Title Page
ABSTRACT
국문 초록
PREFACE
Contents
CHAPTER 1. INTRODUCTION 20
CHAPTER 2. RESULT AND DISCUSSION 24
CHAPTER 3. CONCLUSION 77
REFERENCES 78
SUPPLEMENTARY MATERIALS 81
A. General materials and instruments 81
B. Synthesis 82
C. Quantum yield and determination of fluorescence emission and wavelength maxima 88
D. Spectroscopy in a solvent with increasing viscosity 89
E. Spectroscopy in the presence of Aβ1-42 and potential interferents (metal ions, amino acids, thiols, and albumin) 89
F. Fitting of Aβ saturation titration analysis 90
G. Solubility of ThN, ThB, and ThQ in aqueous solution 90
H. pH dependent absorbance of ThN, ThB, and ThQ 91
I. Photostability 91
J. Aβ1-42 fibril formation[이미지참조] 91
K. Aβ1-40 fibril formation[이미지참조] 91
L. Theoretical Calculations 92
M. Molecular Docking Assays 92
N. Preparation of AD mouse brain tissues 93
O. Fluorescent probe stain and image acquisition 93
P. Image J analysis 94
Table 1. Photophysical properties for ThN, ThB, and ThQ in various organic solvent. 40
Figure 1. ¹H NMR spectrum of compound 1. 26
Figure 2. ¹³C NMR spectrum of compound 1. 26
Figure 3. ¹H NMR spectrum of compound 2. 27
Figure 4. ¹³C NMR spectrum of compound 2. 27
Figure 5. ¹H NMR spectrum of compound 3. 28
Figure 6. ¹³C NMR spectrum of compound 3. 28
Figure 7. ¹H NMR spectrum of compound 4. 29
Figure 8. ¹³C NMR spectrum of compound 4. 29
Figure 9. ¹H NMR spectrum of compound 5. 30
Figure 10. ¹³C NMR spectrum of compound 5. 30
Figure 11. ¹H NMR spectrum of compound 6. 31
Figure 12. ¹³C NMR spectrum of compound 6. 31
Figure 13. ¹H NMR spectrum of compound 7. 32
Figure 14. ¹³C NMR spectrum of compound 7. 32
Figure 15. ¹H NMR spectrum of compound 8. 33
Figure 16. ¹³C NMR spectrum of compound 8. 33
Figure 17. ¹H NMR spectrum of compound 9. 34
Figure 18. ¹³C NMR spectrum of compound 9. 34
Figure 19. ¹H NMR spectrum of compound ThB. 35
Figure 20. ¹³C NMR spectrum of compound ThB. 35
Figure 21. ESI-MS spectrum of ThB. 36
Figure 22. ¹H NMR spectrum of compound ThQ. 36
Figure 23. ¹³C NMR spectrum of compound ThQ. 37
Figure 24. ESI-MS spectrum of ThQ. 37
Figure 25. ¹H NMR spectrum of compound ThN. 38
Figure 26. ¹³C NMR spectrum of compound ThN. 38
Figure 27. ESI-MS spectrum of ThN. 39
Figure 28. Photophysical properties as an intramolecular charge transfer (ICT). (A) The relationship between the maximum of absorbance wavelength of ThN, ThB, and ThQ plotted versus the solvent... 41
Figure 29. The relation between the natural logarithm of the solvent dielectric constant and the fluorescence quantum yield, maximum emission wavelength, or Stokes' shift of (A−C) ThN, (D−F)... 42
Figure 30. Fluorescence emission spectra of A) ThT, B) ThN, C) ThB, and D) ThQ (5 μM) in glycerol-methanol mixtures with different viscosities containing 1% DMSO (slit 5/5, excitation:... 44
Figure 31. Calculated molecular frontier orbitals of ThN, ThB, and ThQ. 45
Figure 32. Solubility in PBS. Absorbance of (A) ThT (B) ThN (C) ThB, and (D) ThQ at various concentrations (0−50 μM) in PBS (1% DMSO). A shift from monomers to soluble aggregates was... 46
Figure 33. pH-dependent absorbance changes. A) Absorbance spectra of ThT, B) ThN C) ThB, and D) ThQ at various pH levels 47
Figure 34. Photostability of ThT, ThN, ThB, and ThQ in DMSO. A 3100 K halogen lamp (Olympus LG-PS2, 12 V, 100 W) was used for irradiation and the absorbance was recorded at 5 min... 49
Figure 35. Absorbance, excitation spectroscopy data and fluorescence spectroscopy data of A) ThT, B) ThN, C) ThB, D) ThQ (5 μM) in the presence of Aβ1-42 fibrils (25 μM) in 10 mM PBS...[이미지참조] 50
Figure 36. Change in the fluorescence intensity of A) ThT, B) ThN, C) ThB, and D) ThQ at 0, 2.5, 5, 10, 17.5, and 25 μM of Aβ1-42 aggregates.[이미지참조] 52
Figure 37. Fluorescence spectroscopy data of A) ThT, B) ThN, C) ThB, D) ThQ (5 μM) in the presence and absence of Aβ1-42 fibrils (25 μM) in 10 mm PBS containing 1% DMSO (slit 5/5,...[이미지참조] 53
Figure 38. Selective fluorescence response assays for A) ThN B) ThB C) ThQ (5 μM) with various potential interferents, a: Aβ1-42 fibrils (25 μM); b: BSA 25μM); c-l: metal ions (1 mM, c: Al3+; d:...[이미지참조] 54
Figure 39. Saturation binding curves of Aβ1-42 fibrils (1 μM) as a function of A) ThT, B) ThN, C) ThB, and D) ThQ (0−1 μM) (n = 3).[이미지참조] 56
Figure 40. Saturation binding curves of Aβ1-40 fibrils (1 μM) as a function of ThN (0−1 μM).[이미지참조] 57
Figure 41. Fluorescence emission spectra of ThT (10 μM) with Aβ fibrils (10 μM) in PBS solution in comparison with varying concentrations of added ThN. A) 440 nm excitation, B) 480 nm excitation... 59
Figure 42. Linear correlation between fluorescence intensity at various concentrations of Aβ1-42 fibrils with A) ThT, B) ThN, C) ThB, and D) ThQ (0.2 μM) in PBS (10 mM, pH = 7.4) containing 1%...[이미지참조] 60
Figure 43. Docking studies of the Aβ1-42 protofibril (with a partial structure of the second protofibril). (a) Aβ1-42 sequence and the position of the essential residues in the cryogenic electron...[이미지참조] 62
Figure 44. Brain slice imaging and quantification data of APP/PS1 mice incubation with ThS (1 μM), ThB (1 μM), ThN (10 μM), and ThQ (10 μM). 64
Figure 45. Brain slice imaging and quantification data of APP/PS1 mice incubation with various concentration of ThB (1, 3, and 10 μM). 65
Figure 46. Brain slice imaging and quantification data of APP/PS1 mice incubation with various concentration of ThN (1, 3, and 10 μM). 66
Figure 47. Brain slice imaging and quantification data of APP/PS1 mice incubation with various concentration of ThQ (1, 3, and 10 μM). 67
Figure 48. Brain slice imaging and quantification data of APP/PS1 mice incubation with ThS (1 μM) and ThT (3 μM). 68
Figure 49. Co-localization between the ThS and ThB in APP/PS1 model mice. 69
Figure 50. ThS/ThB co-staining at compact or diffuse plaque enlarged Hippocampal and cortical images. 70
Figure 51. Co-localization between the ThS and ThN in APP/PS1 model mice. 71
Figure 52. ThS/ThN co-staining at compact or diffuse plaque enlarged Hippocampal and cortical images. 72
Figure 53. Co-localization between the ThS and ThQ in APP/PS1 model mice. 73
Figure 54. ThS/ThQ co-staining at compact or diffuse plaque enlarged Hippocampal and cortical images. 74
Figure 55. Fluorescence images of the brain tissue slice of 20 month-old APP/PS1/TauP301L-BiFC mouse. Magnified fluorescence images of the piriform cortex and hippocampus. white arrows indicate... 76
Scheme 1. Design strategy and Synthetic route for ThN, ThB, and ThQ. The illustration of overview brain slice imaging of Amyloid beta aggregates (Aβ). 23
Scheme 2. Synthetic route for ThT, ThN, ThB and ThQ. 25