing
Scientist
on
March 13, 2024
This installment of Tech Snapshot was written by Immuneed, a Sweden-based CRO providing customers across the globe with physiologically relevant data on the non-clinical safety and efficacy, aiding in selecting their most promising drug candidates.
With the advancement of new technologies, the need for optimal/matching in vitro, in vivo and ex vivo testing in drug development goes hand in hand. The FDA Modernization Act 2.0 and EMA’s recent policy changes have incentivized using non-animal methodologies, including tests based on human cells, thereby encouraging new alternatives to assess a compound’s safety and effectiveness. To demonstrate the analytical power of our non-animal method, ID.Flow®, we present a case study where ID.Flow® simultaneously evaluates the safety and effect of a cytotoxic CD20-targeting antibody.
BACKGROUND
CD20 is highly expressed in malignant lymphoma cells and is a target for cytotoxic antibodies to treat different B-cell malignancies. Antibodies can kill tumour cells via complement-dependent cytotoxicity (CDC) or antibody-dependent cell-mediated cytotoxicity (ADCC). However, the antibodies can also cause off-target effects such as binding and subsequent killing of blood cells, complement activation or severe cytokine release.
CASE STUDY
In this case, a sponsor needed to address several questions, including:
- How effective is the antibody in killing B-cells at different doses?
- Will the antibody induce unwanted effects on other cell types?
- Will it induce complement activation?
- Will it induce cytokine release?
- Is the antibody’s effect different from clinically approved antibodies?
Our innovative ex-vivo system, ID.Flow®, generates samples for evaluating cytokine secretion, complement activation, haemolysis and macroscopic clotting in human whole blood. The process is relevant and efficient, as ID.Flow® simultaneously generates multiple readouts from the same set of healthy volunteers. To start the experiment, fresh human whole blood was incubated immediately after collection with the anti-CD20 antibody and controls in ID.Flow® (see table on test conditions below).
The results showed that the studied anti-CD20 antibody neither displayed any prominent effects on blood cell counts (Figure 1) nor caused macroscopic clotting or haemolysis (data not shown). The negative and positive controls behaved as expected.
The anti-CD20 antibody dose-dependently depleted B-cells but not CD4+ or CD8+ T-cells, monocytes or granulocytes (Figure 2 and data not shown), showing consistency with the expression pattern of its target. From the tested cytokines (IFN-y, IL-2, TNF-α, IL-6 and IL-8) and complement split factors (C3a and C5a), only IL-8 was induced (Figure 3). Further, complement activation was not induced by treatment with the anti-CD20 antibody (Figure 3). From the tested cytokines (IFN-y, IL-2, TNF-α, IL-6 and IL-8) only IL-8 was induced (Figure 3). The test substance selectively depleted B-cells without any unwanted effects on T-cells, platelets or monocytes, most plausibly through antibody-dependent mechanisms and with minimal cytokine secretion.
The ID.Flow® test system is an innovative approach that can simultaneously assess a compound’s safety and efficacy. Immuneed® provides a comprehensive data package to support candidate selection, dose selection and decision-making regarding the drug development of non-clinical and clinical studies. At Immuneed®, we are proud to be part of the revolution in drug testing, and we are confident of the enormous potential of using non-animal methods to transform drug testing, making it more humane, ethical and efficient. As this approach is increasingly being adopted and continues to advance, it will be exciting to see how it transforms drug development and positively impacts human health.