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05.06.2025Technological complementarity at the CCiTUB, key to drug candidate screening
CCiTUB offers a wide range of essential technologies for the screening of molecules with pharmacological potential. The combination of complementary and orthogonal techniques allows for a more complete understanding of molecular characterization, compensating for the individual limitations of each methodology and enabling more robust and precise studies..
A prominent example of this multidisciplinary application is research related to Parkinson’s disease. Recently, Gain Therapeutics has used its computational platform Magellan, formerly known as SEE-Tx (Site-Directed Enzyme Enhancement Therapy), to identify small molecules capable of acting as allosteric modulators of enzymes of interest. These molecules bind to specific sites on enzymes and improve their stability and function without interfering with the natural substrate, which allows for the restoration of defective enzyme activity implicated in genetic diseases.
In this context, Gain Therapeutics has developed the molecule GT-02287, a low molecular weight (LMW) allosteric modulator that is currently in a phase 1b clinical trial for the treatment of Parkinson’s disease. GT-02287 acts on the enzyme glucocerebrosidase (GCase), encoded by the GBA1 gene. Mutations in this gene reduce GCase activity and are associated with an increased risk of Parkinson’s. The drug developed by Gain Therapeutics helps restore enzymatic function, reducing inflammation and the accumulation of damaged proteins in the brain.
To validate the efficacy of these compounds, various technologies available at CCiTUB have been used, which have been essential for characterizing the interactions between candidate molecules and their targets..
Among these, the fluorescent thermal shift assay (FTSA or DSF) technique stands out, implemented at the CCiTUB Genomics laboratory. For this type of experiment, a fluorochrome compatible with the excitation or emission profile of the Light Cycler 480 quantitative PCR (qPCR) configuration was used. The fluorochrome spontaneously binds to the hydrophobic regions of the protein; then, the temperature is gradually increased, and fluorescence is measured at each point, allowing the thermal stability of the protein/LMW complex to be characterized and how it is affected by the presence or absence of the LMW.
Surface Plasmon Resonance (SPR) has also been applied at the CCiTUB Molecular Interaction Analysis laboratory. In these real-time and label-free experiments, the target protein is immobilized on the chip and different concentrations of the compounds (LMW) are passed in soluble phase. Using the appropriate chip design, the association and dissociation constants at equilibrium (KA and KD) can be obtained. In very specific cases, not so typical for LMW, the half-life (t1/2) of the complex can also be calculated.
The Magellan approach to discover non-competitive, pharmacological allosteric regulators for the GCase protein. Schematic workflow of the procedure used to discover new allosteric regulators;
Abbreviations: GCase, glucocerebrosidase; VS, virtual screening; DSF, differential scanning fluorometry.
Use of CCiTUB for GBA1
DSF & SPR:
SPR