Cyclodextrin Scaffolds

CYCLODEXTRIN SCAFFOLDS DESIGNED FOR THE MOLECULAR
RECOGNITION OF ANIONS, CATIONS OR ORGANIC SUBSTRATES
by
Suhash Chandur Harwani
A thesis submitted in partial fulfillment

of the requirements for the Doctor of
Philosophy degree in Chemistry
in the Graduate College of
The University of Iowa
May 2008
Thesis Supervisor: Assistant Professor Jason R. Telford
UMI Number: 3323427

Copyright 2008 by
Harwani, Suhash Chandur
All rights reserved.
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Copyright by
SUHASH CHANDUR HARWANI
2008
All Rights Reserved
To my grandparents and parents

for all their sacrifices and hard work
Action expresses priorities.

Mohandas Ghandi
To My Friends:
If you live to be hundred, I want to live to be a hundred minus one day, so I
never have to live without you.
Winnie The Pooh
In everyone's life, at some time, our inner fire goes out. It is then burst into flame by an
encounter with another human being. We should all be thankful for those people who
rekindle the inner spirit.
Albert Schweitzer
iii
ACKNOWLEDGMENTS
First and foremost, I would like to thank my family for their unwavering support.
No matter how stubborn I can be, I thank my parents and brother for their continued
support. Professor Jason Telford has meant just as much to mein addition to allowing
me to do research in his group, he made sure that we did not lose sight of the important
things in life and always treated us like a part of his own family. I cannot imagine a more
understanding advisorno matter what happened, he was there to support us in our
pursuits. Both Professors Telford and Franklin provided a nurturing environment in
which I was taught to continue to strive and set higher goals. Without both of them I
would not be where I am today or the scientist that I am.
I have made numerous friends throughout graduate school, all of which have had
a profound impact in my life. Mary Daly who made sure I did not lose sight that there
was a world outside of the lab. Ramon Cuellar, a friend who was always there no matter
what time or how much he had on his plate. Nathan Lien whom made the days in lab
(and sometimes the yard ) more light-hearted.
Samantha Soebbing-Nolting ('my
partner in crime'don't fret the t-shirt is coming) was always there when I had a
questionno matter what pain I went through, we went through it together! Koushik
Banerjee for being there when I needed help with organic synthesis, making sure I got out
of the lab and could cook! Asif Rahaman always there to talk to and relax withhe was
not here during the early years of my graduate career, but he made sure I remembered
those days
only to be younger again (old age hits hard and fast you know! ). Sai
Kumar Ramadugu and Harsha Vardhan Reddy Annapureddy for making sure that I
remembered one of the most vital things, making sure we do not lose sight of ourselves
no matter how much there is to accomplishthe right balance of everything always
survives all odds.
iv
I would also like to thank the remaining members of the Franklin and Telford
groups who made lab life much more enjoyable: Dr. Yurndong (David) Jahng, Anthony
Prudden, Heaweon Park, Kinesha Harris, Sui Wah Wong-Deyrup, Oksana Chernyshev,
Chris Lim, and Sarah Brookhart Shields. I would also like to thank Dr. Lynn Teesch for
allowing me to be a research assistant in the HRMSF and, along with Vic Parcell, taught
me more than I would have otherwise learned about mass spectrometry.
In addition to fellow classmates, there have been many professors and support
staff that have taught me and helped through those rough moments of graduate school.
One of the most inspiring is my undergraduate advisor, Dr. Kenneth W. Olsen. No
matter what mistakes I made, he guided my early scientific mind and allowed me to
realize the potential enjoyment that comes from good science. Professor Ned Bowden
who taught me numerous techniques, the process of setting up a lab and always made me
feel like I had immeasurable potential. Professor Louis Messerle, who was always there
when I needed chatno matter how frustrated I was, he provided a soothing voice of
reason. Professors Pienta, Hansen, Pigge, Kohen, Meade, and, of course, Telford who
have helped mold my ability to teach students and ensure that I constantly improved at
teaching. Lastly, I would like to thank all support staffMichele Gerot for dealing with
all my teaching related concerns; Janet Kugley and Sharon Robertson who always
provided reminders/updates and for dealing with important administrative tasks;
Santhana Vellupillai and Rob Brown for their help with NMR experiments and analysis;
Frank Turner for all the favors fixing equipment or crafting things; Gene Hauge and Tim
Koon for their help whenever I needed to order something or could not find something;
and Nelson Reyes-Burgos for those late night chats.
Lastly, I would like to thank my past and present committee members (Professors
Eyman, Jensen, Goff, Kay, Messerle, Quinn, and Telford) for their constructive criticism
and not beating me over my head for all my idiosyncrasies when it came to noun and
verb agreement.
v
Without all of these people I would not be the person that I have become today.
Their influences will last with me and always guide me on my path to strive to be a better
scientist and, more importantly, a better person.
VI
ABSTRACT
The overarching theme reflected herein is the molecular recognition of organic
substrates or environmentally relevant anions and cations. The synthesis of modified
cyclodextrins as molecular scaffolds for the inner- or outer-sphere coordination of Ln m
cations or tetrahedral anions, respectively, is outlined.
Further studies on the
complexation of organic substrates with (3-cyclodextrin and a per-6-modified (3cyclodextrin are also reported.
The outer-sphere coordination of tetrahedral anions (specifically perchlorate,
phosphate, and sulfate) has been targeted. Over the past two decades numerous studies
utilizing ESI-MS for the determination of binding affinities have been reported. The
relatively soft-ionization by ESI-MS provides numerous advantages. Most importantly
are its abilities to model the solution phase closely and provide the unambiguous
stoichiometry of the complex(s) formed. Binding studies of a-cyclodextrin (1) and per-6(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin (6) with perchlorate, phosphate or
sulfate were examined using ESI-MS. The lack of specific binding of a-cyclodextrin is
evidenced with the inclusion of multiple perchlorate ions and competition for binding
with the cations (Li+, Mg2+, K+) of the particular tetrahedral anion. Contrary to this, per6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin (6) shows a preference for binding
anions from solution with modest log Ka values (~ 3.6 - 4.0), and predominantly a 1 : 1
(6 : Tetrahedral Anion) binding stoichiometry.
Further studies report the synthesis of carboxylic acid-functionalized per-6- and
mono-6-modified (3-cyclodextrins for the inner-sphere coordination of the lanthanides
and actinides.
Synthesis of per-6-5'-(thiosalicylic acid)-per-6-deoxy-P-cyclodextrin
exhibited solubility issues below a pH ~ 5. Consequently, potentiometric studies with

this cyclodextrin could not be performed. Additional studies of carboxylic acid mono-6modified P-cyclodextrins were pursued. The synthesis of mono-6-(2-(diethylenetriamine
vii
pentaacetic acid-amino)ethylamino)-mono-6-deoxy-P-cyclodextrin is reported, along

with analysis of binding with europium, terbium, and gadolinium using fluorescence and
ESI-MS.
The last body of this work details host and guest complexes formed between three
small organic molecules with native P-cyclodextrin (2) or per-6-(2-aminoethylamino)per-6-deoxy-(3-cyclodextrin (12). Determination of binding affinities of these three guest
molecules with these cyclodextrins are reported using UV-Vis spectroscopy.
vin
TABLE OF CONTENTS
LIST OF TABLES
xi
LIST OF FIGURES
xii
LIST OF ABBREVIATIONS
xix
CHAPTER
1
OUTER-SPHERE COORDINATION OF METAL COMPLEXES IN

BIOLOGICAL AND CYCLODEXTRIN-BASED SYSTEMS
1.1 Background
1.2 Natural Systems
1.2.1 Bacterial Iron Transport
1.2.2 Mammalian Antibacterial Proteins
1.2.3 Blue Copper Proteins
1.2.4 Recognition of Mg(H 2 0) 6 2+
1.3 Cyclodextrin-based Systems
1.3.1 Host-Guest Chemistry of Metal Complexes
1.3.2 Waste remediation
1.4 Catalysis
1.4.1 Horseradish Peroxidase (HRP) Mimic
1.4.2 Water-soluble Phosphane with Cyclodextrin
1.5 Future Outlook
Notes
1
2
2
3
4
6
7
8
11
12
13
14
14
17
ANION RECOGNITION USING A CYCLODEXTRIN-BASED

SCAFFOLD
21
2.1 Introduction
2.1.1 Oxoanion Recognition Chemistry
2.1.2 Outer-sphere Coordination Using Cyclodextrins
2.1.3 Tetrahedral Anions
2.1.4 Determination of Binding Affinities
2.2 Experimental
2.2.1 ESI-MS
2.2.2 Synthesis
2.2.3 Titrations
2.2.4 Calculation of Binding Affinities
2.2.5 pKa Determination
2.2.6 T\ Inversion Recovery Experiments
2.3 Results and Discussion
2.3.1 Binding mode of Tetrahedral Anions
2.4 Conclusions
Notes
21
21
21
23
23
24
24
25
25
26
27
27
28
35
36
40
CATION RECOGNITION USING ACID-MODIFIED

CYCLODEXTRIN-BASED SCAFFOLDS
42
3.1 Introduction
42
IX
3.1.1 Cation Recognition..

3.1.2 Nuclear Waste Refinement
3.1.3 Lanthanides and Actinides
3.1.4 Supramolecular Host and Choice of Functional Group
3.2 Experimental
3.2.1 Synthesis
3.2.2 Fluorescence Measurements
3.2.3 Mass Spectrometry
3.3.1 Synthesis
3.3.2 Fluorescence
3.4 Conclusions
3.5 Future Work
Notes
42
44
45
46
50
50
62
63
75
75
78
79
80
80
82
ORGANIC SUBSTRATES BINDING WITH NATIVE AND

MODIFIED (3-CYCLODEXTRINS.....
85
4.1 Introduction
4.2 Experimental
4.2.1 Synthesis
4.2.2 UV-Vis Spectroscopy
4.2.3 Crystallization
4.2.4 ESI-MS Measurements
4.4 Conclusions
Notes
85
85
86
87
88
.88
96
97
98
SUMMARY AND FUTURE WORK
99
APPENDIX A DETERMINATION OF BINDING CONSTANTS USING ESIMS
101
A.l Importance and Background of Binding Affinities

A.2 Utility of ESI-MS in Determining Binding Affinities
Notes
102
103
106
APPENDIX B SPECTRA AND RELEVANT DATA FOR THE BINDING OF

TETRAHEDRAL ANIONS WITH 1 AND 6
107
APPENDIX C SPECTRA: MODIFIED CYLODEXTRINS
140
REFERENCES
167
LIST OF TABLES
Table
2.1
Properties of Tetrahedral Anions Studied with Cyclodextrin 6
30
2.2
Sample Calculations for the Titration of 6 with LiC104
31
2.3
Delay Time Prior to ^ - N M R Spectrum Collection for Experiments

Performed to Determine T\ Relaxation Time Perturbations
38
Experimentally Calculated T\ Relaxation Times (sec) for !H-NMR Peaks of

Cyclodextrin 6 Titrated with LiC104
39
3.1
Ionic Radii of the Lanthanides (A)
49
4.1
Log Ka of Guest Molecules with Cyclodextrins 2 or 12
90
B.l
ESI-MS Intensities of Host (IH) and Host / Guest Complex (7#G) for 6 Titrated
withLiC10 4
133
B.2 ESI-MS Intensities of Host (///) and Host / Guest Complex (IHG) for 6 Titrated
withMgS0 4
134
B.3 ESI-MS Intensities of Host (7#) and Host / Guest Complex (IHG) for 6 Titrated
withKH 2 P0 4
135
B.4 Relative JH-NMR Peak Heights for peaks of interest of per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin(6)
136
B.5 Relative 'H-NMR Peak Heights for peaks of interest of per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin (6) titrated with LiC104 (1 :
1.2)
137
B.6 Relative ^ - N M R Peak Heights for peaks of interest of per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin (6) titrated with LiC104 (1 :
4)
138
2.4
B.7 Relative 'H-NMR Peak Heights for peaks of interest of per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin (6) titrated with LiC104 (1 : 8) ....139
XI
LIST OF FIGURES
Figure
1.1
Diagram of (A) the export of a siderophore from bacterial cell, (B)

complexation with Fe +, (C) uptake of the siderophore-Fe3+ complex by an
outer membrane cell surface receptor. (OM = outer membrane, IM = inner
membrane)
1.2
A view of the binding site of Fhu showing hydrogen bonding interactions to

the octahedral iron center (PDB Accession Code: 1QFF)
1.3
Representations of a-, |3-, and y-cyclodextrins. The top structure represents

the full drawn chemical structures of a-, P-, and y-cyclodextrins (b = 1, 2, or
3, respectively). The lower center and right structures are shorthand
representations of the a-, (3-, and y-cyclodextrins (n = 6, 7 or 8, respectively).
Lastly, the lower left-most representation is a cartoon of toroidal shape. The
dispositions of the primary and secondary hydroxyls are presented. The
edges ringed with 1 and 2 hydroxyls are referred to as the lower and upper
rims, respectively
10
Representation of the inclusion of a guest molecule within the cyclodextrin

cavity. Guest molecules could include organic molecules, inorganic metal
complexes, cations, or anions
11
(a) 3-D model of uranyl carbonate showing its D^ symmetry and available
hydrogen bond acceptors (carbon = gray, oxygen = red, uranium = yellow)
(b) a portion of the ^ - N M R spectrum of per-6-(2-aminoethylamino)-per-6deoxy-ot-cyclodextrin titrated with uranyl carbonate shows the clear upfield
shift of proton resonances near the binding site
13
(a) Fe m 2-hydroxy-l-naphthaldehyde thiosemicarbazone (HNT) Schiff-base

complex. This figure shows a three-coordinate Fe; however, the structure is
actually either a square pyramidal or octahedral geometry which utilizes
another HNT ligand to satisfy the additional bonding sites, (b) Representation
of the Schiff-base salicylidene-2-amino-4-phenylthiazole complex (Mn(SAPTS)2)
15
Phosphane ligand for complexation with a metal ion (e.g. Pd[4]3) and
inclusion into the cyclodextrin cavity
16
1.4
1.5
1.6
1.7
2.1
Synthetic transformation for a-cyclodextrin (1) to per-6-iodo-a-cyclodextrin

(5). Removal of the iodides via nucleophilic attack by either of the free
terminal amines of ethylenediamine results in the final per-6-(2-
2.2
aminoethylamino)-per-6-deoxy-a-cyclodextrin (6) product
22
pH-dependent speciation diagram based on estimated P values for the hexaprotonated per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin (6)
32
xn
2.3
2.4
3.1
3.2
3.3
3.4
3.5
3.6
Calculated T\ relaxation times in graphical format for protons of interest in

the 'H-NMR spectrum. The only significant deviation greater than 10% is
seen at 8 3.61 ppm. The yellow line indicates a perturbation greater than 10%
at 5 4.01 ppm; however, no deviation is seen at 8 4.01 ppm with 8 equivalents
of perchlorate indicating that its relaxation time is most likely unaffected by
the perchlorate
33
Generic representation of a tetrahedral anion (TA = C104",SCV",orPCV")

binding to the upper-rim of per-6-(2-aminoethylamino)-per-6-deoxy-acyclodextrin. Depending on the tetrahedral anion binding and charge state of
the species in MS, the charge of the final species in the gas phase will vary,
therefore n would indicate the appropriate charge to get the overall charge on
the species and would be the charge of the each respective tetrahedral anion.
The binding mode seen in this diagram is only assumed as it provides the
most likely electrostatic interaction
34
Structures of 12-crown-4 and 18-crown-6 which are known to bind cations K+

and Na+, respectively
43
Relation between ionic radius and binding free energy (AG) for the trivalent
lanthanide species with the peptide from Imperiali et. al. The table included
provides Kd values for each trivalent lanthanide and their peptide
44
7. Representation of a calix[n]arene with 4-8 monomers. 8. Repesentation of

carbamoylmethylphosphine oxide. 9/10. Representations of calix[4]arene
modified with a CMPO derivative and with the lowers-methyl group also
modified
46
The decrease ('lanthanide contraction') in the ionic radii of the Ln species is

apparent from the above graphical representation where as Z increases, the
radius decreases
48
A generic representation of (a) per-6-modified P-cyclodextrins and (b) mono6-modified P-cyclodextrins. In unmodified natural P-cyclodextrin R = -OH
48
Reaction sequences for the synthesis of per-6-modified P-cyclodextrins (11 15). The arrow for the synthetic route of 12 to 15 is marked with an 'X'
through it to denote that the reaction did not generate the desired product
56
3.7
Synthesis of 16
57
3.8
Syntheses of 17 and 18
58
3.9
Synthesis of 19
59
3.10 Synthesis of 20
3.11 Representations of the functional groups attached to the per-6- or mono-6modified P-cyclodextrins. R is the location where the cyclodextrin group is
attached at C6. For the per-6-modified p-cyclodextrins each of the C6
positions is modified with the labeled group; whereas, for the mono-6modified P-cyclodextrins only one of the seven C6 positions is modified with
the labeled group. Each position for a C-H bond is labeled for ease of NMR
assignments for the ^ - N M R listed in the characterization of each synthesis
xiii
60
61
3.12 Titration of Eu3+ with up to 5 equivalents of 19. The increase in the

fluorescence emission at 615 nm is a result of the exclusion of water from the
first coordination sphere of the Eu +
65
3.13 Relative fluorescence of Eu3+ when titrated with 18, one of the starting
materials for the synthesis of 15. Importantly, there is no increase in the
fluorescence intensity at 615 nm as seen in Figure 3.12
66
3.14 Titration of Tb with up to 5 equivalents of 19. The increase in the

fluorescence emission at 545 nm is a result of the exclusion of water from the
first coordination sphere of the Tb3+
67
3.15 Titration of Tb3+ with up to 5 equivalents of 18. Importantly, there is no

increase in the fluorescence intensity at 545 nm as seen in Figure 3.14
68
3.16 Binding curve generated from fluorescence emission at 615 nm ofEu3+

titrated with 19 is provided in black. The calculated fluorescence emission
based upon fitting to equation 3.1 for determination of Kd is provided in
orange
69
3.17 Binding curve generated from fluorescence emission at 545 nm of Tb

titrated with 19 is provided in black. The calculated fluorescence emission
based upon fitting to equation 3.1 for determination of K<j is provided in
orange
70
3.18 Negative ESI-MS spectrum for the ligand 19. m/z 1533 representative of
[semi-anhydride 19 - H+]"
71
3.19 Positive ESI-MS spectrum for the complexation of Gd + by ligand 19 (ratio

19 / Gd3+ of 1 : 1). m/z 1711 representative of [semi-anhydride 19 - 3H+ +
158
Gd3+ + Na + ] +
,
72
3.20 Positive ESI-MS spectrum for the complexation of Gd3+ by ligand 19 (ratio
19 / Gd3+ of 1 : 3). m/z 1711 representative of rsemi-anhydride 19 - 3H+ +
158
Gd3+ + Na + ] +
73
3.21 Positive ESI-MS spectrum for the complexation of Tb + by ligand 19 (ratio 19

/ Tb3+ of 1 : 1). m/z 1712 representative of [semi-anhydride 19 - 3H+ + Tb3+
+ Na + ] +
74
3.22 Representation of the structure of DTPA dianhydride after nucleophilic attack

by the free amine from either 12 or 18 and hydrolysis of the second anhydride
(15 or 19) and a representation of diethylene triamine pentaacetic acid
dianhydride (DTPA dianhydride, 21). In structure 15 or 19, R is where an
ethylenediamine arm on the modified P-cyclodextrin (either 12 or 18) is
4.1
4.2
attached to the DTPA
77
Structure of guest molecules (22 - 2,3-diaminonaphthalene, 23 - 2,3dihydroxyquinoxaline, 24 - 3 -hydroxy-2-naphthoic acid, 25 - N - ( l pyridylmethyl)-2-aminobenzoic acid, 26 - 2,3-dihydroxynaphthalene) studied
for interaction with native and functionalized P-cyclodextrins
86
UV-Vis absorbance of 22 titrated with 2 from 265 nm - 385 nm. No major

absorbance shifts are noted
91
xiv
4.3
4.4
4.5
4.6
B.l
UV-Vis absorbance of 23 titrated with 2 from 265 nm - 385 nm. No major

absorbance shifts are noted
92
UV-Vis absorbance of 25 titrated with 2 from 265 nm - 385 nm. A blue shift
in the absorption maximum at 340 nm to 332 nm is noted
93
UV-Vis absorbance of 25 titrated with 12 from 265 n m - 385 nm. A blue

shift in the absorption maximum at 340 nm to 332 nm is noted
94
Negative ESI-MS for the analysis of binding of 25 with P-cyclodextrin (2).

At a ratio of 1 : 1 (25 / 2), it is evident that there are both the free forms of 2
and 25 predominant in solution
95
ESI mass spectrum for 1, a-cyclodextrin
108
B.2 ESI mass spectrum for 1, a-cyclodextrin titrated with LiC104 (1 : 3)
109
B.3 ESI mass spectrum for 1, a-cyclodextrin titrated with MgSC>4 (1 : 3)
110
B.4 ESI mass spectrum for 1, a-cyclodextrin titrated with KH2PO4 (1 : 7)
111
B.5 ESI mass spectrum for 6, per-6-(2-aminoethylamino)-per-6-deoxy-acyclodextrin
112
B.6 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin withLiClO4(10 : 1)
113
B.7 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with LiC104 (1 : 1)
114
115
116
B.10 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with MgS04 (10 : 1)
117
B.l 1 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with MgS04 (1 : 1)
118
B.12 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with MgS04(1 : 5)
119
B.l3 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6-
deoxy-a-cyclodextrin with MgS04 (1 : 10)
120
B.14 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrinwithKH2PO4(10 : 1)
121
B.l5 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with KH2PO4 (1 : 1)
122
xv
B.16 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with KH2PO4 (1 : 5)
123
B.17 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with KH 2 P0 4 (1 : 10)
124
B. 18 Linear regression attained by inputing data from the first titration of 6 with
LiC104 to equation A.6. The slope of the line is then used to generate the Kd
value as outlined in Appendix A
125
B.19 Linear regression attained by inputing data from the second titration of 6 with
LiC104 to equation A.6. The slope of the line is then used to generate the Kd
126
B.20 Linear regression attained by inputing data from the third titration of 6 with
LiC104 to equation A. 6. The slope of the line is then used to generate the Kd
127
B.21 Linear regression attained by inputing data from the first titration of 6 with
MgS04 to equation A. 6. The slope of the line is then used to generate the Kd
128
MgSC>4 to equation A.6. The slope of the line is then used to generate the Kd
129
B.23 Linear regression attained by inputing data from the first titration of 6 with
KH2PO4 to equation A.6. The slope of the line is then used to generate the Kd
130
131
B.25 Linear regression attained by inputing data from the third titration of 6 with
132
C.l
Positive ESI-MS spectrum for 6, per-6-(2-aminoethylamino)-per-6-deoxy-acyclodextrin
C.2 'H-NMR spectrum for 6, per-6-(2-aminoethylamino)-per-6-deoxy-acyclodextrin
141
142
C.3 Positive ESI-MS spectrum for 12, per-6-(2-aminoethylamino)-per-6-deoxy-(3cyclodextrin
143
C.4 ^ - N M R spectrum of for 12, per-6-(2-aminoethylamino)-per-6-deoxy-Pcyclodextrin
144
C.5 Positive ESI-MS spectrum of 14, per-6-5'-(thiosalicylic acid)-per-6-deoxy-Pcyclodextrin ([14 + K + ] + = 2125 m/z) along with breakdown products
145
xvi
C.6 ^ - N M R spectrum from 8 6.2 - 8.4 ppm of 14, per-6-5'-(thiosalicylic acid)per-6-deoxy-P-cyclodextrin
146
C.7 ^ - N M R spectrum from 8 2.0 - 5.3 ppm of 14, per-6-5'-(thiosalicylic acid)per-6-deoxy-(3-cyclodextrin
147
C.8 Positive ESI-MS spectrum for 16, mono-6-tosyl-mono-6-deoxy-Pcyclodextrin
148
C.9 'H-NMR spectrum from 8 4.5 - 8.6 ppm of 16, mono-6-tosyl-mono-6-deoxyP-cyclodextrin
149
CIO ^ - N M R spectrum from 8 1.5 - 4.2 ppm of 16, mono-6-tosyl-mono-6-deoxyP-cyclodextrin
150
C.l 1 Positive ESI-MS spectrum of 17, mono-6-iV-(p-aminonicotinic acid)-mono-6deoxy-(3-cyclodextrin
151
C.12 ^ - N M R spectrum from 8 5.9 - 9.0 ppm of 17, mono-6-JV-(p-aminonicotinic

acid)-mono-6-deoxy-P-cyclodextrin
152
C.13 ^ - N M R spectrum from 8 3.2 - 5.3 ppm of 17, mono-6-A^-(p-aminonicotinic

acid)-mono-6-deoxy-p-cyclodextrin
153
C.14 ^ ^ H COSY of 17, mono-6-JV-(p-aminonicotinic acid)-mono-6-deoxy-mono6-deoxy-P-cyclodextrin
154
C.15 'H-^CHMQCofH, mono-6-iV-(p-aminonicotinicacid)-mono-6-deoxy-Pcyclodextrin
155
C.l6 ESI-MS spectrum of 18, mono-6-(2-aminoethylamino)-mono-6-deoxy-Pcyclodextrin
156
C.l7 !H-NMR spectrum of 18, mono-6-(2-aminoethylamino)-mono-6-deoxy-Pcyclodextrin
157
C.l8 Negative ESI-MS spectrum of 19, mono-6-(2-(diethylenetriamine pentaacetic

acid-amino)ethylamino)-mono-6-deoxy-P-cyclodextrin
158
C.l9 ' H - N M R o m mono- mono-6-(2-(diethylenetriamine pentaacetic acidamino)ethylamino)-mono-6-deoxy-P-cyclodextrin
159
C.20 ^ H COSY of 19, mono-6-(2-(diethylenetriamine pentaacetic acidamino)ethylamino)-mono-6-deoxy-P-cyclodextrin.
160
C.21 ^ - ^ C H M Q C o f l ^ mono-6-(2-(diethylenetriamine pentaacetic acid-
amino)ethylamino)-mono-6-deoxy-P-cyclodextrin
161
C.22 Positive ESI-MS spectrum of 20, mono-6-(2-(dansylamino)ethylamino)mono-6-deoxy-P-cyclodextrin

;
162
C.23 ^ - N M R spectrum from 8 7.1 - 9.2 ppm of 20, mono-6-(2(dansylamino)ethylamino)-mono-6-deoxy-P-cyclodextrin
163
xvii
C.24 ^ - N M R spectrum from 8 2.3 - 5.5 ppm of 20, mono-6-(2(dansylamino)ethylamino)-mono-6-deoxy-P-cyclodextrin
164
C.25 ^ H COSY of 20, mono-6-(2-(dansylamino)ethylamino)-mono-6-deoxy-pcyclodextrin
165
C.26 'H-13C HMQC of 20, mono mono-6-(2-(dansylamino)ethylamino)-mono-6deoxy-P-cyclodextrin
166
xvm
LIST OF ABBREVIATIONS
ACN
Acetonitrile, CH3CN
CMPO
Carbamoylmethylphosphine oxide
COSY
Correlation spectroscopy
ddH 2 0
Distilled-deionized water
DEAE
2-(diethylamino)ethyl
DTPA
Diethylene triamine pentaacetic acid
ESI
Electrospray ionization
ESI-MS
Electrospray ionization mass spectrometry
Fhu
Ferric hydroxamate uptake
FRET
Fluorescence resonance energy transfer
HEPES
4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid
HMQC
Heteronuclear multiple quantum coherence
IROMP
Iron responsive outer-membrane protein
/PrOH
Isopropanol
MeOH
Methanol
MS
Mass spectrometry
MWCO
Molecular weight cut off
NGAL
Neutrophil gelatinase-associated lipocalin
NMR
Nuclear magnetic resonance
PBP
Phosphate binding protein
PUREX
Plutonium and uranium recovery extraction
TC
To-contain
TRLFS
Time-resolved laser fluorescence spectroscopy
CHAPTER 1
OUTER-SPHERE COORDINATION OF METAL COMPLEXES IN
BIOLOGICAL AND CYCLODEXTRIN-BASED SYSTEMS
1.1 Background
This background text has been adapted from the following publication: "Outersphere Coordination of Metal Complexes in Biological and Cyclodextrin Systems",
Harwani, S.; Telford, J. R. Chemtracts, 2005,18(8), 437-448.
The use of both inner- and outer-sphere coordination play fundamental roles in
metal-ion reactivity.1"3 Inner-sphere coordination (the first coordination sphere) refers to
the ligands directly bound to a metal; in Nature, this is most often realized by amino acids
and a few other cofactors (i.e. ligands). The inner coordination sphere around metals
have been studied extensively over time; although most instances of metal-bound systems
characterized to date fall within the realm of supra-molecular and molecular recognition
using outer-sphere coordination. Outer-sphere coordination (second coordination sphere)
specifically refers to those interactions outside the inner or primary coordination sphere.
Examples include ordered or disordered solvent and amino acid residues which utilize
hydrogen bonding or dipolar interactions with ligand atoms.
In protein systems, a
receptor provides a binding site with a spatial arrangement of hydrogen bonds or

hydrophobic patches to target a specific substrate. The interactions employed to bind a
ligand in natural protein systems consist mainly of (1) hydrogen bonds, (2) TC-U and other
donor-acceptor interactions, (3) electrostatic interactions, and (4) bonding from the direct
overlap of metal orbitals with outer-sphere ligands.1"4
In addition to naturally occurring systems there are a number of synthetic
achievements in the form of supra-molecular scaffolds designed to bind metal complexes.
These scaffolds have been applied to a variety of tasksmainly in the areas of enzyme
mimics, medicinal chemistry, and environmental waste remediation. Utilizing many of
the same interactions as natural systems, much effort has been directed towards
mimicking biological systems using supramolecular scaffolds such as cyclodextrins.
This review focuses on some recent developments in outer-sphere coordination chemistry
with an emphasis on: (1) natural systems and (2) cyclodextrin-based systems.
1.2
Natural Systems
1.2.1 Bacterial Iron Transport

Recognition, transport, and assimilation of nutrients are essential for bacteria to
satisfy their growth requirements.5'6
This is often accomplished by outer-sphere
recognition of the nutrient substrate. Of specific interest are siderophores, small, ironbinding compounds produced by bacteria, which serve as a paradigm for micro-nutrient
acquisition. Siderophore transport and internalization also provides an example of outersphere recognition of metal complexes (Figure 1.1).
Siderophores play an important role in bacterial and fungal life cycles.7"11 An
important step in the iron uptake pathway is the internalization of the Fe-siderophore
complex. This is accomplished by a variety of outer membrane transport proteins. These
transport proteins are expressed under iron limiting condition, and therefore, are
generally classified as iron responsive outer-membrane proteins (IROMP). IROMPs are
either exceedingly specific or generic in their recognition of substrate. Specific IROMPs
may recognize one or a few substrates, while generic IROMPs recognize a wide variety
of a class of siderophores, such as any iron-tris-hydroxamate complex. This variation in
substrate recognition is one of several ways that bacteria compete for the multitude of
siderophore architectures.
The ferric hydroxamate uptake (Fhu) protein is an IROMP that generically
transports iron-bound hydroxamate siderophores. Recognition is effected by hydrogen
bonding contacts with the octahedral metal center (Figure 1.2) rather than contacts with
the organic siderophore backbone.
The Fhu protein is therefore used as an example for
the general phenomenon of iron uptake, and an interesting example of metal-centric

recognition. It is only after the binding of Fe-siderophore by Fhu that uptake into the cell
can occur.
Figure 1.1 Diagram of (A) the export of a siderophore from bacterial cell, (B)
complexation with Fe 3+ , (C) uptake of the siderophore-Fe3+ complex by an outer
membrane cell surface receptor. (OM = outer membrane, IM = inner membrane).
1.2.2 Mammalian Antibacterial Proteins

Another example of iron-dependent recognition proteins is the lipocalin binding
proteins; however, lipocalins perform a very different function by withholding the Fesiderophore complex from the bacteria. In essence, lipocalins are beating bacteria at their
own game of iron acquisition.
One member of the lipocalin family, neutrophil
gelatinase-associated lipocalin (NGAL), has been implicated in numerous cellular

processes13"15, such as providing an Fe recognition system.
Recent studies have
elucidated the x-ray crystal structure of the protein.16"18 NGAL contains an unusually
shallow and broad binding pocket (calyx) lined with polar and positively charged
residues.
The NGAL calyx has an affinity for small hydrophobic molecules.
In
particular, it has recently been shown that the NGAL calyx has a very high affinity for
Fe-enterobactin (a cyclotriserine-based siderophore), binding and stabilizing the
siderophore from hydrolytic degradation into its monomeric building blocks.19'20
Experimental studies of various substrates binding to the NGAL calyx indicate that the
positive charge and polarity of the residues in the binding pocket are critical for
binding. '
This has also been revealed through additional studies which demonstrate
that NGAL does not bind positively charged iron alone ([Fe(H20)6]3+), suggesting an
electrostatic contribution to the interaction between the negatively charged Feenterobactin complex and the positively charged NGAL calyx.
Overall, it has been
proposed that NGAL and other lipocalins are bacteriostatic agents used by the immune
system in order to fight off bacterial infections more efficiently by removing a vital
bacterial iron source. Other similar mechanisms for iron homeostasis in the body, such as
transferrin and hepicidin-dependent uptake mechanisms (along with NGAL) are
described in a review by Kaplan.21
1.2.3 Blue Copper Proteins
Outer-sphere influences are seen not only through the interaction of two unique
molecules (host and guest), but also intramolecularly through the influences that protein
backbones have on the coordination of metal complexes, as observed in blue copper
proteins. Crystallographic studies of azurin
and plastocyanin
show that the backbone
of the protein is preorganized to bind the copper ion, with halo forms essentially identical
to their respective apo forms. This preorganized structural motif is seen with many blue
copper proteins.
In pseudoazurin, the redox potential of the copper ion is sensitive to an extended

array of amino acid residues. One of the first mutational studies altered the proline (P80)
adjacent to the metal center to alanine or isoleucine. These mutations resulted in an
increase in reduction potential, as the adjacent histidine is not as rigidly enforced in

position nor is it as solvent exposed.24
Recently published data with mutations on
amicyanin give similar results. Mutations were constructed in the binding loop because
these are thought to relax the ligation around Cu1 allowing it to assume its preferred
geometry and thus, increase the reduction potential, .25 The two mutants studied, P94A
and P94F, have a shift in reduction potential by +100 mV, from = 265 mV to 380 mV
and 415 mV, respectively 25
Figure 1.2 A view of the binding site of Fhu showing hydrogen bonding interactions to
the octahedral iron center (PDB Accession Code: 1QFF).1
This interpretation is further corroborated by quantum mechanical calculations by

Li et. al. Their calculations indicate that the dominant influences on the copper reduction
potential in type I blue copper proteins are residues within 6A of the copper center(s) that
contribute H-bonding.26 Their studies also show influences by three key factors: (1) axial
ligand interactions, (2) hydrogen bonding to Scys, and (3) constraint of the inner-sphere
ligand orientation via the protein backbone. These results, along with previous studies,
indicate that the hydrogen bonding network indirectly influences the geometry around the
copper in the active-site(s) and thereby controls the physical properties of the metal. The
experimental data from mutations in the binding loop of the blue copper proteins coupled
with the quantum mechanical data provide additional support that the electronic
properties of the copper center(s) are modulated by the inner-most H-bonding array to the
copper ligands.
1.2.4 Recognition of Mg(H20)62+
Many natural biological systems require the use of [Mg(H20)6]2+ for stability and
or catalytic activity of enzymes. A survey of the Cambridge Structural Database (CSD)
done in 1994 by Bock et. al. revealed the stability of hexaaquamagnesium(II) in many
crystal structures.27
Systems such as topoisomerases, ribonucleases, and bacterial
transport systems, are known to include Mg2+(fli?) as a cofactor.
Magnesium has poorly
defined requirements for coordination, and it is not clear how the ion is recognized or
coordinated in biological systems. In order to probe how Mg +(a?) is coordinated, the
inert hexammine cobalt(III) complex, [Co(NH3)g]3+, has been utilized as a surrogate.
[Co(NH3)6] 3+ is thought to bind in proteins via outer-sphere coordination, because the
amine ligands are kinetically inert, and not readily displaced. Thus, binding to the
complex must occur through the amine ligands, rather than directly to the metal.
Analysis of proteins with [Co(NHs)6]3+ bound to Mg2+(a?) sites suggests that
hexaaquamagnesium(II) also binds via an outer-sphere interaction with the aqua

ligands.28"32
Even though much work has been done in this area, novel methods of studying
the outer-sphere nature of aqueous cations are still being developed.
Computational
studies, along with infrared and Raman spectroscopies, have been used to probe the
binding modes of Mg 2+ and Ca2+ with dimethyl phosphate anion (DMP")an anion used
to mimic the phosphate moiety. These studies indicate that the calcium cationic center
prefers at least one inner-sphere contact with a hard oxygen donor from DMP", while the
magnesium cation prefers outer-sphere binding through the aqua ligands.33
1.3 Cyclodextrin-based Systems
Cyclodextrins (Figure 1.3) can be used in the recognition of different molecules
(or complexes) and have been proposed for a variety of possible applications.
Cyclodextrins are polymeric glucose molecules consisting of 6, 7, or 8 a-(l->4) linked
glucose moieties, known as a-, P-, and y-cyclodextrin, respectively.34'35 The diameter of
the cyclodextrin cavity increases as the number of glucose residues increases from a-, p \
and y-cyclodextrin, with cavity diameters of-5.2, ~6.6, ~8.4 A, respectively. Many areas
exploiting the utility of cyclodextrins have been targeted, including both their host-guest
chemistry and synthetic methods for functionalizing the upper and lower rims of
cyclodextrins.
Host-guest chemistry of cyclodextrins has been studied extensively for the
complexation of hydrophobic molecules for applications, such as drug delivery, chiral
recognition, enzyme mimics, and molecule detection, including secondary detection
methods such as fluorescence enhancement or quenching. "
In order to selectively
complex molecules and increase the solubility of compounds, native cyclodextrins are not
used often due to their modest solubility.
The synthesis of modified cyclodextrins,
therefore, has received much deserved attention. The ability to functionalize one or all of
the primary hydroxyl groups on the upper rim and the secondary hydroxyl groups on the
lower rim of cyclodextrin molecules allows for the development of a wide range of
possible outer-sphere hosts or enzyme mimics, The multitude of possible hosts presents
the ability to target the complexation of certain metal ions or substrates.
Host-guest
chemistry and the synthetic methods for the modification of the upper and lower rims of
cyclodextrin will not be discussed as there are many review articles in these areas.42"45
Cyclodextrin-based chemistry has developed quite extensively since its discovery
in the late nineteenth and early twentieth centuries. Initial work focusing on outer-sphere
coordination of metal complexes with cyclodextrins was done during the middle-to-late
1980's.46"52 Part of the focus in the remaining sections will be recent progress using
cyclodextrins as outer-sphere host architectures.
In addition to the complexation studies, work with cyclodextrins has focused on
their use as catalysts. Much of the research in this area has centered on design of the
cyclodextrin as the catalytic center.
A summary of this area is presented in review
articles by Stoddart et. al, Szejtli, et. al, and Russell, et. a/.3'53'54 Less work has been
done exploring cyclodextrin as an architectural structure to position a catalytically active
metal site. We will present three examples of research that focuses on using cyclodextrin
as a scaffold on which to append a metal-binding center.
1.3.1 Host-Guest Chemistry of Metal Complexes
The ability of cyclodextrins to incorporate molecules within its cavity (Figure
1.4) has been studied extensively; however, analysis of these host-guest complexes
reveals only a few structures with the inclusion of a metal complex. This is surprising as
the inclusion of metal complexes can provide the versatility to mimic biological
enzymes
and create supramolecular assemblies .
A minimalist example of host-guest relationship is provided by Ando et. al.,

wherein
they
were
able
to
study
the
inclusion
complexes
of
[pentaammineruthenium(II)]2+ and [pentacyanoferrate(II)]4" with a-,

cyclodextrin.56
P-, and y-
It is significant to report that it is rare for a charged (specifically,
positively charged) molecule to be included within the cyclodextrin.
This was
accomplished by using the inner sphere ligands, which are hydrophobic and provide the
anchor in the core of the cyclodextrin. The steric bulk of the outer-sphere cyclodextrin
serves to stabilize the monomeric metal center.
10
OH
Lx
HO
yp:<^>y^,
~lS
OH
\K
H
i
^-S
[\|
OH/
I
.>AAoH
\ 0
] (ÔH
HO^
OH-y
\
HO
b = 1(1), 2(2), or 3(3)
OH
upper rim
l-OH
/v.
OH
Â
/ J
lower rim
2-OH
\
\
[i
7/
r /^
/A ^
c 3 v
V
HO-\ ^
OH
ô,
If
*>A
M/l
/n
n = 6, 7 or 8
Figure 1.3 Representations of a-, p-, and y-cyclodextrins. The top structure represents
the full drawn chemical structures of a-, (3-, and y-cyclodextrins (b = 1, 2, or 3,
respectively). The lower center and right structures are shorthand representations of the
a-, P-, and y-cyclodextrins (n = 6, 7 or 8, respectively). Lastly, the lower left-most
representation is a cartoon of toroidal shape. The dispositions of the primary and
secondary hydroxyls are presented. The edges ringed with 1 and 2 hydroxyls are
referred to as the lower and upper rims, respectively.
11
+ Guest
Figure 1.4 Representation of the inclusion of a guest molecule within the cyclodextrin
cavity. Guest molecules could include organic molecules, inorganic metal complexes,
cations, or anions.
1.3.2 Waste remediation

Our group has focused on the remediation of environmental metal complexes,
including the uranyl carbonate anion, [U02(C03)3]4" (Figure 1.5). Uranyl carbonate is
the predominant form of uranium in the environment. The carbonate anion is a powerful
ligand for the Lewis acidic UVI center, and carbonates' ubiquitous presence drives the
formation of the tris-carbonate species in most aquatic systems. It has been shown that
the ligands in uranyl carbonate are not easily displacedthe uranyl tris-carbonato species
has a stability constant, Poo = 10 . "
We have been able to complex uranyl carbonate
by providing a series of complementary hydrogen bond donors via synthetic

modifications to the upper rim of cyclodextrin molecules. In designing a host structure
for uranyl carbonate, both the D^ symmetry and charge of the guest must be considered
as part of the design. One of the first synthetic compounds used as a host was per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin.60
This compound matches the 3-fold
symmetry of [U02(C03)3]4" while providing a pH-dependent ring of positive charges

from protonation of the amines. NMR titrations of per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with uranyl carbonate allowed for determination of the stability
constant (log p = 253 22 M"1) of the cyclodextrin-uranyl carbonate complex in an
12
aqueous environment. It has been shown through complexation induced shifts of the
ethylenediamine protons that the uranyl carbonate moiety is binding through the upperrim.61,62
It was also shown that the complex formed was 1:1 between the synthetic
cyclodextrin and uranyl carbonate anion.

Additional work exploring the effects of outer-sphere ligation of the uranyl
species has been published by Navaza et. al.63 Although the uranyl hydroxide systems
generally form bimetallic complexes, Navaza's work illustrates the ability of a- and pcyclodextrin to stabilize the formation of monomeric benzoatoOHUO2 species within
dimeric cyclodextrins. The mode of binding of the benzoato-OHUO2 species, like the
work by Ando, is again driven by the interaction of ligand and cyclodextrin with
inclusion of the hydrophobic benzoate ligand in the cavity of the p-cyclodextrin dimer.63
1.4 Catalysis
In catalytic processes, it is important to make the distinction between
heterogeneous and homogeneous catalysis.
Typically, the main advantage of
heterogeneous catalysis is that the catalyst can be recycled more easily, and is therefore
more
cost
efficient;
however,
both
cyclodextrin-mediated
homogeneous
and
heterogeneous catalysis historically have not been an area of great focus. In addition to
homogeneous and heterogeneous systems, some work has been done in biphasic systems
where the cyclodextrin acts as an integral part of the catalytic system.64"66 In many cases
of cyclodextrin-mediated catalysis in biphasic systems, the cyclodextrin aids by acting as
a phase transfer agent. Many experimental results have been published with the use of
cyclodextrins in biphasic systems66"69; however, the work represented here will focus on
the use of outer-sphere coordination in homogeneous systems.
13
Figure 1.5 (a) 3-D model of uranyl carbonate showing its D^ symmetry and available
hydrogen bond acceptors (carbon = gray, oxygen = red, uranium = yellow) (b) a portion
of the H-NMR spectrum of per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin
titrated with uranyl carbonate shows the clear upfield shift of proton resonances near the
binding site.
1.4.1 Horseradish Peroxidase (HRP) Mimic

Occasionally, a significant advance can be developed from seemingly unrelated
research areas. It was established in the late 1970's by Tabushi et. al. that P-cyclodextrin
molecules can be linked to create dimers (or polymeric) cyclodextrin molecules, thereby
creating a molecule with two (or more) hydrophobic pockets.
Tang et. al. first showed
the catalytic ability of a Schiff-base metal complex, such as Fe111 2-hydroxy-lnaphthaldehyde thiosemicarbazone (HNT), to be used for the determination of H2O2 and
glucose concentrations.
Building on these studies, Tang et. al. have recently shown that
tethering two cyclodextrins forms dual hydrophobic pockets.
The Schiff-base metal
complex (Figure 1.6) includes as a guest within these pockets, and forms a
supramolecular mimic of HRP.72 Also synthesized were a series of metal complexes
using salicylidene-2-amino-4-phenylthiazole (SAPTS) as a ligand.
These complexes
14
were then investigated as potential superoxide dismutase (SOD) mimics. Of the metals
studied (Cu2+, Zn2+, Ni 2+ , and Co 2+ ), copper was the most active.
The ability of
polymeric-P-CD (P-CDP) to recognize the benzene rings of the Mn-(SAPTS)2 clearly

reveals the binding mode of the complex.
Their work has also provided additional
methodology for the detection of superoxide dismutase (SOD) activity.

1.4.2 Water-soluble Phosphane with Cyclodextrin
Caron et. al. investigated the influences of cyclodextrin on reactivity of a water
soluble Pd-phosphane complex.73
The binding mode of 4 (Figure 1.7) with P-
cyclodextrin was characterized using NMR. The NMR experimental evidence indicates
that the phosphorous is distant enough from the cyclodextrin molecule that the nuclei
from the two moieties do not couple. A palladium-catalyzed cleavage of allyl undecyl
carbonate was tested using [Pd(l)3] in the absence and presence of methylated-Pcyclodextrin.
The results indicate that as the ratio of methylated p-cyclodextrin/4 is
increased the rate of cleavage decreased when the ratio was between -0-2, constant from
-2-8, and increased for ratios from -8-16. Since the catalytic activity of the palladium
complex is similar in the absence and presence of cyclodextrin this signifies that the
cyclodextrin does not electronically or sterically hinder the metal (catalytic) center.73
These studies have illustrated that an organometallic complex in the presence of
cyclodextrin can be used to mimic functional bioenzymes.
Furthermore, with these
synthetic systems, there is no obligate restriction on biologically relevant metals, and thus
a wider variety of metal-chemistry can be explored.
1.5 Future Outlook
Outer-sphere coordination, either in natural or synthetic systems, provides a
versatile
framework
for
recognition
and
control
of
coordination
complexes.
Cyclodextrins particularly have proven to be versatile compounds since their initial use in
host-guest chemistry and have shown great potential for their employment in outer-
15
sphere complexation. The use of cyclodextrins in host-guest chemistry is well studied,

though their full potential has yet to be completely harnessed with outer-sphere metal
complexes. Work aimed toward the development of enzyme mimics, catalysts, and the
remediation of chemical wastes is ongoing.
Figure 1.6 (a) Fe 2-hydroxy-l-naphthaldehyde thiosemicarbazone (HNT) Schiff-base

complex. This figure shows a three-coordinate Fe; however, the structure is actually
either a square pyramidal or octahedral geometry which utilizes another HNT ligand to
satisfy the additional bonding sites, (b) Representation of the Schiff-base salicylidene-2amino-4-phenylthiazole complex (M -(SAPTS)2).
The use of cyclodextrin derivatives as catalysts has great potential due to the
arrangement of cyclodextrin's hydrophobic inner cavity rimmed with outer hydrophilic
functional groups. The recent work by Tang el. al. producing a horseradish peroxidase
mimic has shown the advancement of cyclodextrins in the area of catalysis. The ability
to mimic a specific enzyme has been accomplished and now it remains to be seen if
functional models for other enzymes can be developed.
The ability to modify
cyclodextrin on the upper and lower rims, and possibly attach additional scaffolds to
create an ordered system similar to the tertiary structure of proteins provides many levels
of control over a guest metal complex. The robust nature of the cyclodextrin systems
suggests that they may be suitable for use in environments where proteins are not stable.
16
Nevertheless, the ability to more precisely design supramolecular analogues of natural

systems that utilize outer-sphere coordination hinges on a deeper understanding of the
numerous biological systems still currently being studied.
NaCHB
/
\ f
VSOJNH
Figure 1.7 Phosphane ligand for complexation with a metal ion (e.g. Pd[4]3) and
inclusion into the cyclodextrin cavity.
17
Notes
(1)
Karakhanov, E. E.; Maksimov, A. L.; Runova, E. A.; Kardasheva, Y. S.;

Terenina, M. V.; Buchneva, T. S.; Guchkova, A. Y. Macromol. Symp. 2003, 204,
159-173.
(2)
Colquhoun, H. M.; Stoddart, J. F.; Williams, D. J. Angew. Chem., Int. Ed. Engl.
1983, 25, 487-582.
(3)
Stoddart, J. F.; Zarzycki, R. Reel. Trav. Chim. Pays-Bas 1988,107, 515-528.
(4)
Zamarev, K. New. J. Chem 1994,18, 3-18.
(5)
Yue, W. W.; Grizot, S.; Buchanan, S. K. J. Mol. Biol. 2003, 332, 353-368.
(6)
Telford, J. R.; Raymond, K. N. In Comprehensive Supramolecular Chemistry;

First ed.; Gokel, G. W., Ed.; Pergamon, 1996; Vol. 1, pp 245-266.
(7)
Raymond, K. N., Dertz, Emily A., Kim, Sanggoo S. Proc. Natl. Acad. Sci. U. S.
A. 2003,100, 3584-3588.
(8)
Braun, V. Int. J. Med. Microbiol. 2001, 291, 67-79.
(9)
Neilands, J. B. J. Biol. Chem. 1995, 270, 26723-26726.
(10)
Winkelmann, G. Biometals 2002, 30.
(11)
Challis, G. L. ChemBioChem 2005, 6, 601-611.
(12)
Ferguson, A. D., Welte, Wolfram, Hofmann, Eckhard, Lindner, Buko, Hoist,

Otto, Coulton, James W., Diederichs, Kay.: Structure (London) 8 pp. 592
Structure (London) 2000, 8, 585-592.
(13)
Yang, J.; Goetz, D. H.; Li, J.-Y.; Wang, W.; Mori, K.; Setlik, D.; Du, T.;
Erdjument-Bromage, H.; Tempst, P.; Strong, R. K.; Barasch, J. Mol. Cell 2002,
10, 1045-1056.
(14)
Mishra, J.; Dent, C; Tarabishi, R.; Mitsnefes, M. M.; Ma, Q.; Kelly, C; Ruff, S.
M.; Zahedi, K.; Shoo, M.; Bean, J.; Mori, K.; Barasch, J.; Devarajan, P. Lancet
2005,365, 1231-1238.
(15)
Yang, J.; Mori, K.; Li, J.-Y.; Barasch, J. Am J Physiol Renal Physiol 2003, 285,
F9-F18.
(16)
Bratt, T.; Ohlson, S.; Borregaard, N . Biochim. Biophys. Acta 1999, 1472, 262-
269.
(17)
Coles, M.; Diercks, T.; Muehlenweg, B.; Bartsch, S.; Zolzer, V.; Tschesche, H.;
Kessler, H. J. Mol. Biol. 1999, 289, 139-157.
(18)
Goetz, D. H.; Willie, S. T.; Armen, R. S.; Bratt, T.; Borregaard, N.; Strong, R. K.
Biochemistry 2000, 39, 1935-1941.
18
(19)
Goetz, D. H.; Holmes, M. A.; Borregaard, N.; Bluhm, M. E.; Raymond, K. N.;
Strong, R. K. Mol. Cell 2002,10, 1033-1043.
(20)
O'Brien, I. G.; Gibson, F. Biochim. Biophys. Acta 1970, 215, 393-402.
(21)
Kaplan, J. Cell 2002, 111, 603-606.
(22)
Shepard, W. E. B.; Kingston, R. L.; Anderson, B. F.; Baker, E. N. Acta

Crystallogr. 1993, 49, 331-343.
(23)
Garrett, T. P. J.; Clingeleffer, J. M.; Guss, J. M ; Rogers, S. J.; Freeman, H. C. J.

Biol. Chem. 1984, 259, 2822-2825.
(24)
Libeu, C. A. P.; Kukimoto, M.; Nishiyama, M.; Horinuchi, S.; Adman, E. T.

Biochemistry 1997, 36, 13160-13179.
(25)
Machczynski, M. C ; Gray, H. B.; Richards, J. H. J. Inorg. Biochem. 2002, 88,

375-380.
(26)
Li, H.; Webb, S. P.; Ivanic, J.; Jensen, J. H. J. Am. Chem. Soc. 2004,126, 80108019.
(27)
Bock, C. W.; Kaufman, A.; Glusker, J. P. Inorg. Chem. 1994, 33, 419-427.
(28)
Jou, R.; Cowan, J. A. J. Am. Chem. Soc. 1991,113, 6685-6686.
(29)
Kim, S.; Cowan, J. A. Inorg. Chem. 1992, 31, 3495-3496.
(30)
Kucharski, L. M.; Lubbe, W. J.; Maguire, M. E. J. Biol. Chem. 2000, 275, 1676716773.
(31)
Maguire, M. E.; Cowan, J. A. BioMetals 2002,15, 203-210.
(32)
Gessner, R. V.; Quigley, G. J.; Wang, A. H.-J.; van der Marel, G. A.; van Boom,
J. H.; Rich, A. Biochemistry 1985, 24, 237-240.
(33)
Petrov, A. S.; Funseth-Smotzer, J.; Pack, G. R. Int. J. Quantum Chem. 2005, 102,
645-655.
(34)
Connors, K. A. Chem. Rev. (Washington, DC, U. S.) 1997, 97, 1325-1357.
(35)
Easton, C. J.; Lincoln, S. F. Modified Cyclodextrins: Scaffolds and Templates for

Supramolecular Chemistry; Imperial College Press: London, 1999.
(36)
Hattori, K. Bio Industry 1987, 4, 327-338.
(37)
Bortolus, P.; Grabner, G.; Koehler, G.; Monti, S. Coordination Chemistry

Reviews 1993,125, 261-268.
(38)
Mosinger, J.; Tomankova, V.; Nemcova, I.; Zyka, J. Analytical Letters 2001, 34,
1979-2004.
(39)
D'Souza, V. T.; Lipkowitz, K. B. Chemical Reviews (Washington, D. C.) 1998,

98, 1741-1742.
19
Regiert, M. SOFW Journal 2003,129, 2, 4, 6, 8.
Del Valle, E. M. M. Process Biochem. (Oxford, U. K.) 2004, 39, 1033-1046.
Armspach, D.; Gattuso, G.; Koniger, R.; Stoddart, J. F. Bioorg. Chem.:
Carbohydrates 1999, 458-488, 597-602.
D'Souza, V. T. Supramol. Chem. 2003,15, 221-229.
Khan, A. R.; Forgo, P.; Stine, K. J.; D'Souza, V. T. Chem. Rev. (Washington, DC,
U. S.) 1998, 98, 1977-1996.
Alexeev, Y. E.; Vasilchenko, I. S.; Kharisov, B. I.; Blanco, L. M.; Garnovskii, A.
D.; Zhdanov, Y. A. J. Coord. Chem. 2004, 57, 1447-1517.
Alston, D. R.; Slawin, A. M. Z.; Stoddart, J. F.; Williams, D. J.; Zarzycki, R.
Angew. Chem., Int. Ed. 1988, 27, 1184-1185.
Alston, D. R.; Ashton, P. R.; Lilley, T. H.; Stoddart, J. F.; Zarzycki, R.; Slawin,
A. M. Z.; Williams, D. J. Carbohydr. Res. 1989,192, 259-281.
Harada, A.; Takahashi, S. J. Chem. Soc., Chem. Commun. 1984, 645-646.
Harada, A.; Takahashi, S. J. Chem. Soc, Chem. Commun. 1986, 1229-1230.
Alston, D. R.; Slawin, A. M. Z.; Stoddart, J. F.; Williams, D. J. Angew. Chem.,
Int. Ed. Engl. 1985, 24, 786-787.
Harada, A.; Shimada, M.; Takahashi, S. Chem. Lett. 1989, 275-276.
Harada, A.; Hu, Y.; Yamamoto, S.; Takahashi, S. J. Chem. Soc., Dalton Trans.
1988, 729-732.
Szejtli, J. Starch/Staerke 1990, 42, 444-447.
Russell, N. R.; McNamara, M. In Proceedings of the International Symposium on
Cyclodextrins, 8th: Budapest, 1996, pp 163-169.
Lehn, J.-M. Angew. Chem., Int. Ed. 1988, 27, 89-112.
Ando, I.; Ujimoto, K.; Kurihara, H. Bull. Chem. Soc. Jpn. 2001, 74, 717-721.
Alderighi, L.; Gans, P.; Ienco, D. P.; Sabatini, A.; Vacca, A. Coord. Chem. Rev.
1999, i 4 311-318.
Newton, T. W.; Sullivan, J. C. Actinide Carbonate Complexes in Aqueous
Solution North-Holland, Amsterdam, 1985.
Grenthe, I.; Lagerman, B.Acta. Chem. Scand. 1991, 45, 122-128.
Prudden, A. R.; Lien, N. R.; Telford, J. R. Chem. Commun. (Cambridge, U. K.)
2004, 172-173.
Rudiger, V.; Schneider, H. Chem.-Eur. J. 2000, 6, 3771-3776.
20
(62)
Schneider, H.; Hacket, F.; Volker, R. Chem. Rev. (Washington, DC, U. S.) 1998,
98, 1755-1786.
(63)
Navaza, A.; Iroulart, M. G.; Navaza, J. J. Coord. Chem. 2000, 51, 153-168.
(64)
Cabou, J.; Bricout, H.; Hapiot, F.; Monflier, E. Catal. Commun. 2004, 5, 265-270.
(65)
Bricout, H.; Caron, L.; Bormann, D.; Monflier, E. Catal. Today 2001, 66, 355361.
(66)
Tilloy, S.; Bertoux, F.; Mortreux, A.; Monflier, E. Catal. Today 1999, 48, 245253.
(67)
Smith, G. V.; Cheng, J.; Song, R. Chemical Industries (Dekker) 1996, 68, 479483.
(68)
Mukhopadhyay, S.; Rothenberg, G.; Joshi, A.; Baidossi, M.; Sasson, Y. Adv.
Synth. Catal. 2002, 344, 348-354.
(69)
Strimbu, L.; Liu, J.; Kaifer, A. E. Langmuir 2003, 19, 483-485.
(70)
Tabushi, I.; Kuroda, Y.; Shimokawa, K. J. Am. Chem. Soc. 1979,101, 1614-1615.
(71)
Tang, B.; Du, M.; Sun, Y.; Xu, H.-L; Shen, H.-x. Talanta 1998, 47, 361-366.
(72)
Tang, B.; Zhang, G.-Y.; Liu, Y.; Han, F. Anal. Chim. Acta 2002, 459, 83-91.
(73)
Caron, L.; Bricout, H.; Tilloy, S.; Ponchel, A.; Landy, D.; Fourmentin, S.;
Monflier, E.Adv. Synth. Catal. 2004, 346, 1449-1456.
21
CHAPTER 2
ANION RECOGNITION USING A CYCLODEXTRIN-BASED
SCAFFOLD
2.1 Introduction
2.1.1 Oxoanion Recognition Chemistry
Numerous examples of oxoanion recognition exist.
A relevant example in
relation to these studies is provided by Nature with phosphate binding protein (PBP).
Medveczky et. al. have shown that PBP binds phosphate with a Kd of -0.8 uM.1'2
Binding of phosphate is accomplished with 12 strong hydrogen bonds from the phosphate
binding protein backbone. Encompassed within the hydrogen bonding manifold are 10
hydrogen bonds to three of the phosphate oxygens (01, 02, and 03). 2 Complementing
this are two hydrogen bonds to 04 of the phosphate, one of which is "from an NH of a
backbone peptide unit whose carbonyl oxygen is in turn the recipient of two hydrogen
bonds".2 This set of 12 hydrogen bonds aids in understanding why phosphate binding
protein has such a high affinity for phosphate. Other examples in Nature of oxoanion
binding proteins which exhibit a substrate specificity have also been well characterized
{e.g. sulfate).3"9
2.1.2 Outer-sphere Coordination Using Cyclodextrins
The presence of literature reports for the outer-sphere coordination of metals
using cyclodextrins are lacking. Some notable examples of have been summarized in a
recent review.10 The aim of these studies is on utilizing the oxygen atoms of tetrahedral
anions as 'anchors' for binding to modified cyclodextrins. The focus here remains on the
complexation of perchlorate, phosphate, and sulfate with a-cyclodextrin (1) and per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin (6, Figure 2.1).
DMF, 70C
18h
PPh3,12
72h
45.1% yield
Wc
excess ethylenediamine
:
Figure 2.1 Synthetic transformation for a-cyclodextrin (1) to per-6-iodo-a-cyclodextrin (5). Removal of the iodides via nucleophilic
attack by either of the free terminal amines of ethylenediamine results in the final per-6-(2-aminoethylamino)-per-6-deoxy-acyclodextrin (6) product.
OH
to
to
23
2.1.3 Tetrahedral Anions

Perchlorate, phosphate, and sulfate are prevalent in nature in high concentrations.2
Since 1997 it has been shown that perchlorate levels in drinking water have been steadily
increasing ', more-so in areas related to the testing of solid rocket propellant, companies
synthesizing
perchlorates,
and
companies
utilizing perchlorates.
Furthermore,
experimental data has shown that both perchlorate and pertechnetate can compete with
iodide uptake by the thyroid gland in mammals.9'11 Therefore it is necessary, that prior to
discarding waste, toxic compounds, such as perchlorate, be remediated by the companies
or agencies that produce these wastes. In addition to this, other anions (even though not
targeted here) are of immediate concern. For instance, the heavy use of fertilizers has
created a need for the remediation of nitrates from the environment. ' !
In order to specifically target one tetrahedral anion over another it is important to
pay attention to their size and charge/size ratio (Table 2.1). Here the binding these
anions is first being targeted by the size inherent to the a-cyclodextrin framework,
followed by the modification to the upper (primary hydroxyl) rim of the cyclodextrin. It
is with these upper-rim modifications that targeting a specific charge/size ratio is being
analyzed.
2.1.4 Determination of Binding Affinities
In these studies, the binding of perchlorate has primarily been targeted; however,
it is vital to also investigate the binding of other common tetrahedral anions (e.g.
phosphate and sulfate). Even though numerous methods (NMR, UV-Vis, calorimetric,
etc.) have previously been used for the determination of binding constants, these methods
were not particularly suited to the binding of tetrahedral anions with cyclodextrins.13'14 A
relatively new method for determination of binding affinities using electrosprayionization mass spectrometry (ESI-MS) was employed.15 Numerous studies have shown
that the determination of binding affinities can be accomplished via the intensity ratio of
24
peaks for the uncomplexed and complexed species from ESI-MS spectra.15"17
Of
additional importance, studies indicate that ESI-MS, being a soft-ionization technique,

allows the speciation in the gas phase to mimic that of the solution (aqueous) phase.17
Consequently the mlz ratio will unambiguously indicate the most stable complex, thereby
providing the exact stoichiometry of the host and guest complex formed.15'18"21
Other
groups have shown that making assumptions as to the stoichiometry of the host and guest
complex can provide an erroneous binding constant.
2.2 Experimental
a-Cyclodextrin was obtained from Avocado Research Chemical Ltd (UK) and
was dried under vacuum. Ethylenediamine was obtained from Aldrich Chemical
Company.
LiC104 and MgS04 were obtained from Aldrich Chemical Company.
Potassium phosphate (monobasic, dibasic, and tribasic) was obtained from Fisher
Scientific. These chemicals were dried under vacuum prior to use. All chemicals were
of reagent grade. NMR experiments were performed at 25 C on a Briiker DPX-300,
AVANCE-300, DRX-400 or AVANCE-600 spectrometer using deuterated solvents.
2.2.1 ESI-MS
A Thermo Finnigan LCQ Deca Spectrometer (University of Iowa, High
Resolution Mass Spectrometry Facility, spectrometer purchased with fund from NIH
grant 510-RR13799-01) was used in direct injection mode.
All experiments were
conducted at a capillary temperature of 150 C, spray voltage of 4.5 V, and sample

collection flow rate of 3 uL/min. Prior to each sample, MS solvent (9 : 1 FbO/MeOH,
0.1% NH4OH) was flowed through the system until the detection of relevant mlz species
was not seen. Initial optimization (tuning) on the samples was done, followed by saving
the parameters and using this parameter set for subsequent trials.
Each sample was
allowed to equilibrate into the machine at 10 - 15 uL/min for 1 - 2 minutes prior to

reducing the flow rate to 3 ul/min and collecting data. After adjusting the flow rate, data
25
was collected in positive mode for 2 - 3 minutes from a range of mlz 100 - 1500.
Generally, the initial minute of data collected was discarded, followed by averaging of
100 spectra/sample to obtain intensity values for analysis. Complete spectra for ESI-MS
analysis of each tetrahedral anion with cyclodextrins 1 and 6 are located in Appendix B.
Intensity values (IH and IHG) and concentrations of the host (6) and guests (tetrahedral
anions) are provided in Tables B.l - B.3 in Appendix B.
2.2.2 Synthesis
Per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin (6). The methodology
for this procedure was taken from Ashton et. al. and Prudden et. al. '
Per-6-iodo-per-6-
deoxy-a-cyclodextrin (5) used for the synthesis of 6 was synthesized by Anthony

Prudden of the Telford group from a-cyclodextrin based upon Ashton's previous work.22
Per-6-iodo-per-6-deoxy-a-cyclodextrin
(5,
1.97
g,
1.21
mmol)
was
added
to
ethylenediamine (25 mL, 0.37 mol) under positive argon pressure. After dissolving, the
reaction mixture was heated to 70 C.
The reaction mixture was cooled to room
temperature after 72 h, reduced in volume at reduced pressure to 1 - 2 mL and

precipitated out of -250 mL of cold ethanol. The material was dissolved and reprecipitated two times. The precipitate was then isolated via Buchner funnel and vacuum
dried. Overall yield 1.09 g, 45.1%. 'H NMR (D 2 0, 400 MHz) 5 = 5.09 (6H, br s, HO,
4.01 (12H, br t, H3/H5), 3.67 (6H, br m, H 2 ), 3.56 (6H, br t , H 4 ), 3.21 (6H, br s, HNCH2CH2NH2), 2.65-3.15 (30H, br m, H 6 / -HNCH2CH2NH2).
13
C NMR 23 (D 2 0, 150
MHz) 5 104.2 ( d ) , 86.0 (C4), 75.8 (C3), 74.3 (C2), 73.3 (C5), 53.9 (-NHCH 2 CH 2 NH 2 ),
51.9 (C6), 42.4 (-NHCH 2 CH 2 NH 2 ) ESI-MS: (m/z) 1226 (M + H + ) + , 613 (M + 2H + ) 2+ , 409
(M + 3H+)3+, 205 (M + 6H+)6+2.2.3 Titrations
Concentrated stock solutions (ranging from ~1 - 9 mM) of cyclodextrins 1 and 6,
and the tetrahedral anions were made using volumetric glassware (TC) and then
26
immediately transferred to Corning conical vials that had been pre-rinsed with MS
solvent.
1 rtiL samples for ESI-MS were made from the stock solutions, with a final
[Cyclodextrin] = 90 uM. The desired amounts of each tetrahedral anion were added
ranging from [Cyclodextrin] : [Tetrahedral Anion] from 100 : 1 to 1 : 10 and with the
remaining volume filled to 1 mL with MS solvent.
Titrations with higher guest
concentrations were tested, but at such high salt concentrations the noise in the
instrument provides data that could not be interpreted with any reliability. It is important
to note that samples with ratios of [Cyclodextrin] : [Tetrahedral Anion] ranging from
100:1 to 1:1 generally did not provide any appreciable binding in the case of any
tetrahedral anion.
A set of example calculations for the titration of cyclodextrin 6 with LiC104 is
provided in the following sentences and Table 2.2. Stock solutions were made of 6 (7.96
mM) and LiC104 (7.24 raM). For the example reported here, titrations of the following
molar ratios of [6] : [LiC104] ranging from 100 : 1, 10 : 1, 1 : 1, 1 : 5, and 1 : 1 0 were
performed (see Tables B.l - B.3 in Appendix B for a ratios all titrations). An Eppendorf
tube for each titration point was made, followed by addition of the varying amounts of
each stock solution and MS solvent based on Table 2.2 below. The Eppendorf tubes
were agitated and allowed to stand for 6 - 12 hours to allow for each mixture to reach
equilibrium, followed by ESI-MS analysis of each sample.
2.2.4 Calculation of Binding Affinities
The analysis of ESI-MS data was done using equations A.1 - A.7. Relevant
information for these equations is provided in Appendix A. Experimental values for the
intensity ratios for +1, +2, and +3 charge states of uncomplexed and complexed
cyclodextrins were analyzed and fit into equation A.6.
Initial values of R were
determined, followed by sampling of R' and n values that provided the best linear fit and
conforming to the initial calculated R values. Values used for n were used to correct the
27
y-intercept determined based on the initial concentration of host. The determination of

these values was done by trying to obey trends established by the Hoffmeister with
partition coefficients. The Hoffmeister series indicates the increasing water solubility in
the order of S042" > P043" > > > C104".
2.2.5 pKa Determination
Attempts to determine the pKa values for hexa-protonated cyclodextrin 6 using
potentiometric titrations resulted in large buffer region from which pKa (acid
dissociation) values could not be deconvoluted. Consequently, an estimation of the pH
speciation distribution was created using the program HySS.24 The initial P values were
estimated beginning at 12 and working our way up to accommodate all species. The list
of p values were 12, 24, 33, 41, 48, and 54 for -AH*", AH22+, AH33+, AH44+, AH55+, and
AHg +, respectively. Figure 2.2 clearly indicates the propensity of primary amine system
to retain 2, or possibly more protons in the presence of additional species (tetrahedral
anions in this case). These data are also supported by observance of mainly +1, +2, and
+3 charged states in mass spectra upon binding of tetrahedral anions. This most likely
indicates a protonation state greater than two and less than six, but is highly dependent on
the tetrahedral anion due to their distinct charge states at pH ~ 10.
2.2.6 T\ Inversion Recovery Experiments
T\ inversion recovery experiments were chosen in order to clarify the binding of
the perchlorate, phosphate and sulfate with cyclodextrin 6. Only one tetrahedral anion
was chosen (perchlorate) for these studies due to the time required for each inversion
recovery experiment. Four T\ inversion recovery experiments were performed with
molar ratios of 6 : perchlorate of 1 : 0, 1 : 1.2, 1 : 4, and 1 : 8. Table 2.3 provides the
delay times prior to capturing spectra. Once collected, peak height data for 'H-NMR
peaks of relevance for each titration were noted and are seen in Tables B.4 - B.7 located
in Appendix B. Data analysis was performed by fitting peak height data to equation 2.8.
28
Final proton T\ relaxation data can be seen in Table 2.4 and Figure 2.3, which provides a
graphical version of the data.
Studies of tetrahedral anion binding with cyclodextrin 1 indicate a lack of
specificity.
In the case of 1 with LiC104, spectra indicate the association of Li+ (m/z
979), 2 Li + and C104" (m/z 1085), 3 Li+ and 2 C104" (m/z 1191), or 4 Li+ and 3 C104" (m/z
1298). Similar studies with KH2PO4 indicate that it incorporates mainly K+ (m/z 1011) or
2 H+, 2 K+ and PO43" (m/z 1147). The last studies with sulfate show predominantly the
inclusion of H+, Mg 2+ and SO42" (m/z 1093). In the former two cases, the ability of 1 to
bind the cation of the tetrahedral anion indicates a partial preference toward positivelycharged species. Furthermore, the inclusion of multiple CIO4" moieties indicates the
possibility of multiple association sites. In the cases of PO43" and SO4 " the inclusion of
more than one tetrahedral anion does occur, but to a lesser extent (less than 10% of
relative intensity for these peaks) in the ESI-MS spectrum. In the case of phosphate,
using monobasic, dibasic, or tribasic phosphate could provide differing results. Studies
with monobasic, dibasic and tribasic phosphate were all performed providing similar
results. With the pH of each titration point between 10.1 - 10.5, control of the species
distribution of phosphateno matter what starting phosphate was usedwas controlled
predominantly by the 0.1% NH4OH in the MS solvent.
In the case of cyclodextrin 6, the high propensity of the six pendant
ethylenediamine arms to be protonated provides the most likely area of electrostatic
interaction between the tetrahedral oxyanions and the host (Figure 2.4).
This is an
important distinction with 1, where the primary upper-rim hydroxyl moieties show a
partial preference for the cations of each tetrahedral anion. The estimate of protonation
states for cyclodextrin 6 was illustrated in Figure 2.2. Further support for the estimated
protonation state is provided by the pH of the solutions being studied. As mentioned
29
previously, solution pH measurements indicated valued between 10.1 - 10.5. At this pH

a mixture of mono-, di-, and tri-protonated species should be present (based on pKa
estimates earlier presented). This was further supported by the presence of+1, +2, and
+3 m/z species in the ESI mass spectra.
Given the clear identification of unbound and bound species via ESI-MS,
determination of binding constants for each tetrahedral anion with cyclodextrin 6 was
performed. The log Ka values for each tetrahedral anion can be seen in Table 2.1.
Qualitative analysis of the calculated association constants indicates a small amount of
discrimination, which is almost negligible for the purpose of separating these ions in a
mixture. It also goes without saying that binding for these tetrahedral oxyanions does not
depend on their charge/size ratio in the case of cyclodextrin 6 (Table 2.1).
The analysis of binding constants for perchlorate yielded linear plots with R2
values of 0.76, 0.91, and 0.95. The case of phosphate (either monobasic or dibasic)
yields reproducible results that produce linear plots (R2 values of 0.98, 0.93, and 0.98).
Sulfate also produces two linear plots with good regressions (R2 values of 0.84 and 0.92).
Overall, all three anions provide linear regressions that are within reasonable limits. This
data can be seen in Appendix B, Figures B.18 - B.25.
0.013
3.84 0.02
-8
03
Charge/Size Ratio (A3)

LogKa
R>***
***
15
-4
4.0 0.02
0.050
HPO42""
*** R' x n = R,R defined as response factor in Appendix A.
** Monobasic phosphate (KH2PO4) was used, but at a pH ~ 10 the predominant species of phosphate is HPO42"
-1
3.63 0.05
0.053
S042" *
* two sets of data collected, whereas others were three trial.
C104"
Tetrahedral Anion
Table 2.1 Properties of Tetrahedral Anions Studied with Cyclodextrin 6.
1000
Amount of Cyclodextrin Solution (uL)

Total Volume (uL)
992
1000
991
62
931
1:5
1000 1000
12
981
100: 1 10: 1 1:1
Amount of Perchlorate Solution (uL)
Amount MS Solvent (uL)
Molar ratio of 6 : LiC104
Table 2.2 Sample Calculations for the Titration of 6 with LiC104.
1000
124
869
1 : 10
Figure 2.2 pH-dependent speciation diagram based on estimated p values for the hexa-protonated per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin (6).
3.5
Chemical Shift (ppm)
H
4.5
=1 : 8 ([6]: [Perchlorate]
=1 : 4 ([6]: [Perchlorate]
5.5
=1 : 1.2 ([6]: [Perchlorate]
- = 1 : 0 ([6]: [Perchlorate]
Figure 2.3 Calculated T\ relaxation times in graphical format for protons of interest in the 'H-NMR spectrum. The only significant
deviation greater than 10% is seen at 8 3.61 ppm. The yellow line indicates a perturbation greater than 10% at 5 4.01 ppm; however,
no deviation is seen at 8 4.01 ppm with 8 equivalents of perchlorate indicating that its relaxation time is most likely unaffected by the
perchlorate.
0
25
1
o
0.5
CO
1 5
.2
*? - "
I 2
If 2.5
T\ Relaxation Values: Titration o f Cyclodextrin 6 w i t h LiC10 4
Figure 2.4 Generic representation of a tetrahedral anion (TA = CIO4", SO4 ", or PO4 ") binding to the upper-rim of per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin. Depending on the tetrahedral anion binding and charge state of the species in MS, the
charge of the final species in the gas phase will vary, therefore n would indicate the appropriate charge to get the overall charge on the
species and would be the charge of the each respective tetrahedral anion. The binding mode seen in this diagram is only assumed as it
provides the most likely electrostatic interaction.
35
R'
I HG
\IH
IHG J
1
K,xR'
[H\ x n + r[H\-,
/ HG
[G] 0 x/
<A 6)
Definitions:
/? = response factor = R' x n
[GJo = Initial guest (tetrahedral anion) concentration
[H]o = Initial host (cyclodextrin) concentration
[G] = Amount of free guest (tetrahedral anion) present
[H] = Amount of free host (cyclodextrin) present
IHG = Absolute intensity of HG complex
IH = Absolute intensity of H
M,=M0{l-2xe-").
(2.1)
Definitions:
Mt = peak intensity at a specified x value
Mo = peak intensity at full relaxation
T = delay time before the spectrum taken
T\ = proton T\ relaxation time
2.3.1 B inding mode of Tetrahedral Anions
Even though significant separation of these anions has not been achieved based
upon the association values reported, it remains important to understand the binding
mode(s) of each anion with our cyclodextrin 6. Understanding these interactions is vital
to developing other modified a-cyclodextrins with better separation properties. It has
36
been mentioned that the most likely interaction would be electrostatic via the protonated
pendant ethylenediamine arm(s) on the primary (upper) cyclodextrin surface.
To
determine if this is correct, T\ inversion recovery NMR experiments were performed to

analyze how T\ relaxation times are affected by the tetrahedral anion(s) present. Due to
the 10% error of these measurements, this methodology is not the most sensitive, but
these experiments were chosen, in the absence of crystallographic info, due to their
ability to provide site-specific information with reasonable accuracy.
Since the
experimental log Ka values are close, it was determined that studying cyclodextrin 6 with
CIO4" should be suitable to model all three tetrahedral anions. Figure 2.4 provides a
graph for the relative deviation in T\ relaxation data of cyclodextrin 6 titrated with
LiC104. The peak at 8 3.56 ppm (corresponding to cyclodextrin H4 proton) in the ! HNMR appears to be directly perturbed based on the concentration of tetrahedral anion in
solution.
The fact that the cyclodextrin H4 proton faces outside of the cavity23'25
indicates a possible binding mode outside of the cyclodextrin cavity and extended cavity,
created by the added ethylenediamine arms. It is also possible that perchlorate binding
distorts the cyclodextrin conformation forcing the H4 proton into a different
conformation, interacting with the perchlorate bound in the upper extended cavity formed
the by ethylenediamine arms. The analysis of results from these T\ relaxation studies still
provide some uncertainty due the lack of perturbations in the T\ relaxation times of any
other protons on the cyclodextrin or ethylenediamine arms. In order to fully elucidate the
binding mode a crystal structure would be beneficial; however, attempts towards
crystallization have been unsuccessful to date.
2.4 Conclusions
The binding of tetrahedral anions with both native a-cyclodextrin and

cyclodextrin 6 has been studied using ESI-MS as a probe of thermodynamics and
stoichiometry.
The binding of these ions with a-cyclodextrin indicates the lack of
37
specificity for binding (or competitive aggregation) of species, since both cations (Li ,
Mg , or K ) and the tetrahedral anions have been shown to associate as multiple cations
and/or anions. Studies with cyclodextrin 6 indicate a significant change imparted by the
six pendant ethylenediamine arms. The most prominent of these changes is the binding
of a tetrahedral anion or tetrahedral anion and its counter-ion. The lack of signal for
cyclodextrin 6 binding counter cations reveals that binding for 6 is targeted towards
anionic species. Furthermore, evidence indicates predominantly a 1 : 1 binding ratio
between the tetrahedral anions and cyclodextrin 6, unlike native a-cyclodextrin which
indicates association of up to 4-5 tetrahedral anions/cyclodextrin (depending on the
anion).
Binding analysis based on ESI-MS spectra intensity ratios has provided log Ka
ranging from 3.6 to 4.0, indicating a small amount of discrimination by cyclodextrin 6 for
each of the tetrahedral anions. Even though not useful in the separation of these species,
binding of tetrahedral anions with cyclodextrin 6 is still important for the general removal
of free tetrahedral anions from aqueous solution. In order to gain additional insight into
the binding mode(s) of the tetrahedral anions with cyclodextrin 6, T\ inversion recovery
experiments were performed. These experiments indicated a change in the T\ relaxation
time of the H4 proton from the cyclodextrin proton. These results still provide some
ambiguity in the exact mode of binding since the H4 proton could be perturbed by
binding on the outer suface of the cyclodextrin or through a close proximity if the torsion
on the cyclodextrin is altered upon binding. Consequently, additional studies need to be
pursued to provide a more concrete answer.
38
Table 2.3 Delay Time Prior to 'H-NMR Spectrum Collection for

Experiments Performed to Determine T\ Relaxation Time Perturbations.
Data Point
Delay Time Before Spectrum (sec)
0.0005
0.05
0.10
0.20
0.30
0.40
0.50
0.60
0.70
10
0.80
11
0.90
12
1.00
13
1.10
14
1.20
15
1.40
16
1.60
17
2.00
18
4.00
19
6.00
20
10.00
21
20.00
0.7047
2.341
0.5986
0.6641
0.9427
1.926
0.5963
0.3383
0.7019
5.09
4.01
3.67
3.56
3.21
2.9
0.7782
0.3376
0.8886
1 : 1.2
0.7189
0.3392
0.3399
0.7157
0.6367
2.654
0.948
0.6715
1 :8
0.617
2.521
1.081
0.677
1 :4
[Cyclodextrin 6] :: [LiC104]
1 :0
5 (PPM)
Table 2.4 Experimentally Calculated T\ Relaxation Times (sec) for 'H-NMR Peaks of Cyclodextrin 6 Titrated with LiC104.
40
Notes
(1)
Medveczky, N.; Rosenberg, H. Biochim. Biophys. Acta 1970, 211, 158-168.
(2)
Luecke, H.; Quiocho, F. A. Nature 1990, 347.
(3)
(4)
Quiocho, F. A.; Higgins, C. F. Phil. Trans. R. Soc. 1990, 326, 341-352.
(5)
Pflugrath, J. W.; Quiocho, F. A. Nature 1985, 314, 257-260.
(6)
Pflugrath, J. W.; Quiocho, F. A. J. Mol. Biol. 1988, 200, 163-180.
(7)
Sack, J. S.; Trakhanov, S. D.; Tsigannik, I. H.; Quiocho, F. A. J. Mol. Biol. 1989,
206, 193-207.
(8)
Sack, J. S.; Saper, M. A.; Quiocho, F. A. J. Mol. Biol. 1989, 206, 171-191.
(9)
Wolff, J. Pharmacol. Rev. 1998, 50, 89-102.
(10)
Harwani, S.; Telford, J. R. ChemTracts: Inorganic Chemistry 2005,18, 437-448.
(11)
Team, T. I. T. a. R. C. P. "Perchlorate: Overview of Issues, Status, and Remedial

Options," Environmental Research Institute of the States, 2005.
(12)
Sanchez, C. A. "Preliminary Evaluation of Factors Affecting Perchlorate Uptake,

Accumulation, and Redistribution in Lettuce," Department of Soil, Water, and
Environmental Sciences, Yuma Agricultural Center, The University of Arizona,
2004.
(13)
Connors, K. A.; Pendergast, D. D. Journal of the American Chemical Society

1984,106, 7607-7614.
(14)
(15)
Dotsikas, Y.; Loukas, Y. L. J. Am. Soc. Mass Spectr. 2003,14, 1123-1129.
(16)
Loukas, Y. L.; Vyza, E. A.; Valiraki, A. P. Analyst 1995,120, 533-538.
(17)
Hofstadler, S. A.; Sannes-Lowery, K. A. Nat. Rev. DrugDiscov. 2006, 5, 585595.
(18)
Dotsikas, Y.; Kontopanou, E.; Allagiannis, C ; Loukas, Y. L. J. Pharmaceut.

Biomed. 2000, 23, 997-1003.
41
(19)
Dotsikas, Y.; Loukas, Y. L. /. Biochem. Bioph. Meth. 2002, 52, 121-134.
(20)
Dotsikas, Y.; Loukas, Y. L. J. Pharmaceut. Biomed. 2002, 29, 487-494.
(21)
Veenstra, T. D. Biophys. Chem. 1999, 79, 63-79.
(22)
Ashton, P.; Koniger, R.; Stoddart, J. J. Org. Chem. 1996, 61, 903.
(23)
Prudden, A. R.; Lien, N. R.; Telford, J. R. Chem. Commun. (Cambridge, U. K.)

2004, 172-173.
(24)
Alderighi, L.; Gans, P.; Ienco, A.; Peters, D.; Sabatini, A.; Vacca, A. Coor. Chem.
Rev. 1999,754,311-318.
(25)
Ikeda, Y.; Motoune, S.; Matsuoka, T.; Arima, H.; Hirayama, F.; Uekama, K. J.
Pharm. Sci. 2002, 91, 2390-2398.
42
CHAPTER 3
CATION RECOGNITION USING ACID-MODIFIED
CYCLODEXTRIN-BASED SCAFFOLDS
3.1 Introduction
3.1.1 Cation Recognition
In addition to natural systems that utilize anion recognition, there are numerous
examples of cation recognition in Nature. One notable example is the site-specific
binding of transition metal cations with phosphate, sugar, or base groups of DNA or
RNA. " Furthermore, the development of hosts for the binding of cations has proven
tremendously useful to date. One of the most prominent examples was pioneered by
Pederson at Dupont with the development of crown ethers.5'6 12-Crown-4 and 18-crown6, specifically, have been shown to bind K+ and Na+, respectively. These two crown
ethers have proven their utility in the solvation of ionic compounds into organic phases
which provides reagents in the organic phase that would otherwise be difficult to achieve
(Figure 3.1).7
More detailed investigations into the binding of crown ethers and
modified crown ethers with cations have also been performed.8

Additional studies by Imperiali et. al. have shown the ability to design a peptide
that binds lanthanides.9 In these studies, Imperiali et. al. have designed a 17-mer peptide,
similar to that of an EF-hand protein specific for calcium binding. This small peptide has
shown the ability to bind lanthanides with Kd values ranging from 57 3 nM (Tb +) to
3500 200 nM (La ). Figure 3.2 indicates the dependence that the effective ionic
radius of the Lnm species has upon binding with their peptide.9 This figure clearly
indicates that binding of the largest lanthanide, La +, is the weakest, while intermediate
sizes (including Tb3+, Dy3+, and Er3+) have the highest binding abilities.9 Even though
not specific for one lanthanide ion, the ability to demonstrate a slight selectively towards
lanthanides with middle of the road radii provides an excellent example of the
43
development of a pre-organized binding site containing multiple ligands which can

3+
provide high binding affinities. Additional work in the solid state has shown that U can
out-compete Ce.3+ for the formation of crystals with N-heterocyclic carbenes
/
0
/
10
0
0
<
\\
0
12-crown-4
/
18- crown -6
Figure 3.1 Structures of 12-crown-4 and 18-crown-6 which are known to bind cations
K and Na+, respectively.
44
-30
Metal
ion
la**
Ce3*
-32
Ncf
\*
AG^ /
KJ mo)-'
Ev"
Get3*
TO3*
Dy?*
Er3
-34
36 -
-38
Li/*
A3*3
/(b(nM)
35OO2O0
95tht50
27020
624
846
573
?1S
78*6
100*6
128;fc8
ACe
/
/
j /
jF
-Llf
Oy'3
-40 _
Yb*
fd*l/
Er*3
i"*""^ I Eu*a
TO*3
42
1.00
1.05
110
1.15
Effective' ionic radius/A
* -
Figure 3.2 Relation between ionic radius and binding free energy (AG) for the trivalent
lanthanide species with the peptide from Imperiali et. al. The table included provides Kd
values for each trivalent lanthanide and their peptide.
Source: Nitz, M.; Sherawat, M.; Franz, K. J.; Peisach, E.; Allen, K. N.; Imperiali, B.
Angew. Chem. Int. Ed. 2004, 43, 3682-3685.
3.1.2 Nuclear Waste Refinement
With the relatively high concentration of long-lived half-life radionuclides present
in high-level liquid waste (HLLW), the need for separation of these radionuclides, mainly
actinide cations, for storage purposes is immensely important. The aim of these studies is
for the development of ligands to achieve these separations. In order to develop viable
ligands it is important to look back at some important separation procedures for HLLWs.
Separation of long-lived radionuclides has been given much attention, more so
since World War II and the Manhattan project. Numerous references from over twenty
45
years for the separation of actinides and lanthanides exist11"26 and provide much of this
groundwork.
One of the most useful separation procedures is the Plutonium and
C/ranium .Recovery by .Extraction (PUREX) process which was developed more than 40
years ago. In the PUREX process, U(VI) and Pu(IV) can be extracted from spent fuel
into tributyl phosphate diluted in kerosene, forming ternary complexes U02(N03)2*2TBP
77
and Pu(N03)4*2TBP, respectively.
These complexes, though stable, require the ability
to overcome the entropic barrier of bringing three species together.

Research focusing on the preorganization of ligands (mentioned previously with
work by Imperiali et. al.) exists with the development of supramolecular hosts for
radionuclide separations. The preorganization of ligands aids in relieving the entropic
barrier that is required to bring more than two species together. One such example was
performed by modifying calix[4]arene (Figure 3.3, 7, n=4) with derivatives of
carbamoylmethylphosphine oxide (CMPO, Figure 3.3, 8), a compound known to extract
trivalent cations from acidic solutions. Studies with structures 9 and 10 (Figure 3.3)
have shown an increase in binding of up to three orders of magnitude when focusing on
the size of the lanthanide.28 Even though the ability to bind Lnm or An111 ions from
solution has been shown, the ability to selectively complex one lanthanide or actinide
from mixtures has yielded poor results. Follow-up work to these studies and additional
research with calixarenes and cation binding has also been performed.14'29
3.1.3 Lanthanides and Actinides
With the predominant charge state for the lanthanides and actinides being +3, the
main goal for separation arrises from charge and size, as well as the charge to size ratio.
An effect, referred to as the lanthanide contraction, is key since the size of the lanthanides
5 1
decrease as Z increases, as seen in Table 3.1 and Figure 3.4.

coordination number for the lanthanides is greater than six.
The most common
The ligands utilized for
complexing the lanthanides were based upon Pearson hard-soft acid-base (HSAB) theory.
46
Lanthanides are considered relatively strong lewis acids.30 Being hard acids, Ln m cations
generally prefer hard bases, such as alkoxides, amides, and cyclopentadienyl ligands.30'32
C(CH3)3
NH
K-octyl
Ph
O
J-BlU
N
n = integer value between 4-:
7
-Bu
Ô
9 R = Ph
10 R = C6H13
Figure 3.3 7. Representation of a calix[n]arene with 4-8 monomers. 8. Repesentation of

carbamoylmethylphosphine oxide. 9/10. Representations of calix[4]arene modified with
a CMPO derivative and with the lower />-methyl group also modified.
3.1.4 Supramolecular Host and Choice of Functional

Group
The ability to pre-organize binding sites has been shown with both peptides and
caliaxrenes above.9'14,28'29 In a similar manner to those studies performed, our starting
supramolecular construct begins with the more rigid per-6-modified P-cyclodextrin. The
importance here that cyclodextrins are more rigid is vital to ensure that the ligand atoms
for binding remain within a constrained area. In the case of calixarenes, the increased
flexibility of the host backbone creates a supramolecular construct with an additional
unknown that can potentially be fixed with the cyclodextrin-based systems.
The development of ligands began with natural p-cyclodextrin (2) and initially
focused on substitution of the starting seven primary hydroxyl groups (Figure 3.5).
47
Difficulties synthesizing some of the per-6-modified p-cyclodextrins (where all seven

primary hydroxyls were substituted) led our studies to mono-6-modified (3-cyclodextrin
analogues of those per-6-modified (3-cyclodextrins that were initially desired (Figure
3.5).
The choice of atoms at the preferred binding site plays a more vital role in the
ability of our cyclodextrin hosts to bind lanthanides or actinides. Without atoms at the
preferred binding site that show a preference toward the lanthanides or actinides our
materials would not indicate a preference for binding.
As mentioned above, the
lanthanides generally prefer hard bases that often incorporate negatively charged oxygen
atoms. With this in mind, acid-bearing functional groups were chosen to modify our
cyclodextrin host. In the case of alkoxides (R-O", where R = alkyl chain), the ligands
would be negatively charged, providing an electrostatic interaction.
Of further
importance, it is well-known that lanthanide complexes are often ionic in character. '
This allows our acid-bearing cyclodextrin host to have both oxygen atoms at the
proposed binding site, as well as a negative charge that could aid in electrostatic (ionic)
interactions.
48
Lanthanide Ionic Radii vs. Atomic Number

1.18
1.16
1.14
1.12
i.i
2 1.08
T3
C3
Pi 1.06
'3 1.04
1.02
1
0.98
0.96 -I
55
57
59
61
63
65
67
69
71
73
Z (atomic number)
Figure 3.4 The decrease ('lanthanide contraction') in the ionic radii of the Ln-3+ species
is apparent from the above graphical representation where as Z increases, the radius
decreases.31
Figure 3.5 A generic representation of (a) per-6-modified (3-cyclodextrins and (b) mono6-modified P-cyclodextrins. In unmodified natural P-cyclodextrin R = -OH.
49
Table 3.1 Ionic Radii of the Lanthanides (A).31
Lanthanide
Ln3+ Radius (A)
57
La3+
1.16
58
Ce
3+
1.14
59
Pr3+
1.13
60
3+
1.11
61
Pm
3+
1.09
62
Sm3+
1.08
63
Eu3+
1.07
64
3+
Gd
1.05
65
Tb3+
1.04
66
Dy3+
1.03
67
Ho
3+
1.02
68
Er3+
1.00
Number (Z)
Nd
3+
0.99
70
Yb
3+
0.99
71
Lu3+
0.98
69
Tm
50
3.2 Experimental
P-Cyclodextrin (2) was either from Acros Organics or Cerestar, and was dried
under vacuum at 60 C. Atomic absorption standards of the lanthanides (1000 u,g/mL) in
5% HNO3 were obtained from Alfa Aesar. All other chemicals were obtained either from
Aldrich Chemical Company or Acros Organics. NMR experiments were performed at 25
C on a Bruker DPX-300, AVANCE-300, DRX-400 or AVANCE-600 spectrometer
using deuterated solvents. A Thermo Finnigan LCQ Deca Spectrometer (University of
Iowa, High Resolution Mass Spectrometry Facility, purchased with funds from NIH grant
510-RR13799-01) was used in direct injection mode.
3.2.1 Synthesis
Synthetic routes for compounds in this section are located in Figures 3.6 - 3.10.
For NMR assignments, Figure 3.11 provides labels (used in the characterization of 'HNMR below) for reference of unique protons to the arm(s) attached to p-cyclodextrin
upper-rim.
3.2.1.1 Per-6-modified [3-Cyclodextrins
Per-6-iodo-per-6-deoxy-P-cyclodextrin (11). It is important to note that this
synthesis was performed by Nathan Lien of the Telford lab, and this text is excerpted
from his thesis. The final product 7 was used as provided for subsequent syntheses.
"Triphenylphosphine (40.1 g, 153 mmol) was dissolved in dry DMF (250 mL). Solid I2
(40.7 g, 160 mmol) was carefully added to this solution over 10 min. Dry p-cyclodextrin
(2, 11.6 g, 10.2 mmol) was added to the solution and the temperature was raised to 70 C.
After 20 h, 100 mL of solvent was removed under reduced pressure. A suspension of
NaOMe (10.4 g, 193 mmol) in MeOH (60 mL) was added slowly. The resulting mixture
was stirred for 1 hour, and then poured into chilled MeOH (600 mL). The precipitate was
collected and washed with MeOH (2 x 100 mL). At this point, the procedures for
51
purifying the final product deviate from those listed by Ashton.19 The resulting yellowish
solid was suspended in 300 mL of MeOH and stirred overnight. The suspension was then
filtered and washed with MeOH (2 x 100 mL). The resulting white powder was collected
and dried under vacuum. Yield: 16.79 g, 86%. ! H NMR (300 MHz, CD3SOCD3) 5 =
6.03 (d, J = 5.8 Hz, 7H), 5.95 (s, 7H), 4.98 (d, J = 3.3 Hz, 7H), 3.80 (br d, J = 9.4), 3.533.67 (m, 14H), 3.23-3.49 (m, 21H). The synthesized compound is spectroscopically
identical to that reported by Ashton."
Per-6-7V-(2-aminoethylamino)-per-6-deoxy-P-cyclodextrin
(12). Per-6-iodo-
per-6-deoxy-P-cyclodextrin (11, 1.96 g, 1.02 mmol) was dissolved in freshly distilled

ethylenediamine (50 mL, 0.75 moles) under inert atmosphere. While stirring the reaction
mixture was heated to 70 C. After 36 hours, the reaction mixture was cooled to room
temperature, reduced in volume to ~5 mL, and precipitated by dropwise addition to cold
ethanol (350 mL). This mixture was allowed to stir for 0.5 hour at room temperature,
followed by 2 hours at 4 C. The solid was isolated via Buchner filtration and reprecipitated from cold ethanol again. This suspension was allowed to stand at 4 C
overnight followed by isolation of the off-white precipitate and drying. Overall yield of
54.7%, 1.31 g. 'H NMR (400 MHz, D 2 0) 5 = 5.09 (s, 7H, Hi), 3.94 (br t, J = 8.8 Hz,
14H, H3/H5), 3.63 (m, 7H, H2), 3.53 (br t, J = 9.5 Hz, 7H, H4), 2.62-3.11 (m, 42H, H6/
H2NCH2CH2NH). ESI-MS mlz = 1429 ([M + H + f).
Per-6-5-(thiosalicylic acid)-per-6-deoxy-P-cyclodextrin (14). Per-6-iodo-per-6deoxy-P-cyclodextrin (11, 0.990 g, 0.520 mmol) was dissolved in dry DMF (40 mL)
under argon atmosphere. After dissolving, ah excess of thiosalicylic acid (1.658 g, 10.8
mmol) was added and dissolved within a few seconds. Excess potassium carbonate
(K2CO3, 0.772 g, 5.59 mmol) and sodium dithionite (0.595 g, 3.42 mmol) were added
resulting in a suspension. The reaction mixture was then heated to 70 C. After 36
hours, the reaction mixture was cooled to room temperature, reduced in volume to less
than 1 mL, and precipitated by dropwise addition to ~110 mL of ddHÔ. This suspension
52
was allowed to stir for 0.5 hour, followed by 2 hours at 4 C, with subsequent isolation
via Buchner filtration. The white precipitate was isolated and dried. Overall yield of
0.707 g, 65.1%. Characterization was performed using 'H-NMR and ESI-MS (m/z =
1225 [M + H+]+). Attempts at
unsuccessful.
13
C NMR and COSY spectra were performed, but
This problem was most likely attributed to the fluctuation through
numerous configurations of the thiosalicylate moieties; no further attempts to attain these

spectra were performed. 'H NMR (400 MHz, D 2 0) 8 = 7.59 (d, J = 7.1 Hz, 7H, d),7.\7
(br s, 14H, b/c), 7.02 (br s, 7H, a), 5.04 (br s, 7H, Hi), 4.01 (s, 7H, H5), 3.9 (br t, J = 9
Hz, 7H, H3), 3.66 (m, 14H, H2/H4), 3.22 (br s, 14H, H6).
3.2.1.2 Mono-6-modified |3-cvclodextrins
Mono-6-tosyl-P-mono-6-deoxy-cyclodextrin (16). The procedure reported was
modified from Matsui, Y. and Okimoto, A. Bull Chem SocJpn, 51(10), 3030-3034, 1978.
P-Cyclodextrin (2, 24.1 g, 21.2 mmol) was added to dry pyridine (500 mL) under inert
atmosphere. After dissolving, the round bottom flask was cooled in an ice bath for 15
minutes while stirring. /?-Toluenesulfonyl chloride (3.01 g, 15.8 mmol) was added and
allowed to dissolve. After this point, the procedure deviated from that reported. The
reaction was allowed to proceed in ice bath (slowly warming) for 1-1.5 hours, followed
by addition of ddH20 (300 mL) to quench the reaction. The solvent was removed from
the reaction mixture to yield a white solid on the walls of the round bottom flask. This
solid was dissolved in ddHzO (400 mL) at 95 - 100 C to create a supersaturated
solution. The solution was transferred to a 1 L beaker and cooled to 4 C for 2 hours,
followed by isolation of the white precipitate via Buchner funnel. This was repeated 2
times, followed by isolation and drying. Overall yield of 9.814 g, 35.9%. The
H-NMR
spectrum was analyzed to ensure the presence of peaks for the aromatic protons of the
tosyl group. ESI-MS was performed (m/z 1311.4 [M + Na+]+) to ensure correct product
mass and aid in assessing the purity.
In all syntheses performed of this material,
53
approximately 5 - 10% of un-functionalized P-cyclodextrin (2) and the di-tosyl Pcyclodextrin were present. These materials were more easily separated upon removal of
the tosyl group with other functional groups during later reaction steps. No further
characterization was done as the product was used for the remaining mono-6-modified |3cyclodextrin derivatives. ! H NMR (400 MHz, D 2 0) 5 = 7.72 (d, J = 7.3 Hz, 2H, a), 7.53
(d, J = 7.7 Hz, 2H, b), 4.92-5.08 (m, 7H, HO, 3.75-3.98 (m, 14H, H3/H5), 3.46-3.74 (m,
21H, H2/H4/H6), 3.08-3.43 (m, 7H, H6), 2.55 (s, 3H, c).
Mono-6-Ar-(p-aminonicotinic acid)-mono-6-deoxy-p-cyclodextrin (17). Mono6-tosyl-P-cyclodextrin (16, 0.245 g, 0.19 mmol) was added to dry DMF (20 mL). After
dissolving, jo-aminonicotinic acid (0.352 g, 2.55 mmol) was added, followed by addition
of potassium carbonate (~ 0.1 g, ~ 0.7 mmol). The suspension was heated to 80 C.
After 72 hours, the reaction mixture was cooled to room temperature and the solvent was
removed to yield a white solid. The solid was taken up in minimal ddH20 and dialyzed
against ddH20 using a 500 MWCO dialysis cellulose ester membrane (Spectrum
Laboratories). The ddH20 was changed 3 times over 36 hours. Overall yield was 0.195
g, 81.6%. 'H NMR (400 MHz, D 2 0) 5 = 8.6 (s, 1H, c), 7.97 (dd, J = 9 Hz / 1.8 Hz, 1H,
b), 6.63 (d, J = 8.7 Hz, 1H, a), 5.11 (d, J = 4 Hz, 1H, Hi), 5.09 (m, 5H, Hi), 5.01 (d, J =
3.5 Hz, 1H, Hi), 3.81-3.97 (m, 21H, H3/H5), 3.63-3.72 (m, 14H, H2/H4), 3.56-3.6 (m, 7H,
H6). ESI-MS: m/z of 1255 ([M + H+]+).
Mono-6-(2-aminoethylamino)-mono-6-deoxy-p-cyclodextrin (18).
Mono-6-
tosyl-mono-6-deoxy-P-cyclodextrin (16, 4.532 g, 3.52 mmol) was added to the

ethylenediamine (150 mL, 2.24 mol) while stirring under inert atmosphere. The reaction
mixture was heated to 75 C. After 48 - 72 hours, the solvent was removed from the
reaction mixture to yield an off-white solid. This solid was dissolved in minimal ddH20
(-15 mL) and precipitated from cold ethanol (-500 mL). The solid was isolated via
Buchner funnel and re-precipitated from cold ethanol (-500 mL) again. The product was
vacuum dried and characterized. Overall yield was 3.81 g, 92%. ! H NMR (400 MHz,
54
D 2 0) 5 = 5.09 (br s, 7H, Hi), 3.85-4.02 (m, 28H, H3/H5/H2/H4), 3.65-3.68 (m, 7H, H 6 ),
3.60 (t, J = 9 Hz, 7H, H 6 ), 2.85 (m, 2H, a), 2.74 (t, J = 6.4 Hz , 2H, b).
Positive ESI-
MS: m/z of 1177 ([M + H+]+).

A second methodology, where the solid was dissolved in minimal ddH20 and
chromatographed on an LH-20 column was also used. After lyophilization, yields up to
83.8% were realized; however, the previous methodology was utilized due to its
simplicity and better yields.
Mono-6-(2-(diethylenetriamine pentaacetic acid-amino)ethylamino)-mono-6deoxy-p-cyclodextrin
(19).
Mono-6-(2-aminoethylamino)-mono-6-deoxy-P-
cyclodextrin (18, 0.46 g, 0.39 mmol) was dissolved in 10 mL of dry DMSO under
positive argon pressure. Diisopropyl ethyl amine (DIEA, 0.2 mL, 1.2 mmol) was added
and the mixture was allowed to stir for 1 hour.
To this clear, yellow solution,
diethylenetriamine pentaacetic acid dianhydride (21, DTPA dianhydride, 0.16 g, 0.45

mmol) was added. Addition of the DTPA dianhydride (21) resulted in a cloudy, yellow
solution that became clear, yellow within 1 hour. The reaction was allowed to proceed
for a total of 5 hours. Crude product analysis via ESI-MS indicated product formation
with a modest yield.
The solvent was removed from the reaction mixture at reduced pressure and 40
C. The solid was dissolved in 1 mL ddH20 and applied to a DEAE A-25 column (3.8
cm x 6.4 cm). A light yellowish band remained on the column. The column was washed
with five column volumes (-1500 mL) of ddHiO. The column was washed with three
column volumes (-900 mL) of 0.5 M NaCl. Fractions were concentrated by blowing air
over the solution heated at 70C. When 100 mL remained, the solution was cooled to
room temperature, concentrated and dialyzed in an Amicon filtration cell with a YM1
membrane (1000 MWCO, Millipore).
The solution was concentrated to 2 - 3 mL,
diluted with ddH 2 0 to 50 mL, and concentrated again to 2 - 3 mL. This was done until
-150 - 180 mL of ddH 2 0 had passed through the sample. The remaining 2 - 3 mL were
55
then reacted with 3 equivalents NaOH, dialyzed against 100 mL ddH20 via Amicon
filtration cell with a YM1 membrane, and lyophilized to give a white solid. Overall yield
of 0.505 g, 83.19%. ! H NMR (400 MHz, D 2 0) 5 = 5.09 (s, 7H, Hi), 3.84-4.85 (m, 21H,
H3/H5/H4), 3.64-3.77 (m, 14H, He), 3.55-3.63 (m, 7H, H2), 3.37 (s, 10H, c/d/g/j/k), 2.943.28 (m, 12H, a/b/e/f/h/i). Negative ESI-MS m/z = 1533 (in semi-anhydride form [M H+D, 766 ([M - 2H+]272).
Mono-6-(2-(dansylamino)ethylamino)-mono-6-deoxy-P-cyclodextrin
(20).
Mono-6-(2-aminoethylamino)-mono-6-deoxy-p-cyclodextrin (18, 0.430 g, 0.37 mmol)

was added to dry DMF (50 mL) under inert atmosphere. After dissolving, dansyl
chloride (0.540 g, 2.00 mmol) was added and allowed to dissolve followed by
triethylamine (3 mL, 0.022 mol). The reaction mixture was heated 85 C. After 72
hours, the reaction mixture was cooled to room temperature and the solvent removed to
yield a brown solid. The solid was dissolved in minimal ddH20 and chromatographed on
Sephadex G-10.
Fractions containing product were lyophilized.
Any fractions
containing dansyl sulfonate were dialyzed against ddH20 for 72 hours while changing the
water three times. The dialyzed material was analyzed to ensure the removal of dansyl
sulfonate and then combined with the remaining earlier product. Overall yield of product
was 0.406 g, 78.8%. Fluorescence excitation and emission spectra provided Agitation =
355 nm and ^missl0n = 537 nm, respectively. ]H NMR (400 MHz, D 2 0) 5 = 8.77 (d, J =
8.5 Hz, lH,y), 8.45 (d, J = 8.8 Hz, 1H, c), 8.29 (t, J = 7.3 Hz, 1H, e), 7.9 (t, J = 8.6 Hz,
1H, d), 7.7 (t, J = 7.9 Hz, 1H, g), 7.59 (d, J = 7.9 Hz, 1H, h), 4.90-5.15 (m [numerous
doublets], 7H, Hi), 3.83-4.02 (m, 14H, H3/H5), 3.58-3.7 (m, 14H, H2/H4), 3.55 (t, J = 7.8
Hz, 7H, He), 3.44 (m, 4H, a/b), 3.35-3.4 (m, 7H, H6), 2.98 (s, 6H, /). Positive ESI-MS:
m/z = 1410 ([M + H+]+), 1432 ([M+Na+]+), 1460 ([M + H+ + H 2 0 + CH3OH]+), 1519
([M+ H+ + H 2 0 + CH3OH + Na+ + Cl"]+).
13
HN^V
^ X O O H
H,N
11
14
12
15
DTPA dianhydride "
excess ethylenediamine
70C
''
72h
62% yield
K2C03, NaS204
SH
X X O O H DMF, 70C
72h
K2C03
kj
65.1% yield
DMF, 70C
72h
1
COOH /
PPh3) I2
DMF, 70C
18h
OH
OH O . /
OH
Figure 3.6 Reaction sequences for the synthesis of per-6-modified P-cyclodextrins (11 - 15). The arrow for the synthetic route of 12
to 15 is marked with an 'X' through it to denote that the reaction did not generate the desired product.
OH
NH, + r
W \HO-
^^-"\
Figure 3.7 Synthesis of 16.
HO-\*.
HO
OH
bH
^-o
V
/6
35.9% yield
2.H 2 0
0.25h
l.~leqTs-Cl
pyridine, <5C
lh
m^
16
^
HO-
sb=
OH
Figure 3.8 Syntheses of 17 and 18.
HO-X-
r\^ \
,NH,
excess ethyienediamine
'6
70C
48-60h
8 5 - 9 2 % vield
HO-\.
r^" " ^ N
\
OH
K 2 C0 3
DMF. 70C, I8h
81.6% yield
C:- A\
u
HO-
/,
OHI
COOH
BO--\-----,,,,,>0\
mi
COOH
00
34.2% yield
1. ~1 eq. DTPA dianhydride

DIEN
DMSO, RT
4-5 hours
2. 2-3 eq. NaOH
Ih
0"Na +
O^
ÔNa+
^
^
18
OH
\HO-
HO\^-
nc^- A
HN""'
/NH 2
^
/6
K 2 C0 3
DMF, 70C
18h
78.8% yield
excess dansyl chloride
/ ?H
20
\HO-V-.*
-Â AX
Jr
-A
HOX
HN
A
/6
as
o
COOH
0"
16
.so 2
Figure 3.11 Representations of the functional groups attached to the per-6- or mono-6-modified P-cyclodextrins. R is the location
where the cyclodextrin group is attached at C6. For the per-6-modified P-cyclodextrins each of the C6 positions is modified with the
labeled group; whereas, for the mono-6-modified P-cyclodextrins only one of the seven C6 positions is modified with the labeled
group. Each position for a C-H bond is labeled for ease of NMR assignments for the 'H-NMR listed in the characterization of each
synthesis.
12 or 18
V^w
COOH
ON
62
3.2.2 Fluorescence Measurements

Fluorescence measurements were performed on an Aminco Boman Series 2
fluorimeter with a scan rate ranging from 4 - 6 nm/sec with an average of 3 scans and a
cell with a 1 cm path length. Due to the fluorescence bestowed by the dansyl moiety onto
P-cyclodextrin, mono-6-(2-(dansylamino)ethylamino)-mono-6-deoxy-P-cyclodextrin (20)
is fluorescent. Studies analyzing the ability of tryptophan fluorescence emission (Agitation
= 290 nm, Xemjssion = 350 nm) to excite the dansyl group (Agitation = 355 nm) in phosphate
buffer (pH 7.4) were performed with no energy transfer evidenced (data not shown).
Additional studies analyzing the fluorescence quenching of 20 at 537 nm by up to 5
equivalents of D-(+)-glucose or Af-(l-pyridylmethyl)-2-aminobenzoic acid (25, see
Figure 4.1) were performed. No fluorescence quenching of 20 was seen by the addition
of either D-(+)-glucose or AL(l-pyridylmethyl)-2-aminobenzoic acid (25, data not
shown).
The binding affinity
of mono-6-(2-(diethylenetriamine
pentaacetic acid-
amino)ethylamino)-mono-6-deoxy-[3-cyclodextrin (19) with Lnm ions was studied (Lnm

= Tb3+ or Eu3+) in 0.01 M HEPES buffer (pH 7.4). In the case of Lnm (mainly Eu, Sm,
and Tb), the vibrational modes of water provide an efficient electronic relaxation mode
for the triplet excited state(s), resulting in no fluorescence. On the other hand, in the
presence of a ligand that excludes water from the first coordination sphere of the Ln10
metal center, the resulting hydrophobic environment around the metal center yields a
longer lived triplet state, allowing for fluorescence enhancement. "
50 uL of standard
solutions (1000 ug/uL) of Tb3+ or Eu3+ were added to 1750 uL ddH20 and 200 uL 0.1 M
HEPES buffer (pH 7.4).
The solution was then titrated with up to five equivalents
(relative to europium or terbium) of 18 or 19 (Figures 3.12 - 3.15). Tb3+ and Eu3+

excitations were performed at 350 nm, and emission monitored from 370 nm - 680 nm
(specifically analyzing 545 nm forTb 3+ and615 nm for Eu3+ for fluorescence emission
63
signals). Binding curves generated from the fluorescence enhancement measurements of

19 and Eu3+ (at 615 nm) or 19 and Tb3+ (at 545 nm) are seen in Figures 3.16 and 3.17,
respectively. Both figures indicate a linear increase in fluorescence up to 1 equivalent of
19, followed by a leveling off of fluorescence emission above 1 equivalent, indicating the
formation of 1 : 1 complex between 19 an Tb3+ or Eu3+.
The magnitudes of the
fluorescence emission for Eu3+ (at 615 nm) and Tb3+ (at 545 nm) were used for
determination of Kd values. These fluorescence measurements were modeled using
equation 3.1 based upon literature for specific and non-specific binding.38 The nonspecific binding term, C, was expected to be 0. Using Sigmaplot 8.0 (Systat Software,
Inc.), iterative fitting to equation 3.1 generated Kd values of 19 with Eu3+ and Tb3+ of
13.8 2.1 uM and 28.2 uM, respectively. Simulated fluorescence emission spectra
(overlayed in Figures 3.16 and 3.17), determined during the calculation of Kd from
equation 3.1, for the complexation of with Eu3+ or Tb3+ with 19 agree well with
experimental data (R2 > 0.99 in all cases).
AC AC
Mr +[M]T +Kd -J([P]T +[M]r +Kdf - 4 x [ P ] r x[M]7
X
*
r-j^
+ Cx[M]T
W =A 5
(3.1)
z x yr\T
Definitions:
ASmax maximum fluorescence change

AS = fluorescence signal change
[p] r = Total host concentration
[M]T = Total metal concentration
Kd = Dissociation constant
C = Contribution from non-specific binding
Samples were diluted to final volume of 1 mL in 1:1 I-bO/MeOH. Titration of 19

with Gd3+ indicated increasing complexation from the unbound semi-anhydride form of
64
19 (m/z 1532/1533 [semi-anyhydride 19 - lH+]"and 766 [semi-anyhydride 19 - 2H+]272)
until what appears to be near complete complexation (m/z 1711 [semi-anyhydride 19 3H+ +
158
Gd3+ + Na+]+) with ratios of ligand to metal greater than 1 : 3 (Figures 3.18,
3.19, and 3.20). A mass spectrum of the complex between Tb3+ with 19 (1 : 1) was also
performed, providing a m/z of 1712 ([semi-anyhydride 19 - 3H+ + Tb3+ + Na+]+) along
with other multiply charged peaks (Figure 3.21). Unlike the case of gadolinium, the
complex between 19 and Tb appears to be unstable, possibly due to temperature effects;
however, extensive characterization of all the peaks in the mass spectrum has not been
completed due to peaks that have eluded identification. Analysis of samples with 19 and
^4-
EuJX (1 : 1) was performed, but solubility issues in 1 : 1 ACN/H2O provided data that did
not provide useful information. Studies in 1 : 1 MeOH/tkO or another more suitable
solvent were not performed.
0.5
1.5
580
590
600
620
Wavelength (nm)
610
630
640
Oeq. 19
650
Figure 3.12 Titration of Eu.3+ with up to 5 equivalents of 19. The increase in the fluorescence emission at 615 nm is a result of the
exclusion of water from the first coordination sphere of the Eu3+.
o
C
u
o
o
2.5
Fluorescence Emission of Eu Titrated with 19
580
0.6
600
620
640
Wavelength (nm)
660
680
Figure 3.13 Relative fluorescence of Eu3+ when titrated with 18, one of the starting materials for the synthesis of 15. Importantly,
there is no increase in the fluorescence intensity at 615 nm as seen in Figure 3.12.
>
&
3+
Fluorescence Titration of Eu with 18
520
530
540
550
Wavelength (nm)
Titrated with 19
3+ ,
560
570
J1
Figure 3.14 Titration of Tb-3+
~ with up to 5 equivalents of 19. The increase in the fluorescence emission at 545 nm is a result of the
exclusion of water from the first coordination sphere of the Tb3+.
u>
S-H
a
a
o
64
Fluorescence Emission of Tb
Figure 3.15 Titration of Tb with up to 5 equivalents of 18. Importantly, there is no increase in the fluorescence intensity at 545 nm
as seen in Figure 3.14.
0.5
1.5
k.f
4
4
x- -
"
^r
""* -"
Experimental Calculated
,_,, ^
Titrated with 19
Equivalents of 19
"
3+
Figure 3.16 Binding curve generated from fluorescence emission at 615 nm of Eu3+ titrated with 19 is provided in black,
calculated fluorescence emission based upon fitting to equation 3.1 for determination of Kd is provided in orange.
13
i-c
o
a
aa
a
2.5
Binding Curve for Eu
</
",
1 l
Experimental
- " *
Equivalents of 19
Titrated with 19
Calculated
<
Figure 3.17 Binding curve generated from fluorescence emission at 545 nm of Tb titrated with 19 is provided in black,
calculated fluorescence emission based upon fitting to equation 3.1 for determination of Kj is provided in orange.
ti 2 -
>; 3 -
1 4^
i-4
<u
o
<u 5 o
/->
3 6-
7 -
Binding Curve of Tb
71
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200
400
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658.26
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600
616.22 llilji
509.21
718.33
800
790.34
m/z
1000
1200
1123.43
1204.59
11 1237.20
1042.40
961.34
880.38
1400
1376.33
:
1600
1609.28
1800
2000
1780.19 i 868.53
1712.82
Figure 3.21 Positive ESI-MS

spectrum
for the complexation of Tb3+ by ligand 19 (ratio 19 / Tb3+ of 1 : 1). m/z Y1Y1 representative
+
3+
+ +
of [semi-anhydride 19 - 3H + Tb + Na ] .
0^
5:
10: 144.74
15:
20-
25
30
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60
to
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75
80:
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90
95
100
75

3.3.1 Synthesis
Synthesis of per-6-iodo-(3-cyclodextrin (11) was performed in our lab based on
previous literature methodology.
For the remaining per-6-modified p-cyclodextrins,
nucleophilic substitution of the iodides with desirable ligands yielded modified

cyclodextrins.
For the synthesis of
per-6-(2-aminoethylamino)-per-6-deoxy-P-
cyclodextrin (12), excess ethylenediamine (H2NCH2CH2NH2) was used as both a

nucleophile and a base for neutralization of HI formed. Characterization of 12 using HNMR indicated the per-6-modified product based upon the lack of multiple H-l protons
seen right at 5 5.09 ppm. Additional characterization of 12 indicated a m/z of 1429 ([M +
H+]+) which is consistent with that calculated. Synthesis of per-6-5'-(thiosalicylic acid)per-6-deoxy-P-cyclodextrin (14) was carried out using a procedure similar to that
outlined by Adam et al. and Guillo et al.40'41 Unlike the 12, the synthesis of 14 required
the nucleophile, thiosalicylic acid, and an additional base, carbonate (CO32"). Initial
characterization of this species by ESI-MS indicated m/z of 2108.9 ([M + H+]+) which is
in good agreement with the calculated value (m/z 2109). Additional evidence of product
formation was provided by ^-NMR with the appearance of aromatic peaks between 5
6.8 - 7.8 ppm and a single peak for the H-l cyclodextrin protons, indicating that all seven
iodides had been substituted. Attempts to obtain COSY and HMQC data by NMR
facility staff were unsuccessful; the difficulty with obtaining these spectra was attributed
to the likely numerous conformations and the 'floppy' nature of the seven thiosalicate
moieties. This was further supported by the broad peaks seen in the 'H-NMR for the
aromatic protons. Initial studies to determine the binding constants of the Ln111 species
with 14 revealed that the solubility of 14 below a pH of 5 was poor. Consequently,
stability constants of 14 with the Ln1 species were not determined.
Due to the
76
insolubility of 14, the per-6-modified/>-aminonicotinic acid (3-cyclodextrin analogue (13)

was not further pursued as a potential ligand.
In order to append the acid-bearing functional group(s) to the upper-rim of the
cyclodextrin, the seven free primary amines in 12 (from the ethylenediamine arms) could
be used for nucleophilic attack on diethylenetriamine pentaacetic acid dianhydride (21),
synthesizing 15. Attempts to append seven DTPA dianhydride (21) molecules onto 12
were unsuccessful. We attributed this lack of reactivity to the sterics of attaching seven
bulky DTPA groups in close proximity. ESI-MS analysis of the crude product indicated
the presence of mono-, di-, and tri-DTPA (3-cyclodextrin derivatives. Attempts to isolate
these products using column chromarography were unsuccessful.
Further efforts to
control the stoichiometry of the reaction and trying to append only one DTPA
dianhydride (21) molecule onto one of the pendant ethylenediamine arms were also
unsuccessful due to similar purification issues of mainly mono- and di-DTPA 0cyclodextrin derivatives.
With attempts at synthesizing desirable per-6-modified P-cyclodextrins proving
difficult, focus turned to mono-6-modified p-cyclodextrin derivatives. The synthesis of
mono-6-tosyl-mono-6-deoxy-p-cyclodextrin (16) is crucial to these derivatives due to the
ease with which the tosyl group can be displaced. With a stoichiometric amount of tosyl
chloride (1 - 1.1 eq.) the synthesis of 16 was possible after reaction with p-cyclodextrin
in pyridine and recrystallizing the product three times.
Pyridine is critical to this
synthesis since it acts both a proton scavenger (base) and aids in blocking the cavity from
including the tosyl chloride within it, making it non-reactive.
H-NMR spectrum of 16
confirmed the appearance peaks for the four aromatic protons (5 7.5 - 7.8 ppm, two
equivalent pairs). The 'H-NMR spectrum also exhibited peaks for 7 non-equivalent (5
4.85 - 5.25 ppm) H-l protons, which indicate an unsymmetrical substitution on the
cyclodextrin. Further evidence was provided by ESI-MS with the presence of a mlz of
1311 ([M + Na + ] + ). The synthesis of 16 also yielded less than 10% of unreacted and di-
77
tosylated products; however, these were more easily separated during subsequent reaction
steps. Cyclodextrin 16 was then reacted with /?-aminonicotinic acid or ethylenediamine
to yield 17 and 18, respectively. Based upon the peaks for H-l protons (three doublets)
in the ^ - N M R spectrum of 17, an unsymmetrical substitution pattern is evident.
Additional evidence is provided by the aromatic peaks present between 8 6.5-8.5 ppm
and positive ESI-MS of 1255 ([M + H+]+).
OH
OH HO
\ /
^-jsT
X
'
AA\
X
^ ^
L.OH
J0
19
>
O
^~/ % ) H
^^Y
^x
^^
O
21
Figure 3.22 Representation of the structure of DTPA dianhydride after nucleophilic

attack by the free amine from either 12 or 18 and hydrolysis of the second anhydride (15
or 19) and a representation of diethylene triamine pentaacetic acid dianhydride (DTPA
dianhydride, 21). In structure 15 or 19, R is where an ethylenediamine arm on the
modified P-cyclodextrin (either 12 or 18) is attached to the DTPA.
The last set of cyclodextrin derivatives was synthesized by nucleophilic attack of

DTPA dianhydride (21) and dansyl chloride by the free primary amine of the
ethylenediamine arm from 18, to yield 19 and 20, respectively. The 'H-NMR for 19
shows signals for the 10 alpha methylene protons and the 8 methylene protons of the
ethylenediamine arms of DTPA (these protons are coupled with the 4 protons from the
mono-6-(2-aminethylamino)- arm). Molecule 20 is inherently fluorescent due to the
dansyl moiety.
The 'H-NMR shows signals from the 6 aromatic protons and the 6
78
protons from the N, iV-dimethyl groups. Additional characterization of both 19 and 20

via ESI-MS was also performed.
Of additional relevance are
13
C-NMR, COSY, and HMQC spectra that were
11
collected.
C- and HMQC-NMR data provided mixed results even when summing scans
for over 24 hours.
The cyclodextrin backbone contributes 42 carbons which easily
outweighs the contribution from the carbons from the added functional groups in
molecules 17, 19 and 20 (6, 14, 16 carbons, respectively). Additional contributions from
the difficulty in resolving quaternary carbons and the 'floppy' nature of the added
functional groups in 17, 19 and 20 are thought to be governing the relatively poor
13
C-
NMR spectra (also impacting the HMQC). Consequently, those data are included, but
not of great utility.
COSY spectra agree well with data expected for the mono-
functionalized P-cyclodextrins synthesized.
Spectra for all per- and mono-modified
cyclodextrins derivatives can be seen in Appendix C.

3.3.2 Fluorescence
Excitation at 355 nm of the dansyl moiety on cyclodextrin 20 and emission at 537
nm served as a probe in binding studies with 20.
Initial studies to determine an
interaction between tryptophan and 20 were performed using FRET. The ability to excite
tryptophan (Agitation = 290 nm, Amission = 350 nm) resulted in no excitation at 355 nm for
molecule 20 in phosphate buffer (pH 7.4).
The studies indicate that there is not
significant interaction between 20 and tryptophan. Additional fluorescence quenching

studies indicate that there is no close interaction between D-(+)-glucose and
pyridylmethyl)-2-aminobenzoic acid.
JV-(1-
The lack of interactions with possible guest
molecules and with tryptophan FRET supports literature evidence of the encapsulation of
the dansyl group within the cyclodextrin cavity.42
Mono-6-(2-(diethylenetriamine
pentaacetic
acid-amino)ethylamino)-mono-6-
deoxy-P-cyclodextrin (19) is not naturally fluorescent, however as previously mentioned,
79
the removal of water from the first coordination sphere for Eu
(and Tb ) yields
fluorescence enhancement. As Eu3+ is titrated with. 19, fluorescence enhancement is seen

at 615 nm (Figure 3.12). In order to ensure that this enhancement could not be achieved
with cyclodextrin-based intermediate 18, utilized to synthesize 19, this compound was
also tested. Figure 3.13 provides the fluorescence data when 18 was titrated into a
solution containing Eu3+, clearly indicating that the enhancement seen by 19 is most
likely provided by the binding of Eu3+ to the DTPA arm.
Similar spectra for Tb 3+
titratated with 18 and 19 also provided this enhancement evidenced at 545 nm (Figures
3.15 and 3.14, respectively).
Binding curves generated from these fluorescence
enhancement studies at 615 or 545 nm of Eu3+ or Tb 3+ titrated with 19 (Figures 3.16 and
3.17, respectively) indicate a linear increase in fluorescence emission up to the addition 1
equivalent of 19. This data indicates a 1 : 1 binding stoichiometry between 19 and Ln in
ions (Ln = Eu, Tb).
Calculation of dissociation constants (Kd) indicate modest
complexation of Eu3+ or Tb 3+ by 19 with values of 13.8 2.1 uM and 28.2 uM,

respectively.
Promising results from fluorescence enhancement studies urged us to consider
additional methods to identify complexes for with 19 and Ln m cations. Consequently,
ESI-MS studies with gadolinium and terbium were performed. The binding of Gd3+ with
19 provides indicate the ability to achieve near complete complexation (m/z 1711) with a
ratio as low as 1 : 3 (19 / Gd +, Figure 3.14). MS analysis of the complexation of Tb
by 19 indicates a 1 : 1 ligand/metal complex as well (m/z 1712). The complex with

terbium appears to be less stable in the ESI-MS instrument either producing multiply
charged species, or causing the degradation of the cyclodextrin sugar backbone due to the
low intensity of the m/z 1712 peak, along with the appearance of peaks at m/z 790, 880,
and 961. Identification of these three latter peaks has been unsuccessful still providing
80
more uncertainty as to the complex formed with Tb
and 19. Similar to fluorescence
binding curves generated, MS data suggests a 1 : 1 binding stoichiometry between 19 and

Ln in cations (Ln = Gd or Tb).
3.4 Conclusions
The development of acid-based cyclodextrins as ligands for the lanthanides and
actinides has been attempted. Initial developments with per-6-modified p-cycldoextrins
proved useful, but provided difficulty with appending multiple pendant acid aromatic
arms bearing multiple chelation sites. Additional per-6-modified p-cyclodextrins have
also been synthesized, however testing their ability to binding lanthanides has been
limited due to solubility issues with 14. Due to these difficulties, mono-6-modified Pcycldoextrins were pursued. The synthesis of 16 was accomplished based on previous
literature reports.
The ability to easily substitute the tosyl group of 16 provided a
versatile scaffold, which could be built upon by the attachment of multidentate ligands.
To this end, diethylenetriamine pentaacetic acid which is known to bind Gd3+
strongly '44,
was
attached
to
mono-6-(2-aminoethylamino)-mono-6-deoxy-p-
cyclodextrin (18). Fluorescence studies indicate that 19 binding with Ln111 (Ln = Eu or
Tb) based upon induction of the metal center into a more hydrophobic environment.
These fluorescence enhancement studies, along with ESI-MS data, indicate a 1 : 1
stoichiometry between 19 and Ln111 (Ln = Gd, Tb, Eu) cations. Additional studies with
cyclodextrin 20 have shown the inability to quench the fluorescence from the dansyl
group. This is attributed to, and further supports literature reports, of the inclusion of the
dansyl arm into the cyclodextrin cavity.
3.5 Future Work
The development of additional multidentate ligands on the mono-6-modified pcyclodextrin scaffold is still desired. In order to see if discrimination of Ln111 species is
possible by 19, determination of the binding affinities for all the Ln111 ions with 19 need
81
to be measured. The acid dissociation constants of the four acid chelation sites also
remain to be determined, but should be similar to unmodified DTPA. Additional studies
for determining the coordination environment of bound Lnm species with 14 and 19
would also need to be probed using Time-Resolved Laser Fluorescence Spectroscopy
(TRLFS) methods.
82
Notes
(1)
Davey, C. A.; Richmond, T. J. P. Natl. Acad. Sci. USA 2002, 99, 11169-11174.
(2)
Tereshko, V.; Wilds, C. J.; Minasov, G.; Prakash, T. P.; Maier, M. A.; Howard,
A.; Wawrzak, Z.; Manoharan, M.; Egli, M. Nucleic Acids Res. 2001, 29, 12081215.
(3)
Chiu, T. K.; Dickerson, R. E. J. Mol. Biol. 2000, 301, 915-945.
(4)
Howerton, S. B.; Sines, C. C ; VanDerveer, D.; Williams, L. D. Biochemistry

2001,40,10023-10031.
(5)
Pederson, C. J. J. Am. Chem. Soc. 1967, 89, 7017-7036.
(6)
(7)
Carey, F. A. Organic Chemistry; 6th ed.; McGraw-Hill, 2006.
(8)
Bradshaw, J. S.; Izatt, R. M.; Bordunov, A. V.; Zhu, C. Y.; Hathaway, J. K. In

Molecular Recognition: Receptors for Cationic Guests; Gokel, G. W., Ed.;
Pergamon: New York, 1996; Vol. 1, pp 35-152.
(9)
Nitz, M.; Sherawat, M.; Franz, K. J.; Peisach, E.; Allen, K. N.; Imperiali, B.
Angew. Chem. Int. Ed. 2004, 43, 3682-3685.
(10)
Mehdoui, T.; Berthet, J.-C; Thuery, P.; Ephritikhine, M. Chem. Comm. 2005,
2860-2862.
(11)
Bhattacharyya, A.; Mohapatra, P. K.; Manchanda, V. K. Solvent. Extra. Ion Exc.

2006,24, 1-17.
(12)
Bhattacharyya, A.; Mohapatra, P. K.; Manchanda, V. K. Solvent. Extra. Ion Exc.

2007, 25, 27-39.
(13)
Chiarizia, R.; McAlister, D. R.; Herlinger, A. W. Separ. Sci. Technol. 2005, 40,
69-90.
(14)
Delmau, L. H.; Simon, N.; Schwing-Weill, M.-J.; Arnaud-Neu, F.; Dozol, J.-F.;
Eymard, S.; Tournois, B.; Gruttner, C ; Musigmann, C ; Tunayar, A.; Bohmer, V.
Separ. Sci. Technol. 1999, 34, 863-876.
(15)
Draye, M.; Thomas, S.; Cote, G.; Favre-Reguillon, A.; LeBuzit, G.; Guy, A.;
Foos, J. Separ. Sci. Technol. 2005, 40, 611-622.
(16)
Fuks, L.; Majdan, M. Min. Pro. Ext. Met. Rev. 2000, 21, 25-48.
(17)
Geist, A.; Weigl, M.; Gompper, K. Separ. Sci. Technol. 2002, 37, 3369-3390.
(18)
Ionova, G.; Ionov, S.; Rabbe, C ; Hill, C ; Madic, C ; Guillaumont, R.; Krupa, J.
C. Solvent. Extra. Ion Exc. 2001,19, 391-414.
83
(19)
Jenkins, I. L. Solvent. Extra. Ion Exc. 1984, 2, 1-27.
(20)
Kumar, M. Analyst 1994,119, 2013-2023.
(21)
Nash, K. L.; Rickert, P. G. Separ. Sci. Technol. 1993, 28, 25-41.
(22)
Nash, K. L.; Choppin, G. R. Separ. Sci. Technol. 1997, 32, 255-21 A.
(23)
Wilmarth, W. R.; Rosencrance, S. W.; Nash, C. A.; Edwards, T. B. Separ. Sci.

Technol. 2001, 36, 1283-1305.
(24)
Cassidy, R. M.; Knight, C. H.; Recoskie, B. M.; Elchuk, S.; Miller, F. C ; Green,
L. W. In Actinide Separations; Navratil, J. D., Schulz, W. W., Eds.; World
Scientific Publishers: Singapore, 1988, p 1-18.
(25)
Nash, K. L.; Jensen, M. P. Separ. Sci. Technol. 2001, 36, 1257-1282.
(26)
Robards, K.; Clarke, S.; Patsalides, E. Analyst 1988,113, 1757-1779.
(27)
May, I.; Taylor, R. J.; Denniss, I. S.; Wallwork, A. L. Czech. J. Phys. 1999, 49.
(28)
Delmau, L. H.; Simon, N.; Schwing-Weill, M.-J.; Arnaud-Neu, F.; Dozol, J.-F.;
Eymard, S.; Tournois, B.; Bohmer, V.; Griittner, C ; Musigmannc, C ; Tunayarc,
A. Chem. Comm. 1998, 1627-1628.
(29)
Mckervey, M. A.; Schwing-Weill, M.-J.; Arnaud-Neu, F. In Molecular

Recognition: Receptors for Cationic Guests; Gokel, G. W., Ed.; Pergamon: New
York, 1996; Vol. 1, pp 537-603.
(30)
Anwander, R. In Lanthanides: Chemistry and Use in Organic Synthesis;

Kobayashi, S., Ed.; Springer, 1999; Vol. 2, p 305.
(31)
Housecroft, C. E.; Sharpe, A. G. In Inorganic Chemistry; 2nd ed.; Pearson

Education Limited: England, 2005.
(32)
Lien, N. R. (2007). Cyclodextrins as molecular scaffolds for anion and cation

recognition. Chemistry. Iowa City, University of Iowa: 245.
(33)
Peter, S.; Panigrahi, B. S.; Viswanathan, K. S.; Mathews, C. K. Anal. Chim. Acta
1992,260,135-141.
(34)
Hemmila, I. Anal. Chem. 1985, 57, 1676-1681.
(35)
Panigrahi, B. S.; Gajendran, N.; Suryamurthy, N. Spectrochim Acta A 2003, 59,

1905-1910.
(36)
Taketatsu, T. Talanta 1982, 29, 397-400.
(37)
Xu, Y.-Y.; Hemmila, I. A. Anal. Chim. Acta 1992, 256, 9-16.
(38)
Ye, Y.; Lee, H.-W.; Yang, W.; Shealy, S.; Yang, J. J. J. Am. Chem. Soc. 2005,
127, 3743-3750.
(39)
84
(40)
Guillo, F.; Hamelin, B.; Jullien, L.; Canceill, J.; Lehn, J.-M.; De Robertis, L.;
Driguez, H. Bull. Soc. Chim. Fr. 1995,132, 857-866.
(41)
Adam, J. M.; Bennett, D. J.; Bom, A.; Clark, J. K.; Feilden, H.; Hutchinson, E. J.;
Palin, R.; Prosser, A.; Rees, D. C; Rosair, G. M.; Stevenson, D.; Tarve, G. J.;
Zhang, M.-Q.J. Med. Chem. 2002, 45, 1806-1816.
(42)
Corradini, R.; Dossena, A.; Marchelli, R.; Panagla, A.; Sartor, G.; Saviano, M.;
Lombardi, A.; Pavone, V. Chem.-Eur. J. 2006, 2, 373-381.
(43)
Weiner, E. C; Brechbiel, M. W.; Brothers, H.; Magin, R. L.; Gansow, O. A.;

Tomalia, D. A.; Lauterbur, P. C. Magn. Reson. Med. 1994, 31, 1-8.
(44)
Weinmann, H. J. Characteristics ofGd-DTPA dimeglumine in Magnevist

monograph; Blackwell Scientific Publications: Oxford, 1994.
85
CHAPTER 4
ORGANIC SUBSTRATES BINDING WITH NATIVE AND MODIFIED
p-CYCLODEXTRINS
4.1 Introduction
The ability of supramolecular hosts, such as cyclodextrins, to include within its
cavity and bind numerous targets has envisioned a wide-array of applications. These
applications include, but are not limited to, drug delivery, solubility in distinct phases,
contrast agents, catalysis, etc.1 An example of a similar system that has had tremendous
impact on organic synthesis is the solubility of salts in the organic phase by crown
ethers.2 Often times the hydrophobic interactions are thought to be driven by entropic
gains, however in the case of cyclodextrins the enthalpic gains outweighs the entropic
loss.3 For cyclodextrins, the hydrophobic cavity of the cyclodextrin plays a vital role in
binding organic substrates due to the release of water from within the cavity. Therefore,
initial studies focused on the binding of organic substrates with either native or modified
cyclodextrins. It was with hope that the host-guest complexes formed could be combined
with metals, creating a catalytically active metal center.
UV-Vis spectroscopy has been shown to provide information with regards to the
inclusion complex formed. Further data (e.g. association constants) can be elucidated
from analysis of the increase or decrease and red or blue shift evidenced from the UV-Vis
spectra. Consequently, our studies conclude with studying cyclodextrin-based host-guest
complexes utilizing predominantly UV-Vis spectroscopy.
4.2 Experimental
P-cyclodextrin (2) used was either from Acros Organics or Cerestar, and was
dried under vacuum at 60 C. All other chemicals were either obtained from Aldrich
Chemical Company or Acros Organics. Relevant substrates for inclusion studies are seen
in Figure 4.1.
86
Figure 4.1 Structure of guest molecules (22 - 2,3-diaminonaphthalene, 23 - 2,3dihydroxyquinoxaline, 24 - 3-hydroxy-2-naphthoic acid, 25 - JV-(l-pyridylmethyl)-2aminobenzoic acid, 26 - 2,3-dihydroxynaphthalene) studied for interaction with native
and functionalized p-cyclodextrins.
4.2.1 Synthesis
Per-6-iodo-per-6-deoxy-P-cyclodextrin (7). See synthesis previously performed
for acid scaffold synthesis (Chapter 3).
Per-6-(2-aminoethylamino)-per-6-deoxy-(3-cyclodextrin (12).
See synthesis
previously performed for acid scaffold synthesis (Chapter 3).

iV-(l-pyridyl-methyI)-2-amino-benzoic acid (25).
The synthesis of 25 was
performed by Visting Professor David Jahng (College of Pharmacy, Yeungnam

University, Kyongsan 712-749, Korea) in the Telford Research group based upon
procedures previously reported by Steiner and Schetty.4'5 This procedure is based upon
Professor Jahng's laboratory notes.
"A mixture of 6.48 g (0.06 mol) of 2-
aminomethylpyridine, 5.06 g (0.02 mol) of 2-iodobenzoic acid, and 3.40 g (0.025 mol) of
87
potassium carbonate (K2CO3) was heated to 130 C for 12 h. The reaction mixture turned
into a gel. The reaction mixture was dissolved in 20 mL of water and acidified with 1 N
HC1 (pH ~ 3) to yield a white precipitate. Recrystallization from CH3OH afforded white
o
needles, m.p. 153 - 154 C." H-NMR yielded a spectrum consistent with the structure.
UV-Vis spectroscopy indicate two main absorption bands at X - 250 nm and X = 340 nm.
4.2.2 UV-Vis Spectroscopy
UV-Vis experiments were performed on an Agilent 8453 diode array
spectrophotometer using 1 cm path length quartz cuvette. The spectrophotometer was
equipped with an HP 89090A temperature controller (at 25 C for these studies) and
stirring apparatus. Initial solutions for absorbance spectroscopy were prepared in a
suitable solvent (generally ddfbO, except for 25 which was in 1 : 1 ddtÔ / /PrOH). In
all studies performed the UV-Vis spectroscopic handle was present on the guest
molecule, since cyclodextrins have no strong absorbance characteristics. Preliminary
studies were performed by titrating the guest molecule into a solution containing the host
(cyclodextrin); the results from these titrations often provided data outside the useful
range of equilibrium binding from which stability constant calculations would not
converge. Therefore, initial solutions of the guest were then titrated with the cyclodextrin
of interest, ensuring that the region of 20 - 80% complexation was analyzed. Absorbance
spectra analysis using HyperQuad 2000/2006 software package was performed.
Absorbance measurements of a standard solution of the guest were taken in order to
calculate extinction coefficients at different wavelengths of interest. Analysis of the
complete absorbance spectra (280 nm - 800 nm) was not performedgenerally, only
peaks indicating evident perturbations (e.g. red or blue shifts) upon titration with the
cyclodextrin of interest were used for analysis.

Data was initially analyzed with a log Ka ~ 3 and allowing the software to
iteratively fit and refine the value. The log Ka (log p value) was refined based upon
88
fitting simulated absorbance spectra using the Hyperquad software package. This was
continued until the value generated was within the 95% confidence level and refinement
produced no change in the log Ka value. The three main ligands of interest were
diaminonaphthalene (22), dihydroxyquinoxaline (23), and iV-(l-pyridylmethyl)-2aminobenzoic acid (25).
Titrations of 22 and 23 with 2 yielded the spectra shown in Figures 4.2 and 4.3,
respectively (representative spectra from one trial shown).
Absorbance maxima at
wavelengths 280 nm and 337 nm were analyzed for binding studies with 22.
Complexation studies with 23 focused on the absorbance maximum at 258 nm. The
titrations involving 22 and 23 reveal no significant shifts in absorption maxima upon
binding. Spectra of titrations of 25 with cyclodextrin 2 and 12 are shown in Figures 4.4
and 4.5, respectively. Both titrations reveal a complexation-induced blue shift from 340
nm to 332 nm (the peak analyzed for the determination of each association constant).
Log Ka values for the guest molecules along with the cyclodextrins 2 and 12 are provided
in Table 4.1.
4.2.3 Crystallization
Attempts at crystallization of structures 22 - 25 were attempted using solvent
diffusion crystallization and slow cooling methods. All crystallization attempts resulted
in crystals without guest associated, crystals not suitable for diffraction or no crystals at
all.
Initial studies of crystals formed between 24 with p-cyclodextrin exhibited
fluorescence properties of the 3-hydroxynaphthoic acid, however, no inclusion or

association product was evident by X-ray analysis.
This suggests that either the
concentration of the 3-hydroxynaphthoic acid was very low, or both low and disordered.
4.2.4 ESI-MS Measurements

Thermo Finnigan LCQ Deca Spectrometer (University of Iowa, High Resolution
Mass Spectrometry Facility, purchased with funds from NIH grant 510-RR13799-01)
89
was used in direct injection mode with a capillary temperature of 150 C and a flow rate
of 3 ul/min for data collection. Analysis of binding of 25 with cyclodextrin 2 was
performed and can be seen in Figure 4.6. The mixture was preparing in 1 : 1 MeOH /
H2O using a 1 : 1 mole ratio of 2 / 25. Solutions of 2 were prepared in ddFÔ and 25 in
z'PrOH. A final solution of 90uM host and guest were prepared in 1:1 MeOH/H20 with a
final volume of 1 mL.
Table 4.1 Log Ka of Guest Molecules with Cyclodextrins 2 or 12.
Compound
Cyclodextrin
Log Ka
22
2.7 0.1
23
4.7 0.1
25
3.1 0.1
25
12
5.4 0.1
26*
3.0 0.35
6
* work reported by Heaweon Park in the Telford lab
285
305
325
Wavelength (nm)
345
365
385
Figure 4.2 UV-Vis absorbance of 22 titrated with 2 from 265 nm - 385 nm. No major absorbance shifts are noted.
265
Titration of 22 with 2
245
250
260
265
Wavelength (nm)
255
270
275
280
Figure 4.3 UV-Vis absorbance of 23 titrated with 2 from 265 nm - 385 nm. No major absorbance shifts are noted.
240
to
275
Wavelength (nm)
325
375
425
15eq.2
~12eq.2
8eq. 2
4 eq. 2
leq. 2
~ 0.2 eq. 2
Oeq. 2
Figure 4.4 UV-Vis absorbance of 25 titrated with 2 from 265 nm 385 nm. A blue shift in the absorption maximum at 340 nm to
332 nm is noted.
225
255
275
295
315
335
355
Wavelength (nm)
375
395
415
Figure 4.5 UV-Vis absorbance of 25 titrated with 12 from 265 nm - 385 nm. A blue shift in the absorption maximum at 340
332 nm is noted.
235
95
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96

Chemical studies focused on catalytic activity in biological and chemical systems
are prevalent. The development of catalysts targeting specific chemical transformations
has received much deserved attention. Our efforts toward developing a catalytic center
for the synthesis of chiral products or hydrolysis of esters, though limited, has provided
rigid, small models upon which might have utility in further investigations.
The
inclusion of naphthalene and naphthalene derivatives has shown the ability to mimic
enzyme systems. One example of this were attempts to mimic catechol dioxygenase by
Heaweon Park in our own lab.6 The inclusion of 22, 23, and 25 is expected due to the
suitability of the cavity size of p-cyclodextrin (6.0 - 6.5 A) and the length of these guests
(22 / 23 / 25 ~ 5 A [distance between para protons (H1-H4) of the aromatic ring]). In the
cases of 22 and 25, binding with native P-cyclodextrin (2) appears to be modest at best,
with a log Ka ~ 3. 23 with P-cyclodextrin (2) has -100 fold increase in binding affinity.
This increased affinity is interesting since the main difference between 22 and 23 is
hydroxyl or amine groups appended to the ring. Similarly 25 binding with 2 indicates
modest binding with a log Ka ~ 3. Evaluation of these binding affinities indicate that
molecules having the proper fit with cyclodextrin, but the possibility of being charged in
aqueous solutionhere, the positive protonated amine for 22 and the negative
depronatonated acid of 25have a lower affinity with p-cyclodextrin.
A direct comparison of 25 binding with cyclodextrin 12 indicates that 25 has
~100 times increase in affinity for cyclodextrin 12 over 2. The largest perturbation
driving this increase in affinity is tripling the number of H-bonding sites between 2 (7
total -OH) and 12 (7 -NHR 2 and 7 -NH 2 ). This, coupled with the elongation of the
hydrophobic cavity and possible electrostatic interactions (with the deprotonated acid and
protonated cyclodextrin amine), are the most probable explanation for this increased
affinity.
97
4.4 Conclusions
It is well-established that hydrophobic, electrostatic, and H-bonding interactions
play a vital role in the binding of guest molecules with cyclodextrins.1 The affinities of
the molecules reported here agree well with literature reports for similar molecules.1
With the UV-Vis spectroscopy studies presented here it is evident that guest molecules
22, 23, and 25 exhibit binding with our cyclodextrins, presumably due to these
interactions mentioned above. The log Ka values provide for both 23 with 2 and 25 with
12 indicate a relatively strong binding when compared to those for 22 and 25 with 2.
Here it is evident that guest molecules that have a higher propensity of being charged (22
and 25) in aqueous solution, have lower binding affinities with uncharged, native Pcyclodextrin, 2. Further support for this is provided by the higher affinity for the
negatively-charged 25 with positively-charged cyclodextrin 12. In order to analyze the
exact contribution of each of these interactions additional studies (e.g. X-ray
crystallography) are still required.
98
Notes
(1)
(2)
Gokel, G. W.; Leevy, W. M.; Weber, M. E. Chem. Rev. 2004,104, 2723-2750.
(3)
Schneider, H.-J. In Macrocyclic Chemistry: Current Trends and Future

Perspectives.; Gloe, K., Ed., 2005, pp 277-289.
(4)
Steiner, E.; Schetty, G. Helv. Chim. Acta 1973, 56, 368-374.
(5)
Jahng, Y.; Telford, J. R. B. Kor. Chem. Soc. 2005, 26, 1614-1616.
(6)
Park, H. (2007). Biomimetic Models of Intradiol Catechol Dioxygenases:

Structures and Properties of Inclusion Complexes of Fe(III) Centered Metal
Complexes With beta-Cyclodextrin Derivatives. Chemistry. Iowa City, University
of Iowa: 319.
99
CHAPTER 5
SUMMARY AND FUTURE WORK
The utility of cyclodextrin for a wide variety of applications have been reported,
along with the synthesis of modified upper- and low-rim cyclodextrins. Within this body
of work, the ability to synthesize novel per-6- and mono-6-modified a- and Pcyclodextrins has been reported. The ability to use ESI-MS for the detection of binding
constants for the second-sphere coordination of tetrahedral oxoanions has been shown.
While our studies are viable based upon recent literature reports, corroboration of another
series of molecules using a similar methodology is required. Even though separation of
these tetrahedral anions based on binding with per-6-(2-aminoethylamino)-a-cyclodexrrin
(6) was desired, no such results were found. It is clear, however, that the size inherent to
the a-cyclodexrrin architecture provides the approximate size needed for binding
tetrahedral anions. With this in mind, additional studies with other functionalized acyclodextrins are desired to see if better separation between the tetrahedral anions studied
can be achieved. Further studies would also encompass a larger range of tetrahedral
anions, such as pertechnetate and chromate.
Even though analysis of binding with the trivalent lanthanides and actinides is
limited herein, synthesis of the per-6- and mono-6-modified P-cyclodextrins has been
performed to target Lnm and An111 cations. Initial potentiometric analysis of binding the
lanthanides with per-6-5'-(thiosalicylic acid)-per-6-deoxy-P-cyclodextrin (14) provided
difficulty because of the poor solubility of 14 below a solution pH of 5. This made it
evident that below a pH of 5, the seven acid groups of 14 were being protonated
generating a neutral hydrophobic molecule.
Additional studies focusing on mono-6-
modified P-cyclodextrins were also pursued due to difficulties synthesizing desirable per6-modified P-cyclodextrins. Initial syntheses focused on the synthesis of mono-6-(2aminoethylamino)-P-cyclodextrin (18) from which a larger framework could be
100
constructed by reaction with the free primary amine of 18. The most promising of these
results
was
provided
by
mono-6-(2-(diethylenetriamine
pentaacetic
acid-
amino)ethylamino)-mono-6-deoxy-P-cyclodextrin (19) where fluorescence enhancement

of the Eu3+ fluorescence emission was seen at 615 nm. Further analysis of 19 for
determination of the four acid pKa (-log [acid dissociation]) value and log Ka (log
[association constant]) values are still required. Additional studies using time-resolved
fluorescence of these complexes with the lanthanides (e.g. Tb3+, Eu3+) are also desired for
determination of the environment around the Lnm metal center.
Further studies with
mono-6-(2-(dansylamino)ethylamino)-mono-6-deoxy-P-
cyclodextrin indicate that it provides no fluorescence resonance energy transfer from

tryptophan. This is corroborated by reports that the dansyl group from such a derivative
is included within the cavity of the p-cyclodextrin and should exhibit a decrease in the
fluorescence of the dansyl moiety upon its release from cavity (no decrease was seen
except due to concentration effects).
The inclusion of guest molecules diaminonaphthalene (22), dihydroxyquinoxaline
(23), and Ar-(l-pyridylmethyl)-2-amino-benzoic acid (25) with P-cyclodextrin (2), and 25
with per-6-(2-aminoethylamino)-per-6-deoxy-P-cyclodextrin (8) illustrate the strength of
association of these guests with the respective cyclodextrins. For the naphthalene based
structures (22 and 23) the values of log Ka ~ 3 are close to literature based values for
similar molecules. The main goals of this work were to expand literature reports of
cyclodextrin host and guest chemistry, and explore cyclodextrin as a potential candidate
for catalyst architecture.
Additional studies pursuing the binding mode(s) (and
orientation) of the guests molecules with or within the cyclodextrins need to be

addressed.
In addition, further studies with regards to their catalytic affects and
association with other derivatized cyclodextrins need to be pursued.
101
APPENDIX A
DETERMINATION OF BINDING CONSTANTS USING ESI-MS
102
A.l Importance and Background of Binding Affinities

The ability to determine the strength of interactions between multiple species is
evident in biological systems.
One of the most prominent examples is the case of
myoglobin and hemoglobin. Here the ability to quantify these interactions indicates that
CO binding to the heme in myoglobin 200 times better than O2. Another example of this
is provided by RNA and DNA.
In the case of DNA and RNA, the binding of
Mg(H20)62+ is vital to its function and structure.1"5 Along these same lines, without the
recognition of specific amino acids, the efficiency of /RNAs during translation would be
dramatically less. With the innumberable number of biologically related compounds and
processes this list continues on and on.
Biological systems provide the inspiration upon which chemists continue to build.
Literature reports of developing binding sites and quantifying their ability to target a
specific species are of major importance in chemical and environmental systems. As
previously mentioned, cyclodextrins have found utility for analogous applications.6
Studies using cyclodextrins as a supramolecular host targeted for different applications
were performed within this body of work. Understanding the interactions and binding
affinities between our cyclodextrins and the guest molecules is of key importance. The
majority of studies in our lab have utilized widely accepted methods (UV-Vis, NMR, Xray crystallography, etc.) to study these interactions. X-ray crystallography provides a
concrete answer; however, the ability to generate suitable crystals is difficult. It has also
been shown that obtaining the exact stoichiometry of binding is often problematic, and
sometimes produces erroneous results.
These problems, along with difficulties
encountered in studying cyclodextrins with tetrahedral anions required a more suitable

methodology for determining these binding affinities. In this case, ESI-MS has provided
data that was easily attainable and more suited to the system being analyzed.
103
A.2 Utility of ESI-MS in Determining Binding Affinities

Reports using ESI-MS for the determination of binding affinities were initially
met with skepticism, but have become more common over the last two decades.8 The
ability of ESI-MS to provide the stoichiometry of binding unambiguously, and utilize
micromolar concentrations makes it an ideal method for use in our studies.7'9"12 ESI-MS
provides a relatively soft-ionization into the gas phase that has been shown to take
solution based species into the gas phase. With this knowledge, an understanding of the
binding of perchlorate, phosphate or sulfate with a-cyclodextrin (1) and per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin (6) was pursued using ESI-MS. Initial
measurements analyzing the intensity of both the cyclodextrin and tetrahedral anions
proved difficult due to the positive charge state preference for the cyclodextrin and the
negative charge of the tetrahedral anions. Data encompassing the intensity of both 1 and
6 in positive or negative mode of ESI-MS would not be possible. In order to circumvent
this problem, an indirect method of achieving this was derived and provided in equations
A.1-A.7.
The determination of binding constants began with equations A.1 and A.2.
Additional guidance based upon literatures reports of binding constants determination via
ESI-MS was also utilized. '
Equation A.3 follows from the concept that the percent
bound is related to intensity of the host and guest complex (IHG) divided by the total
amount of host present. A correction (response) factor, R, is used since the intensity
ratios from the ESI-MS data should correlate in a linear manner, but not necessarily
based upon the same magnitude change in the concentrations. Since different molecular
species may interact with the mass spectrometer to differing extents, a response factor
was incorporated for each intensity ratio. Therefore, each intensity value is given its own
response factor, and all of these response factors are pulled out into one consolidated
response factor. The equations were then corrected to intensity ratios of host and guest
by utilizing the ESI-MS data to provide equation A.3. Further simplification of equation
104
A.3 to contain all variables into either the x or y terms yields equation A.4. The right side
of A.4 is then split into two terms, one constant term (b) and a term variable upon the
intensity ratios of IHG I IH, which is encompassed with the x value. Once separated, the R
(correction factor located in the b term), was then split into two terms, R' and n. Splitting
R into these two terms allow for correction of both the slope and ^-intercept of the linear
plot created, respectively. The values of R' and n are arbitrary and can vary, however,
their product, R, should remain the same for titrations with the same species involved.
The value of Kd (and therefore Ka = 1 / Kj) can be determined from the slope of the linear
line obtained, based on equation A.6.
105
K; = [H]x[G]
[HG]
(A.1)
G\
^ [G\-[G] = H -Kd [HG]/[H]
%Bound = H ound
\otal
[Hl
[#]o
(
I HG
%Bound = Rx
V tf
Rx
(
\IH
/ HG
\IH
(A.3)
HG J
_[G]0-RxKd(lHG/IH)
+
[Hi
IHG J
I HG
(A.2)
>
\G\
+ IHG J [H\xR
^d
(A.4)
*HG
[H\
IH
(A.5)
Note: Here R' is allowed to encompass the negative sign.

n = arbitrary correction factor (part of response factor) for y-intercept.
R =
R'*\n\
f
x
[G\
HG
V lH + IffG J
[H\xn
[H]0
HG
[G\xIIt
y=b+mxx
Definitions:
R = response factor = R' x n
[G]o - Initial guest (tetrahedral anion) concentration
[H]o = Initial host (cyclodextrin) concentration
[G] = Amount of free guest (tetrahedral anion) present
[H] = Amount of free host (cyclodextrin) present
IHG = Absolute intensity of HG complex
IH~ Absolute intensity of H
^
(A.7)
106
Notes
(1)
(2)
(3)
(4)
Kucharski, L. M.; Lubbe, W. J.; Maguire, M. E. J. Biol. Chem. 2000, 275, 1676716773.
(5)
Maguire, M. E.; Cowan, J. A. BioMetals 2002,15, 203-210.
(6)
Connors, K. A. Chem. Rev. (Washington, DC, U. S.) 1997,97,1325-1357.
(7)
(8)
Hofstadler, S. A.; Sannes-Lowery, K. A. Nat. Rev. Drug Discov. 2006, 5, 585595.
(9)
Dotsikas, Y.; Kontopanou, E.; Allagiannis, C; Loukas, Y. L. J. Pharmaceut.

Biomed. 2000, 23, 997-1003.
(10)
(11)
Dotsikas, Y.; Loukas, Y. L. J. Biochem. Bioph. Meth. 2002, 52, 121-134.
(12)
(13)
Loukas, Y. L. J. Pharm. Pharmacol. 1997, 49, 944-948.
107
APPENDIX B
SPECTRA AND RELEVANT DATA FOR THE BINDING OF
TETRAHEDRAL ANIONS WITH 1 AND 6
108
o
_ LO
o
T- -
ooo-
w
*
T -
140
Si
CO
1CO
_
,
296.
130
: o
en
CN
CO
*~
CO
00
^
T
~
-L
O)
1001.
990.9
J9.94
CO
.
CN
I 100
CO
CO
00
1100
CO
o
o
o
01
f^
en
CN
CM
rO)
006
05 O)
0 0 CO
T^ CN;
03
"""
S3
o
co co
O
", - O
N
>
CO C
S
o
00
: o
-o
gs
8 S i-
CD
CO
: o
: co
CM
- O
CO
i O
CO CO
f ^ OO
CO CO
*,*'
hCO
T
-a
-s
-
LO.
o
CO
CO
*r
400
384.
r-
LO
. t
m
i
CM
3"
CO
CO
i-
CM
CO
h1"-;
-o
co
CO
CM
CO
O
Vi
:r
VJ
Vi
200
d
Tf
CO
t>0
csi~~
CO
O
o
LO
en
o
en
to
oo
Q
oo
LO
1*-
CD
o
<o
LO
LO
o
in
m
^-
o
fl-
eouepunqv eAêiey
1 l|
LO
CO
II
LO
CN
O
CN
LO
to
c3 ^~
CO
0^T
100
10
15
20
25
30f
35
40
221.09
240.10
331 14
412.35 486,81
ll ,L
550.11
I 549.13
655.23
I 656.26
759.92
m/z
8O0
81734
Figure B.2 ESI mass spectrum for 1, a-cyclodextrin titrated with LiC104 (1 : 3).
0)
I 50^
I 55-
$ i
60
65-
70-
75
80^
85-
90
95
100^
985.35
989.90
1088.15
1091.20
1092.21
1197.06
1194.04
1191.02
1193.02
1351.32
1351.3: 1404.87
S0
5
0^J
100
10d
15
20t
25-
30
35
40-
J I 202.63
147.03
| I .
247.68
1
417.11
509.13
489.10 558.25
670 6
810 .85
785.33 t
811.87
g0286
972.O8
rrVz
îL.jiiiJfiJklJLMW'
800
728.00
Figure B.3 ESI mass spectrum for 1, a-cyclodextrin titrated with MgSC>4 (1 : 3).
H 45-
I 50^
>
c
a 55c
" _
65-
70z
75f
80-
85-
90
95
100
1
t
1100
1156.55
, 1220.55
il
1274.16 133423
1470.65
^ ^ J J i u a i ^ f-i "-- - ' L u u . ; ^ , r . , 7 . - , r J , . , . . . ^ i . . , n
1200
1300
1400
1500
k
r i
| 1105.18
L.I. l . L
162.76
272.66
240.08
174.86 212.91
408.57
400
i l l I, I,
348.68
622.58
680.52
892.42
m/z
800
iLiJii,Liliii.L.ii,jLI.
816.34
Figure B.4 ESI mass spectrum for 1, a-cyclodextrin titrated with KH2PO4 (1 : 7).
5^
10-
15
20^
25-
30
35
4CH
1 45
>
I 50
I 55
65r
70-E
75z
80z
85z
90^
,,,1 J V " ? ' 8 2
I 1186.98
1360.93
1320.89 ^35891
"I , I
1456.80
112
o
o
to
co
Tf
00^
CO
o)
-J;
X
ID
en
CN
o
<D
o
a,
o
i t
^
<D
o
c
'e
s-
a,
1-4
,o
3
-*
-t->
m
o
K
O
ID
OH
205
200
l!
11111
11 111
m
CO
M
o
CO
11111
11 11
11 [i i
m
CO
to
,:
m
1
o
GDuepunqy 8A|ie|oy
-*
i i i i i
m
co
i 11 11
CO
i i I i i i i I i i 111 11 11 l i ii
m
o in
o
CN
OJ
11 11 11
tn
CO
w
ID
PQ
0)
Ml
113
~5
l
CO , _
o> to
Q>
CN
eouepunqvsAiiBjsy
T-
T-
-za.
10z
15]
20]
15-_
30i
35z
40-
200
LL
34
9.12
369.29
375.33
300
400
.JL1.-..,J_.U,.II.-,L.L1,L ,LIJ!
247.06
205.07
389.35
395.36
442.69
409.35
500
583.40
600
700
800
m/z
900
1000
1123.37
LiLlL,
1100
1353.15
1400
1500
U.LLLILLIllkgfl,.^^
1283.27
1225.44
1183.41
11 i i k l
1105.38
979,32 1045.30
kj,lllJ.I.,lll:^J.Ui.L,,.,.LL_.i.L..,[.LJ.LL...I,i.J.L,L .;1..,...L.
677.27
mm,
613.41
592.39
Figure B.7 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin with LiC104 (1 : 1).
a.
1 45
I 50
1 55
0 60
o
65
70
75
80
85^
90z
95
100
5f
10
15-
20
25
30
35
40
400
,-J. ,i_iiJMJill
300
200
355.16
375,30
ii-iiJ
i 247.06
205.05
389.38
409.29
532.34
500
592.39
600
583.40 !
uiyiMill
442.67
700
663.37
642.32
979.28
1123.37
1063.32
m/z
900
1000
1100
ILl,..,lj.Ll.lN.Li.:..l.,...,.J.. l |.. l .,.l,>...l,L..,l.li.Lj.^.L,.tl 1 .J.J. l ..
775.18 817.20
1200
1183.43
1425.22
1300
1400
1500
hlhb]iiJli,MllLl,.,Jlllfel,,u,...-.
1325.28
1283.25
cc
I 55^
I 50^
I 45
a, 60o
65z
70^
75^
80
85
90
95
100
10d
15
20z
25-
30
35
40-
45_
169.02
I i
206.07
205.05
307
-12n
n
i, .
409.27
i
395.38
;1.."JDLMJIII
367
532.36
523.28
lilllliiii
428.63
422.73
m/z
817.09
901.06
1300
1400
^j.L.j.^n.iu.,il.itUL>t^.tyt->JJnix..L.-^:U.l,
1283.23
1123.33 11S3-34 1225.31 I
1325.23 13B3.16
1100
iLli.LL.ybLjtlUjLlt.JbLj^laj
979.30 1021.25
<D
I 50
I 55-
60
65d
70
75f
80
85
90f
95d
100-
55^
200
_i.J,,
247.08
205.0S
300
^J.I.-JJLLLUI
349.09
369.32
375.32
389.35
395.37
553.34
562.36
500
LUJillkLklk
600
77516
817.23
700
m/z
800
900
1000
1100
1200
UJJUii,
1300
1253.39
1225.45
1183.43
1165.41
1105.34
LILJJLJJLUL
979.30 1045.30
m i L i L L . j LLJJiîi,...L1.L..i.u,iJ_..l.,..j.i.jj
bU^.ô
RQ5
627.46
656.24
613.39
583.41
532.35
523.36
511.21
451.88
409.36
592.40
1400
1353.06
1500
Figure B.10 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin with MgSC>4 (10:1).
10
15f
20f
25^
30f
35-]
I 4Sf
a.
40z
fl>
I 50^
o 60f
65^
70^
75T
80q
85
90H
95
100
0:
5:
10:
15:
20-
'I
30^
35:
40^
200
! 247.07
-LlL-1..-
205.05
532,39
553.40
562,35
583.42
500
:l.J(l...J....l..l.,JtHl,l
; 523.31
; 511.26 ,,
451.84
400
- ?
-ulL-.J-IU, nil.
355 1
369.31
375.29
389.35
395.37
409.37
77
5-21
817.18
700
m/z
900
1000
1200
! 1225.46
1183.42
1165.40
1100
1063.31
979,28 1045.26
,U,l.1ij,...Ll..,l.l,.,,ll.,..J.,..I....L,...l,..l.. ,J
| 692.30
677.16
662.39
613.41
592.41
1310.98
1400
1383.35
1500
1481.27
Figure B.ll ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin with MgSC>4 (1 : 1).
<i>
<D
I 50^
1 55^
0 60o
65^
70^
75z
80
85
90
95
100
119
PQ
CI
120
(M
T-
1-
aouepunqv sAjiepy
s
ISA
5
04,
10
15
20
25
30^
35z
40
451.82
523.37
532.33
678 07
-
775
'26
817.27
[
925.29
979.33 1045.36
I I
1105.39
LluL
1063.37
1165.43
ilJU
1141.41
LLlU l l . I i . l , l
1311.05 1353.07
| 1354.10
13 17
1429.16
Figure B.14 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin with KH2P04 (10 : 1).
1 45
o
I 50^
>
S 60
I 55
65z
70z
75
80
85
90-
95
100
122
i
o
c
<N
a,
o
S3
S3
a
i i
GO
sS
5^
10
15
20f
25-
30
35-
40^
205.05
151.05
389.33
2,r/,.l,i,.Jl,lij4il4^ji;î;k^:4lî
375.30
571.36
553,38
700
m JL
i 613.42
592.43
800
1400
1324.18
1353.09
1354.09
UtDo.To 14.'i? 73
1500
Figure B.16 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin with KH2PO4 (1 : 5).
I 45
I 50
S 55
c
8 -
65
70
75f
80
85
90
95-
100
80
205.03
.86 J
613.42
592.43
1400
1429.34
1500
Figure B.17 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin with KH2PO4 (1 : 10).
5-E
10z
15-E
20f
25r
30E
35z
I 45^
I 5ol
I 55H
65-
70^
75
95
100^
K>
4^
* -15000
3800
.^^^
3900
4100
IHG'(!HX[G]0)
4000
4200
CM"1)
4300
^ ^ ^ ^ ^ ^ ^
" ""-^^^^^^
4400
4500
y = -12.812x +37193
R2 = 0.7552
4600
Figure B.18 Linear regression attained by inputing data from the first titration of 6 with LiC104 to equation A.6. The slope of the line
is then used to generate the Kd value as outlined in Appendix A.
3700
/
xc\nnr\
-JUUUU I
-25000
iJ? -20000
-10000
-5000 -
n
u
Linear Regression 1 - Cyclodextrin 6 with LiC104
3000
-JUUUU
incinn
-25000 -
-20000
3500
N .
*
^ s .
N.
4000
[G] 0 )
(NT1)
4500
^ V .
^ ^ \ ^
IHGIVH*
N .
5000
y = -12.508x + 37037
R2 = 0.9138
5500
Figure B.19 Linear regression attained by inputing data from the second titration of 6 with LiC104 to equation A.6. The slope of the
line is then used to generate the K<i value as outlined in Appendix A.
*, -15000
-10000 -
-5000 -
Linear Regression 2 - Cyclodextrin 6 with LiC104
3900
4100
IHGKIH*
4300
[G]0)
-K
(M-)
4500
4700
4900
y = -12.298x + 37076
R2 = 0.9594
5100
Figure B.20 Linear regression attained by inputing data from the third titration of 6 with LiOC>4 to equation A.6. The slope of the
line is then used to generate the K<j value as outlined in Appendix A
-30500
3700
-25500
-20500
g -15500
10500
-5500
Linear Regression 3 - Cyclodextrin 6 with LiC10 4
-2000
2400
-3500
-3000
2500
2600
2800
-i>
IHGI{IH x [G]0) (M-)
2700
2900
3000
y = -2.8086x + 5578.9
R2 = 0.9294
3100
Figure B.21 Linear regression attained by inputing data from the first titration of 6 with MgSC>4 to equation A.6. The slope of the
line is then used to generate the Kd value as outlined in Appendix A.
1 -2500
1500
1000
Linear Regression 1 - Cyclodextrin 6 with MgS0 4
^nn
2700
-JUUU
^nnn
-2500
-2000
2800
"1
^ ^
3000
IHG/(Iffx[G]0)
2900
(M"1)
3100
3200
y = -2.3889x +5555.6
R2 = 0.8489
3300
Figure B.22 Linear regression attained by inputing data from the second titration of 6 with MgSC>4 to equation A.6. The slope of the
line is then used to generate the IQ value as outlined in Appendix A.
1 -1500 -
-1000
-JUU
Linear Regression 2 - Cyclodextrin 6 with MgS0 4
-3500
200
^v
400
^ v
N.
600
IHG/(IHx[G]0)
^s.
(IvT1)
800
1000
\ v
1200
^s.
y = -4.3466x +743.8
R2 = 0.9509
1400
Figure B.23 Linear regression attained by inputing data from the first titration of 6 with KH2PO4 to equation A.6. The slope of the
line is then used to generate the Kj value as outlined in Appendix A.
()
-5000 -
-4500
-4000
-3000
* -2500 -
1g -2000
-1500
-1000 -
-500
Linear Regression 1 - Cyclodextrin 6 with KH 2 P0 4
-3500
-3000
400
500
700
IHG/(IHx[G]0)
600
1
(M"-u
)
800
900
y = -4.2808x + 740.74
R2 = 0.9383
1000
Figure B.24 Linear regression attained by inputing data from the second titration of 6 with KH2PO4 to equation A.6. The slope of the
line is then used to generate the Kd value as outlined in Appendix A.
^T
1 -2500
g -2000
-1500
1000
2.
-2000
-1500 -
400
^ \ .
500
N .
700
^
^
800
IHGI{IH x [G]0) (M"1)
600
^"\#
900
^"v^
y = -4.5445x +741.51
R2 = 0.9898
1000
Figure B.25 Linear regression attained by inputing data from the third titration of 6 with KH2PO4 to equation A.6. The slope of the
line is then used to generate the Kj value as outlined in Appendix A.
Afififi
-tUUU
-3500
J? -3000
* -2500 -
innn
-1UUL;
10: 1
8.99066E-05
1.04887E-05
545152939
0
100 : 1
Ratio 6 : LiC104
[6](M)
8.99066E-05
3.49624E-06
Trial 3 [LiC10 4 ](M)
464004601
IH
0
IHG
Trial 2
10:1
0.00009
0.000018
644137403
23137057
100: 1
0.00009
0.0000009
664580981
0
IHG
latio 6 : LiC104
[6](M)
[LiC10 4 ](M)
10: 1
8.96239E-05
8.60526E-06
998485199
0
100 : 1
Ratio 6 : LiC104
[6](M)
8.96239E-05
Trial 1 [LiC10 4 ](M)
2.86842E-06
1086434834
IH
0
IHG
Table B.l ESI-MS Intensities of Host (IH) and
1 :5
0.00009
0.00045
196108077
368705095
1 :10
0.00009
0.0009
119831027
354568636
1: 1
1 :3
1 :5
8.99066E-05 8.99066E-05 8.99066E-05
0.000104887 0.000272707 0.000451015
361493206
262677727
127213542
189575933
314205511
224141592
1: 1
0.00009
0.00009
376604237
172452712
1: 1
1 :5
1 : 10
8.96239E-05 8.96239E-05 8.96239E-05
8.89211E-05 0.000447474 0.000899537
948492891.2 352768338.4 151795774.5
614443815
518874176.4
381948924
1 :7
8.991E-05
0.0006293
93731613
229219563
/ Guest Complex (IHG) for 6 Titrated with LiC104
IH
IHG
Ratio 6 : MgS0 4
[6](M)
Trial 2 [ M g S 0 4 ] ( M )
100: 1
0.00009
0.0000009
602698222
0
10: 1
0.00009
0.000009
528198373
0
1 :1
0.00009
7.82609E-05
386088730
98415945
1 :1
Ratio 6 : MgS0 4
100: 1
10: 1
[6](M)
8.96239E-05 8.96239E-05 8.96239E-05
Trial 1 [ M g S 0 4 ] ( M )
1.02961E-06 9.06061E-06 8.99883E-05
1177956595 1157617645 1008793205
IH
0
0
69290633.99
IHG
1 :5
0.00009
0.00045
108043567
140249053
1 : 10
0.00009
0.0009
44996966
113561190
1 : 10
1 :5
8.96239E-05 8.96239E-05
0.000449942 0.000899883
469192360.9 222694035.9
144558588.3 124504491.8
Table B.2 ESI-MS Intensities of Host (IH) and Host / Guest Complex (IHG) for 6 Titrated with MgS04
10: 1
8.99066E-05
8.15343E-06
509601418
0
100: 1
Ratio 6 : KH 2 P0 4
[6](M)
8.99066E-05
Trial 3 [ K H 2 P 0 4 ] ( M )
2.03836E-06
682513081
h
0
IHG
IH
I\\G
10: 1
0.00009
0.000009
1442349414
0
100: 1
0.00009
0.0000009
1160716738
0
Ratio 6 : KH 2 P0 4
[6](M)
[ KH2PO4 ] (M)
100: 1
10: 1
Ratio 6 : KH 2 P0 4
[6](M)
8.9623 9E-05 8.96239E-05
Trial 1 [ KH 2 P0 4 ] (M)
1.7459E-06 8.72952E-06
2671456992 2766224870
IH
0
0
IHG
1 :1
8.99066E-05
8.96877E-05
332173905
27780680
1 :1
0.00009
0.00009
1235021841
99345727
1 : 1
8.96239E-05
9.0787E-05
1331809151
152309250.7
1 :3
8.99066E-05
0.000269063
213960149
39391500
1 :5
0.00009
0.00045
573119721
197308807
1 :5
8.96239E-05
0.000450443
902340435.4
159888330.6
1 :5
1 :7
8.99066E-05 8.991E-05
0.000450477 0.0006299
108609574
81703276
23857996
24075217
1 : 10
0.00009
0.0009
256589742
114003292
1 : 10
8.96239E-05
0.000900886
411584482
95756599
Table B.3 ESI-MS Intensities of Host (IH) and Host / Guest Complex (Im) for 6 Titrated with KH2P04
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1.1
1.2
1.4
1.6
2
4
6
10
20
0.00005
0.05
Delay Time (sec)

5.09
-147287
-127300
-107905
-70710.9
-38900.5
-11176.1
14470.5
35523.81
54435.81
70456.09
84344.09
96860.19
107473.7
117334.6
133002.5
144703.8
160952
183339.8
185641.5
185477.1
186788.8
4.01
-162746
-147234
-132648
-104121
-78029.9
-54554
-33091.9
-13331.2
681.38
40600
50735.66
62810.5
73704.19
83276
99869.69
113079.9
138608.9
190376.5
199646.5
202188.8
203614
3.67
-84117.3
-79505.3
-74684.2
-65887.9
-58035.5
-50266.8
-43860
-37743.8
-31480.4
-26162.3
-21379.5
-16771.5
-12151.1
-8203.62
-950.25
5232
16828.44
49866.59
67633.2
87837.83
107565.6
3.56
-47313.7
-39757.9
-32409.3
-15022.5
-9033.09
648.81
9213.69
15891.78
21854.5
26943.75
31661.59
35294.31
38921.22
41830.12
46462.03
49713.75
54730.92
60864.62
61511.44
61909.39
61714.81
5 (PPM)
-64810.8
-46808.3
-31079.5
-5606.84
14003.28
29334.25
40601.47
49251.34
56336.47
61101.66
65328.34
68443.34
70778.78
72433.97
74861.12
76159.81
77690.31
78545.38
78212.09
78265.69
78235.88
3.21
-138238
-111906
-88194.6
-49830.2
-19599.8
5021.66
23860.75
38958.69
51599.91
61958
70310.19
77461.66
83944.56
89173.84
97646.53
104699.7
115469.8
146200.2
161046.1
173634.1
178811.4
2.9
Table B.4 Relative H-NMR Peak Heights for peaks of interest of per-6-(2-aminoethylamino)per-6-deoxy-a-cyclodextrin (6).
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1.1
1.2
1.4
1.6
2
4
6
10
20
0.00005
0.05
Delay Time (sec)

5.09
-409538
-350222
-294617
-194750
-107716
-32946.8
36449.62
-91677.6
141210
184490.4
223970.5
256115.3
284502.5
310674.1
352692.1
385085.9
428359.6
488225.8
493200.6
495133.4
495890.5
4.01
-456316
-406169
-360544
-272301
-192984
-122375
-59129
-574.5
61543.5
98380.62
140287
178537.8
213094.2
244730.4
297980.2
342496.8
410781.6
534488.8
552977.1
558180.3
559439.5
3.67
-204753
-363342
-188278
-171036
-154180
-138941
-123169
-109789
-98562.6
-86777.1
-74738.5
-66127.1
-55914
-45899.3
-30337.9
-16240.2
12387.62
99278.64
148818.1
210559.5
262796.6
3.56
-126811
-105691
-86010.9
-51301
-22442.1
2372.38
23583.62
42195.75
58060.75
71411.62
82706.25
93191.88
101482.8
109899.6
120820.6
130207.8
142604.9
159255.9
160819.6
161103.2
161167.5
5 (PPM)
3.21
-162392
-116715
-77727.9
-13484.8
36007.12
72716.62
100900.5
123160.4
139920
152400.8
163602.6
170148.4
175632.1
180251.9
186049.6
189615.8
193659.1
196057
195555.2
194668.5
194660.3
-385511
-320761
-259673
-158949
-80192.1
-17199.8
33988.88
76285.38
112743.4
139224.5
163749
183616.3
202051
217586
243116.3
264038.8
295053.5
392854.3
437659.6
474687.5
487438.8
2.9
Table B.5 Relative ^-NMR Peak Heights for peaks of interest of per-6-(2-aminoethylamino)per-6-deoxy-a-cyclodextrin (6) titrated with LiC104 (1 : 1.2).
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1.1
1.2
1.4
1.6
2
4
6
10
20
0.00005
0.05
Delay Time (sec)

5.09
-380623
-324671
-274051
-183205
-101080
-30315.1
29154.25
83735
129083.1
169699.3
205685.6
236663.8
262202.4
286905.9
326809.9
355436.8
398550.1
457768.4
461175.3
463571.5
-380623
4.01
-395634
-354635
-317807
-244425
-180706
-122542
-681615
-21167.8
22104.25
68087
102228.9
134618.6
165079.9
191316.5
239100.1
277886.3
339916.2
458814.1
478633.5
482862.2
-395634
3.67
-169997
-163151
-156350
-142811
-126978
-114785
-102888
-94111.4
-81840.5
-70548.4
-66503.8
-61591.3
-55060.8
-49893.4
-34491.5
-19160.9
-2499
55145.66
107565.2
166835.4
-169997
3.56
-132262
-109643
-89931.3
-54881.9
-27230.8
1029.5
23027.75
39728.12
54940.38
69838.25
82398.12
92990
104021.4
110007.7
122739.8
132534.4
146915.3
164897
167375.1
167406.6
-132262
8 (PPM)
3.21
-145391
-104566
-69956.6
-12712.6
30966.38
64403.25
91719.38
109773.6
124440
135386.3
143980
152586.3
158131.8
160838.1
167440.9
170295.8
174787.8
176656.7
175923.9
175189
-145391
-376896
-313324
-257240
-160042
-83764.5
-20528.1
30633.75
68655.25
312239.6
134019.5
155712.3
173553.4
191445.3
260324.6
273759.8
282765.8
297087.8
377724.4
416554.6
460586.3
-376896
2.9
Table B.6 Relative 'H-NMR Peak Heights for peaks of interest of per-6-(2-aminoethylamino)per-6-deoxy-a-cyclodextrin (6) titrated with LiC104 (1 : 4).
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1.1
1.2
1.4
1.6
2
4
6
10
20
0.00005
0.05
Delay Time (sec)

5.09
-348358
-297631
-251395
-166593
-92683.5
-28363
29560
76546.81
118502.4
156244
189070.8
216798.7
242839.8
264105.4
299148
327774.4
365054.8
415077.8
420271.3
419724.9
421820.8
4.01
-320786
-288941
-258839
-201593
-149404
-102556
-58812.6
-20258.9
15835.25
76678.62
77918.69
105682,5
130219.5
153068.5
191372.4
223851.6
273240.9
368574.2
384704.4
388115.1
388721.9
3.67
-120731
-125099
-120265
-114546
-102771
-94498.1
-86087.1
-77360.3
-69138.4
-62373.5
-55794.4
-49360.9
-44668.5
-36149.6
-28366.6
-20526.1
-5412
47786.08
82184.79
124107
164717.3
3.56
-132337
-111810
-92345.5
-58629
-28467.3
-4579
18716.75
38842.31
55606.31
68519.88
82645.56
91625.94
101904.4
109981.3
123884.4
134316.4
147168.4
168214.2
169893.2
171319.2
172381.8
5 (PPM)
3.21
-127682
-92580
-61626.1
-13610
27992.38
56234.5
79182.12
95030.69
108919.9
119242
126119.6
132459.1
136962.1
140358.6
144964.8
147803.4
150479.3
151680
151714.7
151643.1
151078.4
-404684
-288891
-189737
-28801.6
93693.12
183869.4
256368.1
309393
352448.6
384433.7
409678.4
429127.2
445336.4
459246
475432.9
487423.8
499713.1
508866.6
510596.8
510850.4
507204.5
2.9
Table B.7 Relative !H-NMR Peak Heights for peaks of interest of per-6-(2-aminoethylamino)per-6-deoxy-a-cyclodextrin (6) titrated with LiC104 (1 : 8)
140
APPENDIX C
SPECTRA: MODIFIED CYLODEXTRINS
375.31
200
300
j.lLl,û^. LLJ.LJ,MI1
247.06 349.11
205.07
395.37
409.37
583.41
613.4-1
"35233"
900
1000
1100
-r-T-j-T-
1045.31
961.28
JjJiLiL Ld
1105.37
1123.39
1165.43
1200
1400
1500
"1 T T I
1383.29 1458.31
I I . i - i ^.ilu.,...-.
1300
AJ.JI.J
I 1310.97
1253.43
1225.48
1183.44
Figure C.l Positive ESI-MS spectrum for 6, per-6-(2-aminoethylamino)-per-6-deoxy-a-cyclodextrin.
0:
10;
20;
30H
"5 40
50i
60-
70-
80-
90-
100-
142
1000
1046.06
1200
124902,
1309.88
1400
m/z
662.09
1600
in Lin 1 5 2 7 0 9
1800
2000
2200
1899.72 2030.63 2189.31
Figure C.3 Positive ESI-MS spectrum for 12, per-6-(2-aminoethylamino)-per-6-deoxy-p-cyclodextrin.
800
779.83
818.39
1430.26
2400
2384.80
>.
OJ
.4.5
-i
\
1
r-
"I
WV.
'
'
'
Figure C.4 H-NMR spectrum of for 12, per-6-(2-aminoethylamino)-per-6-deoxy-P-cyclodextrin.
5.0
A
I
'
>"
V\
H
W
v.
yJiyilJiiili
800
812.89
1000
:-
985.73
i,:
893.67
1200
1073.28
liljiUiilLilii,!
1400
1352.68
1190.97
I 1214.08
1650.37
m/z
1600
ilUlUMyi
1489.06
1800
1786.19
1811.86
1837.50
1948.52
2000
1974.23
2200
2400
I ....;. I I ; ! . . 3, k i l l .nJllliii
2298.18 2428.72
2178.82
2108.93
Figure C.5 Positive ESI-MS spectrum of 14, per-6-5'-(thiosalicylic acid)-per-6-deoxy-p-cyclodextrin ( [10 + K]+ = 2125 m/z) along
with breakdown products.
QjllljJiilliU
53
10^
15f
20^
25
30
35
40
45
50^
55
60
65f
70^
75
80l
85^
90
95a
100-
r-
3.0
-i
r-
AA-"1*^
7.6
T
7.4
r-
v._v
7.2
70
-i
6.E
V t f "
.6
r-
-i
6.4
PFH
** H'"'V"lWVW>^"î*.'^MI4AlVi l 'J/vJ*^ f 'M t ^"i'*>Ji tf> *"*l
Figure C.6 H-NMR spectrum from 8 6.2 - 8.4 ppm of 14, per-6-5'-(thiosalicylic acid)-per-6-deoxy-P-cyclodextrin.
L j M p , J - ^ l u l t | . , | ^ H , . *V>iAW^t
OS
5.D
<~
">
r~
-|
40
3.5
V.
.:
J \
^'UJ
-i
JjÂ/
1
J. .U
r-
Figure C.7 H-NMR spectrum from 5 2.0 - 5.3 ppm of 14, per-6-5'-(thiosalicylic acid)-per-6-deoxy-(3-cyclodextrin.
~"
It
PPM
70^
80
90^
o-
10^
20^
30^
200
400
298.47 416.11
478 54
675.25
600
...mil >. : JllljiiilUltiUi.il

' " I " 1
l - ' " l ' "
L;
800
825
19
1000
m/z
891.00 1020.91
1200
,l I .,., ,
[1177.42
1139.45
1400
J..,
1600
1505.19 1627.54
1465.32
i.jrillt.iiillti^.iBiii
1311.43
Figure C.8 Positive ESI-MS spectrum for 16, mono-6-tosyl-mono-6-deoxy-P-cyclodextrin.
DC
1 40-
_>
Abundance
100-
en
o
m
o
1911.88
1800
2000
II . | I I I I | M I I | l II l | I I I I )
00
149
x
<u
o
_o
o
>>
o
I
CO.
I
o
o
o
o
c
o
e
I
o
vo
"
'
3.5
3.0
^ ^ \ y ^ v ^ - ^""imfriiftî^M^m^ n/n^JlUMîUjw^jt^ft^î^/f
*[wlw/f
2.5
^** f,A ^V | iw^fA^vOV*Ârfy/ V p ^ t d y ^ ^ i i f r y , ^ , ^ ^ ^
Figure CIO H-NMR spectrum from 8 1.5- 4.2 ppm of 16, mono-6-tosyl-mono-6-deoxy-P-cyclodextrin.
4.0
Ay
Vu /W%
->
PPM
fry%gjj^*jvi*W-'^'W*S<^
70
80^
9I
20:
30^
200
28
400
^ 9 4 431.88 [
445.04
600
800
791.28
769.09
587.29 ] [688.31
60
1000
[ 948.92
93^3
m/z
^ ^
1200
|L
..h
1400
IW*L
1375.26
1277.44
1255.2?
1600
1605.16 1696.43
2000
1851.95 1956.78
1800
Figure C.ll Positive ESI-MS spectrum of 17, mono-6-./V-(/>amirionicotinic acid)-mono-6-deoxy-P-cyclodextrin.
Relative Abundance
100-
I I I I I
8.5
8.0
r-
7.5
-1
7.0
6.5
<
6.0 PPM
Figure C.12 'H-NMR spectrum from 5 5.9 - 9.0 ppm of 17, mono-6-Ar-(p-aminonicotinic acid)-mono-6-deoxy-p-cyclodextrin.
9.0
153
6JD
Figure C.14 'rl-'H COSY of 17, mono-6-iV-(/7-aminonicotinic acid)-mono-6-deoxy-mono-6-deoxy-p-cyclodextrin.
^
4^
155
156
o
o
t~-
COIL
cri't
^
CM!^
cni
< :p
oof
lOt
LOiF
n t
o
COih CD
t - IF OO
t
coir
.
if
CD If-"
m
CD
5;
CO
cn 1
CM j
CO i
ool
* !
^~~
393.
5
CS1
CM
CD
1200
1199.6
co
CM
CD
CO
140
CD
CO
r-:
in
"*""
*m'
in
T
CD
'S
O
0
<=
>a *
co
LO
00
:
1--
co
10
r~,-i
00
O
OO
r L
co
CD
CD
*
_
O
"
I D _, :
CD^
O
CD
r
O
O
<
CO
CM
ID
CM
CM
S ' 0
CM C : <*
r^
:
CD
CO
-
:
h:O
LD
CD
CM
CD
O
1 1 1 1 | !
CD
CO
1 1
o
oo
o
r~-
o
to
o
in
o
<*
eouepunqv eApiey
o
co
o
CM
-'
CM
1 1 | 1 1 1 1 |
CD
CD
T
M5
u
s-
WD
157
x
-a
_o
o
o
I
ca
>?
x
o
o
~o
\o
o
a
o
c
fi
+-
ID
1
I
ô
I
o
c
o
s
e
a
o
OH
tzi
a:
I.
342.22 I
374.27
527.24
800
m/z
1021.91
1000
927.82
886.59
736.36
781.59
649.81
0 : JLJII i.^^liiiiyilj, i,hft liifai

200
400
600
10: 161.03
20
30-
766.32
1200
ill
1160.47
1333.541
1400
1432.23
1354.70
1800
1644.82 1773.84
,
i ij",,
1600
1533.00
2000
1984.30
Figure C.18 Negative ESI-MS spectrum of 19, mono-6-(2-(diethylenetriamine pentaacetic acid-amino)ethylamino)-mono-6-deoxy-pcyclodextrin.
t.
50;
>
to
40
60 :
70-
80-
90
100
_^-"
^
r~
4.0
2.5
3.0
Figure C.19 H-NMR of 19, mono- mono-6-(2-(diethylenetriamine pentaacetic acid-amino)ethylamino)-mono-6-deoxy-Pcyclodextrin.
5.0
- 1
PPV
...
'v'Û.
:
2
ppo*
-6,8
-8.0
4,5
-4.0
3,0
-2.S
a.o
1.S
1.0
-as
-0.0
ppfs
ITT 1
Figure C.20 'H-'H
COSY of 19, mono-6-(2-(diethylenetriaminepentaacetic acid-amino)ethylamino)-mono-6-deoxy-p-cyclodextrin.
"1
hiV
f i *jjf-
it!
~7""
T 13/Figure C.21 I THC HMQC of 19, mono-6-(2-(diethylenetriamine pentaacetic acid-amino)ethylamino)-mono-6-deoxy-Pcyclodextrin.
200
280.11
238.86
400
466.02
600
569.08 661.95
628.02
800
790.01
m/z
:...':
1114.01
1000
951.99
1200
1400
; la
1432.45
1273.01
. : : ; ' .
1157.46
1600
liiul jiiiiii
1636.11
1578.08
1519.45
1460.51
1800
1751.27
Figure C.22 Positive ESI-MS spectrum of 20, mono-6-(2-(dansylamino)ethylamino)-mono-6-deoxy-p-cyclodextrin.
10
15
20
25
30
35^
40
45
50
55
60
65
70
75^
80
85
90
95
100
1872.79
2000
<-
8.5
'
<~
8.0
<-
7.5
r-
""
<~
7.0 PPM
Figure C.23 H-NMR spectrum from 5 7.1 - 9.2 ppm of 20, mono-6-(2-(dansylamino)ethylamino)-mono-6-deoxy-p-cyclodextrin.
9.0
164
GO.
TT 1
Figure C.25 I'H-'H
COSY of 20, mono-6-(2-(dansylamino)ethylamino)-mono-6-deoxy-P-cyclodextrin.
ON
Figure C.26 'H-13C HMQC of 20, mono mono-6-(2-(dansylamino)ethylamino)-mono-6-deoxy-(3-cyclodextrin.
g^
167
REFERENCES
Adam, J. M.; Bennett, D. J.; Bom, A.; Clark, J. K.; Feilden, H.; Hutchinson, E. J.; Palin,
R.; Prosser, A.; Rees, D. C; Rosair, G. M.; Stevenson, D.; Tarve, G. J.; Zhang,
M.-Q. J. Med. Chem. 2002, 45, 1806-1816.
Alderighi, L.; Gans, P.; Ienco, A.; Peters, D.; Sabatini, A.; Vacca, A. Coor. Chem. Rev.
1999, 7S4, 311-318.
Alexeev, Y. E.; Vasilchenko, I. S.; Kharisov, B. I.; Blanco, L. M.; Garnovskii, A. D.;
Zhdanov, Y. A.J. Coord. Chem. 2004, 57, 1447-1517.
Alston, D. R.; Slawin, A. M. Z.; Stoddart, J. F.; Williams, D. J. Angew. Chem., Int. Ed.
Engl. 1985, 24, 786-787.
Alston, D. R.; Slawin, A. M. Z.; Stoddart, J. F.; Williams, D. J.; Zarzycki, R. Angew.
Chem., Int. Ed. 1988, 27, 1184-1185.
Alston, D. R.; Ashton, P. R.; Lilley, T. H.; Stoddart, J. F.; Zarzycki, R.; Slawin, A. M. Z.;
Williams, D. J. Carbohydr. Res. 1989,192, 259-281.
Ando, I.; Ujimoto, K.; Kurihara, H. Bull. Chem. Soc. Jpn. 2001, 74, 717-721.
Anwander, R. In Lanthanides: Chemistry and Use in Organic Synthesis; Kobayashi, S.,
Ed.; Springer, 1999; Vol. 2, p 305.
Armspach, D.; Gattuso, G.; Koniger, R.; Stoddart, J. F. Bioorg. Chem.: Carbohydrates
1999, 458-488, 597-602.
Bhattacharyya, A.; Mohapatra, P. K.; Manchanda, V. K. Solvent. Extra. Ion Exc. 2006,
24, 1-17.
Bhattacharyya, A.; Mohapatra, P. K.; Manchanda, V. K. Solvent. Extra. Ion Exc. 2007,
25, 27-39.
Bortolus, P.; Grabner, G.; Koehler, G.; Monti, S. Coord. Chem. Rev. 1993,125, 261-268.
Bradshaw, J. S.; Izatt, R. M.; Bordunov, A. V.; Zhu, C. Y.; Hathaway, J. K. In Molecular
Recognition: Receptors for Cationic Guests; Gokel, G. W., Ed.; Pergamon: New
York, 1996; Vol. 1, pp 35-152.
Bratt, T.; Ohlson, S.; Borregaard, N. Biochim. Biophys. Acta 1999, 1472, 262-269.
Braun, V. Int. J. Med. Microbiol. 2001, 291, 67-79.
Bricout, H.; Caron, L.; Bormann, D.; Monflier, E. Catal. Today 2001, 66, 355-361.
Cabou, J.; Bricout, H.; Hapiot, F.; Monflier, E. Catal. Commun. 2004, 5, 265-270.
168
Carey, F. A. Organic Chemistry; 6th ed.; McGraw-Hill, 2006.
Caron, L.; Bricout, H.; Tilloy, S.; Ponchel, A.; Landy, D.; Fourmentin, S.; Monflier, E.
Adv. Synth. Catal. 2004, 346, 1449-1456.
Cassidy, R. M ; Knight, C. H.; Recoskie, B. M.; Elchuk, S.; Miller, F. C ; Green, L. W. In
Actinide Separations; Navratil, J. D., Schulz, W. W., Eds.; World Scientific
Publishers: Singapore, 1988, p 1-18.
Challis, G. L. ChemBioChem 2005, 6, 601-611.
Chiarizia, R.; McAlister, D. R.; Herlinger, A. W. Separ. Sci. Technol. 2005, 40, 69-90.
Chiu, T. K.; Dickerson, R. E. J. Mol. Biol. 2000, 301, 915-945.
Coles, M.; Diercks, T.; Muehlenweg, B.; Bartsch, S.; Zolzer, V.; Tschesche, H.; Kessler,
H. J. Mol. Biol. 1999, 289, 139-157.
Colquhoun, H. M.; Stoddart, J. F.; Williams, D. J. Angew. Chem., Int. Ed. Engl. 1983, 25,
487-582.
Connors, K. A.; Pendergast, D. D. Journal of the American Chemical Society 1984,106,
7607-7614.
Corradini, R.; Dossena, A.; Marchelli, R.; Panagla, A.; Sartor, G.; Saviano, M.;
Lombardi, A.; Pavone, V. Chem.-Eur. J. 2006, 2, 373-381.
D'Souza, V. T.; Lipkowitz, K. B. Chem. Rev. (Washington, DC, U. S.) 1998, 98, 17411742.
D'Souza, V. T. Supramol. Chem. 2003, 75, 221-229.
Davey, C. A.; Richmond, T. J. P. Natl. Acad. Sci. USA 2002, 99, 11169-11174.
Del Valle, E. M. M. Process Biochem. (Oxford, U. K.) 2004, 39, 1033-1046.
Delmau, L. H.; Simon, N.; Schwing-Weill, M.-J.; Arnaud-Neu, F.; Dozol, J.-F.; Eymard,
S.; Tournois, B.; Bohmer, V.; Griittner, C ; Musigmannc, C.; Tunayarc, A. Chem.
Comm. 1998, 1627-1628.
Delmau, L. H.; Simon, N.; Schwing-Weill, M.-J.; Arnaud-Neu, F.; Dozol, J.-F.; Eymard,
S.; Tournois, B.; Gruttner, C.; Musigmann, C ; Tunayar, A.; Bohmer, V. Separ.
Sci. Technol. 1999, 34, 863-876.
Dotsikas, Y.; Kontopanou, E.; Allagiannis, C ; Loukas, Y. L. J. Pharmaceut. Biomed.
2000, 23, 997-1003.
Dotsikas, Y.; Loukas, Y. L.J. Biochem. Bioph. Meth. 2002, 52, 121-134.
169
Draye, M.; Thomas, S.; Cote, G.; Favre-Reguillon, A.; LeBuzit, G.; Guy, A.; Foos, J.
Separ. Sci. Technol. 2005, 40, 611-622.
Easton, C. J.; Lincoln, S. F. Modified Cyclodextrins: Scaffolds and Templates for
Supramolecular Chemistry; Imperial College Press: London, 1999.
Ferguson, A. D., Welte, Wolfram, Hofmann, Eckhard, Lindner, Buko, Hoist, Otto,
Coulton, James W., Diederichs, Kay.: Structure (London) 8 pp. 592 Structure
(London) 2000, 8, 585-592.
Fuks, L.; Majdan, M. Min. Pro. Ext. Met. Rev. 2000, 21, 25-48.
Garrett, T. P.; Clingeleffer, D. J.; Guss, J. M.; Rogers, S. J.; Freeman, H. C. J. Biol.
Chem. 1984, 259, 2822-2825.
Geist, A.; Weigl, M.; Gompper, K. Separ. Sci. Technol. 2002, 37, 3369-3390.
Gessner, R. V.; Quigley, G. J.; Wang, A. H.-J.; van der Marel, G. A.; van Boom, J. H.;
Rich, A. Biochemistry 1985, 24, 237-240.
Goetz, D. H.; Willie, S. T.; Armen, R. S.; Bratt, T.; Borregaard, N.; Strong, R. K.
Biochemistry 2000, 39, 1935-1941.
Goetz, D. H.; Holmes, M. A.; Borregaard, N.; Bluhm, M. E.; Raymond, K. N.; Strong, R.
K.Mol. Ce//2002,10, 1033-1043.
Gokel, G. W.; Leevy, W. M.; Weber, M. E. Chem. Rev. 2004,104, 2723-2750.
Grenthe, I.; Lagerman, B.Acta. Chem. Scand. 1991, 45, 122-??
Guillo, F.; Hamelin, B.; Jullien, L.; Canceill, J.; Lehn, J.-M.; De Robertis, L.; Driguez, H.
Bull. Soc. Chim. Fr. 1995,132, 857-866.
Harada, A.; Takahashi, S. J. Chem. Soc, Chem. Commun. 1984, 645-646.
Harada, A.; Takahashi, S.J. Chem. Soc, Chem. Commun. 1986, 1229-1230.
Harada, A.; Hu, Y.; Yamamoto, S.; Takahashi, S. J. Chem. Soc, Dalton Trans. 1988,
729-732.
Harada, A.; Shimada, M.; Takahashi, S. Chem. Lett. 1989, 275-276.
Harwani, S.; Telford, J. R. ChemTracts: Inorganic Chemistry 2005,18, 437-448.
Hattori, K. Bio Industry 1987, 4, 327-338.
Hemmila, I. Anal. Chem. 1985, 57, 1676-1681.
Hofstadler, S. A.; Sannes-Lowery, K. A. Nat. Rev. DrugDiscov. 2006, 5, 585-595.
Housecroft, C. E.; Sharpe, A. G. In Inorganic Chemistry; 2nd ed.; Pearson Education
Limited: England, 2005.
170
Howerton, S. B.; Sines, C. C ; VanDerveer, D.; Williams, L. D. Biochemistry 2001, 40,
10023-10031.
Ikeda, Y.; Motoune, S.; Matsuoka, T.; Arima, H.; Hirayama, F.; Uekama, K. J. Pharm.
Sci. 2002, 91, 2390-2398.
Ionova, G.; Ionov, S.; Rabbe, C ; Hill, C ; Madic, C ; Guillaumont, R.; Krupa, J. C.
Solvent. Extra. Ion Exc. 2001,19, 391-414.
Jahng, Y.; Telford, J. R. B. Kor. Chem. Soc. 2005, 26, 1614-1616.
Jenkins, I. L. Solvent. Extra. Ion Exc. 1984, 2, 1-27.
Kaplan, J. Cell 2002, 111, 603-606.
Karakhanov, E. E.; Maksimov, A. L.; Runova, E. A.; Kardasheva, Y. S.; Terenina, M. V.;
Buchneva, T. S.; Guchkova, A. Y. Macromol. Symp. 2003, 204, 159-173.
Khan, A. R.; Forgo, P.; Stine, K. J.; D'Souza, V. T. Chem. Rev. (Washington, DC, U. S.)
1998,95, 1977-1996.
Kucharski, L. M.; Lubbe, W. J.; Maguire, M. E. J. Biol. Chem. 2000, 275, \6161-\6111>.
Kumar, M. Analyst 1994,119, 2013-2024.
Lehn, J.-M. Angew. Chem., Int. Ed. 1988, 27, 89-112.
Li, H.; Webb, S. P.; Ivanic, J.; Jensen, J. H. J. Am. Chem. Soc. 2004,126, 8010-8019.
Libeu, C. A. P.; Kukimoto, M.; Nishiyama, M.; Horinuchi, S.; Adman, E. T.
Biochemistry 1997, 36, 13160-13179.
Lien, N. R. (2007). Cyclodextrins as molecular scaffolds for anion and cation
recognition. Chemistry. Iowa City, University of Iowa: 245.
Loukas, Y. L.; Vyza, E. A.; Valiraki, A. P. Analyst 1995,120, 533-538.
Loukas, Y. L. J. Pharm. Pharmacol. 1997, 49, 944-948.
Luecke, H.; Quiocho, F. A. Nature 1990, 347.
Machczynski, M. C ; Gray, H. B.; Richards, J. H. J. Inorg. Biochem. 2002, 88, 375-380.
Maguire, M. E.; Cowan, J. A. BioMetals 2002, 75, 203-210.
May, I.; Taylor, R. J.; Denniss, I. S.; Wallwork, A. L. Czech. J. Phys. 1999, 49.
Mckervey, M. A.; Schwing-Weill, M.-J.; Arnaud-Neu, F. In Molecular Recognition:
Receptors for Cationic Guests; Gokel, G. W., Ed.; Pergamon: New York, 1996;
Vol. l , p p 537-603.
171

Mehdoui, T.; Berthet, J.-C; Thuery, P.; Ephritikhine, M. Chem. Comm. 2005, 2860-2862.
Mishra, J.; Dent, C ; Tarabishi, R.; Mitsnefes, M. M.; Ma, Q.; Kelly, C ; Ruff, S. M.;
Zahedi, K.; Shoo, M.; Bean, J.; Mori, K.; Barasch, J.; Devarajan, P. Lancet 2005,
365, 1231-1238.
Mosinger, J.; Tomankova, V.; Nemcova, I.; Zyka, J. Anal. Lett. 2001, 34, 1979-2004.
Mukhopadhyay, S.; Rothenberg, G.; Joshi, A.; Baidossi, M.; Sasson, Y. Adv. Synth.
Catal. 2002, 344, 348-354.
Nash, K. L.; Rickert, P. G. Separ. Sci. Technol. 1993, 28, 25-41.
Nash, K. L.; Choppin, G. R. Separ. Sci. Technol. 1991,32, 255-21 A.
Nash, K. L.; Jensen, M. P. Separ. Sci. Technol. 2001, 36, 1257-1282.
Navaza, A.; Iroulart, M. G.; Navaza, J. J. Coord. Chem. 2000, 51, 153-168.
Neilands, J. B. J. Biol. Chem. 1995, 270, 26723-26726.
Newton, T. W.; Sullivan, J. C. Actinide Carbonate Complexes in Aqueous Solution
North-Holland, Amsterdam, 1985.
Nitz, M.; Sherawat, M.; Franz, K. J.; Peisach, E.; Allen, K. N.; Imperiali, B. Angew.
Chem. Int. Ed. 2004, 43, 3682-3685.
O'Brien, I. G.; Gibson, F. Biochim. Biophys. Acta 1970, 215, 393-402.
Panigrahi, B. S.; Gajendran, N.; Suryamurthy, N. Spectrochim Acta A 2003, 59, 19051910.
Park, H. (2007). Biomimetic Models of Intradiol Catechol Dioxygenases: Structures and
Properties of Inclusion Complexes of Fe(III) Centered Metal Complexes With
beta-Cyclodextrin Derivatives. Chemistry. Iowa City, University of Iowa: 319.
Peter, S.; Panigrahi, B. S.; Viswanathan, K. S.; Mathews, C. K. Anal. Chim. Acta 1992,
260, 135-141.
Petrov, A. S.; Funseth-Smotzer, J.; Pack, G. R. Int. J. Quantum Chem. 2005,102, 645655.
Pflugrath, J. W.; Quiocho, F. A. Nature 1985, 314, 257-260.
Pflugrath, J. W.; Quiocho, F. A. J. Mol. Biol. 1988, 200, 163-180.
172
Prudden, A. R.; Lien, N. R.; Telford, J. R. Chem. Commun. (Cambridge, U. K.) 2004,
172-173.
Quiocho, F. A.; Higgins, C. F. Phil. Trans. R. Soc. 1990, 326, 341-352.
Raymond, K. N., Dertz, Emily A., Kim, Sanggoo S. Proc. Natl. Acad. Sci. U. S. A. 2003,
100, 3584-3588.
Regiert, M. SOFWJournal 2003,129, 2, 4, 6, 8.
Robards, K.; Clarke, S.; Patsalides, E. Analyst 1988,113, 1757-1779.
Rudiger, V.; Schneider, H. Chem.-Eur. J. 2000, 6, 3771-3776.
Russell, N. R.; McNamara, M. In Proceedings of the International Symposium on
Cyclodextrins, 8th: Budapest, 1996, pp 163-169.
Sack, J. S.; Trakhanov, S. D.; Tsigannik, I. H.; Quiocho, F. A. J. Mol. Biol. 1989, 206,
193-207.
Sack, J. S.; Saper, M. A.; Quiocho, F. A. J. Mol. Biol. 1989, 206, 171-191.
Sanchez, C. A. "Preliminary Evaluation of Factors Affecting Perchlorate Uptake,
Accumulation, and Redistribution in Lettuce," Department of Soil, Water, and
Environmental Sciences, Yuma Agricultural Center, The University of Arizona,
2004.
Schneider, H.; Hacket, F.; Volker, R. Chem. Rev. (Washington, DC, U. S.) 1998, 98,
1755-1786.
Schneider, H.-J. In Macrocyclic Chemistry: Current Trends and Future Perspectives.;
Gloe, K, Ed., 2005, pp 277-289.
Shepard, W. E. B.; Kingston, R. L.; Anderson, B. F.; Baker, E. N. Acta Crystallogr.
1993,49,331-343.
Smith, G. V.; Cheng, J.; Song, R. Chemical Industries (Dekker) 1996, 68, 479-483.
Steiner, E.; Schetty, G. Helv. Chim. Acta 1973, 56, 368-374.
Stoddart, J. F.; Zarzycki, R. Reel. Trav. Chim. Pays-Bas 1988,107, 515-528.
Strimbu, L.; Liu, J.; Kaifer, A. E. Langmuir 2003,19, 483-485.
Szejtli, J. Starch/Staerke 1990, 42, 444-447.
Tabushi, I.; Kuroda, Y.; Shimokawa, K. J. Am. Chem. Soc. 1979, 101, 1614-1615.
Taketatsu, T. Talanta 1982, 29, 397-400.

Tang, B.; Du, M.; Sun, Y.; Xu, H.-l.; Shen, H.-x. Talanta 1998, 47, 361-366.
Tang, B.; Zhang, G.-Y.; Liu, Y.; Han, F. Anal. Chim. Acta 2002, 459, 83-91.
173
Team, T. I. T. a. R. C. P. "Perchlorate: Overview of Issues, Status, and Remedial
Options," Environmental Research Institute of the States, 2005.
Telford, J. R.; Raymond, K. N. In Comprehensive Supramolecular Chemistry; First ed.;
Atwood, J. L., Davies, J. E. D., Macnicol, D. D., Vogtle, F., Gokel, G. W., Eds.;
Pergamon, 1996; Vol. 1, pp 245-266.
Tereshko, V.; Wilds, C. J.; Minasov, G.; Prakash, T. P.; Maier, M. A.; Howard, A.;
Wawrzak, Z.; Manoharan, M.; Egli, M. Nucleic Acids Res. 2001, 29, 1208-1215.
Tilloy, S.; Bertoux, F.; Mortreux, A.; Monflier, E. Catal. Today 1999, 48, 245-253.
Weiner, E. C ; Brechbiel, M. W.; Brothers, H.; Magin, R. L.; Gansow, O. A.; Tomalia, D.
A.; Lauterbur, P. C. Magn. Reson. Med. 1994, 31, 1-8.
Weinmann, H. J. Characteristics ofGd-DTPA dimeglumine in Magnevist monograph;
Blackwell Scientific Publications: Oxford, 1994.
Wilmarth, W. R.; Rosencrance, S. W.; Nash, C. A.; Edwards, T. B. Separ. Sci. Technol.
2001,36,1283-1305.
Winkelmann, G. Biometals 2002, 30.
Wolff, J. Pharmacol. Rev. 1998, 50, 89-102.
Xu, Y.-Y.; Hemmila, I. A. Anal. Chim. Acta 1992, 256, 9-16.
Yang, J.; Goetz, D. H.; Li, J.-Y.; Wang, W.; Mori, K.; Setlik, D.; Du, T.; ErdjumentBromage, H.; Tempst, P.; Strong, R. K.; Barasch, J. Mol. Cell 2002,10, 10451056.
Yang, J.; Mori, K.; Li, J.-Y.; Barasch, J. Am J Physiol Renal Physiol 2003, 285, F9-F18.
Ye, Y.; Lee, H.-W.; Yang, W.; Shealy, S.; Yang, J. J. J. Am. Chem. Soc. 2005,127, 37433750.
Yue, W. W.; Grizot, S.; Buchanan, S. K. J. Mol. Biol. 2003, 332, 353-368.
Zamarev, K. New. J. Chem 1994,18, 3-18.

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CYCLODEXTRIN SCAFFOLDS DESIGNED FOR THE MOLECULAR

RECOGNITION OF ANIONS, CATIONS OR ORGANIC SUBSTRATES

A thesis submitted in partial fulfillment

UMI Number: 3323427

All rights reserved.

To my grandparents and parents

Action expresses priorities.

Samantha Soebbing-Nolting ('my

Further studies on the

Synthesis of per-6-5'-(thiosalicylic acid)-per-6-deoxy-P-cyclodextrin

exhibited solubility issues below a pH ~ 5. Consequently, potentiometric studies with

pentaacetic acid-amino)ethylamino)-mono-6-deoxy-P-cyclodextrin is reported, along

OUTER-SPHERE COORDINATION OF METAL COMPLEXES IN

ANION RECOGNITION USING A CYCLODEXTRIN-BASED

CATION RECOGNITION USING ACID-MODIFIED

3.1.1 Cation Recognition..

ORGANIC SUBSTRATES BINDING WITH NATIVE AND

SUMMARY AND FUTURE WORK

APPENDIX A DETERMINATION OF BINDING CONSTANTS USING ESIMS

A.l Importance and Background of Binding Affinities

APPENDIX B SPECTRA AND RELEVANT DATA FOR THE BINDING OF

APPENDIX C SPECTRA: MODIFIED CYLODEXTRINS

Properties of Tetrahedral Anions Studied with Cyclodextrin 6

Sample Calculations for the Titration of 6 with LiC104

Delay Time Prior to ^ - N M R Spectrum Collection for Experiments

Experimentally Calculated T\ Relaxation Times (sec) for !H-NMR Peaks of

Ionic Radii of the Lanthanides (A)

Log Ka of Guest Molecules with Cyclodextrins 2 or 12

B.4 Relative JH-NMR Peak Heights for peaks of interest of per-6-(2aminoethylamino)-per-6-deoxy-a-cyclodextrin(6)

Diagram of (A) the export of a siderophore from bacterial cell, (B)

A view of the binding site of Fhu showing hydrogen bonding interactions to

Representations of a-, |3-, and y-cyclodextrins. The top structure represents

Representation of the inclusion of a guest molecule within the cyclodextrin

(a) Fe m 2-hydroxy-l-naphthaldehyde thiosemicarbazone (HNT) Schiff-base

Synthetic transformation for a-cyclodextrin (1) to per-6-iodo-a-cyclodextrin

aminoethylamino)-per-6-deoxy-a-cyclodextrin (6) product

Calculated T\ relaxation times in graphical format for protons of interest in

Generic representation of a tetrahedral anion (TA = C104",SCV",orPCV")

Structures of 12-crown-4 and 18-crown-6 which are known to bind cations K+

7. Representation of a calix[n]arene with 4-8 monomers. 8. Repesentation of

The decrease ('lanthanide contraction') in the ionic radii of the Ln species is

3.12 Titration of Eu3+ with up to 5 equivalents of 19. The increase in the

3.14 Titration of Tb with up to 5 equivalents of 19. The increase in the

3.15 Titration of Tb3+ with up to 5 equivalents of 18. Importantly, there is no

3.16 Binding curve generated from fluorescence emission at 615 nm ofEu3+

3.17 Binding curve generated from fluorescence emission at 545 nm of Tb

3.19 Positive ESI-MS spectrum for the complexation of Gd + by ligand 19 (ratio

3.21 Positive ESI-MS spectrum for the complexation of Tb + by ligand 19 (ratio 19

3.22 Representation of the structure of DTPA dianhydride after nucleophilic attack

attached to the DTPA

UV-Vis absorbance of 22 titrated with 2 from 265 nm - 385 nm. No major

UV-Vis absorbance of 23 titrated with 2 from 265 nm - 385 nm. No major

UV-Vis absorbance of 25 titrated with 12 from 265 n m - 385 nm. A blue

Negative ESI-MS for the analysis of binding of 25 with P-cyclodextrin (2).

ESI mass spectrum for 1, a-cyclodextrin

B.2 ESI mass spectrum for 1, a-cyclodextrin titrated with LiC104 (1 : 3)

B.3 ESI mass spectrum for 1, a-cyclodextrin titrated with MgSC>4 (1 : 3)

B.4 ESI mass spectrum for 1, a-cyclodextrin titrated with KH2PO4 (1 : 7)

B.5 ESI mass spectrum for 6, per-6-(2-aminoethylamino)-per-6-deoxy-acyclodextrin

B.6 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin withLiClO4(10 : 1)

B.7 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with LiC104 (1 : 1)

B.8 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with LiC104 (1 : 5)

B.l 1 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with MgS04 (1 : 1)

B.12 ESI mass spectrum for titration of 6, per-6-(2-aminoethylamino)-per-6deoxy-a-cyclodextrin with MgS04(1 : 5)