1. Philip A. Gale
Transmembrane anion transport mediated by
calix[4]pyrrole
Nanjing
June 2014

2. Cystic fibrosis transmembrane conductance
regulator CFTR
• Cystic fibrosis
CFTR found in the epithelial cells of many tissues including
those in the intestine, lung, reproductive tracts, pancreatic
ducts and sweat glands.
CF is caused by a mutation in a single gene and affects 1 in
2000-2500 people in Northern Europe and the US.
Disruption of chloride and bicarbonate transport through
epithelial cell membranes.
Lipid bilayer anion transport
• Cancer
Serratia marcescens
• Induction of tumour cell apoptosis
• Immunosuppression
• Toxicity against bacteria, protozoa, fungi and the
malaria parasite
• Resensitization of cancer cells to drugs

3. Hofmeister series
organic anions > ClO4
- > SCN- > I- > NO3
- > Br- > Cl- > HCO3
-> H2PO4
- > F-, SO4
2- > HPO4
2-
...orders anions in terms of their hydrophobicity
F. Hofmeister, Arch. Exp. Patol. Phamakol. 1888, 24, 247.
Hydration energy
Anion ΔGh(X-) (kJmol-1)
NO3
- -314
Cl- -338
HCO3
- -388
SO4
2- -1103
B.A. Moyer et al. in Supramolecular Chemistry of Anions, Eds. A. Bianchi, K.
Bowman-James and E. García-España, Wiley, 1997.

4. Lipids and bilayer membranes
7
+
3
POPC
Dissolve in CH2Cl2 and then
evaporate to make a film
Add a solution of buffer and
NaCl in water and sonicate
Nine freeze thaw cycles
Pass suspension through a
200nm membrane 29 times

5. NaCl NaCl
buffered solution
Vesicles
7:3 POPC:cholesterol
200 nm
NaNO3
Dialysis

6. buffered solution
Antiport
7:3 POPC:cholesterol
Cl-
NO3
-

7. buffered solution
Symport
7:3 POPC:cholesterol
Cl-
M+

8. 455. Adolf von Baeyer: Uber ein
Condensationsproduct von Pyrrol mit Aceton
Berichte 1886, 19, 2184
White crystalline material

9. Calix[4]pyrrolePorphyrinogen

10. P.A.Gale,J.L.Sessler,V.KrálandV.Lynch,J.Am.Chem.Soc.1996,118,
5140.
Calix[4]pyrrole
TEA Cl Ka = 2.2 x 105 M-1 in
acetonitrile

11. Caesium inclusion
R. Custelcean, L.H. Delmau, B.A. Moyer, J.L. Sessler, W.-S. Cho, D. Gross, G.W. Bates, S.J. Brooks, M.E. Light and P.A. Gale,
Angew. Chem. Int. Ed. 2005, 44, 2537 .
J.L. Sessler, D.E. Gross, W.-S. Cho, V.M. Lynch, F.P. Schmidtchen, G.W. Bates, M.E. Light and P.A. Gale,
J. Am. Chem. Soc. 2006, 128, 12281-12288.
D.E. Gross, F.P. Schmidtchen, W. Antonius, P.A. Gale, V.M. Lynch and J.L. Sessler,
Chem. Eur. J., 2008, 14, 7822-7827

12. Calixpyrrole: an old yet new
membrane transport agent?
C.C. Tong, P.A. Gale, J.L. Sessler and R. Quesada, Chem. Commun. 2008,
6321.

13. Calixpyrrole: an old yet new
membrane transport agent?
NaNO3
phosphate buffer, pH 7.2
POPC vesicle
C.C. Tong, P.A. Gale, J.L. Sessler and R. Quesada, Chem. Commun. 2008,
6321.

14. Calixpyrrole: an old yet new
membrane transport agent?
POPC vesicle
NaNO3
phosphate buffer pH 7.2
C.C. Tong, P.A. Gale, J.L. Sessler and R. Quesada, Chem. Commun. 2008,
6321.

15. Strapped calix[4]pyrrole

16. Synthesis
M.G. Fisher, P.A. Gale, J.R. Hiscock, M.B. Hursthouse, M.E. Light, F.P. Schmidtchen and C.C. Tong, Chem. Commun. 2009, 3017-3019.

17. Crystal structure
M.G. Fisher, P.A. Gale, J.R. Hiscock, M.B. Hursthouse, M.E. Light, F.P. Schmidtchen and C.C. Tong, Chem. Commun. 2009, 3017-3019.

18. NMR titrations: Cl- in CD3CN
TEA Cl Ka = 1.3
x 106 M-1 in
acetonitrile

19. Chloride transport
POPC vesicle
NaNO3 or Na2SO4
phosphate buffer pH 7.2
M.G. Fisher, P.A. Gale, J.R. Hiscock, M.B. Hursthouse, M.E. Light, F.P. Schmidtchen and C.C. Tong, Chem. Commun. 2009, 3017-3019.

20. Octafluorocalix[4]pyrrole
P.A. Gale, C.C. Tong, C.J.E. Haynes, O. Adeosun, D.E. Gross E. Karnas, E. Sedenburg, R. Quesada and J.L. Sessler , J. Am. Chem. Soc.
2010,132, 3240.

21. Octafluorocalix[4]pyrrole
Chloride efflux promoted by 0.04 molar equivalents of 1 across unilamellar POPC
vesicles loaded with 489 mM cesium chloride and by 2 in unilamellar POPC vesicles
loaded with 489 mM sodium , potassium, rubidium and cesium chloride salts buffered to
pH 7.2 with 5mM phosphate. The vesicles were dispersed in 489 mM NaNO3 buffered to
pH 7.2 with 5 mM phosphate. Each point represents the average of three trials.
POPC vesicle
NaNO3
phosphate buffer pH 7.2
P.A. Gale, C.C. Tong, C.J.E. Haynes, O. Adeosun, D.E. Gross E. Karnas, E. Sedenburg, R. Quesada and J.L. Sessler , J. Am. Chem. Soc.
2010,132, 3240.

22. Octafluorocalix[4]pyrrole
POPC vesicle
Na2SO4
phosphate buffer pH 7.2
Chloride efflux promoted by 0.04 molar equivalents of 1 across unilamellar POPC
vesicles loaded with 489 mM cesium chloride and by 2 in unilamellar POPC vesicles
loaded with 489 mM sodium (), potassium, rubidium and cesium chloride salts buffered
to pH 7.2 with 5mM phosphate. The vesicles were dispersed in 162 mM Na2SO4
buffered to pH 7.2 with 5 mM phosphate. Each point represents the average of three
trials.
P.A. Gale, C.C. Tong, C.J.E. Haynes, O. Adeosun, D.E. Gross E. Karnas, E. Sedenburg, R. Quesada and J.L. Sessler , J. Am. Chem. Soc.
2010,132, 3240.

23. Octafluorocalix[4]pyrrole
POPC vesicle
Na2SO4
phosphate buffer pH 7.2
Chloride efflux promoted by 0.04 molar equivalents of 1 and 2 across unilamellar
POPC vesicles loaded with 489 mM NaCl buffered to pH 7.2 with 20 mM phosphate
upon addition of a NaHCO3 pulse, to make the extravesicular bicarbonate
concentration 40 mM. The vesicles were dispersed in 162 mM Na2SO4 buffered to pH
7.2 with 20 mM phosphate. Each point represents the average of three trials.
NaHCO3
P.A. Gale, C.C. Tong, C.J.E. Haynes, O. Adeosun, D.E. Gross E. Karnas, E. Sedenburg, R. Quesada and J.L. Sessler , J. Am. Chem. Soc.
2010,132, 3240.

24. A dual-host approach to
transmembrane transport
S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Chem. Commun. 2011, 47, 689-691.

25. valinomycin
CsCl co-transport only
Cl-/NO3
- antiport
M. Yano, C.C. Tong, M.E. Light, F.P.
Schmidtchen and P.A. Gale, Org. Biomol.
Chem. 2010, 8, 4356.
K+ transport
S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Chem. Commun. 2011, 47, 689-691.
M.G. Fisher, P.A. Gale, J.R. Hiscock, M.B.
Hursthouse, M.E. Light, F.P. Schmidtchen and
C.C. Tong, Chem. Commun. 2009, 3017-3019.
C.C. Tong, P.A. Gale, J.L.
Sessler and R. Quesada, Chem.
Commun. 2008, 6321.
A dual-host approach to
transmembrane transport

26. Crystal structure
M. Yano, C.C. Tong, M.E. Light, F.P. Schmidtchen and P.A. Gale, Org. Biomol. Chem. 2010, 8, 4356.

27. POPC vesicle
Na2SO4
phosphate buffer, pH 7.2
lucigenin
KCl pulse
A dual-host approach to
transmembrane transport
Na2SO4
phosphate buffer
pH 7.2
S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Chem. Commun. 2011, 47, 689-691.

28. Chloride influx (each carrier at 2%
molar carrier to lipid loading)
Fluorescence studies
S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Chem. Commun. 2011, 47, 689-691.

29. Chloride influx (each carrier at 2%
molar carrier to lipid loading)
Fluorescence studies
S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Chem. Commun. 2011, 47, 689-691.

30. S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Chem. Commun. 2011, 47, 689-691.
A dual-host approach to
transmembrane transport

31. Synergy in anion antiport?
A1 =
A2 =
S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Dalton Trans. 2011, 40, 12017-12020.

32. POPC vesicle
Na2SO4
phosphate buffer, pH 7.2
NaHCO3 pulseNaCl
Synergy in anion antiport?
HCO3
-
Cl- Cl-
HCO3
-
S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Dalton Trans. 2011, 40, 12017-12020.

33. Synergy in anion antiport?
A1 =
A2 =
Chloride efflux from unilamellar POPC vesicles containing 489mM sodium chloride,
buffered to pH 7.2 with 20mM sodium phosphate salts and suspended in 167mM
sodium sulfate, buffered to pH 7.2 with 20mM sodium phosphate salts, upon addition
of DMSO solutions of 1 (2 mol%), 4 (2 mol%) and both 1 (2 mol%) and 4 (2 mol%).
The sum of 1 (2 mol%) and 4 (2 mol%) is shown for comparison. A sodium
bicarbonate pulse was added at t = 60 s such that the external bicarbonate
concentration was 40mM. Each point represents an average of three trials.
1
4
S.J. Moore, M.G. Fisher, M. Yano, C.C. Tong and P.A. Gale, Dalton Trans. 2011, 40, 12017-12020.

34. Conclusions
“From anion receptors to
transporters”
P.A. Gale, Acc. Chem. Res.
2011, 44, 216-226.
meso-Octamethylcalixpyrrole functions as a cesium chloride co-transporter.
If the affinity of the calixpyrrole is enhanced by introducing a strap then the macrocycle
becomes capable of chloride ‘uniport.’
Anion transport processes by different carriers can be coupled to enhance salt co-
transport.
“Small molecule lipid bilayer anion
transporters for biological
applications” N. Busschaert and
P.A. Gale,
Angew. Chem. Int. Ed., 2013, 52,
1374-1382.
“Anion transporters and biological
systems”
P.A. Gale, R. Pérez-Tomás and
R. Quesada, Acc. Chem. Res.
2013, 46, 2801-2813.

35. Steve Moore
Matt Fisher
Masafumi Yano
Christine C. Tong
Cally E. Haynes
Nola Adeosun
Robert Quesada
Dustin E. Gross
Elizabeth Karnas
E. Sedenburg
Jonathan L. Sessler
Franz P. Schmidtchen
Acknowledgements

36. CM1005
Supramolecular
Chemistry in Water
The Erskine Programme
University of Canterbury