Table of Contents
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Table of Contents
Table of Contents _________________________________ pag. I
List of Abbreviations ______________________________ pag. V
Preface ___________________________________________ pag. IX
Chapter 1.: The PI3K/Akt/PDK1 pathway ___________ pag. 1
1.1. The PI3K/Akt/PDK1 pathway pag. 11.1.1 Physiological role pag. 1
1.1.2 Oncogenic role pag. 3
1.1.3 Ras/Raf/MEK/MAPK and PI3K/Akt/PDK1
signalling network pag. 4
1.2. PI3K pag. 6
1.2.1 Structure pag. 6
1.2.2 Physiological role pag. 7
1.2.3 PI3K in cancer pag. 8
1.2.4 PI3K inhibitors pag. 9
1.3. Akt/PKB kinase pag. 11
1.3.1 Structure pag. 11
1.3.2 Physiological role pag. 13
1.3.3 Akt in cancer pag. 15
1.3.4 Akt inhibitors pag. 15
1.4. PDK1, the master kinase of AGC pag. 18
1.4.1 Structure pag. 18
1.4.2 Physiological role pag. 19
1.4.3 PDK1 in cancer pag. 22
1.4.4 PDK1 inhibitors pag. 24
1.5. mTOR pag. 28
1.5.1 Structure pag. 28
1.5.2 Physiological role pag. 29
1.5.3 mTOR in cancer pag. 30
1.5.4 mTOR inhibitors pag. 32
1.5.5 PI3K/mTOR inhibitors pag. 34
1.6. Therapeutic Strategies & Future Perspectives pag. 35
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Chapter 2.: OXIDs _________________________________ pag. 41
2.1. Story of OXIDs pag. 41
2.2. OX1 pag. 48
2.2.1 OX1 in NSCLC: preliminary studies pag. 48 2.2.2 OX1 in NSCLC: Western Blot analysis pag. 49 2.2.3 OX1 in MDA-‐MB-‐231 cell: preliminary study pag. 51 2.2.4 OX1: evaluation in AsPC1 cell line and
kinases assay pag. 52
2.3. Preliminary evaluation of OX2 derivatives pag. 54 2.4. Preliminary evaluation of OX3 derivatives pag. 56 2.5. Preliminary evaluation of OX4 derivatives pag. 58 2.6. Preliminary evaluation of OX5 derivatives pag. 60 2.7. OXIDs in GBM pag. 61
2.7.1 Importance of PI3K/PDK1/ Akt pathway
in GBM pag. 61
2.7.2 Selecting the best compound pag. 62 2.7.3 Physicochemical properties calculation pag. 63 2.7.4 G51: in vitro assays on GBM pag. 64 2.7.5 G51: select screening profile kinases pag. 66 2.8. OXIDs in GBM cancer stem cells? pag. 70 2.8.1 FC85 in U87MG cells pag. 70 2.8.2 FC85 in U87-‐derived cancer stem cells pag. 72
2.9. Conclusions pag. 75
2.10. References pag. 77
Chapter 3.: OXID-‐Pyridonyl Hybrids ________________ pag. 81
3.1. MP7 pag. 82
3.1.1 Development of a selective PDK1 inhibitor pag. 82
3.1.2 Synthesis pag. 87
3.2. OXID-‐pyridon-‐1-‐yl hybrids pag. 88 3.2.1 Design and synthesis of OXID-‐pyridon-‐1-‐yl
hybrids pag. 88
3.2.2 Computational studies pag. 94
3.2.3 Biological evaluation pag. 101
3.3. Med-‐Chem optimization pag. 101
3.4. Conclusions pag. 105
3.5. References pag. 108
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Chapter 4.: Materials & Methods __________________ pag. 109
4.1. Chemistry pag. 109
4.1.1 General details pag. 109
4.1.2 Synthetic procedures and characterization
data pag. 111
4.2. Biology & Biochemistry pag. 150
4.2.1 General details pag. 150
4.2.2 Cell lines and cell culture pag. 151 4.2.3 Cell seeding for amplification pag. 151 4.2.4 Cell proliferation assay pag. 151
4.2.5 MTT assay pag. 152
4.2.6 Cell lysates and sample preparation pag. 152 4.2.7 Protein quantification pag. 153 4.2.8 Gel electrophoresis and Western Blot
Analysis pag. 153
4.2.9 Band densitometry pag. 154 4.2.10 Statistical analysis pag. 154 4.2.11 Kinase-‐specific assay pag. 154
