ABOUT TASPPI


With estimates between 130,000 and 650,000 Protein-Protein Interactions (PPIs) in the human body, pharmacological exploitation of these interactions as drug targets significantly enlarges the “drugable genome”. Whereas successful examples of disrupting PPIs have been reported , the targeted stabilization of PPIs with small molecules has not been realized in a systematic way yet. However, the fortuitous finding that the immunosuppressant drugs rapamycin and FK506 confer their activity by stabilizing the protein complexes mTOR/FKBP12 or calcineurin/FKBP12, respectively, illustrates the feasibility of this approach .
An example where PPI stabilizing molecules have been (a) found by High-Throughput-Screening (HTS), (b) crystallized and (c) shown physiological activity was reported by the group of the coordinator of this ETN . In the project proposed here, ESRs will develop novel drug-like molecules that mediate their activity by stabilizing PPIs of the adapter protein 14-3-3 with partner proteins important in the development of human diseases. The principal advantages of PPI stabilizers over conventional inhibitors are their uncompetitive nature and high specificity due to binding to preformed, composite pockets established by two proteins at the rim of their binding interface. The binding interfaces of protein complexes show a much less conserved structural architecture than, for example, active sites of enzymes. Hence the chances to develop a specific molecule binding to such an interface pocket should be higher than with “traditional” inhibitors. This higher specificity has the potential to lead to markedly reduced toxicity of these compounds and consequently less adverse effects.
The early stage scientists will work on a diverse set of PPIs implicated in Neurodegeneration, Cancer Development, Inflammation, Pulmonary Diseases, and Metabolic Disorders. This wide and diverse field of applications illustrates the general utility of this technology platform for the Pharma and Biotech Industry. The objectives of this consortium project therefore serve to address problems in Horizon 2020 key areas “Ageing Population” (Alzheimer’s Disease, Parkinson’s Disease, Cancer), and “Increased Burden of Chronic Diseases” (Metabolic Disorders, Inflammation, Pulmonary Diseases).

The ESRs will employ High-Throughput-Screening (HTS), Fragment-Based Ligand Discovery (FBLD), and Virtual Screening (VS) to identify initial stabilizers of the adapter protein class 14-3-3 interacting with a number of partner proteins (Tau, p53, Gab2, GR, NFκB, BAD, BAX, ASK1). Working on a number of different 14-3-3 targets will enable us to cross-validate specificity of hit compounds already with the assay format of the initial screening. A variety of biophysical and structural analysis methods will subsequently be used to characterize and validate the molecules stabilizing activity towards their respective 14-3-3 PPI complexes. In a third step, these validated molecules will be optimized by chemical synthesis. In the last phase of the project the optimized compounds will undergo pharmacological and cell biology evaluations to produce a list of optimized structures for further drug development.

TASPPI is a prime example of a consortium where both the academic partners (innovative idea, de-risking by profound basic research) and the industry part (highly sophisticated process of drug discovery) play hand in hand to train early stage researchers in applying a disruptive technology solution at the interface of academic and industrial research that will ensure their future employability.
This ETN will train the scientists in diverse scientific disciplines such as structural biology, medicinal chemistry, chemical biology, biophysical, biochemical and cellular analyses as well as pharmacology. Our long-term scientific goal is to initiate a paradigm-shift in drug development, complementing the development of active site and PPI inhibitors with small molecules stabilizing protein-protein interactions.

The combination of knowledge from academia and industry will maximize the synergy and the output potential of the project. To demonstrate the universal applicability of our technology platform we will distribute the 13 PhD projects among a number of therapeutic indication fields. The unifying theme in all these PPIs is the involvement of the 14-3-3 class of adapter proteins. These 14-3-3 proteins (7 isoforms in humans) are involved in a wide array of regulatory functions in diverse signalling pathways, cell cycle control and apoptosis 14- 3-3 proteins exert their physiological function by directly binding to other proteins. As a consequence, their target proteins are modulated in their enzymatic activity, their subcellular localization or their ability to further bind other protein partners. In this sense 14-3-3 proteins are switchable docking modules controlling the biological activity of their target proteins.

To date there are more than 200 protein partners of 14-3-3 proteins known, some of which play prominent roles in cancer development (e.g. p53) , neurodegeneration (e.g. Tau) , diabetes (e.g. BAD) , Pulmonary Diseases (e.g. GR10) , and inflammation(e.g. NFκB) . The following section will describe molecular targets for the development of drug candidates to address each of these themes. In addition to the prospect of identifying drug candidates the small molecules will serve the consortium’s goals as Chemical Biology tool compounds which will facilitate greater understanding of 14-3-3 signalling biology.

ABOUT TASPPI


With estimates between 130,000 and 650,000 Protein-Protein Interactions (PPIs) in the human body, pharmacological exploitation of these interactions as drug targets significantly enlarges the “drugable genome”. Whereas successful examples of disrupting PPIs have been reported , the targeted stabilization of PPIs with small molecules has not been realized in a systematic way yet. However, the fortuitous finding that the immunosuppressant drugs rapamycin and FK506 confer their activity by stabilizing the protein complexes mTOR/FKBP12 or calcineurin/FKBP12, respectively, illustrates the feasibility of this approach .

An example where PPI stabilizing molecules have been (a) found by High-Throughput-Screening (HTS), (b) crystallized and (c) shown physiological activity was reported by the group of the coordinator of this ETN . In the project proposed here, ESRs will develop novel drug-like molecules that mediate their activity by stabilizing PPIs of the adapter protein 14-3-3 with partner proteins important in the development of human diseases. The principal advantages of PPI stabilizers over conventional inhibitors are their uncompetitive nature and high specificity due to binding to preformed, composite pockets established by two proteins at the rim of their binding interface. The binding interfaces of protein complexes show a much less conserved structural architecture than, for example, active sites of enzymes. Hence the chances to develop a specific molecule binding to such an interface pocket should be higher than with “traditional” inhibitors. This higher specificity has the potential to lead to markedly reduced toxicity of these compounds and consequently less adverse effects.

The early stage scientists will work on a diverse set of PPIs implicated in Neurodegeneration, Cancer Development, Inflammation, Pulmonary Diseases, and Metabolic Disorders. This wide and diverse field of applications illustrates the general utility of this technology platform for the Pharma and Biotech Industry. The objectives of this consortium project therefore serve to address problems in Horizon 2020 key areas “Ageing Population” (Alzheimer’s Disease, Parkinson’s Disease, Cancer), and “Increased Burden of Chronic Diseases” (Metabolic Disorders, Inflammation, Pulmonary Diseases).

The ESRs will employ High-Throughput-Screening (HTS), Fragment-Based Ligand Discovery (FBLD), and Virtual Screening (VS) to identify initial stabilizers of the adapter protein class 14-3-3 interacting with a number of partner proteins (Tau, p53, Gab2, GR, NFκB, BAD, BAX, ASK1). Working on a number of different 14-3-3 targets will enable us to cross-validate specificity of hit compounds already with the assay format of the initial screening. A variety of biophysical and structural analysis methods will subsequently be used to characterize and validate the molecules stabilizing activity towards their respective 14-3-3 PPI complexes. In a third step, these validated molecules will be optimized by chemical synthesis. In the last phase of the project the optimized compounds will undergo pharmacological and cell biology evaluations to produce a list of optimized structures for further drug development.

TASPPI is a prime example of a consortium where both the academic partners (innovative idea, de-risking by profound basic research) and the industry part (highly sophisticated process of drug discovery) play hand in hand to train early stage researchers in applying a disruptive technology solution at the interface of academic and industrial research that will ensure their future employability.

This ETN will train the scientists in diverse scientific disciplines such as structural biology, medicinal chemistry, chemical biology, biophysical, biochemical and cellular analyses as well as pharmacology. Our long-term scientific goal is to initiate a paradigm-shift in drug development, complementing the development of active site and PPI inhibitors with small molecules stabilizing protein-protein interactions.

The combination of knowledge from academia and industry will maximize the synergy and the output potential of the project. To demonstrate the universal applicability of our technology platform we will distribute the 13 PhD projects among a number of therapeutic indication fields. The unifying theme in all these PPIs is the involvement of the 14-3-3 class of adapter proteins. These 14-3-3 proteins (7 isoforms in humans) are involved in a wide array of regulatory functions in diverse signalling pathways, cell cycle control and apoptosis 14- 3-3 proteins exert their physiological function by directly binding to other proteins. As a consequence, their target proteins are modulated in their enzymatic activity, their subcellular localization or their ability to further bind other protein partners. In this sense 14-3-3 proteins are switchable docking modules controlling the biological activity of their target proteins.

To date there are more than 200 protein partners of 14-3-3 proteins known, some of which play prominent roles in cancer development (e.g. p53) , neurodegeneration (e.g. Tau) , diabetes (e.g. BAD) , Pulmonary Diseases (e.g. GR10) , and inflammation(e.g. NFκB) . The following section will describe molecular targets for the development of drug candidates to address each of these themes. In addition to the prospect of identifying drug candidates the small molecules will serve the consortium’s goals as Chemical Biology tool compounds which will facilitate greater understanding of 14-3-3 signalling biology.