Periodic Reporting for period 1 - Cytodesign (Novel engineered cytokines for human therapy)
Période du rapport: 2022-07-01 au 2024-06-30
The direct use of recombinant bacteria, or indirectly using bacteriophages, producing and delivering in situ a therapeutic molecule of interest has been extensively described. Many such efforts have also shown high efficacy in animal models but have yet to yield clinical successes, in part because of difficulties in achieving and maintaining sufficiently effective concentrations of the therapeutic molecule at the site of disease. Irrespective of the vector used, a recurring problem to be solved is that of the dose administered of either recombinant bacteria or phage which should be as small as possible, in order to ensure clinical efficacy and lack of toxic side effects. This is an issue since many of the biotherapeutic molecules to be expressed by bacteria are from human origin, and therefore can have folding problems, low stability, low serum half-life, and adverse effects due to multiple cell targets.
To solve these problems, we have developed the concept of foldikines. Foldikines are uniquely modified cytokines with improved properties, and are more amenable to delivery by a therapeutic microorganism. Amongst such improved properties, foldikines show increased stability, higher affinity and specificity for their target receptors, and reduced toxicity, all of which shall translate into greater safety and efficacy in treating human patients.
We have initial evidence (both in vitro and in vivo) of the superiority of foldikines over conventional cytokines, and in this project, we will aim to fully validate the concept and lay the basis for the creation of a new start-up company.
To solve these problems, we have developed the concept of foldikines. Foldikines are uniquely modified cytokines with improved properties, and are more amenable to delivery by a therapeutic microorganism. Amongst such improved properties, foldikines show increased stability, higher affinity and specificity for their target receptors, and reduced toxicity, all of which shall translate into greater safety and efficacy in treating human patients.
We have initial evidence (both in vitro and in vivo) of the superiority of foldikines over conventional cytokines, and in this project, we will aim to fully validate the concept and lay the basis for the creation of a new start-up company.
Within this ERC PoC grant we have validated the concept of Foldikine and filed two patent applications (wo2023144393a1) and ((PCT/EP2023/052202 Priority: EP 22382073.9 therapeutic cytokines and methods) licensed in exclusivity to the Start-up company (Orikine, https://www.orikine.bio/(s’ouvre dans une nouvelle fenêtre)) created as a result of this grant.
We have achieved the following milestones:
1-Engineered a Foldikine IL22 with four times better EC50 in HEK reporters for IL-22R
2-Designed a group of point mutations that improve EC50 of IL22 between 2 to four times and that in combination should improve it by around 10 times.
3-Engineered an IL22 cytokine resistant to degradation by P. aerugynosa protease, as well as not inhibited by the antagonistic soluble receptor of IL22
3-Engineered a Foldikine IFNg which is significantly superior to WT IFNg in different cell assays,
4-Developed a Foldikine IL4 and a hybrid IFNl/IL22 and IL2/IL4 cytokines
5-Develop protein modelling approaches to simulate cytokines binding to receptors. By integrating RNA quantification into an ODE-based model, our IL-2 signaling model accurately predicts responses across different cell types and mutants on in-vitro dose-response curves and in-vivo assays.
We have achieved the following milestones:
1-Engineered a Foldikine IL22 with four times better EC50 in HEK reporters for IL-22R
2-Designed a group of point mutations that improve EC50 of IL22 between 2 to four times and that in combination should improve it by around 10 times.
3-Engineered an IL22 cytokine resistant to degradation by P. aerugynosa protease, as well as not inhibited by the antagonistic soluble receptor of IL22
3-Engineered a Foldikine IFNg which is significantly superior to WT IFNg in different cell assays,
4-Developed a Foldikine IL4 and a hybrid IFNl/IL22 and IL2/IL4 cytokines
5-Develop protein modelling approaches to simulate cytokines binding to receptors. By integrating RNA quantification into an ODE-based model, our IL-2 signaling model accurately predicts responses across different cell types and mutants on in-vitro dose-response curves and in-vivo assays.
Our work has shown how using protein design tools and the Foldikine concept we can engineer Cytokines with superior properties in terms of stability, specificity, as well as combing the functional properties of two different cytokines. We have used our protein design tools FoldX and ModelX to engineer new linkers between two cytokines, as well as mutations that improve stability and binding to receptors.
Based on these results we have secured seed funding to incorporate a new company, Orikine, to develop the new Cytokines for human therapy.
Based on these results we have secured seed funding to incorporate a new company, Orikine, to develop the new Cytokines for human therapy.