Periodic Reporting for period 2 - CypTox (Training next level scientists and researchers to develop highly selective and safe insecticides)
Berichtszeitraum: 2023-01-01 bis 2025-06-30
Some of the most acute challenges that the world faces now and in the foreseeable future are caused by insects and mites that seriously threaten human health and food security. Their control is primarily achieved by the use of insecticides: for example, malaria prevalence has halved since 2000, saving 660 million lives, with 80% of the reduction being attributable to the use of insecticides. However, both the limited availability of low risk insecticides and insecticide resistance represent a major threat, and there is an urgent need to develop new insecticides. CypTox will apply biotechnology excellence to exploit the cytochrome P450 (CYP) metabolic/detoxification pathway of target and non-target organisms, to develop novel insecticides, efficient against selected insect & mite major pests and vectors, but are highly selective and safe for mammals, pollinators and the environment. Outcomes will also include biotechnology-based platforms (high-throughput cell/enzyme- screening assays and in silico pipelines), in line and beyond Pharma state of the art, that will advance research capabilities, for future developments of low risk insecticides. CypTox will provide excellent research training within a creative and flexible environment that actively promotes the integration of academic rigor and commercial pragmatism through mobility between sectors and focused training events. The size and balance of a highly motivated consortium (6 academics/6 industrial in EU and 3 TC partners; 300 well distributed secondments), the involvement of experienced PIs in Horizon 2020 and ERC projects and the modern communication, dissemination and exploitation approaches of CypTox will ensure efficient implementation and impact, at several levels. Enhanced career perspectives for scientists and researchers will be achieved by building on a sustainable multidisciplinary and inter-sectoral network in a wide range of fields, with the integration of world leading researchers and stakeholders.
The overarching objective of CypTox is to exploit the primary P450-based metabolic pathway of target and non-target organisms, for the development of highly selective, safe insecticides and new resistance breaking formulations, to improve the efficiency and sustainability of pests and vector control.
The overarching objective of CypTox is to exploit the primary P450-based metabolic pathway of target and non-target organisms, for the development of highly selective, safe insecticides and new resistance breaking formulations, to improve the efficiency and sustainability of pests and vector control.
During the second and final reporting period (RP2), the CypTox consortium significantly accelerated implementation and completed the majority of planned activities, successfully meeting its scientific, training, and innovation objectives. Secondments increased substantially (108.9 months in RP2), bringing the total mobility effort to 152 months. This active exchange between academia and industry enhanced collaboration, knowledge transfer, and cross-sectoral integration.
Objective 1. All coordination, reporting, and amendment tasks were successfully completed, alongside the preparation of the Final Ethics Report. Communication and dissemination were intensified, with strong participation in international events, workshops, multiple publications, and outreach initiatives. The website was maintained and updated, while the second career workshop and training sessions were delivered as planned.
Objective 2. A curated CYP sequence database was developed for key pest and pollinator species, complemented by gene expression profiling under xenobiotic exposure. RP2 activities included in vivo RNAi assays and dual-luciferase reporter systems to validate CYP roles in resistance and selectivity. Field-collected mosquito and pest populations were screened, revealing novel detoxification-linked CYPs.
Objective 3. The project established scalable bacterial and insect expression systems for arthropod CYPs. Functional enzymes were produced and tested using model substrates. Stable insect and mammalian cell lines expressing detoxification genes and targets were developed for toxicity assays of pro-drugs and synergists. High-throughput CYP assays and ligand-fishing methods were also refined.
Objective 4. In silico docking and molecular dynamics were applied to key pest P450s and ligands. Synergists such as curcumin and synthetic leads were identified via DSF and enzyme assays. CYP profiling confirmed in vivo activation or detoxification of pro-drugs in pests versus beneficials, supporting the design of selective, safer insecticidal chemistries.
Objective 5. Controlled-environment and mosquito bioassays tested candidate compounds on important disease vectors and agricultural pests. Dual-species toxicity testing with engineered cell lines showed favorable selectivity for certain compounds. Efficacy enhancements and lower non-target toxicity were demonstrated for lead candidates.
Objective 6. All planned training, secondments, and workshops were delivered. An e-learning capsule by Syngenta is publicly available and being expanded. Training covered enzyme biotechnology, resistance, molecular diagnostics, and rational pesticide use. Participation in INNODAYS, Researcher’s Night, and stakeholder events broadened the project’s knowledge-transfer impact.
Finally, the consortium completed its draft Exploitation Plan, identifying three main exploitable results—CYP screening platforms, recombinant enzyme systems, and synergist compounds—with commercialization or licensing potential. The IMCC continues to advise on IP and market valorization strategies.
Objective 1. All coordination, reporting, and amendment tasks were successfully completed, alongside the preparation of the Final Ethics Report. Communication and dissemination were intensified, with strong participation in international events, workshops, multiple publications, and outreach initiatives. The website was maintained and updated, while the second career workshop and training sessions were delivered as planned.
Objective 2. A curated CYP sequence database was developed for key pest and pollinator species, complemented by gene expression profiling under xenobiotic exposure. RP2 activities included in vivo RNAi assays and dual-luciferase reporter systems to validate CYP roles in resistance and selectivity. Field-collected mosquito and pest populations were screened, revealing novel detoxification-linked CYPs.
Objective 3. The project established scalable bacterial and insect expression systems for arthropod CYPs. Functional enzymes were produced and tested using model substrates. Stable insect and mammalian cell lines expressing detoxification genes and targets were developed for toxicity assays of pro-drugs and synergists. High-throughput CYP assays and ligand-fishing methods were also refined.
Objective 4. In silico docking and molecular dynamics were applied to key pest P450s and ligands. Synergists such as curcumin and synthetic leads were identified via DSF and enzyme assays. CYP profiling confirmed in vivo activation or detoxification of pro-drugs in pests versus beneficials, supporting the design of selective, safer insecticidal chemistries.
Objective 5. Controlled-environment and mosquito bioassays tested candidate compounds on important disease vectors and agricultural pests. Dual-species toxicity testing with engineered cell lines showed favorable selectivity for certain compounds. Efficacy enhancements and lower non-target toxicity were demonstrated for lead candidates.
Objective 6. All planned training, secondments, and workshops were delivered. An e-learning capsule by Syngenta is publicly available and being expanded. Training covered enzyme biotechnology, resistance, molecular diagnostics, and rational pesticide use. Participation in INNODAYS, Researcher’s Night, and stakeholder events broadened the project’s knowledge-transfer impact.
Finally, the consortium completed its draft Exploitation Plan, identifying three main exploitable results—CYP screening platforms, recombinant enzyme systems, and synergist compounds—with commercialization or licensing potential. The IMCC continues to advise on IP and market valorization strategies.
CypTox has advanced the state of the art by developing novel tools and methods that improve the understanding and application of cytochrome P450s (CYPs) in insecticide selectivity and resistance. It established robust expression systems for key P450s in E. coli and insect cells, enabling reliable metabolic studies and the development of functional, scalable enzyme screening platforms.
High-throughput fluorescent and chemiluminescent assays were developed to screen compound metabolism by CYPs, while synergist discovery through in silico and in vitro methods identified potent natural enhancers (e.g. curcumin). Furthermore, dual-cell assays using insect and mammalian systems were established to assess compound selectivity—supporting safer pesticide development.
Results include ready-to-use CYP assay platforms, identified synergists for crop protection, and new mechanistic insights into resistance. These outcomes are relevant for regulatory testing, agrochemical R&D, and sustainable pest management. The project also delivered e-learning modules and workshops to build capacity on low-risk pesticide use.
Overall, CypTox results contribute to EU Green Deal goals by enabling safer, targeted insecticide strategies, reducing reliance on broad-spectrum chemicals, and supporting biodiversity protection. Impact was further ensured through coordinated training, stakeholder engagement, and a draft exploitation plan, setting the foundation for licensing and commercial follow-up.
High-throughput fluorescent and chemiluminescent assays were developed to screen compound metabolism by CYPs, while synergist discovery through in silico and in vitro methods identified potent natural enhancers (e.g. curcumin). Furthermore, dual-cell assays using insect and mammalian systems were established to assess compound selectivity—supporting safer pesticide development.
Results include ready-to-use CYP assay platforms, identified synergists for crop protection, and new mechanistic insights into resistance. These outcomes are relevant for regulatory testing, agrochemical R&D, and sustainable pest management. The project also delivered e-learning modules and workshops to build capacity on low-risk pesticide use.
Overall, CypTox results contribute to EU Green Deal goals by enabling safer, targeted insecticide strategies, reducing reliance on broad-spectrum chemicals, and supporting biodiversity protection. Impact was further ensured through coordinated training, stakeholder engagement, and a draft exploitation plan, setting the foundation for licensing and commercial follow-up.