Title Page

Contents

Ⅰ. Research Objectives 15

Ⅱ. Design and Development of small molecule anticancer agents and cell imaging probes 17

Abstract 17

1. Introduction 18

2. Materials and Methods 21

2.1. Materials and Instrumentation 22

2.2. General procedure for the synthesis of BE7-1~BE7-7 and BE-C1, O7-1~O7- 7 and O-C1, C7, C7-1~C7-7 and C-C1, B1-C1~BA-C1. 23

2.3. in vitro Assay 51

2.4. Molecular Docking 56

2.5. Predicted ADME properties 57

3. Result & Discussion 58

3.1. Design and Synthesis of Benzimidazole derivatives 58

3.2. in vitro Assay 62

3.3. Molecular docking 74

3.4. In silico ADME analysis-based prediction of drug-likeness 78

4. Conclusion 86

5. Reference 87

Ⅲ. Fluorescence 'turn-on' probe for nanomolar Zn(II) detection in living cells and environmental samples 92

Abstract 92

1. Introduction 93

2. Materials and Methods 95

2.1. Chemicals and instruments 95

2.2. Synthesis of the receptor (FHE) 96

2.3. FHE specificity determination by spectral measurements 97

2.4. Effect of pH, reversibility, and quantum yield of FHE and FHE-Zn²⁺ complex 100

2.5. Computational insight into the structures of FHE and FHE-Zn²⁺ complex 101

2.6. Cell culture, anti-proliferation activity, and cell imaging assay 101

3. Results & Discussion 102

3.1. Synthesis and characterization of probe FHE 102

3.2. FHE demonstrates high selectivity for Zn²⁺ ions 106

3.3. Binding stoichiometry and limit of detection 107

3.4. Mechanism for detection of Zn²⁺ by FHE 112

3.5. Application of FHE for detecting Zn²⁺ in the living cells and environmental samples 116

4. Conclusions 119

5. Reference 120

Ⅳ. A dual-function fluorescence 'turn-on' probe that allows Zn (II) bioimaging and quantification of water in the organic solvent 125

Abstract 125

1. Introduction 126

2. Materials and Methods 128

2.1. Chemicals and instruments 128

2.2. Synthesis of the fluorescence turn-on probe FLHE 129

2.3. FLHE specificity determination by spectral measurements 132

2.4. Computational calculations for the HOMO-LUMO energies of FLHE and FLHE-Zn²⁺ complex 134

2.5. Effect of pH, reversibility, and quantum yield of FLHE and FLHE-Zn²⁺ complex 134

2.6. Cell culture, cell cytotoxicity, and cell imaging assay 135

2.7. FLHE-based paper strip (FLHE-PS) assay for quantification of Zn²⁺ ions in water 136

2.8. FLHE-based paper strip (FLHE-PS) assay for determination of water content in the organic solvent 137

3. Results and Discussion 138

3.1. Synthesis probe FLHE 138

3.2. Photophysical properties of FLHE 140

3.3. FLHE allows highly selective recognition of Zn²⁺ ions 142

3.4. Binding stoichiometry, association constant, and detection mechanism 144

3.5. Competition assay, pH effect, LOD, and quantum yield 148

3.6. Detecting Zn²⁺ in the live cells and environmental samples 153

3.7. FLHE-PS allows quantifying the water content in the organic solvent 157

4. Conclusions 159

5. Reference 160

Ⅴ. Appendix 166

Korean Abstract 236

English Abstract 238