From the identification of active compounds in known medicines to finding new drugs through high-throughput screening, fragment-based screening, and virtual screening, drug discovery has shifted. Developing effective small molecule therapeutics and finding out their MOA are the biggest hurdles in the stroke drug discovery process. What MOA studies try to do is to define the chemical interactions between molecules and their targets to see how molecules act on their targets, and how physiological concentrations of natural substrates affect that activity. Identifying toxicity and potential side-effects of a compound, as well as justifying phenotypic effects, is only possible with an understanding of its biological mechanism. This information builds faith in a promising compound prior to its trial entry.

Fig.1. Molecular mechanisms of stroke. (Shehjar, et al., 2023)

Ace Therapeutics offers various approaches to help clients analyze the mechanism of action of stroke drugs.

Related Services

• Pathogenic Mechanism Analysis Services for Stroke
• ADME/DMPK Services for Stroke Drugs

The Importance of Mechanism of Action in Stroke Drug Discovery

A clear understanding of the MOA of drug compounds is necessary for stroke drug development and prevents failure at a later stage in the approval process. MOA studies help determine clinical safety and act as a benchmark for new stroke drugs before their release in the market.

Here are some of the benefits of MOA research:

• Cellular response insights: These studies help identify how cells respond to specific drug compounds, including potential off-target pathways activated by the drug.
• Dosing optimization: Monitoring drug pathways and reactions allows for improved dosing strategies.
• Combination therapy development: MOA studies can identify effective drug combinations, reducing the likelihood of drug resistance by targeting multiple pathways simultaneously.
• Selectivity and drug design: Understanding how drug compounds interact with receptors enables the development of new drugs with similar selective actions.

Approaches to Understanding Mechanism of Action in Stroke Drug Discovery

Testing the Bioactivity of Compounds to Targets

The list of assays for measuring the bioactivity of compounds to specific targets is vast and covers:

• Direct Biochemical Approaches: These tests measure the molecular bonding between a compound and the target. There are enzyme assays (the amount of activity an enzyme is measured in the presence of a compound) and binding assays (the amount of affinity a compound has with its target using, for example, surface plasmon resonance or radiolabeled binding studies).
• Genetic Interaction Methods: These techniques use genetic code to see how compounds interact with cells. RNA interference (RNAi) or CRISPR gene editing can knock out or delete particular genes so that when the expression of a target is manipulated, the change in bioactivity can be measured.

Cell Imaging Assays

Cell high-throughput imaging tests provide the morphological changes in cells (size and shape of organelles) that are due to modulated signaling and modulation of cytoskeletal protein activity. Connecting changes in cell structure to certain signals (e.g., growth factor receptor stimulation or signal inhibition), scientists are able to explain how cells react to drugs or to the environment.

Transcriptomics for Mechanism of Action Studies

Transcriptomics involves the analysis of the complete set of RNA transcripts produced by the genome under specific circumstances. Techniques such as microarray analysis and RNA sequencing (RNA-seq) are commonly used to study changes in gene expression, providing insights into how signaling pathways and transcription factor activities are modulated.

Proteomics for Mechanism of Action Studies

Scientists use mass spectrometry to find proteins that bind to small molecules, a critical part of drug mechanisms and creating novel therapeutics. Mass spectrometry can also give quantitative information about interactions between small molecules and protein-bound objects. In the presence of intense ions, chemists can determine binding affinities and how well a tiny molecule bounds to a protein.

Metabolomics for Mechanism of Action Studies

Metabolomics is the comprehensive study of metabolites, which are small molecules produced during metabolism. If we can identify changes in metabolite abundance, we can know how chemicals regulate signaling networks and metabolism. There are two main techniques of metabolomics: Nuclear Magnetic Resonance (NMR) and Liquid Chromatography-Mass Spectrometry (LC-MS).

Computational Analysis of Mechanism of Action

The computational interpretation of the mechanism of action helps to aggregate all the different data from different experimental methods. The combination of historical screening results from sites such as ChemBank and current findings can help us get a better grasp on compound functions for better drug discovery and development. Computational tools and robust data visualizations enable us to glean more insight about mechanism of action of stroke drugs and aid decision making in therapy development.

1. Shehjar, F., et al. (2023). Stroke: Molecular mechanisms and therapies: Update on recent developments. Neurochemistry international, 162, 105458.