Introduction of reacting partners
□ 31
Jan Novotný
Recommended procedure
O Assignment of free receptor: /3-cyklodextrine (ID H, DQF-COSY) O Assignment of free ligand: doxepine (ID 1H, DQF-COSY, NOESY)
• Identification of proton resonances of ring A and B
• Determination of major and minor conformation of doxepin
O ID NMR titration: rearrangement of /3-cyclodextrin resonaces upon
interaction =>* identification of inner protons O ID NMR titration: rearrangement of doxepin resonaces upon complexation
=>* estimation of binding mode Q ROESY spectrum of complex: ROE intermolecular contacts O Fitting the titration isotherm
ID ľH of ^fcyclodextrin in D 20
DQF-COSY of ^yclodextrin in D 20
3.5
4.0-
E
Q. Q_
4.5
5.0-
Jan Novotný
f
5.0
5.0
4.5
e c,d b,a
4.0 3.5
m Ti
4.5 4.0 2-^ (ppm)
3.5
4.0
4.5
5.0
3.5
May 3, 2017       5 / 17
ID XH of doxepin in D20
ID XH of doxepin in D20
NOESY 700ms of doxepin in D20
NOESY 700ms of doxepin in D20
7.5    /   A \
M   L K J
C02 - H (ppm)
Jan Novotný
May 3, 2017        10 / 17
ID ľH NMR titration: ^fcyclodextrin
H NMR titration: doxepin
ID *H NMR titration: doxepin
1.9605 1.6154
1.1021
doxepin :(3CD 5:1
3,75:1
1,75:1 1:1
0,75:1
0,2:1 0:1
1.0035
K,L J   | '
1394
0.7815
H-A2
7.4
7.2
7.0
6.8
A
6.6
[ppm]
Find the most perturbed resonances and estimate the time regime of interaction
Jan Novotný
May 3, 2017        14 / 17
ROESY 300ms - doxepin:/iyclodextrin=5:l