Page 26 - I. Chemical biology and drug discovery

[I. Chemical biology and drug discovery-14]

HCS-based discovery of mitophagy modulators using Small

Molecule Natural Products

Minjeong Kim¹, EunJu Jin¹, Dao Thi Minh Tam¹, Jeongho Kwon¹, Dongryeol Ryu¹

¹Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine , Suwon 16419, Republic of

Korea

Mitophagy, one most famous selective autophagy, is crucial to eliminate damaged and dysfunctional mitochondria.
Mitophagy-enhancing drugs could have the potential to cure a variety of human diseases such as metabolic, age-

associated, neuromuscular, and cardiovascular disease. To find the optimal screening method for drugs modulating

mitophagy, we performed a comparative assessment based on probability, efficiency, and accuracy. First, we tested
with synergy Neo is a High-throughput screening (HTS) multi-mode microplate reader. However, we failed to detect
the shifted fluorescence response to a positive control such as oligomycin plus antimycin A (OA) as well as a

negative control such as Bafilomycin A (BFA). Then, we evaluated with a High Content Screening (HCS) system. The
HCS-based tests were done with both IN Cell analyzer 2200 (GE Healthcare) and Cytation 5 (BioTek). Unlike the HTS,

both HCS analysis systems distinguished signals from the basal, OA-treated positive, and BFA-combined negative
control. To discover a mitophagy enhancing or modulating drug, we evaluated the bioactivity of 640 small molecules,

isolated from Korean plants. Finally, we got 14 hits (11 mitophagy enhancers and 3 blockers).