Discovery of Small-Molecule Modulators of Protein-RNA Interactions for Treating Cancer and COVID-19

Chapter 1. Introduction............................................................................ 1

1.1. Protein-RNA interaction.............................................................. 1

1.2. Therapeutic potentials of modulating PRI.................................... 1

1.3. Chemical biology approaches for the discovery of PRI modulator

............................................................................................................. 3

1.4. Aims and scope of the thesis........................................................ 5

1.5. References.................................................................................... 5

Chapter 2. Identification of Small-Molecule Inhibitors of Oncogenic Lin28-Let-7 Interaction 8

2.1. Discovery of a small-molecule inhibitor of protein-microRNA interaction using binding assay with a site-specifically labeled Lin28..................................................................... 8

2.1.1. Introduction..................................................................... 8

2.1.2. Results and discussion................................................... 11

2.1.3. Conclusion..................................................................... 26

2.1.4. Experimental section..................................................... 27

2.1.5. References..................................................................... 37

2.2. Restoring let-7 microRNA biogenesis using a small-molecule inhibitor of the protein-RNA interaction................................................................................................................... 41

2.2.1. Introduction................................................................... 41

2.2.2. Results and discussion................................................... 43

2.2.3. Conclusion..................................................................... 53

2.2.4. Experimental section..................................................... 53

2.2.5. References..................................................................... 58

Chapter 3. Discovery of Small-Molecule Modulators of Protein-RNA Interactions by Fluorescence Intensity-Based Binding Assay............................................................... 61

3.1. Introduction................................................................................ 61

3.2. Results and discussion................................................................ 63

3.3. Conclusion.................................................................................. 76

3.4. Experimental section.................................................................. 76

3.5. References.................................................................................. 85

Chapter 4. Identification of Protein-RNA Interaction Modulators for Treating COVID-19 88

4.1. Harnessing stress granule formation by small molecules to inhibit the cellular replication of SARS-CoV-2......................................................................................................... 88

4.1.1. Introduction................................................................... 88

4.1.2. Results and discussion................................................... 89

4.1.3. Conclusion................................................................... 103

4.1.4. Experimental section................................................... 104

4.1.5. References.................................................................... 110

4.2. Discovery of a small-molecule degrader of SARS-CoV-2 Nsp1 by autophagic pathway