VDP-21 [Funding] Discovery of novel mitophagy activators w/ Evandro Fang

Identification of novel mitophagy inducers for healthy brain ageing and longevity.

• Project PI: Evandro Fang
• Project Page on Molecule

Simple Summary

My group was one of the very first research teams to propose defective mitophagy as a key driver in AD initiation and progression and to demonstrate its causative role; we have demonstrated the effectiveness of mitophagy induction in inhibiting memory loss in multiple AD animal models (Trends Neurosci 2017; Nature Neurosci 2019; Cell Metabolism 2019; Nature Biomedical Engineering 2022)3-5, 8. Previous work has highlighted how impaired mitophagy coincides with the behavioural and pathological development and progression of AD. Genetic and pharmacological promotion of mitophagy rescued cognitive decline in AD models, and inhibited the phosphorylation of Tau (p-Tau) in both human cell lines and 3xTgAD mice4. However, robust neuronal mitophagy inducers with clinical potential, that is to induce mitophagy but without causing mitochondrial damage at the same dose, is sparse. Very recently, we have established an artificial intelligence (named Fang-AI) plus wet lab validation platform (Figs. 1 and 2), enabled us successful identification of two lead compunds as drug candidates for AD8, 9. This proposal aims to use the ‘Fang-AI’ plus our wet lab platform (C. elegans, mice and iPSCs) to swiftly identify new drug candidates for AD.

Background

Alzheimer’s disease (AD) affects over 35 million people worldwide and causes formidable economic challenges1. Since 2003, over 250 drug candidates, predominantly targeting two pathological proteins, amyloid-β (Aβ) and pathological Tau, have been tested in clinical trials for AD, but most of them have failed2. There is a need to pursue new mechanistic studies in order to better understand the underlying causes of AD, and to discover new drug targets.

Mitochondria operate as cellular “powerhouses” and play a pivotal role in neuroplasticity and memory, thus dysfunction in mitochondria can impair neuronal function and trigger neurodegeneration3. The homeostatic maintenance of functional mitochondria is crucial for neuronal health and survival. Mitochondria are constantly exposed to stress and damage; to cope, dysfunctional mitochondria must be specifically and efficiently eliminated via a cellular self-clearance system, “mitophagy”. Mitophagy is impaired in the elderly, leading to the accumulation of damaged mitochondria4. The impaired energy production brought about by these dysfunctional mitochondria manifests in the senescence, inflammation, and neuronal loss seen in the elderly, and in AD patients3-5. Large-scale omics and functional studies using post-mortem brain tissues and iPSCs from AD identified impaired mitochondrial function and impairment of mitochondria-related pathways as major changes in AD4, 6, 7.

My group was one of the very first research teams to propose defective mitophagy as a key driver in AD initiation and progression and to demonstrate its causative role; we have demonstrated the effectiveness of mitophagy induction in inhibiting memory loss in multiple AD animal models (Trends Neurosci 2017; Nature Neurosci 2019; Cell Metabolism 2019; Nature Biomedical Engineering 2022)3-5, 8. Previous work has highlighted how impaired mitophagy coincides with the behavioural and pathological development and progression of AD. Genetic and pharmacological promotion of mitophagy rescued cognitive decline in AD models, and inhibited the phosphorylation of Tau (p-Tau) in both human cell lines and 3xTgAD mice4. However, robust neuronal mitophagy inducers with clinical potential, that is to induce mitophagy but without causing mitochondrial damage at the same dose, is sparse. Very recently, we have established an artificial intelligence (named Fang-AI) plus wet lab validation platform (Figs. 1 and 2), enabled us successful identification of two lead compunds as drug candidates for AD8, 9. This proposal aims to use the ‘Fang-AI’ plus our wet lab platform (C. elegans, mice and iPSCs) to swiftly identify new drug candidates for AD.

As defective mitophagy is likely a shared cause of different neurodegenerative diseases (such as Parkinson’s disease/PD, Huntington’s disease/HTT, and Amyotrophic Lateral Sclerosis/ALS)10, turning up mitophagy via pharmaceutically approaches could be a druggable target for broad neurodegenerative diseases.

Proposed Work Packages

WP1: To use our established Fang-AI + wet lab validation platform to screen new mitophagy inducers.

∙ To use our established Fang-AI + wet lab validation platform (Figs. 1 and 2)8to screen new mitophagy inducers from the Finnish Institute of Molecular Medicine (FIMM), consisting 140,000 compounds (U. Helsinki, with access). We will perform AI screening and wet lab verification as detailed elsewhere8. We have already identified 26 molecules from the FIMM library (AI score ≥0.8) such as FIMM136359, 104757, 106153, etc.

WP2: AI-based structure modifications for drug candidates EFF-AA and EFF-BA, and wet lab validation experiments:

Unpublished data from the Fang group have identified two structurally similar natural compunds EFF-AA and EFF-BA. They are robust mitophagy inducers with high translational potential as EFF-AA and EFF-BA induce mitophagy at 10% dose as used for other compunds (e.g, compunds K and R8), and without detectible toxicity in the bioactive doses in cells and C. elegans.

For the purpose of further optimization and patent, we aim:

∙ To use machine learning to provide suggestions on structure modifications on current known mitophagy inducers (EFF-AA, EFF-BA), aiming to increase ‘solubility’, ‘activity’, ‘bioavailability’ and to reduce ‘toxicity’;

∙ To work with chemists and pharmacists to select from the ‘AI-suggested list’, and to synthesize new compunds (the first two steps will be done via paid service to the experienced medchem CRO companies, such as Spirochem or Symeres); and

∙ To use wet lab techniques (majorly cell culture and C. elegans) to validate bioactivity and anti-AD function of the new compunds.

∙ Patent applications (note, background IP will apply)

Note:

∙ The Fang group can hold the EFF-AA and EFF-BA data confidential until 30th Dec 2022. After that, the data will be submitted in international peer-reviewed journal (s); we will have a deal with the target journal and to make sure it will not published until the end of 30th March 2023. The Ph.D. student will need to get her Ph.D by the fall of 2023 in the precondition of getting her Ph.D. studies to be published by May 2023.

Proposed Milestones

∙ Milestone 1 (M01-03): Toxicity assays for FIMM hits - release of 50% of healthspan, lifespan, mitophagy monitoring budget

∙ Milestone 2 (M01-03): Toxicity assays for EFF-AA/EFF-BA, EC50 EFF AA/EFF-BA mitophagy assays (quantitative criteria tbd.); selected AI

suggested EFF-AA/EFF-BA compound variants and confirmation of chemical feasibility (quantitative criteria tbd.) - release of chemical synthesis budget and release of 50% of healthspan, lifespan, mitophagy monitoring budget

∙ Milestone 3 (M03-07): Positive outcome of mitophagy assays of FIMM hits (quantitative criteria tbd.) - release of 50% of IHC/ICC, proteomics and RNA seq budget

∙ Milestone 4 (M03-08): Successful chemical synthesis of EFF-AA/EFF-BA analogues -release of 50% of IHC/ICC, proteomics and RNA-seq budget

Evandro F. Fang (Oslo)

The Fang team will have their best effort to get all the milestones achieved as proposed. Due to the COVID-19 (impact the delivery of agents and dedication of the scientists in the lab) and for any unforeseen difficulties, if an extention is needed for a specific milestone, Fang will be reaching Molecule/VitaDao to get a new agreed deadline.

The total cost of the project would be +$250 - $400k and the DAO would own 100% of the project and associated IP. We could also explore an option where we pay less and co-own the IP with the university.

For more information, please see the phase 2 proposal and discussion on Discourse.