Periodic Reporting for period 1 - SmallDrugRheuma (Discovery of Immune Therapeutic Targets and Immunomodulators for the Development of Novel Therapies in Rheumatoid Arthritis.)
Okres sprawozdawczy: 2018-06-29 do 2020-06-28
We developed an in vitro inflammation model in human primary immune cells, and tested on it 3,000 different small molecules (including both approved and novel lead-like drugs), covering the whole chemical space. We identified a portfolio of small molecules capable of reducing the secretion of proinflammatory cytokines while do not altering cell viability (n=25). We validated them in several donors and at different concentrations. The extensive screening effort led to 2 strong candidates (repurposing drugs), which allow for 30-50% reduction of IL-1β and/or TNF-α secretion by inflamed PBMCs. Further, we validated this reduction in cytokine secretion in PBMCs from newly diagnosed RA patients which have not yet received any treatment, and identified the intracellular pathways being targeted. In summary, we identified novel compounds capable of reducing IL-1β and TNF-α secretion in both in vitro inflamed HD PBMCs and PBMCs from naïve RA patients.
We validated ex vivo the top-5 candidates at several concentrations in several donors and repeated the assays, to confirm the results: we observed that compound #6 consistently reduced IL-1β secretion by the in vitro inflammation model, while compound #13 reduced both IL-1β and TNF-α secretion. We validated these findings through an orthogonal assay, including this time also cells from RA patients which have not yet received any treatment (naïve). To interrogate the mechanism of action of these compounds, we treated cells from both HD and naïve RA patients with compound #6 and #13, and analysed them by flow cytometry. Compound #6 impeded the secretion of IL-1β, while compound #13 interfered with its production. Regarding TNF-α, compound #13 interfered with TNF-α production, upstream of secretion.
Therefore, we generated a portfolio of novel immunomodulatory compounds, whose characterization and validation for the treatment of human diseases will follow up in further projects, and identified two drugs capable of reducing the secretion of two key components of inflammation processes (IL-1β and TNF-α), which we validated in vitro for the treatment of RA. Regarding the latter, we are preparing two patents that will be submitted soon, together with a publication that will follow the patent applications.
The work carried out enhance the innovation capacity of the ERA as a whole, as the anti-inflammatory properties of the identified drugs are completely novel and, once protected, will open up a new way for their potential use in RA patients. The results of the project will also step up the development of novel therapies, increasing the potential of citizens to live longer in good health, pushing Europe 2020 objectives. Equally, these results will contribute in the mid-long term towards lessening the impact of RA on EU economy and business, as RA has been recognised by the WHO as a Non-Communicable Disease that can lead to substantial costs for European healthcare.
The results also contribute to Innovation Union and ERA priorities, because addresses the challenge “Health, demographic change and wellbeing” by developing novel drugs for a chronic disease. In particular, these results increase European excellence and benefit the scientific community and society at different levels: (i) Contribution towards science, innovation and generation of knowledge, by providing valuable insights into novel drugs will be useful for specialists in Immunology, but also for those working in a broad range of disciplines and diseases (e.g. cancer); (ii) Contribution to public health and welfare, through the identification of novel drugs, opening the doors to preclinical/clinical trials in chronic diseases and by improving bench-to-bed translation, contributing to the Europe 2020 challenge “Health, demographic change and wellbeing”; (iii) Quick transfer of results to the market through the host’s experience in PPP and IPR planning, bringing rapid benefits to citizens and competitiveness gains.