Supporting Information · i) Binding data from NMR titration experiments a) NMR titration data for...
Transcript of Supporting Information · i) Binding data from NMR titration experiments a) NMR titration data for...
Supporting Information
© Copyright Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, 2005
Novel Enantioselective Receptors for N-Protected Glutamate and Aspartate
Andrea Ragusa,[a] Sara Rossi,[a] Joseph M. Hayes,[b] Matthias Stein[b] and Jeremy D. Kilburn,*[a]
[a] A. Ragusa, S. Rossi, Prof. J. D. Kilburn, School of Chemistry, University of Southampton, Southampton, SO17 1BJ,
U.K.; Fax: (+44)-2380-596805; E-mail: [email protected]
[b] Dr. J. M. Hayes, Dr. M. Stein, Anterio Consult & Res GmbH, Augustaanlage 26, D-68165 Mannheim, Germany
i) Binding data from NMR titration experiments
a) NMR titration data for N-Boc-L-Glu with macrocycle 1 in CD3CN
b) NMR titration data for N-Boc-D-Glu with macrocycle 1 in CD3CN
c) NMR titration data for N-Boc-L-Glu with macrocycle 1 in d6-DMSO
d) NMR titration data for Acetate with macrocycle 1 in CD3CN
e) NMR titration data for N-Ac-L-Glu with macrocycle 1 in CD3CN
f) NMR titration data for N-Ac-D-Glu with macrocycle 1 in CD3CN
g) NMR titration data for N-Ac-L-Glu with macrocycle 1 in d6-DMSO
h) NMR titration data for N-Ac-D-Glu with macrocycle 1 in d6-DMSO
i) NMR titration data for N-Boc-L-Asp with macrocycle 1 in CD3CN
j) NMR titration data for N-Boc-L-Asp with macrocycle 1 in d6-DMSO
at 363 K
k) NMR titration data for DiBz-L-tartrate with macrocycle 1 in d6-
DMSO
l) NMR titration data for DiBz-D-tartrate with macrocycle 1 in d6-
DMSO
ii) Binding data from isothermal calorimetry experiments
a) ITC data for N-Boc-L-Glu with macrocycle 1 in CH3CN
b) ITC data for N-Boc-D-Glu with macrocycle 1 in CH3CN
c) ITC data for N-Boc-L-Glu with macrocycle 1 in DMSO
d) ITC data for Acetate with macrocycle 1 in CH3CN
e) ITC data for N-Ac-L-Glu with macrocycle 1 in CH3CN
f) ITC data for N-Ac-D-Glu with macrocycle 1 in DMSO
g) ITC data for N-Boc-L-Glu with macrocycle 2 in CH3CN
h) ITC data for N-Boc-D-Glu with macrocycle 2 in CH3CN
i) ITC data for N-Boc-L-Glu with macrocycle 2 in DMSO
j) ITC data for N-Boc-L-Asp with macrocycle 2 in CH3CN
k) ITC data for N-Boc-D-Asp with macrocycle 2 in CH3CN
l) ITC data for N-Boc-L-Glu with macrocycle 3 in DMSO
m) ITC data for N-Boc-D-Glu with macrocycle 3 in DMSO
i) Binding data from NMR titration experiments
The binding constants were calculated by fitting the titration
data, using NMRTit HG software, kindly provided by Prof C. A.
Hunter, University of Sheffield (A. P. Bisson, C. A. Hunter, J. C.
Morales, K. Young, Chem. Eur. J. 1998, 4, 845). Data was recorded
using standard NMR titration protocols, recording the shift for
various 1H signals on addition of aliquots of guest. An association
constant was determined using the data from each separate 1H
signal, and averaged to give a final association constant. The
rather large discrepancies between association constants obtained
from the data for different 1H signals in CD3CN, in the same
titration experiment, is not surprising in view of the fact that
the data gives binding constants at the upper limit of
detectability using NMR titrations (104 - 105 M-1) (L. Fielding,
Tetrahedron 2000, 56, 6151; C. S. Wilcox, Frontiers in
Supramolecular Organic Chemistry and Photochemistry (Eds.: H.-J.
Schneider, H. Dürr), VCH, Weinheim, 1991, pp. 123-143; G. Weber,
S. R. Anderson, Biochemistry, 1965, 4, 1942.) In these cases it
can be concluded that Ka > 104 M-1, but more precise values cannot
be reliably obtained from the NMR titration data.
a) NMR titration data for N-Boc-L-Glu with macrocycle 1 in CD3CN
Starting volume of host solution: 600µL
Concentration of host solution: 1.5 mM
Concentration of guest solution: 13.6 mM
Association constant: Ka1:1 = 1.5×105 M-1 ie Ka
1:1 > 104 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm CH proton / ppm 0 9.1229 8.2484 5.3583 20 9.3901 8.5985 5.5302 40 9.6823 9.0186 5.7108 60 9.9558 9.4214 5.8587 80 10.1239 9.6020 5.9303 100 10.1728 9.6647 5.9454 120 10.1791 9.6810 5.9478 140 10.1816 9.6798 5.9492 170 10.1829 9.6861 5.9492 200 10.1854 9.6798 5.9492
Titration curves for N-Boc-L-Glu with macrocycle 1 in CD3CN
Boc-L-glu in MeCN (amide protons)
9.12
9.32
9.52
9.72
9.92
10.12
10.32
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03
[guest]
ppm
Boc-L-glu in MeCN (thiourea protons)
8.2
8.4
8.6
8.8
9
9.2
9.4
9.6
9.8
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03
[guest]
ppm
Boc-L-glu in MeCN (CH protons)
5.35
5.45
5.55
5.65
5.75
5.85
5.95
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03
[guest]
ppm
b) NMR titration data for N-Boc-D-Glu with macrocycle 1 in CD3CN
Starting volume of host solution: 600µL
Concentration of host solution: 1.5 mM
Concentration of guest solution: 13.0 mM
Association constant (guest:host): Ka2:1 = 1.4×105 M-1 ie Ka
2:1 > 104 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm CH proton / ppm 0 9.1153 8.2473 5.3521 20 9.2470 8.4154 5.4512 40 9.3424 8.5809 5.6117 60 9.5117 8.7980 5.6882 80 9.6397 8.9773 5.7535 100 9.7589 9.1216 5.8099 120 9.8768 9.2772 5.8501 140 9.9545 9.3788 5.8802 170 10.0336 9.4779 5.8839 200 10.0662 9.5155 5.8839
Titration curves for N-Boc-D-Glu with macrocycle 1 in CD3CN
Boc-D-glu in MeCN (amide protons)
9.1
9.3
9.5
9.7
9.9
10.1
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03
[guest]
ppm
Boc-D-glu in MeCN (thiourea protons)
8.2
8.4
8.6
8.8
9
9.2
9.4
9.6
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03
[guest]
ppm
Boc-D-glu in MeCN (CH protons)
5.4
5.5
5.6
5.7
5.8
5.9
6.0
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03
[guest]
ppm
c) NMR titration data for N-Boc-L-Glu with macrocycle 1 in d6-DMSO
Starting volume of host solution: 600µL
Concentration of host solution: 2.35 mM
Concentration of guest solution: 94.2 mM
Association constant: Ka1:1 = 3.7×103 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.471 8.395 20 9.496 8.447 40 9.555 8.548 60 9.603 8.653 80 9.656 8.754 100 9.713 8.856 140 9.803 9.093 180 9.886 9.251 220 9.946 9.359 260 9.989 9.444 300 10.011 9.483 360 10.033 9.510 420 10.048 9.537 480 10.050 9.556 540 10.058 9.556 600 10.068 9.564
Titration curves for N-Boc-L-Glu with macrocycle 1 in d6-DMSO
L-Boc Glu in DMSO (amide protons)
9.45
9.55
9.65
9.75
9.85
9.95
10.05
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03 4.0E-03 4.5E-03 5.0E-03
[guest]
ppm
L-Boc Glu in DMSO (thiourea protons)
8.3
8.5
8.7
8.9
9.1
9.3
9.5
9.7
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03 4.0E-03 4.5E-03 5.0E-03
[guest]
ppm
d) NMR titration data for Acetate with macrocycle 1 in CD3CN
Starting volume of host solution: 600µL
Concentration of host solution: 3.1 mM
Concentration of guest solution: 27.9 mM
Association constant (guest:host): Ka2:1 = 1.12×104 M-1 ie Ka
2:1 > 104 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.0731 8.2417 10 9.1560 8.3567 20 9.2678 8.5170 30 9.3460 8.6789 40 9.4359 8.8267 50 9.4994 8.9167 60 9.5454 8.9933 70 9.5619 9.0269 80 9.5798 9.0598 90 9.6017 9.0833 100 9.6205 9.1075 110 9.6386 9.1239 120 9.6479 9.1364 130 9.6644 9.1552 140 9.6777 9.1724 150 9.6941 9.1842 160 9.7066 9.1982 170 9.7121 9.2076 180 9.7270 9.2240 190 9.7356 9.2350 200 9.7489 9.2514 220 9.7731 9.2694
Titration curves for Acetate with macrocycle 1 in CD3CN
Acetate in MeCN (amide protons)
9.0
9.1
9.2
9.3
9.4
9.5
9.6
9.7
9.8
0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03 6.0E-03 7.0E-03
[guest]
ppm
Acetate in MeCN (thiourea protons)
8.2
8.4
8.6
8.8
9.0
9.2
0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03 6.0E-03 7.0E-03
[guest]
ppm
e) NMR titration data for N-Ac-L-Glu with macrocycle 1 in CD3CN
Starting volume of host solution: 500µL
Concentration of host solution: 1.0 mM
Concentration of guest solution: 5.2 mM
Association constant (guest:host): Ka1:1 = 8.97×104 M-1 ie Ka
1:1 > 104 M-1
Ka2:1 = 5.1×103 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.0953 8.2141 5 9.1380 8.2731 10 9.1832 8.3309 15 9.2460 8.4049 20 9.2811 8.4765 25 9.3301 8.5543 30 9.3903 8.6334 35 9.4481 8.7187 40 9.5146 8.7941 45 9.5611 8.8732 50 9.6176 8.9359 60 9.6900 9.0500 70 9.7556 9.1129 80 9.7808 9.1481 100 9.8448 9.2321 120 9.8962 9.2899 150 9.9678 9.3539 200 10.0180 9.4104 250 10.0419 9.4280 300 10.0569 9.4418
Titration curves for N-Ac-L-Glu with macrocycle 1 in CD3CN
Ac-L-Glu in MeCN (amide protons)
9.0
9.2
9.4
9.6
9.8
10.0
0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03
[guest]
ppm
Ac-L-Glu in MeCN (thiourea protons)
8.2
8.4
8.6
8.8
9.0
9.2
9.4
0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03
[guest]
ppm
f) NMR titration data for N-Ac-D-Glu with macrocycle 1 in CD3CN
Starting volume of host solution: 500µL
Concentration of host solution: 0.8 mM
Concentration of guest solution: 5.6 mM
Association constant (guest:host): Ka1:1 = 4.5×103 M-1
Ka2:1 = 1.67×105 M-1 ie Ka
2:1 > 104 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.0916 8.2204 5 9.1142 8.2467 10 9.1518 8.3120 15 9.1995 8.3861 20 9.3025 8.5191 25 9.4079 8.6384 30 9.4845 8.7777 35 9.5937 8.8895 40 9.6828 9.0250 45 9.7494 9.0891 50 9.8322 9.1945 60 9.9490 9.3037 70 10.0055 9.3401 80 10.0318 9.3665 100 10.0469 9.3853 120 10.0544 9.3866 150 10.0645 9.3866 200 10.0594 9.3878 250 10.0632 9.3979 350 10.0607 9.3941
Titration curves for N-Ac-D-Glu with macrocycle 1 in CD3CN
Ac-D-Glu in MeCN (amide protons)
9.0
9.2
9.4
9.6
9.8
10.0
0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03
[guest]
ppm
Ac-D-Glu in MeCN (thiourea protons)
8.2
8.4
8.6
8.8
9.0
9.2
9.4
0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03
[guest]
ppm
g) NMR titration data for N-Ac-L-Glu with macrocycle 1 in d6-DMSO
Starting volume of host solution: 500µL
Concentration of host solution: 0.6 mM
Concentration of guest solution: 4.6 mM
Association constant (guest:host): Ka1:1 = 4.0×104 M-1 ie Ka
1:1 > 104 M-1
Ka2:1 = 1.0×103 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.4256 8.3435 5 9.4419 8.3812 10 9.4708 8.4352 15 9.5085 8.5117 20 9.5499 8.5871 25 9.6064 8.6737 30 9.6553 8.7666 35 9.6900 8.8500 40 9.7432 8.9473 45 9.7809 9.0189 50 9.8198 9.0980 60 9.8762 9.1782 70 9.9139 9.2549 80 9.9441 9.3202 100 9.9891 9.3978 120 9.9968 9.4256 150 10.0156 9.4746 200 10.0345 9.5047 300 10.0520 9.5286
Titration curves for N-Ac-L-Glu with macrocycle 1 in d6-DMSO
Ac-L-Glu in DMSO (amide protons)
9.4
9.5
9.6
9.7
9.8
9.9
10.0
10.1
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03 4.0E-03 4.5E-03
[guest]
ppm
Ac-L-Glu in DMSO (thiourea protons)
8.3
8.5
8.7
8.9
9.1
9.3
9.5
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03 4.0E-03 4.5E-03
[guest]
ppm
h) NMR titration data for N-Ac-D-Glu with macrocycle 1 in d6-DMSO
Starting volume of host solution: 500µL
Concentration of host solution: 0.5 mM
Concentration of guest solution: 4.7 mM
Association constant (guest:host): Ka1:1 = 8.0×102 M-1
Ka2:1 = 1.68×104 M-1 ie Ka
2:1 > 104 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.4269 8.3410 5 9.4369 8.3699 10 9.4620 8.4251 15 9.4809 8.4678 20 9.5160 8.5256 25 9.5400 8.6100 30 9.5913 8.6975 35 9.6240 8.7540 40 9.6541 8.8331 45 9.6892 8.8896 50 9.7219 8.9674 60 9.7730 9.0578 70 9.8035 9.1444 80 9.8361 9.1846 100 9.8725 9.2813 120 9.9026 9.3302 150 9.9227 9.3842 200 9.9378 9.4068 350 9.9554 9.4557
Titration curves for N-Ac-D-Glu with macrocycle 1 in d6-DMSO
Ac-D-Glu in DMSO (amide protons)
9.4
9.5
9.6
9.7
9.8
9.9
10.0
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03 4.0E-03 4.5E-03
[guest]
ppm
Ac-D-Glu in DMSO (thiourea protons)
8.3
8.5
8.7
8.9
9.1
9.3
9.5
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03 3.5E-03 4.0E-03 4.5E-03
[guest]
ppm
i) NMR titration data for N-Boc-L-Asp with macrocycle 1 in CD3CN
Starting volume of host solution: 600µL
Concentration of host solution: 1.2 mM
Concentration of guest solution: 13.4 mM
Association constant (guest:host): Ka1:1 = 2.1×105 M-1 ie Ka
1:1 > 104 M-1
Ka2:1 < 100 M-1
Volume added / µL Aromatic protons / ppm CH proton / ppm 0 7.625 5.325 10 7.638 5.369 20 7.645 5.423 30 7.667 5.479 40 7.678 5.516 50 7.690 5.625 60 7.702 5.817 70 7.710 5.850 80 7.710 5.862 90 7.709 5.872 100 7.708 5.877 110 7.708 5.914 120 7.708 5.901 130 7.707 5.927 140 7.706 5.895 150 7.706 5.887
Titration curves for N-Ac-L-Asp with macrocycle 1 in CD3CN
Boc-L-Asp in MeCN (aromatic protons)
7.62
7.63
7.64
7.65
7.66
7.67
7.68
7.69
7.70
7.71
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03
[guest]
ppm
Boc-L-Asp in MeCN (CH protons)
5.32
5.37
5.42
5.47
5.52
5.57
5.62
5.67
5.72
5.77
5.82
5.87
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03
[guest]
ppm
j) NMR titration data for N-Boc-L-Asp with macrocycle 1 in d6-DMSO at 363 K
Starting volume of host solution: 600µL
Concentration of host solution: 3.1 mM
Concentration of guest solution: 27.9 mM
Association constant (guest:host): Ka1:1 = 1.2×103 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.2826 8.1808 20 9.3595 8.3035 40 9.4307 8.5045 60 9.506 8.5931 80 9.5223 8.6850 100 9.5475 8.7365 120 9.5826 8.7658 140 9.5972 8.8163
Titration curves for N-Ac-L-Asp with macrocycle 1 in d6-DMSO at 363 K
Boc-L-Asp in DMSO (amide protons)
9.28
9.33
9.38
9.43
9.48
9.53
9.58
0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03
[guest]
ppm
Boc-L-Asp in DMSO (thiourea protons)
8.18
8.23
8.28
8.33
8.38
8.43
8.48
8.53
8.58
8.63
8.68
8.73
8.78
0.0E+00 1.0E-03 2.0E-03 3.0E-03 4.0E-03 5.0E-03
[guest]
ppm
k) NMR titration data for DiBz-L-tartrate with macrocycle 1 in d6-DMSO
Starting volume of host solution: 600µL
Concentration of host solution: 1.3 mM
Concentration of guest solution: 12.6 mM
Association constant (guest:host): Ka2:1 = 211 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.4428 8.3590 20 9.4478 8.3766 40 9.4554 8.4292 60 9.4591 8.4793 80 9.4641 8.5221 100 9.4641 8.5446 120 9.4654 8.5760 140 9.4704 8.5998 170 9.4654 8.6248 200 9.4679 8.6325
Titration curves for DiBz-L-tartrate with macrocycle 1 in d6-DMSO
DiBz-L-tartrate in DMSO (amide protons)
9.44
9.45
9.46
9.47
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03
[guest]
ppm
DiBz-L-tartrate in DMSO (thiourea protons)
8.35
8.40
8.45
8.50
8.55
8.60
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03
[guest]
ppm
l) NMR titration data for DiBz-D-tartrate with macrocycle 1 in d6-DMSO
Starting volume of host solution: 600µL
Concentration of host solution: 1.3 mM
Concentration of guest solution: 12.6 mM
Association constant (guest:host): Ka2:1 = 103 M-1
Volume added / µL Amide proton / ppm Thiourea proton / ppm 0 9.4428 8.3590 20 9.4491 8.3777 40 9.4529 8.4118 60 9.4591 8.4982 80 9.4641 8.5396 100 9.4654 8.5797 120 9.4679 8.6061 140 9.4717 8.6324 170 9.4704 8.6788 200 9.4729 8.6751
Titration curves for DiBz-D-tartrate with macrocycle 1 in d6-DMSO
DiBz-L-tartrate in DMSO (amide protons)
9.44
9.45
9.46
9.47
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03
[guest]
ppm
DiBz-L-tartrate in DMSO (thiourea protons)
8.35
8.40
8.45
8.50
8.55
8.60
8.65
0.0E+00 5.0E-04 1.0E-03 1.5E-03 2.0E-03 2.5E-03 3.0E-03
[guest]
ppm
ii) Binding data from ITC titration experiments
All binding experiments were performed on an isothermal titration
calorimeter from Microcal Inc. (Northampton, MA). In a typical
experiment a 1 - 2 mM receptor solution is added to the
calorimetric cell. A 48 - 78 mM solution of TBA acetate is
introduced in 50 injections of 5 µL, for a total of 250µL of added
guest. Such high concentrations of guests are important to
generate sharp curves, necessary for acceptable curve fitting. The
solution is continuously stirred to ensure rapid mixing and kept
at 25°C, through the combination of an external cooling bath and an
internal heater. Dilution effects are determined by performing a
blank experiment by adding the same guest solution into the pure
solvent and subtracting this from the raw titration to produce the
final binding curve. Association parameters are found by applying
either one-site or two-sites models, using the Origin software
provided. These methods rely on standard nonlinear least-squares
regression (Levenberg-Marquard method) to fit the curves, taking
into account the change in volume that occurs during the
calorimetric titration (W. H. Press, S. A. Teukolsky, W. T.
Vetterling, B. P. Flannery, Numerical recipes in C, 2nd ed.,
Cambridge University Press, Cambridge, 1988).
a) Data for N-Boc-L-Glu and macrocycle 1 in CH3CN (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
-189.249 -153.764 -48.8784 -38.8221 -39.0044 -37.446
-28.8245 -27.3975 -19.1454 -3.40689 23.05663 68.71635 123.3233 155.6574 182.2862 185.8849 195.1904 195.5603 198.4545 200.9079 197.6934 201.0834 199.4598 199.4112 199.9179 197.5559 204.3711 197.0166 198.3899 199.3671 408.6134 397.7125 397.2287 392.9763 387.8707 383.2565 380.2648 378.5055 373.7875 371.6359 368.181 362.3546 358.7509 355.7086 353.4957 345.1381 341.4609 337.9775 336.817 330.3869 328.342 325.628 324.4354 319.7499
2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5
0 0.10922 0.21825 0.3271 0.43576 0.54423 0.65251 0.76061 0.86852 0.97624 1.08378 1.19113 1.29829 1.40527 1.51206 1.61866 1.72507 1.8313 1.93734 2.04319 2.14886 2.25434 2.35963 2.46474 2.56966 2.67439 2.77893 2.88329 2.98746 3.09144 3.19524 3.40227 3.60856 3.81409 4.01888 4.22292 4.42621 4.62876 4.83055 5.0316 5.2319 5.43145 5.63025 5.82831 6.02562 6.22218 6.41799 6.61305 6.80737 7.00094 7.19376 7.38583 7.57715 7.76773
1.29 1.2878 1.28559 1.2834 1.2812 1.27901 1.27683 1.27464 1.27247 1.27029 1.26812 1.26595 1.26379 1.26163 1.25947 1.25732 1.25517 1.25302 1.25088 1.24874 1.24661 1.24448 1.24235 1.24022 1.2381 1.23599 1.23387 1.23176 1.22966 1.22755 1.22545 1.22126 1.21709 1.21293 1.20878 1.20465 1.20053 1.19642 1.19233 1.18825 1.18419 1.18013 1.17609 1.17207 1.16806 1.16406 1.16007 1.1561 1.15213 1.14819 1.14425 1.14033 1.13642 1.13252
0.08481 0.16977 0.25487 0.34011 0.42551 0.51104 0.59672 0.68255 0.76852 0.85464 0.9409 1.0273 1.11385 1.20055 1.28739 1.37438 1.46151 1.54878 1.6362 1.72377 1.81148 1.89933 1.98733 2.07548 2.16377 2.2522 2.34078 2.42951 2.51838 2.6074 2.78586 2.96491 3.14453 3.32473 3.50552 3.68688 3.86883 4.05135 4.23446 4.41814 4.6024 4.78725 4.97267 5.15867 5.34526 5.53242 5.72016 5.90848 6.09739 6.28687 6.47693 6.66757 6.85879 7.0506
-1162.51 -1234.32
-- -- -- --
-621.702 -625.842 -603.787 -494.202 -355.264 -60.2823 262.9994 484.5682 634.4697 671.4336 714.1357 707.7618 736.8489 746.3467 728.3377 744.327 736.6176 732.5924 736.027 722.5011 764.1504 722.6998 734.2305 737.2481 747.1079 724.165 727.0855 717.4262 698.1583 689.2948 684.52 683.213 671.6182 668.813 657.3806 644.8344 637.9774 632.1624 626.5861 606.4587 595.0101 590.042 588.205 573.8739 570.6078 566.5464 561.2056 553.1474
317.4505 313.8304 308.8789 306.1113 304.1368 302.4893 297.8454 295.0179 292.5818 291.6189 287.6484
5 5 5 5 5 5 5 5 5 5 5
7.95756 8.14664 8.33497 8.52255 8.70939 8.89548 9.08082 9.26541 9.44925 9.63235 9.8147
1.12864 1.12476 1.1209 1.11706 1.11322 1.1094 1.10559 1.10179
1.098 1.09422 1.09046
7.24298 7.43594 7.62948 7.8236 8.0183 8.21358 8.40944 8.60588 8.8029 9.0005 9.19868
548.6419 541.9815 530.8968 525.0446 521.322 518.0835 505.7222 500.6646 495.67
494.4923 486.2193
N-Boc-L-Glu and macrocycle 1 in CH3CN
Data fitted using two binding site model:
Ka1:1 = 27270 ± 4820 M-1
∆H = -4.49 ± 0.18 kJmol-1,
∆S = 70.48 Jmol-1K-1)
(Ka2:1 = 25 ± 4 M-1)
0 2 4 6 8 10
-1.0
-0.5
0.0
0.5
1.0
LGluMeCNC_NDH LGluMeCNC_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
b) Data for N-Boc-D-Glu and macrocycle 1 in CH3CN (data corrected by subtracting the background dilution data for an injection of
the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
-163.719 -196.252 -188.462 -183.107 -186.473 -185.276 -178.878 -186.245 -173.145 -171.419 -162.417 -131.26
-94.9491 -61.1531 -42.0515 -28.0974 -20.2937 -13.1429 -8.54055 -4.22314 -0.45496 1.15022 2.37363 4.78172 7.65869 8.14733 9.06747 9.9707
12.13907 13.29957 29.02517 31.09795 34.02882 36.58593 38.94935 40.09813 40.88099 42.01826 43.33526 43.74343 45.50059 45.81537 46.65492 47.17971 47.51448 47.64501 48.34218 49.1942 49.67137 48.73941 49.47997 49.06059 49.83424 50.39214
2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5
0 0.10905 0.21791 0.32659 0.43507 0.54338 0.65149 0.75942 0.86716 0.97472 1.08208 1.18927 1.29626 1.40307 1.50969 1.61612 1.72237 1.82843 1.93431
2.04 2.1455 2.25081 2.35594 2.46088 2.56563 2.6702 2.77458 2.87878 2.98278 3.08661 3.19024 3.39695 3.60291 3.80812 4.01259 4.21631 4.41928 4.62151 4.82299 5.02372 5.22371 5.42295 5.62144 5.81919 6.01619 6.21244 6.40795 6.60271 6.79672 6.98998 7.1825 7.37427 7.5653 7.75557
0.94 0.93839 0.93679 0.93519 0.93359 0.93199 0.9304 0.92881 0.92722 0.92564 0.92406 0.92248 0.9209 0.91933 0.91775 0.91618 0.91462 0.91306 0.91149 0.90994 0.90838 0.90683 0.90528 0.90373 0.90218 0.90064 0.8991 0.89756 0.89603 0.8945 0.89297 0.88991 0.88687 0.88384 0.88082 0.87781 0.8748 0.87181 0.86883 0.86586 0.8629 0.85994
0.857 0.85407 0.85114 0.84823 0.84532 0.84243 0.83954 0.83666 0.8338 0.83094 0.82809 0.82525
0.11621 0.23261 0.34922 0.46602 0.58302 0.70023 0.81763 0.93522 1.05302 1.17102 1.28921 1.4076 1.52619 1.64498 1.76397 1.88316 2.00254 2.12213 2.24191 2.36189 2.48207 2.60245 2.72303 2.84381 2.96478 3.08595 3.20733 3.3289 3.45067 3.57263 3.81717 4.06249 4.30861 4.55553 4.80324 5.05174 5.30104 5.55113 5.80202 6.0537 6.30618 6.55945 6.81352 7.06838 7.32403 7.58048 7.83772 8.09576 8.35459 8.61422 8.87464 9.13586 9.39787 9.66068
-- -- --
-1953 -1994.29 -2042.53 -1990.68 -2074.57 -1970.88 -1964.27 -1927.97 -1713.25 -1516.77 -1283.92 -1183.63 -1085.22 -1019.05 -999.216 -951.25
-936.936 -903.327 -890.811 -876.484 -849.993 -844.294 -823.989 -826.177 -814.06 -791.97
-799.035 -787.032 -755.91
-735.573 -718.604 -700.216 -688.415 -675.368 -662.838 -649.707 -641.149 -623.003 -611.197 -607.127 -594.563 -583.136 -577.195 -561.819 -550.084 -539.863 -535.845 -522.743 -517.462 -509.725 -500.277
50.46469 50.54157 50.91086 51.19185 50.30102 50.31416 50.05067 49.99682 50.63248 50.94008 50.28977
5 5 5 5 5 5 5 5 5 5 5
7.9451 8.13389 8.32193 8.50922 8.69576 8.88156 9.06661 9.25091 9.43447 9.61727 9.79934
0.82242 0.8196 0.81678 0.81398 0.81118 0.8084 0.80562 0.80285 0.80009 0.79734 0.7946
9.92427 10.18867 10.45386 10.71984 10.98662 11.25419 11.52256 11.79172 12.06167 12.33242 12.60397
-493.883 -482.286 -473.826 -465.492 -459.291 -453.488 -447.322 -446.568
-432.4 -425.134 -423.894
N-Boc-D-Glu and macrocycle 1 in CH3CN
Data fitted using two binding site model:
Ka2:1 = 49150 ± 675 M-1
∆H = -6.73 ± 0.067 kJmol-1,
∆S = 67.58 Jmol-1K-1
(Ka1:1 = 38 ± 2 M-1)
0 2 4 6 8 10 12
-1.5
-1.0
-0.5
0.0
DGluMeCNB_NDH DGluMeCNB_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
c) Data for N-Boc-L-Glu and macrocycle 1 in DMSO (data corrected by subtracting the background dilution data for an injection of
the guest solution into the cell containing neat DMSO).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
-619.179 -304.269 -222.763 -183.857 -85.3283 1.27622 53.82913 81.51115 96.5027 111.7939 124.4743 139.3054 152.0669 162.0422 172.2156 178.1855 184.0366 189.3605 194.1516 194.4186 195.8559 199.1482 200.2779 201.7373 200.8781 200.9426 202.1667 201.1533 199.376 199.1672 198.4799 196.0795 196.3583 194.5739 193.5206 193.2806 190.5462 190.9743 190.7106 189.3373 187.1899 185.095 183.6648 182.276 180.1428 179.9729 177.0212 176.2865 175.6264 173.5617
5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5
0 0.21791 0.43507 0.65149 0.86716 1.08208 1.29626 1.50969 1.72237 1.93431 2.1455 2.35594 2.56563 2.77458 2.98278 3.19024 3.39695 3.60291 3.80812 4.01259 4.21631 4.41928 4.62151 4.82299 5.02372 5.22371 5.42295 5.62144 5.81919 6.01619 6.21244 6.40795 6.60271 6.79672 6.98998 7.1825 7.37427 7.5653 7.75557 7.9451 8.13389 8.32193 8.50922 8.69576 8.88156 9.06661 9.25091 9.43447 9.61727 9.79934
1.4 1.39522 1.39045 1.3857 1.38097 1.37625 1.37155 1.36687 1.3622 1.35754 1.35291 1.34828 1.34368 1.33909 1.33451 1.32995 1.3254 1.32087 1.31636 1.31186 1.30737 1.3029 1.29844
1.294 1.28958 1.28516 1.28077 1.27638 1.27201 1.26766 1.26332 1.25899 1.25468 1.25038 1.24609 1.24182 1.23757 1.23332 1.22909 1.22488 1.22067 1.21648 1.21231 1.20815
1.204 1.19986 1.19574 1.19163 1.18753 1.18345
0.15618 0.3129 0.47015 0.62794 0.78625 0.9451 1.10449 1.26441 1.42486 1.58584 1.74736 1.90941
2.072 2.23512 2.39877 2.56295 2.72767 2.89293 3.05871 3.22503 3.39188 3.55927 3.72719 3.89564 4.06463 4.23415 4.4042 4.57479 4.74591 4.91756 5.08975 5.26247 5.43573 5.60951 5.78383 5.95869 6.13408
6.31 6.48645 6.66344 6.84096 7.01902 7.19761 7.37673 7.55639 7.73657 7.9173 8.09855 8.28034 8.46266
-2204.11 -1685.22
-1561 -1469.19 -1171.09 -893.42
-714.804 -622.951 -561.644 -499.076 -447.928 -392.581 -342.44
-303.265 -258.999 -231.477 -200.229 -175.519 -149.353 -135.855 -122.111 -103.635 -91.2995 -78.2756 -71.4866 -62.8806 -52.1727 -48.3605 -44.8698 -38.0242 -28.0197 -33.5746 -22.1267 -26.6567 -10.2245 -7.21463 -14.1484 -5.07739 1.56024 3.18772 4.32944 1.88594 1.28534 4.43816 0.89774 3.44965 2.04426 2.53264 5.15434 4.17212
N-Boc-L-Glu and macrocycle 1 in DMSO
Data fitted using two binding site model:
Ka1:1 = 2281 ± 258 M-1
∆H = -10.73 ± 0.32 kJmol-1,
∆S = 28.56 Jmol-1K-1
(Ka2:1 = 28 ± 1 M-1)
0 2 4 6 8 10
-2.0
-1.5
-1.0
-0.5
0.0
srLbocglu4_NDH srLbocglu4_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
d) Data for Acetate and macrocycle 1 in CH3CN (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
27.48395 5 0 0.7 0.10771 -- 353.1491 5 0.07514 0.69761 0.21579 --
534.4007 5 0.15003 0.69523 0.32424 --
537.5806 5 0.22465 0.69285 0.43306 4840.67
414.798 5 0.29902 0.69049 0.54224 3793.619
232.6422 5 0.37313 0.68813 0.6518 2153.909
79.75142 5 0.44699 0.68578 0.76172 856.0021
-7.07607 5 0.52058 0.68343 0.872 46.55876
-54.8458 5 0.59392 0.6811 0.98266 -391.044
-80.1968 5 0.667 0.67877 1.09368 -689.01
-94.1475 5 0.73983 0.67645 1.20508 -790.584
-100.88 5 0.81239 0.67414 1.31684 -802.869
-105.2 5 0.8847 0.67184 1.42896 -876.027
-107.712 5 0.95675 0.66954 1.54146 -816.877
-115.521 5 1.02855 0.66725 1.65432 -924.35
-117.849 5 1.10008 0.66497 1.76755 -964.503
-118.827 5 1.17136 0.6627 1.88115 -997.75
-115.41 5 1.24238 0.66044 1.99512 -949.577
-117.851 5 1.31315 0.65818 2.10946 -948.094
-116.813 5 1.38365 0.65593 2.22416 -955.266
-114.247 5 1.4539 0.65369 2.33923 -924.585
-114.628 5 1.52389 0.65145 2.45467 -945.067
-112.597 5 1.59362 0.64922 2.57048 -932.944
-110.41 5 1.6631 0.647 2.68665 -914.888
-109.113 5 1.73232 0.64479 2.80319 -886.63
-110.926 5 1.80128 0.64258 2.9201 -920.155
-107.61 5 1.86998 0.64038 3.03738 -872.229
-105.783 5 1.93843 0.63819 3.15503 -833.541
-101.07 5 2.00662 0.63601 3.27304 -792.678
-100.063 5 2.07455 0.63383 3.39142 -813.583
-97.5315 5 2.14222 0.63166 3.51017 -766.788
-94.2183 5 2.20964 0.6295 3.62929 -744.404
-92.4658 5 2.27679 0.62734 3.74878 -696.03
-90.7906 5 2.3437 0.62519 3.86863 -701.474
-88.4277 5 2.41034 0.62305 3.98885 -646.497
-87.3031 5 2.47672 0.62091 4.10944 -678.056
-83.7412 5 2.54285 0.61878 4.2304 -669.866
-81.3188 5 2.60872 0.61666 4.35172 -660.036
-78.6804 5 2.67434 0.61455 4.47342 -611.159
-75.8436 5 2.73969 0.61244 4.59548 -586.745
-76.3901 5 2.80479 0.61034 4.71791 -599.614
-71.3492 5 2.86963 0.60824 4.8407 -549.858
-70.1711 5 2.93421 0.60615 4.96387 -522.438
-67.4958 5 2.99854 0.60407 5.0874 -509.733
-65.4191 5 3.06261 0.602 5.2113 -465.177
-63.6086 5 3.12642 0.59993 5.33557 -442.424
-60.634 5 3.18997 0.59787 5.4602 -443.242
-56.662 5 3.25326 0.59581 5.58521 -391.451
-56.8592 5 3.3163 0.59377 5.71058 -421.343
-56.7613 5 3.37908 0.59172 5.83632 -372.347
Acetate and macrocycle 1 in CH3CN
Data fitted using two binding site model:
Ka1:1 = 222 ± 28 M-1
∆H = -2.73 ± 0.12 kJmol-1,
∆S = 35.21 Jmol-1K-1
Ka2:1 = 5100 ± 127 M-1
∆H = -5.81 ± 0.22 kJmol-1,
∆S = 50.33 Jmol-1K-1
0 1 2 3 4 5 6
0
2
4
6
S4166641_NDH S4166641_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
e) Data for N-Ac-L-Glu and macrocycle 1 in CH3CN (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
278.2733 5 0 1 0.17479 -- 505.0682 5 0.17419 0.99658 0.35017 --
415.5797 5 0.34779 0.99318 0.52616 --
126.969 5 0.52078 0.98979 0.70274 --
-62.9556 5 0.69318 0.98641 0.87991 547.8061
-105.289 5 0.86499 0.98304 1.05769 376.2888
-114.218 5 1.0362 0.97968 1.23606 319.3029
-116.969 5 1.20681 0.97633 1.41503 277.1702
-125.705 5 1.37682 0.973 1.59459 220.7007
-131.88 5 1.54623 0.96967 1.77475 183.1136
-133.597 5 1.71505 0.96636 1.95551 165.5847
-138.31 5 1.88327 0.96306 2.13687 132.3266
-137.51 5 2.0509 0.95977 2.31882 136.332
-142.527 5 2.21793 0.95649 2.50137 106.3029
-146.147 5 2.38436 0.95322 2.68452 93.79639
-148.146 5 2.55019 0.94996 2.86826 76.54938
-150.154 5 2.71543 0.94672 3.0526 62.64744
-151.878 5 2.88007 0.94348 3.23754 52.17631
-151.003 5 3.04411 0.94025 3.42307 54.4694
-148.424 5 3.20756 0.93704 3.6092 67.57537
-150.496 5 3.3704 0.93384 3.79593 60.86384
-151.141 5 3.53266 0.93064 3.98326 43.50492
-151.133 5 3.69431 0.92746 4.17118 41.20537
-151.176 5 3.85537 0.92429 4.3597 36.72307
-150.306 5 4.01583 0.92113 4.54882 35.76104
-150.652 5 4.17569 0.91797 4.73853 33.70992
-151.162 5 4.33496 0.91483 4.92884 22.9673
-149.708 5 4.49363 0.9117 5.11975 25.51959
-150.493 5 4.6517 0.90858 5.31125 22.06272
-148.197 5 4.80918 0.90547 5.50335 27.76487
-149.277 5 4.96606 0.90237 5.69605 10.22058
-149.934 5 5.12234 0.89928 5.88935 8.28937
-149.934 5 5.27802 0.8962 6.08324 9.80645
-149.448 5 5.43311 0.89313 6.27773 7.7555
-147.57 5 5.5876 0.89007 6.47282 10.73838
-145.108 5 5.7415 0.88702 6.6685 15.13758
-148.479 5 5.89479 0.88398 6.86478 -3.36891
-147.121 5 6.04749 0.88094 7.06166 2.00785
-143.684 5 6.1996 0.87792 7.25913 12.50763
-144.889 5 6.3511 0.87491 7.45721 11.03006
-144.921 5 6.50201 0.87191 7.65587 1.81596
-146.556 5 6.65232 0.86892 7.85514 -3.21853
-144.01 5 6.80204 0.86593 8.055 -2.27012
-141.361 5 6.95116 0.86296 8.25546 3.86182
-144.622 5 7.09968 0.86 8.45652 -14.0371
-141.213 5 7.2476 0.85704 8.65817 -0.95278
-143.384 5 7.39493 0.8541 8.86042 -6.07434
-140.788 5 7.54166 0.85116 9.06327 -5.25443
-139.69 5 7.68779 0.84824 9.26672 2.33798
-137.23 5 7.83333 0.84532 9.47076 6.82351
N-Ac-L-Glu and macrocycle 1 in CH3CN
Data fitted using two binding site model:
Ka1:1 = 10210 ± 428 M-1
∆H = 2.37 ± 0.23 kJmol-1,
∆S = 76.75 Jmol-1K-1
Ka2:1 = 1201 ± 89 M-1
∆H = 2.04 ± 0.18 kJmol-1,
∆S = 64.73 Jmol-1K-1
0 2 4 6 8 10
0.0
0.1
0.2
0.3
0.4
0.5
0.6
S4166304ar_NDH S4166304ar_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
f) Data for N-Ac-D-Glu and macrocycle 1 in CH3CN (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
-221.759 5 0 0.75 0.11424 -- -194.953 5 0.08539 0.74744 0.22887 --
-114.827 5 0.17048 0.74489 0.34389 --
-63.746 5 0.25529 0.74234 0.45931 --
-33.403 5 0.3398 0.73981 0.57511 --
-18.1534 5 0.42401 0.73728 0.6913 --
-11.341 5 0.50794 0.73476 0.80788 1090.615
-18.7213 5 0.59157 0.73225 0.92485 1017.931
-32.5127 5 0.67491 0.72975 1.04222 903.7822
-52.1443 5 0.75796 0.72726 1.15997 727.2042
-71.839 5 0.84071 0.72477 1.27811 560.8852
-88.5154 5 0.92317 0.72229 1.39664 431.4605
-111.364 5 1.00534 0.71983 1.51557 237.9472
-126.367 5 1.08722 0.71737 1.63488 124.4941
-132.915 5 1.1688 0.71492 1.75459 52.96839
-134.707 5 1.25009 0.71247 1.87468 37.90549
-139.959 5 1.33109 0.71004 1.99516 -6.59837
-136.838 5 1.4118 0.70761 2.11604 13.16458
-139 5 1.49221 0.70519 2.2373 2.10895
-142.273 5 1.57233 0.70278 2.35896 -30.8697
-142.456 5 1.65216 0.70038 2.481 -22.5812
-143.224 5 1.73169 0.69798 2.60344 -40.4059
-140.522 5 1.81094 0.69559 2.72626 -10.9662
-141.54 5 1.88989 0.69322 2.84948 -26.3939
-143.953 5 1.96854 0.69084 2.97308 -40.4583
-146.427 5 2.04691 0.68848 3.09708 -67.9596
-140.416 5 2.12498 0.68612 3.22146 -12.957
-143.147 5 2.20276 0.68378 3.34624 -36.597
-143.409 5 2.28025 0.68144 3.47141 -34.5234
-145.673 5 2.35744 0.6791 3.59696 -45.0446
-149.585 5 2.43434 0.67678 3.72291 -76.9401
-144.709 5 2.51095 0.67446 3.84925 -46.7871
-146.672 5 2.58727 0.67215 3.97598 -56.3195
-147.891 5 2.66329 0.66985 4.10309 -65.2876
-146.631 5 2.73902 0.66755 4.2306 -55.8779
-145.113 5 2.81446 0.66526 4.3585 -49.0303
-147.3 5 2.8896 0.66298 4.48679 -75.9782
-143.058 5 2.96446 0.66071 4.61546 -22.6095
-147.337 5 3.03902 0.65844 4.74453 -74.1015
-143.766 5 3.11329 0.65618 4.87399 -37.0838
-140.751 5 3.18726 0.65393 5.00384 -14.4072
-143.133 5 3.26094 0.65169 5.13408 -39.5481
-142.913 5 3.33433 0.64945 5.26471 -27.1402
-141.842 5 3.40743 0.64722 5.39573 -24.9534
-146.204 5 3.48023 0.645 5.52714 -84.8392
-142.435 5 3.55275 0.64278 5.65894 -36.8902
-141.836 5 3.62496 0.64057 5.79113 -18.9316
-141.518 5 3.69689 0.63837 5.92371 -13.7272
-141.077 5 3.76852 0.63618 6.05668 -21.9565
-138.016 5 3.83987 0.63399 6.19004 4.48711
N-Ac-D-Glu and macrocycle 1 in CH3CN
Data fitted using two binding site model:
Ka1:1 = 3112 ± 132 M-1
∆H = -5.92 ± 0.27 kJmol-1,
∆S = 46.30 Jmol-1K-1
Ka2:1 = 16013 ± 1228 M-1
∆H = 12.51 ± 0.48 kJmol-1,
∆S = 120.45 Jmol-1K-1
0 2 4 6
0.0
0.5
1.0
1.5
S4166322ar_NDH S4166322ar_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
g) Data for N-Ac-D-Glu and macrocycle 1 in DMSO (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat DMSO).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
-79.8204 5 0 0.6 0.1885 -- -142.007 5 0.11271 0.59795 0.37764 --
-164.055 5 0.22504 0.59591 0.56742 --
-135.572 5 0.33698 0.59387 0.75785 -1064.75
-71.4047 5 0.44853 0.59184 0.94893 -712.889
-23.6634 5 0.5597 0.58982 1.14064 -439.618
7.8213 5 0.67048 0.58781 1.333 -264.35
29.66179 5 0.78087 0.5858 1.52601 -150.246
44.30953 5 0.89088 0.5838 1.71966 -72.0762
54.38499 5 1.0005 0.5818 1.91395 -19.4201
61.95552 5 1.10974 0.57982 2.10888 18.12689
67.80546 5 1.21859 0.57784 2.30446 45.65264
73.5768 5 1.32705 0.57586 2.50069 67.65807
75.91952 5 1.43513 0.57389 2.69755 73.6753
78.11185 5 1.54282 0.57193 2.89507 60.34269
82.10485 5 1.65012 0.56998 3.09322 61.64274
86.52682 5 1.75704 0.56803 3.29202 63.15863
87.50279 5 1.86357 0.56609 3.49146 50.68085
88.75868 5 1.96972 0.56415 3.69155 51.92872
89.2455 5 2.07548 0.56222 3.89228 54.91998
89.06273 5 2.18085 0.5603 4.09365 56.27225
87.27867 5 2.28584 0.55839 4.29567 51.52905
87.24718 5 2.39044 0.55648 4.49833 59.18711
86.24453 5 2.49465 0.55457 4.70164 61.48504
84.71813 5 2.59848 0.55268 4.90559 51.47595
84.98468 5 2.70192 0.55078 5.11018 65.3158
82.89713 5 2.80497 0.5489 5.31542 50.35923
81.33911 5 2.90764 0.54702 5.5213 42.58948
81.92646 5 3.00993 0.54515 5.72782 57.17032
80.22306 5 3.11182 0.54328 5.93499 50.75474
78.22755 5 3.21333 0.54142 6.1428 31.07314
77.95998 5 3.31446 0.53957 6.35126 33.79863
77.44351 5 3.41519 0.53772 6.56036 36.90357
76.14324 5 3.51554 0.53588 6.7701 27.65998
76.33149 5 3.61551 0.53404 6.98049 33.58349
76.864 5 3.71509 0.53221 7.19152 33.07593
76.34105 5 3.81428 0.53039 7.4032 33.76892
74.51271 5 3.91308 0.52857 7.61552 22.04331
76.31938 5 4.0115 0.52675 7.82848 28.49482
74.58309 5 4.10954 0.52495 8.04208 18.76371
74.89448 5 4.20718 0.52315 8.25634 20.52174
75.63293 5 4.30444 0.52135 8.47123 26.74503
75.51682 5 4.40132 0.51956 8.68677 28.88892
70.79126 5 4.49781 0.51778 8.90295 0.35948
71.23486 5 4.59391 0.516 9.11978 4.7571
71.61063 5 4.68962 0.51423 9.33724 3.82284
72.24823 5 4.78495 0.51246 9.55536 6.74236
71.0056 5 4.8799 0.5107 9.77412 1.59857
71.33121 5 4.97445 0.50894 9.99352 7.72336
70.65046 5 5.06862 0.50719 10.21356 2.94226
N-Ac-D-Glu and macrocycle 1 in DMSO
Data fitted using two binding site model:
Ka1:1 = 503 ± 52 M-1
∆H = 3.48 ± 0.19 kJmol-1,
∆S = 62.41 Jmol-1K-1
Ka2:1 = 24993 ± 1309 M-1
∆H = -6.38 ± 0.37 kJmol-1,
∆S = 61.74 Jmol-1K-1
0 2 4 6 8 10
-1.2
-1.0
-0.8
-0.6
-0.4
-0.2
0.0
0.2
S4166631_NDH S4166631_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
h) Data for N-Boc-L-Glu and macrocycle 2 in CH3CN (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
-395.261 5 0 1.31 0.16691 -4738.71 -18.0526 5 0.21791 1.30553 0.3344 -4110.07
217.1694 5 0.43507 1.30107 0.50245 -3976.36
218.5506 5 0.65149 1.29662 0.67108 -3679.89
345.9632 5 0.86716 1.29219 0.84027 -3213.71
401.026 5 1.08208 1.28778 1.01004 -2998.11
413.8809 5 1.29626 1.28338 1.18037 -2918.83
424.3791 5 1.50969 1.279 1.35127 -2814.75
429.1041 5 1.72237 1.27463 1.52275 -2749.86
437.1479 5 1.93431 1.27027 1.69479 -2646.95
439.5955 5 2.1455 1.26593 1.86741 -2588.65
446.2682 5 2.35594 1.26161 2.04059 -2516.02
441.9686 5 2.56563 1.2573 2.21435 -2488.01
420.4742 5 2.77458 1.253 2.38867 -2485.92
455.0899 5 2.98278 1.24872 2.56357 -2321.26
479.042 5 3.19024 1.24445 2.73903 --
444.6691 5 3.39695 1.2402 2.91507 --
442.6216 5 3.60291 1.23596 3.09168 -2195.86
438.0617 5 3.80812 1.23173 3.26885 -2278.91
433.8685 5 4.01259 1.22752 3.4466 --
429.6047 5 4.21631 1.22333 3.62491 -2105.85
433.8393 5 4.41928 1.21914 3.8038 -2046.33
426.4315 5 4.62151 1.21497 3.98326 -2021.4
423.8645 5 4.82299 1.21082 4.16328 -1984.1
416.5014 5 5.02372 1.20667 4.34388 -1966
415.6812 5 5.22371 1.20255 4.52504 -1927.89
412.9732 5 5.42295 1.19843 4.70678 -1894.87
404.246 5 5.62144 1.19433 4.88909 -1875.51
402.3044 5 5.81919 1.19024 5.07196 -1849.63
400.1466 5 6.01619 1.18617 5.25541 -1824.11
394.5112 5 6.21244 1.1821 5.43943 -1801.69
392.9762 5 6.40795 1.17806 5.62402 -1753.18
386.5491 5 6.60271 1.17402 5.80917 -1738.29
383.1872 5 6.79672 1.17 5.9949 -1720.64
381.7875 5 6.98998 1.16599 6.1812 -1677.53
375.9513 5 7.1825 1.16199 6.36806 -1657.43
374.7868 5 7.37427 1.15801 6.5555 -1630.41
372.7234 5 7.5653 1.15404 6.74351 -1603.5
366.7657 5 7.75557 1.15008 6.93209 -1588.96
363.1843 5 7.9451 1.14613 7.12124 -1566.56
362.5511 5 8.13389 1.1422 7.31095 -1538.1
360.4525 5 8.32193 1.13828 7.50124 -1522.61
355.0395 5 8.50922 1.13437 7.6921 -1492.78
353.0595 5 8.69576 1.13048 7.88353 -1467.15
346.3298 5 8.88156 1.1266 8.07553 -1463.05
347.2566 5 9.06661 1.12273 8.26809 -1423.82
335.6471 5 9.25091 1.11887 8.46123 -1433.31
340.3261 5 9.43447 1.11502 8.65494 -1388.2
328.4469 5 9.61727 1.11119 8.84922 -1396.16
333.6669 5 9.79934 1.10737 9.04407 -1344.24
N-Boc-L-Glu and macrocycle 2 in CH3CN
Data fitted using two binding site model:
Ka1:1 = 1804 ± 1082 M-1
∆H = -21.84 ± 2.74 kJmol-1,
∆S = -11.25 Jmol-1K-1
(Ka2:1 = 11 ± 5 M-1)
0 2 4 6 8 10
-5
-4
-3
-2
-1
SRmacroLgl_NDH SRmacroLgl_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
i) Data for N-Boc-D-Glu and macrocycle 2 in CH3CN (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
-95.1721 5 0 1.25 0.17849 -- -178.098 5 0.22235 1.24573 0.35759 -2644.15
105.351 5 0.44394 1.24148 0.5373 -1762.82
193.9395 5 0.66477 1.23724 0.71762 -1497.53
233.9105 5 0.88483 1.23301 0.89855 -1350.73
257.3857 5 1.10413 1.2288 1.08009 -1268.58
284.3982 5 1.32267 1.2246 1.26223 -1161.79
299.4787 5 1.54045 1.22042 1.44499 -1093.66
305.9875 5 1.75747 1.21625 1.62836 -1037.23
297.7314 5 1.97372 1.21209 1.81233 -1038.24
307.8205 5 2.18921 1.20795 1.99692 -969.666
317.3467 5 2.40394 1.20382 2.18212 -915.321
320.6082 5 2.61791 1.19971 2.36792 -870.234
309.3379 5 2.83112 1.19561 2.55434 -881.594
317.4429 5 3.04356 1.19153 2.74136 -836.684
317.6478 5 3.25524 1.18745 2.929 -804.291
317.6342 5 3.46616 1.1834 3.11724 -775.894
318.3204 5 3.67632 1.17935 3.3061 -742.89
310.0876 5 3.88572 1.17532 3.49556 -743.057
312.2355 5 4.09435 1.1713 3.68563 -714.017
308.4497 5 4.30222 1.1673 3.87632 -686.694
310.6111 5 4.50933 1.1633 4.06761 -666.212
305.2017 5 4.71568 1.15932 4.25951 -640.607
293.0352 5 4.92127 1.15536 4.45202 -677.271
298.8794 5 5.12609 1.15141 4.64514 -616.282
299.5859 5 5.33015 1.14747 4.83888 -608.516
295.8911 5 5.53345 1.14354 5.03322 -594.949
299.2872 5 5.73599 1.13963 5.22817 -564.103
293.7691 5 5.93776 1.13573 5.42373 -557.338
289.7578 5 6.13878 1.13184 5.6199 -549.435
287.7317 5 6.33903 1.12796 5.81668 -530.913
283.8299 5 6.53852 1.1241 6.01406 -516.642
288.7942 5 6.73724 1.12025 6.21206 -480.116
280.8526 5 6.93521 1.11641 6.41067 -490.786
284.7717 5 7.13241 1.11258 6.60989 -456.533
280.4919 5 7.32885 1.10877 6.80972 -448.627
281.7936 5 7.52453 1.10497 7.01015 -437.801
281.7778 5 7.71945 1.10118 7.2112 -414.914
274.9901 5 7.9136 1.0974 7.41286 -409.693
268.7288 5 8.107 1.09364 7.61512 -423.917
272.9118 5 8.29963 1.08989 7.818 -399.117
263.376 5 8.49149 1.08615 8.02148 -398.862
264.1807 5 8.6826 1.08242 8.22558 -387.223
267.5108 5 8.87295 1.0787 8.43028 -354.269
266.1047 5 9.06253 1.075 8.6356 -344.118
263.7852 5 9.25135 1.0713 8.84152 -337.559
258.847 5 9.43941 1.06762 9.04806 -341.548
257.3103 5 9.6267 1.06395 9.2552 -327.827
256.6965 5 9.81324 1.06029 9.46295 -319.989
252.5119 5 9.99901 1.05665 9.67131 -318.538
N-Boc-D-Glu and macrocycle 2 in CH3CN
Data fitted using two binding site model:
Ka2:1 = 2071 ± 1136 M-1
∆H = -11.73 ± 3.41 kJmol-1,
∆S = 24.16 Jmol-1K-1
(Ka1:1 = 28 ± 17 M-1)
0 2 4 6 8 10
-2.5
-2.0
-1.5
-1.0
-0.5
SRmacroDgl_NDH SRmacroDgl_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
j) Data for N-Boc-L-Glu and macrocycle 2 in DMSO (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat DMSO).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
-108.462 5 0 1.2 0.18221 -514.989 -66.8496 5 0.21791 1.1959 0.36505 -617.304
-26.5499 5 0.43507 1.19182 0.54851 -507.419
3.31264 5 0.65149 1.18775 0.73259 -430.309
19.273 5 0.86716 1.18369 0.9173 -396.834
20.00068 5 1.08208 1.17965 1.10262 -411.167
21.08862 5 1.29626 1.17562 1.28857 -424.426
30.42911 5 1.50969 1.1716 1.47514 -393.692
48.33094 5 1.72237 1.1676 1.66233 -347.208
65.99387 5 1.93431 1.16361 1.85015 -299.62
78.27402 5 2.1455 1.15963 2.03859 -270.706
86.32697 5 2.35594 1.15567 2.22765 -242.885
94.31854 5 2.56563 1.15172 2.41733 -218.614
100.9725 5 2.77458 1.14779 2.60764 -197.917
107.7744 5 2.98278 1.14387 2.79856 -164.019
111.3553 5 3.19024 1.13996 2.99011 -154.228
114.1639 5 3.39695 1.13606 3.18229 -144.25
116.0986 5 3.60291 1.13218 3.37508 -133.559
119.223 5 3.80812 1.12831 3.5685 -117.498
116.3975 5 4.01259 1.12445 3.76254 -121.807
119.6677 5 4.21631 1.1206 3.9572 -104.092
121.3333 5 4.41928 1.11677 4.15248 -95.9757
118.7878 5 4.62151 1.11295 4.34839 -99.1466
117.6171 5 4.82299 1.10914 4.54492 -97.977
121.9495 5 5.02372 1.10535 4.74207 -76.9825
120.6552 5 5.22371 1.10157 4.93984 -78.9524
118.3217 5 5.42295 1.0978 5.13824 -80.4157
119.2518 5 5.62144 1.09404 5.33725 -72.5174
120.548 5 5.81919 1.0903 5.53689 -58.7343
119.318 5 6.01619 1.08656 5.73716 -57.958
119.2199 5 6.21244 1.08284 5.93804 -54.9158
117.1565 5 6.40795 1.07913 6.13955 -59.6967
114.9613 5 6.60271 1.07544 6.34168 -57.1836
117.1894 5 6.79672 1.07175 6.54443 -46.6266
115.7506 5 6.98998 1.06808 6.74781 -42.9217
114.6413 5 7.1825 1.06442 6.9518 -49.1772
114.0004 5 7.37427 1.06077 7.15642 -42.6776
113.5383 5 7.5653 1.05713 7.36166 -41.0047
111.8289 5 7.75557 1.05351 7.56753 -42.7212
112.1663 5 7.9451 1.04989 7.77402 -35.909
110.696 5 8.13389 1.04629 7.98112 -38.4326
108.7372 5 8.32193 1.0427 8.18886 -42.7354
108.4469 5 8.50922 1.03912 8.39721 -39.6221
107.687 5 8.69576 1.03555 8.60618 -34.0158
106.374 5 8.88156 1.032 8.81578 -31.3372
106.6299 5 9.06661 1.02845 9.026 -31.2879
107.4048 5 9.25091 1.02492 9.23685 -23.8
104.5279 5 9.43447 1.02139 9.44831 -28.9614
100.2885 5 9.61727 1.01788 9.6604 -36.6289
101.1653 5 9.79934 1.01438 9.87311 -33.2215
N-Boc-L-Glu and macrocycle 2 in DMSO
Data fitted using two binding site model:
Ka1:1 = 427 ± 28 M-1
∆H = 7.84 ± 0.25 kJmol-1,
∆S = 24.07 Jmol-1K-1
(Ka2:1 = 21 ± 5 M-1)
0 2 4 6 8 10
-0.6
-0.5
-0.4
-0.3
-0.2
-0.1
0.0
SRLgluDMSO_NDH SRLgluDMSO_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
k) Data for N-Boc-L-Asp and macrocycle 2 in CH3CN (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
113.4196 5 0 1.24 0.17634 -- -21.4013 5 0.21791 1.23576 0.35328 --
272.1399 5 0.43507 1.23154 0.53082 -2404.4
345.9685 5 0.65149 1.22734 0.70896 -2150.45
395.3356 5 0.86716 1.22315 0.88771 -1945.51
454.1679 5 1.08208 1.21897 1.06705 -1708.36
509.3942 5 1.29626 1.2148 1.247 -1480.67
563.2501 5 1.50969 1.21065 1.42756 -1244.94
588.7566 5 1.72237 1.20652 1.60871 -1125.81
607.0309 5 1.93431 1.2024 1.79047 -1016.9
617.8467 5 2.1455 1.19829 1.97283 -930.758
614.8726 5 2.35594 1.19419 2.15579 -891.685
614.8521 5 2.56563 1.19011 2.33935 -831.687
608.2634 5 2.77458 1.18605 2.52352 -823.315
602.1144 5 2.98278 1.18199 2.70829 -784.84
595.1951 5 3.19024 1.17795 2.89366 -766.977
589.2062 5 3.39695 1.17393 3.07963 -741.659
578.0951 5 3.60291 1.16992 3.26621 -736.005
569.1938 5 3.80812 1.16592 3.45338 -715.961
561.8611 5 4.01259 1.16193 3.64116 -688.488
555.2768 5 4.21631 1.15796 3.82955 -666.282
547.9678 5 4.41928 1.154 4.01853 -641.222
538.8672 5 4.62151 1.15005 4.20812 -632.995
535.2448 5 4.82299 1.14612 4.39831 -606.457
527.0428 5 5.02372 1.1422 4.5891 -604.919
519.5921 5 5.22371 1.13829 4.78049 -583.471
513.5225 5 5.42295 1.13439 4.97249 -571.177
506.6473 5 5.62144 1.13051 5.16508 -557.617
498.3341 5 5.81919 1.12664 5.35829 -543.514
491.9654 5 6.01619 1.12278 5.55209 -525.359
488.6812 5 6.21244 1.11894 5.74649 -497.417
480.4525 5 6.40795 1.11511 5.9415 -490.119
473.565 5 6.60271 1.11129 6.13711 -485.469
465.2013 5 6.79672 1.10748 6.33332 -484.55
462.7295 5 6.98998 1.10368 6.53014 -456.465
454.6407 5 7.1825 1.0999 6.72755 -449.007
450.4784 5 7.37427 1.09613 6.92557 -425.768
444.4835 5 7.5653 1.09237 7.12419 -423.489
439.1584 5 7.75557 1.08863 7.32342 -413.13
434.8225 5 7.9451 1.08489 7.52324 -393.938
429.1925 5 8.13389 1.08117 7.72367 -384.541
422.2736 5 8.32193 1.07746 7.9247 -379.857
417.3219 5 8.50922 1.07376 8.12633 -367.34
410.0866 5 8.69576 1.07007 8.32857 -360.917
405.0114 5 8.88156 1.0664 8.5314 -347.406
401.9393 5 9.06661 1.06273 8.73484 -331.246
394.5031 5 9.25091 1.05908 8.93888 -331.818
390.8598 5 9.43447 1.05544 9.14353 -325.969
384.9848 5 9.61727 1.05181 9.34877 -313.307
379.6274 5 9.79934 1.04819 9.55462 -314.613
N-Boc-L-Asp and macrocycle 2 in CH3CN
Data fitted using two binding site model:
Ka1:1 = 3919 ± 1773 M-1
∆H = -14.73 ± 1.44 kJmol-1,
∆S = 19.42 Jmol-1K-1
(Ka2:1 = 24 ± 16 M-1)
0 2 4 6 8 10-3.0
-2.5
-2.0
-1.5
-1.0
-0.5
SRmacroLas_NDH SRmacroLas_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
l) Data for N-Boc-D-Asp and macrocycle 2 in CH3CN (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat CH3CN).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
1727.611 5 0 1.24 0.17744 -- 1774.855 5 0.21928 1.23576 0.35549 --
1784.452 5 0.4378 1.23154 0.53414 -11.2114
1772.393 5 0.65558 1.22734 0.7134 4.38203
1749.548 5 0.8726 1.22315 0.89327 8.7899
1776.496 5 1.08887 1.21897 1.07374 168.3662
1781.112 5 1.30439 1.2148 1.25482 248.98
1804.072 5 1.51915 1.21065 1.43651 414.7873
1823.299 5 1.73317 1.20652 1.6188 543.8012
1835.088 5 1.94644 1.2024 1.80169 654.4388
1837.472 5 2.15895 1.19829 1.9852 710.7435
1832.122 5 2.37071 1.19419 2.1693 754.9333
1818.242 5 2.58172 1.19011 2.35402 755.1242
1810.272 5 2.79198 1.18605 2.53934 765.3966
1794.855 5 3.00149 1.18199 2.72527 787.7359
1781.873 5 3.21024 1.17795 2.9118 796.8801
1753.841 5 3.41824 1.17393 3.09894 796.999
1734.489 5 3.6255 1.16992 3.28668 794.3373
1714.629 5 3.832 1.16592 3.47504 806.7799
1699.205 5 4.03775 1.16193 3.66399 798.313
1681.575 5 4.24275 1.15796 3.85356 790.2168
1662.684 5 4.44699 1.154 4.04372 782.0844
1649.271 5 4.65049 1.15005 4.2345 773.7968
1634.683 5 4.85323 1.14612 4.42588 777.1863
1615.328 5 5.05522 1.1422 4.61787 759.87
1601.738 5 5.25646 1.13829 4.81046 750.5448
1580.689 5 5.45695 1.13439 5.00366 --
1567.068 5 5.65669 1.13051 5.19747 728.9108
1551.534 5 5.85567 1.12664 5.39188 725.5576
1539.26 5 6.05391 1.12278 5.5869 737.5146
1520.831 5 6.25139 1.11894 5.78252 715.9899
1500.055 5 6.44812 1.11511 5.97875 --
1491.178 5 6.6441 1.11129 6.17559 721.8724
1479.314 5 6.83933 1.10748 6.37303 714.6847
1462.409 5 7.03381 1.10368 6.57108 709.4065
1450.789 5 7.22753 1.0999 6.76973 710.4806
1443.856 5 7.42051 1.09613 6.96899 --
1427.553 5 7.61273 1.09237 7.16886 700.4468
1415.443 5 7.8042 1.08863 7.36933 706.1012
1401.768 5 7.99492 1.08489 7.57041 698.2735
1389.129 5 8.18488 1.08117 7.77209 694.0906
1372.148 5 8.3741 1.07746 7.97438 683.2207
1353.013 5 8.56257 1.07376 8.17728 648.6628
1341.485 5 8.75028 1.07007 8.38078 655.6341
1329.359 5 8.93724 1.0664 8.58489 648.6679
1323.152 5 9.12345 1.06273 8.78961 659.2498
1306.038 5 9.30891 1.05908 8.99493 --
1303.291 5 9.49362 1.05544 9.20085 661.5095
1289.66 5 9.67757 1.05181 9.40739 651.9963
1279.571 5 9.86077 1.04819 9.61453 643.9757
N-Boc-D-Asp and macrocycle 2 in CH3CN
Data fitted using two binding site model:
Ka2:1 = 2125 ± 633 M-1
∆H = -8.44 ± 0.98 kJmol-1,
∆S = 10.55 Jmol-1K-1
(Ka1:1 = 8 ± 4 M-1)
0 2 4 6 8 10
-0.2
0.0
0.2
0.4
0.6
0.8
SRDAspMeCN_NDH SRDAspMeCN_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
m) Data for N-Boc-L-Glu and macrocycle 3 in DMSO (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat DMSO).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
74.50041 2.5 0 0.96 0.11379 -- 97.52066 2.5 0.10905 0.95836 0.22777 --
-29.821 2.5 0.21791 0.95672 0.34194 -1249.02
-70.8563 2.5 0.32659 0.95509 0.45631 -1397.78
-65.0043 2.5 0.43507 0.95345 0.57088 -1244.89
-59.6385 2.5 0.54338 0.95182 0.68564 -1231.21
-53.3412 2.5 0.65149 0.9502 0.80059 -1118.21
-44.5306 2.5 0.75942 0.94857 0.91574 -1092.06
-33.7178 2.5 0.86716 0.94695 1.03108 -1005.43
-16.6089 2.5 0.97472 0.94533 1.14662 -875.893
0.39347 2.5 1.08208 0.94372 1.26235 -780.586
17.38409 2.5 1.18927 0.9421 1.37828 -636.947
26.74415 2.5 1.29626 0.94049 1.4944 -617.174
32.39921 2.5 1.40307 0.93889 1.61071 -533.351
41.73516 2.5 1.50969 0.93728 1.72722 -489.626
45.87701 2.5 1.61612 0.93568 1.84393 -445.421
52.99478 2.5 1.72237 0.93408 1.96082 -385.822
55.71731 2.5 1.82843 0.93248 2.07792 -370.74
57.81987 2.5 1.93431 0.93089 2.19521 -342.512
60.84859 2.5 2.04 0.9293 2.31269 -335.504
60.13929 2.5 2.1455 0.92771 2.43036 -319.769
65.0282 2.5 2.25081 0.92612 2.54823 -287.322
65.15118 2.5 2.35594 0.92454 2.6663 -272.984
65.50691 2.5 2.46088 0.92296 2.78456 -262.728
67.2987 2.5 2.56563 0.92138 2.90301 -261.128
65.78315 2.5 2.6702 0.9198 3.02166 -270.504
68.09184 2.5 2.77458 0.91823 3.14051 -254.368
66.02349 2.5 2.87878 0.91666 3.25954 -260.327
67.38673 2.5 2.98278 0.91509 3.37878 -228.833
68.78892 2.5 3.08661 0.91353 3.4982 -228.674
134.0012 5 3.19024 0.91196 3.73764 -227.93
130.8076 5 3.39695 0.90885 3.97786 -210.406
131.3317 5 3.60291 0.90574 4.21885 -192.208
129.2559 5 3.80812 0.90264 4.46062 -190.157
129.3438 5 4.01259 0.89956 4.70317 -177.236
128.4375 5 4.21631 0.89648 4.9465 -174.287
127.2875 5 4.41928 0.89342 5.1906 -165.535
127.3219 5 4.62151 0.89036 5.43548 -157.21
129.2325 5 4.82299 0.88732 5.68115 -139.966
131.1952 5 5.02372 0.88428 5.92758 -124.587
130.9191 5 5.22371 0.88125 6.1748 -112.607
130.8043 5 5.42295 0.87824 6.4228 -103.942
130.1433 5 5.62144 0.87523 6.67157 -98.0994
129.0193 5 5.81919 0.87224 6.92112 -92.5438
128.8926 5 6.01619 0.86925 7.17145 -86.7879
127.217 5 6.21244 0.86627 7.42255 -81.5758
126.6934 5 6.40795 0.86331 7.67444 -75.9408
124.353 5 6.60271 0.86035 7.9271 -76.611
123.5786 5 6.79672 0.8574 8.18054 -71.1053
122.6288 5 6.98998 0.85446 8.43476 -68.3219
N-Boc-L-Glu and macrocycle 3 in DMSO
Data fitted using two binding site model:
Ka1:1 = 27450 ± 8173 M-1
∆H = -5.93 ± 0.16 kJmol-1,
∆S = 65.09 Jmol-1K-1
Ka2:1 = 393 ± 38 M-1
∆H = -8.64 ± 0.34 kJmol-1,
∆S = 20.69 Jmol-1K-1
0 2 4 6 8 10 12 14
-1.4
-1.2
-1.0
-0.8
-0.6
-0.4
-0.2
0.0
GKBLGlu1_NDH GKBLGlu1_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t
n) Data for N-Boc-D-Glu and macrocycle 3 in DMSO (data corrected by subtracting the background dilution data for an injection of the guest solution into the cell containing neat DMSO).
area
(µcal)
injection
(µL)
[guest] (mM)
(before inj.)
[host] (mM)
(before inj.)
[host] / [guest]
(after inj.)
cal / mol of
injected guest
62.91751 2.5 0 0.96 0.11379 -- 51.89695 2.5 0.10905 0.95836 0.22777 --
-81.1643 2.5 0.21791 0.95672 0.34194 -1700.32
-118.208 2.5 0.32659 0.95509 0.45631 -1717.81
-123.3 2.5 0.43507 0.95345 0.57088 -1673.49
-113.732 2.5 0.54338 0.95182 0.68564 -1549.1
-92.9804 2.5 0.65149 0.9502 0.80059 -1374.42
-77.6178 2.5 0.75942 0.94857 0.91574 -1274.09
-51.2993 2.5 0.86716 0.94695 1.03108 -1101.81
-26.3541 2.5 0.97472 0.94533 1.14662 -950.899
-3.06833 2.5 1.08208 0.94372 1.26235 -784.097
21.84286 2.5 1.18927 0.9421 1.37828 -608.811
46.14084 2.5 1.29626 0.94049 1.4944 -467.679
52.79022 2.5 1.40307 0.93889 1.61071 -409.025
60.69648 2.5 1.50969 0.93728 1.72722 -361.994
63.13243 2.5 1.61612 0.93568 1.84393 -342.981
70.30955 2.5 1.72237 0.93408 1.96082 -293.15
79.02114 2.5 1.82843 0.93248 2.07792 -243.001
77.30496 2.5 1.93431 0.93089 2.19521 -228.703
84.52519 2.5 2.04 0.9293 2.31269 -193.837
83.26833 2.5 2.1455 0.92771 2.43036 -196.865
89.45713 2.5 2.25081 0.92612 2.54823 -153.039
88.53462 2.5 2.35594 0.92454 2.6663 -157.001
85.01858 2.5 2.46088 0.92296 2.78456 -170.776
86.70741 2.5 2.56563 0.92138 2.90301 -188.376
89.02892 2.5 2.6702 0.9198 3.02166 -160.371
93.22999 2.5 2.77458 0.91823 3.14051 -125.588
88.2301 2.5 2.87878 0.91666 3.25954 -149.696
94.04049 2.5 2.98278 0.91509 3.37878 -104.3
93.30024 2.5 3.08661 0.91353 3.4982 -118.132
184.2897 5 3.19024 0.91196 3.73764 -121.358
178.8933 5 3.39695 0.90885 3.97786 -118.409
179.6489 5 3.60291 0.90574 4.21885 -94.2429
181.234 5 3.80812 0.90264 4.46062 -85.9075
178.9776 5 4.01259 0.89956 4.70317 -88.1921
176.2317 5 4.21631 0.89648 4.9465 -91.3095
175.3992 5 4.41928 0.89342 5.1906 -80.7888
175.6947 5 4.62151 0.89036 5.43548 -65.6817
173.7684 5 4.82299 0.88732 5.68115 -64.0699
174.2266 5 5.02372 0.88428 5.92758 -56.2853
170.9364 5 5.22371 0.88125 6.1748 -58.5747
168.7411 5 5.42295 0.87824 6.4228 -59.2024
169.737 5 5.62144 0.87523 6.67157 -51.0549
170.5513 5 5.81919 0.87224 6.92112 -42.6537
168.1541 5 6.01619 0.86925 7.17145 -43.7751
168.948 5 6.21244 0.86627 7.42255 -37.3792
167.0643 5 6.40795 0.86331 7.67444 -35.4893
164.4968 5 6.60271 0.86035 7.9271 -41.4772
164.3187 5 6.79672 0.8574 8.18054 -37.5788
162.3624 5 6.98998 0.85446 8.43476 -38.2072
N-Boc-D-Glu and macrocycle 3 in DMSO
Data fitted using two binding site model:
Ka1:1 = 262 ± 46 M-1
∆H = -4.37 ± 0.27 kJmol-1,
∆S = 31.03 Jmol-1K-1
Ka2:1 = 19380 ± 2061 M-1
∆H = -7.97 ± 0.10 kJmol-1,
∆S = 55.34 Jmol-1K-1
0 2 4 6 8 10 12 14-2.0
-1.5
-1.0
-0.5
0.0
GKBDGlu2_NDH GKBDGlu2_Fit
Molar Ratio
kcal
/mol
e of
inje
ctan
t