Studies Aimed at the Synthesis of Anti-Infective Agents

216 Pages·2017·7.99 MB·English

UUnniivveerrssiittyy ooff SSoouutthh FFlloorriiddaa
DDiiggiittaall CCoommmmoonnss @@ UUnniivveerrssiittyy ooff
SSoouutthh FFlloorriiddaa
USF Tampa Graduate Theses and Dissertations USF Graduate Theses and Dissertations
April 2018
SSttuuddiieess AAiimmeedd aatt tthhee SSyynntthheessiiss ooff AAnnttii--IInnffeeccttiivvee AAggeennttss
Ankush Kanwar
University of South Florida, [email protected]
Follow this and additional works at: https://digitalcommons.usf.edu/etd
Part of the Chemistry Commons
SScchhoollaarr CCoommmmoonnss CCiittaattiioonn
Kanwar, Ankush, "Studies Aimed at the Synthesis of Anti-Infective Agents" (2018). USF Tampa Graduate
Theses and Dissertations.
https://digitalcommons.usf.edu/etd/7176
This Dissertation is brought to you for free and open access by the USF Graduate Theses and Dissertations at
Digital Commons @ University of South Florida. It has been accepted for inclusion in USF Tampa Graduate Theses
and Dissertations by an authorized administrator of Digital Commons @ University of South Florida. For more
information, please contact [email protected].
Studies Aimed at the Synthesis of Anti-Infective
Agents

by

Ankush Kanwar

A dissertation submitted in partial fulfilment
of the requirements for the degree of
Doctor of Philosophy
Department of Chemistry
College of Arts and Sciences
University of South Florida

Major Professor: James W. Leahy, Ph.D.
Bill Baker, Ph.D.
Jianfeng Cai, Ph.D.
Sandy Westerheide, Ph.D.

Date of Approval:
April 5, 2018

Keywords: Infectious diseases, Leishmania donovani, promastigotes, amastigotes, radicicol,
Plasmodium, xanthurenic acid, gametogenesis

Copyright © 2017, Ankush Kanwar
Acknowledgments
Finishing PhD is truly a marathon event and it would have not been possible for me to finish
this long and interesting journey without the help of countless people that were around me in last
five years. I have been lucky enough to have Dr. James Leahy as my mentor, who has helped me
to cope up with all the trials and tribulations of graduate school.
Firstly, I would like to thank Dr. James Leahy for giving me the opportunity to work under
his supervision and pursue my dream. He constantly keeps motivating me by giving suggestions
personally and professionally and believing in me to work independently and helping me in
times of need with his guidance. I will always be indebted to him for his guidance and
exceptional support.
I would sincerely thank my committee members Dr. Bill Baker, Dr. Jianfeng Cai, Dr. Sandy
Westerheide and Dr. Dennis Kyle for their valuable time and suggestions during these years. I’m
lucky to have very cheerful and friendly graduate and undergraduate students who worked with
me. I’m thankful to Linda Barbeto, Benjamin Eduful, Zachary Schultz, Jeanine Yacoub, Andrea
Lemus and Elena Bray for having wonderful science discussions and fun during weekends. It
always feels great to have good friends especially when staying far away from family and I’m
grateful to everyone. My fellow undergraduate students: Christian, Bernard and Piero: I am very
grateful for all your dedication, assistance and friendly talks during our shared time.
I am also grateful to our collaborators: Prof. Dennis Kyle and his team (Dr. Brian Vesely, Ala
Azhari, Tina Mutka, Tommmy McGaha and Vivian Padin), for getting the biological data for our
projects. I’m thankful for my friends outside lab. Dr. Shri Krishna Nimmagadda who helped me
during my early years of grad school with science discussions and patiently answering my
questions. Gaurav Verma, Chetan Joshi, Aneel and Shaq for having great life discussions, fun
weekends and cricket.
Lastly, I would like to thank my parents, especially my dad who keeps encouraging me in
everything, giving me confidence during bad times and for all his sacrifices in life. I would also
like to thank my brother Amit Kanwar for being there all the time for me and taking care of
parents back home. I would also thank my family members Sandeep Kanwar, Mohan Pal,
Avdesh Saroch, Vivek Tomar, Subart Mohaparta for their continuous support.

Thank you!
Table of Contents

List of Abbreviations...................................................................................................................... iii

List of Tables .................................................................................................................................. vi

List of Figures ............................................................................................................................... vii

List of Schemes .............................................................................................................................. ix

Abstract ............................................................................................................................................x

Chapter 1 Introduction ..............................................................................................................1
1.1 Infectious diseases ...................................................................................................1
1.2 Leishmaniasis ...........................................................................................................2
1.2.1 Life cycle of Leishmania ................................................................................3
1.2.2 Leishmaniasis manifestations and epidemiology............................................5
1.3 Current treatments against leishmaniasis .................................................................7
1.4 Heat Shock Protein 90 (Hsp 90) ..............................................................................9
1.4.1 Heat shock protein 90 (Hsp 90) in Leishmania ............................................ 11
1.5 High throughput Screening ....................................................................................13
1.6 Biological assays ....................................................................................................14
1.7 References ..............................................................................................................14

Chapter 2 Tetrahydro-indazole and tetrahydro-pyrazolo analogs ..........................................20
2.1 Drug design strategies ............................................................................................20
2.1.1 Docking and activity studies .........................................................................21
2.2 Synthesis of initial tetrahydro-indazole analog......................................................24
2.3 Synthesis of initial tetrahydro-pyrazolo pyridine analog .......................................26
2.3.1 Synthesis of substituted tetrahydro-pyrazolo pyridinyl analogs ...................28
2.4 Results and discussions ..........................................................................................30
2.5 Experimental ..........................................................................................................33
2.6 References ..............................................................................................................52

Chapter 3 Radicicol hybrid analog .........................................................................................55
3.1 Radicicol as Hsp 90 inhibitor .................................................................................55
3.2 Second generation Hsp 90 inhibitors .....................................................................57
3.3 NVP-AUY922(Luminespib) ..................................................................................58
i
3.4 Drug design strategy ..............................................................................................59
3.5 Synthesis of radicicol analog .................................................................................60
3.6 Results and discussions ..........................................................................................62
3.7 Experimental ..........................................................................................................62
3.8 References ..............................................................................................................68

Chapter 4 Introduction to Malaria ..........................................................................................72
4.1 Malaria ..................................................................................................................72
4.2 Life cycle of Plasmodium ......................................................................................73
4.3 Current treatments against malaria ........................................................................74
4.4 Role of xanthurenic acid in mosquitoes .................................................................77
4.5 Role of xanthurenic acid in Plasmodium ...............................................................78
4.6 References ..............................................................................................................79

Chapter 5 Synthesis of xanthurenic acid analogs ...................................................................83
5.1 Drug design strategies ............................................................................................84
5.2 Synthesis of 4-amino-8-hydroxyquinoline-2-carboxylic acid analog....................83
5.3 Synthesis of 8-amino-4-hydroxyquinoline-2-carboxylic acid analog....................85
5.4 Synthesis of 8-substituted amino-8-hydroxyquinoline-2-carboxylic
acid analog .............................................................................................................86
5.5 Synthesis of 6-(substituted 4-8-hydroxyquinoline-2-carboxylic acid analogs ......87
5.6 Synthesis of biotinylated xanthurenic acid analog .................................................89
5.7 Synthesis of clickable xanthurenic acid analog .....................................................92
5.8 Results and Discussion ..........................................................................................94
5.9 Experimental ..........................................................................................................97
5.10 References ............................................................................................................122

Appendix A. Biological Assays .................................................................................................125
Appendix B. NMR Spectra ........................................................................................................130
Appendix C. Copyright Permission ...........................................................................................200
Appendix D. Copyright Permission ...........................................................................................201

ii
List of Abbreviations

AA Axenic amastigote
AcOH Acetic acid
ATP Adenosine triphosphate
Bn Benzyl
CL Cutaneous leishmaniasis
DCM Dichloromethane (methylene chloride)
DIPEA Diisopropylethylamine
DMAD Dimethyl acetlyenedicarboxylate
DMAP 4-dimethylaminopyridine
DMF Dimethylformamide
DMSO Dimethyl sulfoxide
DNA Deoxyribonucleic acid
DPPF 1,1'-Bis(diphenylphosphino)ferrocene
ESI Electrospray ionization
Et N Triethylamine
3
EtOAc Ethyl acetate
EtOH Ethanol
FDA Food and Drug Administration
GA Geldanamycin
iii
h Hour(s)
H O Hydrogen peroxide
2 2
HCl Hydrochloric acid
HCS High content screen
HPLC High pressure liquid chromatography
Hsp 90 Heat shock protein 90
HTS High throughput screen
KOAc Potassium acetate
IM Infected macrophages
LCMS Liquid chromatography mass spectrometry
MeOH Methanol
min Minute(s)
ML Mucocutaneous leishmaniasis
mW microwave irradiation
NaOH Sodium hydroxide
NaOtBu Sodium tert-butoxide
NMR Nuclear magnetic spectroscopy
NTDs Neglected tropical diseases
Pyr Pyridine
Pd(OAc)2 Palladium(II) acetate

Pd (dba) Bis (dibenzylideneacetone)palladium (0)
2
Pd(dppf)Cl [1,1´-Bis(diphenylphosphino)ferrocene
2
Pd(PPh ) Tertakis(triphenylphosphine)-palladium (0)
3 4
iv
rt Room temperature
SAR Structure activity relationships
TBAF Tetrabutyl ammonium fluoride
tBuOK Potassium tert-butoxide
TEA Triethylamine
TFA Trifluoroacetic acid
THF Tetrahydrofuran
TLC Thin Layer Chromatography
USF University of South Florida
VL Visceral leishmaniasis
WHO World Health Organization
XA Xanthurenic acid

v
List of Tables

Table 2.1: Biological data of tetrahydro-indazole and tetrahydro-pyrazolo pyridinyl analogs .....32

vi