Part I: Synthesis of Cyclobutyl Nucleoside Analogs That Mimic AZT for Inhibition of the K65R HIV-1 Reverse Transcriptase Mutant Part II: Synthesis and Evaluation of Truncated Triptolide Analogs to Suppress Chronic Inflammation Público
Bluemling, Gregory Richard (2011)
Abstract
Part I of this dissertation describes the synthesis and
biological activity of novel
3'-azido cyclobutyl nucleoside analogs. These analogs were
developed for inhibition of
both the WT HIV RT and the K65R HIV RT mutant. The nucleoside
analogs were
prepared from cyclobutenones which were generated via a [2+2]
cycloaddition reaction.
The azide was installed through a Michael reaction, and both
thymine and adenine were
coupled to the cyclobutyl ring through a SN2 reaction via a
brosylate. The nucleosides
GB119, GB120, GB123, and GB124 did not
show any anti-HIV activity in human PBM
cells and were not toxic to PBM, CEM, and Vero cells up to 100
μM.
Part II of this dissertation describes the synthesis and biological
activity of
truncated analogs of the diterpenoid triepoxide natural product
triptolide. Triptolide
possesses potent anti-inflammatory activity but is highly
cytotoxic. In an effort to retain
the desired anti-inflammatory properties of triptolide while
reducing cytotoxicity, a series
of truncated analogs were synthesized from phenols. Triptolide
showed cytotoxicity at
50 nM in Jurkat cells. None of the analogs tested showed
cytotoxicity in PMBCs or
Jurkat cells up to 1000 nM. The analogs GB67B, GB97,
and GB187D were screened in
a carrageenan edema model in mice. GB67B was found to reduce
TNF-α levels and
reduce paw inflammation. To further study this compound,
GB67B at 10 mg/kg and 30
mg/kg was given orally to rats once daily for 28 consecutive days
in an adjuvant arthritis
model. GB67B was found to give a dose-dependent reduction of
paw volume and
arthritic score. A luciferase reporter gene assay showed that
unlike triptolide, GB67B did
not inhibit NF-κB activation. However, GB67B was found
to inhibit TNF-α levels from
LPS stimulated mouse splenocytes in a dose-dependent manner
supporting the carrageenan edema model results. Triptolide is known
to be lethal to mice and rats at a single dose of 1 mg/kg. In order
to observe toxicity of the truncated analogs, GB67B
was
dosed at 30, 90, and 180 mg/kg in rats for 7 days. While no animals
died during the
study there were signs of clinical intolerance for GB67B in
the 180 mg/kg group.
Table of Contents
Table of Contents
Part I: Synthesis of Cyclobutyl Nucleoside Analogs That Mimic AZT for Inhibition of the K65R HIV-1 Reverse Transcriptase Mutant
1.1 Statement of purpose ……………………………………….. 1
1.2 Introduction ………………………………………....……… 4
1.2.1 Current status of the HIV/AIDS pandemic ………………...... 4
1.2.2 The HIV replication cycle …………………………………... 6
1.2.3 FDA approved anti-HIV therapeutics ………………….…… 10
1.2.4 Drugs currently in clinical trials …………………………...…. 15
1.2.5 Additional classes of anti-viral therapies ………………...…... 19
1.2.6 HIV RT ……………………………………………...…….. 22
1.2.7 NRTI mechanism of action …………………………….…… 26
1.2.8 HIV RT NRTI resistance mechanisms …………….…….….. 28
1.3 Background ……………………………………..……..…... 33
1.3.1 3'-Azido-2',3'-dideoxynucleoside analogues ......................... 33
1.3.2 Syntheses of AZT and AZA …………………………….…. 34
1.3.3 Anti-viral natural product oxetanocin ……………….....…… 34
1.3.4 Syntheses of oxetanocin A and cyclobut-A ……..……..…… 36
1.3.5 Synthesis of cyclobutenones ……………………..…....…… 38
1.3.6 Methods for the Michael addition of azide ………...…..…… 41
1.3.7 Methods for coupling nucleoside bases to a cyclobutyl
ring ………………………………………...………….….. 46
1.3.8 Methods for synthesizing nucleoside triphosphates ……..….. 50
1.3.9 Synthesis and anti-HIV activity of 3'-hydroxymethyl
cyclobutyl nucleosides ……………………………...…….. 53
1.4 Results and discussion ……………………………...…….. 56
1.4.1 Design and synthesis of 3'-azido-3'-hydroxyethyl
cyclobutyl adenine ………………………………..……… 57
1.4.2 Design and synthesis of 3'-azido-3'-hydroxyethyl
cyclobutyl thymine………………………………..…….…. 60
1.4.3 Anti-viral activity…………………………………..……... 62
1.5 Conclusion …………………………………………….… 63
1.6 Experimental …………………………………….………. 64
1.7 References ………………………………………….…… 96
Part II: Synthesis and Evaluation of Truncated Triptolide Analogs to Suppress Chronic Inflammation
2.1 Statement of purpose ……………………………………… 103
2.2 Introduction ……………………………………………….. 106
2.2.1 Current status of autoimmune and inflammatory diseases ........ 106
2.2.2 Overview of the immune system ……………………..…….. 107
2.2.3 Autoimmunity ……………………………………………... 121
2.2.4 Inflammation …………………………………….………… 124
2.2.5 Drugs used to treat autoimmune/inflammatory diseases .....…. 128
2.3 Background ………………………………………...…….. 136
2.3.1 Triptolide as a potent immunosuppressant ……….…..…….. 136
2.3.2 Inhibition of the NF-kB signaling pathway by triptolide ......... 136
2.3.3 Synthesis of triptolide …....................................................... 140
2.3.4 Design and synthesis of monocyclic truncated
triptolide analogs…………………………………………… 143
2.3.5 Methods for o-formylation of phenols ……….………..…… 144
2.3.6 Methods for oxidative dearomatization ………………....….. 146
2.3.7 Epoxidation of electron deficient olefins …………….…....…. 148
2.4 Results and discussion ……………………………….……. 150
2.4.1 Synthesis of 2,5-substituted phenols ………………..….... 150
2.4.2 Synthesis of salicylic alcohols ……………………..…..…... 152
2.4.3 Oxidative dearomatization of salicylic alcohols ……….....…. 153
2.4.4 Synthesis of diepoxide analogs ………………………..….... 154
2.4.5 Triepoxide formation ………………….……………….…. 156
2.4.6 Deprotection of the triisopropylsilyl protected analogs …....…158
2.4.7 Design and synthesis of bicyclic truncated triptolide
analogs …………………………………………….......…. 158
2.4.8 Minimization of reactive functional groups …..………......…. 162
2.4.9 Synthesis of deuterated analogs for use as biological
standards …………………………………………..….…. 169
2.4.10 Biological evaluation of truncated triptolide analogs ….....…. 171
2.5 Conclusion …………………………………………..….. 206
2.6 Experimental ………………………………………......... 208
2.7 References ………………………………………..…….. 368
3.1 Appendix …………………………………………..…… 376
About this Dissertation
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Primary PDF
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Part I: Synthesis of Cyclobutyl Nucleoside Analogs That Mimic AZT for Inhibition of the K65R HIV-1 Reverse Transcriptase Mutant Part II: Synthesis and Evaluation of Truncated Triptolide Analogs to Suppress Chronic Inflammation () | 2018-08-28 11:12:03 -0400 |
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Supplemental Files
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MD-3-3-093-1100_FinalReport_Jurkat_AppendixB.pdf () | 2018-08-28 11:16:33 -0400 |
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Triptolide Final Report (11-19-09)_AppendixL.pdf () | 2018-08-28 11:17:42 -0400 |
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MD-31-012-0013 Final Report_AppendixD.pdf () | 2018-08-28 11:19:26 -0400 |
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AC13JU-JV.801.BTL Final Report_AppendixG.pdf () | 2018-08-28 11:20:53 -0400 |
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MD-3-3-093-1053_FinalReport_AppendixA.pdf () | 2018-08-28 11:24:03 -0400 |
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MD-3-3-093-1100_FinalReport_Splenocyte_AppendixC.pdf () | 2018-08-28 11:25:23 -0400 |
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0440RE27.002 Final Report_Appendix K.pdf () | 2018-08-28 11:28:03 -0400 |
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AC13JU-JV.850.BTL Final Report_AppendixF.pdf () | 2018-08-28 11:29:16 -0400 |
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MD-3-3-093-1052_FinalReport_AppendixE.pdf () | 2018-08-28 11:30:39 -0400 |
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0440RE27.001.prt_AppendixJ.pdf () | 2018-08-28 11:31:53 -0400 |
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024347 FINAL REPORT_AppendixI.pdf () | 2018-08-28 11:33:18 -0400 |
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Entelos_Summary_Report_AppendixH.pdf () | 2018-08-28 11:35:13 -0400 |
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