594
CFTR pharmacology and its role in intestinal fluid secretion
Jay R Thiagarajah and AS Verkman
The cystic fibrosis transmembrane conductance regulator (CFTR)
is a cAMP-activated Cl
channel expressed in epithelial cells in
the airways, pancreas, intestine and other fluid-transporting
tissues. Cystic fibrosis is caused by mutations in the CFTR,
resulting in impaired Cl
transport and plasma membrane
targeting. CFTR is expressed in the lumenal membrane of
enterocytes, where it functions as the principal pathway for
secretion of Cl
and fluid in enterotoxin-induced secretory
diarrheas such as cholera. Small-molecule CFTR inhibitors reduce
enterotoxin-induced intestinal fluid secretion in animal models.
CFTR inhibition might also reduce intestinal fluid losses in cholera
and possibly in other infectious and non-infectious diarrheas.
Addresses
Departments of Medicine and Physiology, Cardiovascular Research
Institute, University of California, San Francisco, CA 94143-0521, USA
Correspondence: Alan S Verkman; e-mail: verkman@itsa.ucsf.edu;
http://www.ucsf.edu/verklab
Current Opinion in Pharmacology 2003, 3:594–599
This review comes from a themed issue on
Gastrointestinal pharmacology
Edited by David Grundy and Wendy Winchester
1471-4892/$ – see front matter
ß 2003 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.coph.2003.06.012
Abbreviations
ABC ATP-binding cassette
CaCC Ca2þ-activated Cl
channel
CFTR cystic fibrosis transmembrane conductance regulator
NBD nucleotide-binding domain
NPPB 5-nitro-2-(3-phenylpropylamino)-benzoic acid
SUR sulfonylurea receptor
Introduction
The gene encoding the cystic fibrosis transmembrane
conductance regulator (CFTR) was identified in 1989 as
the genetic basis of the hereditary lethal disease cystic
fibrosis. CFTR is a cAMP-activated Cl
channel expressed
in epithelial cells in the airways, sweat duct, testis, pan-
creas, intestine and other fluid-transporting tissues. In
cystic fibrosis, defective epithelial cell fluid transport
produces chronic lung infection and a slow deteriora-
tion in lung function, as well as pancreatic insufficiency,
meconium ileus and male infertility [1]. CFTR is a large
transmembrane glycoprotein containing two six-helix
membrane-spanning domains, each followed by a nucleo-
tide-binding domain (NBD), with a regulatory domain
linking the first NBD and the second membrane-spanning
domain (Figure 1a). CFTR activation involves ATP bind-
Current Opinion in Pharmacology 2003, 3:594–599
ing and hydrolysis at NBDs, and phosphorylation of multi-
ple regulatory domain sites (reviewed in [2]), although
many details of CFTR regulation and gating remain
unknown. CFTR has homology with members of the
ATP-binding cassette (ABC) family of membrane proteins
(including the multi-drug resistance protein-1 and sulfo-
nylurea receptor [SUR]) in their membrane-spanning
domains and NBDs; however, unlike other ABC proteins,
CFTR contains a regulatory domain.
CFTR is expressed at the apical membrane of entero-
cytes in intestine, and is thought to be the primary
pathway for Cl
and hence fluid secretion into the intest-
inal lumen in cAMP-mediated diarrheas such as cholera
[3]. Secretory diarrheas caused by Vibrio cholera and
enterotoxogenic Escherichia coli, in which intestinal fluid
hypersecretion leads to a massive loss of fluid and elec-
trolytes, are a major cause of morbidity and mortality
worldwide [4]. An antisecretory drug that acts directly on
Cl
secretion might complement oral fluid replacement,
which has been the mainstay of diarrheal therapy [5]. This
review is focused on small-molecule activators and inhi-
bitors of CFTR function, and on the role that CFTR plays
in intestinal fluid secretion.
CFTR activators
CFTR activation can be accomplished in many ways, the
most important being elevation of cytosolic cAMP (pro-
moting CFTR phosphorylation), inhibition of phospha-
tase activity (blocking CFTR dephosphorylation) and
direct interaction [6]. Small molecules that activate
CFTR by direct interaction include flavones/isoflavones
(e.g. genistein [7]), benzo[c]quinoliziniums [8], xanthines
[9] and benzimidazolones [10]. Figure 1b shows the
structures of IBMX, a xanthine that activates CFTR both
by direct and indirect (phosphodiesterase inhibition)
mechanisms; NS004, a benzimidazolone that was origin-
ally characterized as a Kþ channel opener; and UCCF-
029, a benzoflavone that was discovered from the screen-
ing of a flavone library [11,12]. In a recent systematic
search for small-molecule CFTR activators, 60 000 com-
pounds were screened using a cell-based fluorescence
assay [13]. More than a dozen compounds with novel
structures were identified with submicromolar potency
for activation of human wild-type CFTR. The compounds
activated CFTR without elevating cAMP or inhibiting
phosphatase activity, suggesting direct CFTR binding.
The structures of two such compounds (CFTRact-09
and -12), which also correct the DF508–CFTR gating
defect, are shown in Figure 1b. Activation of wild-type
CFTR by drug-like small molecules is relatively easy
compared with activation of cystic fibrosis-causing CFTR
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 CFTR pharmacology and its role in intestinal fluid secretion Thiagarajah and Verkman 595

Figure 1

(a) (b) CFTR activators

Cell surface

IBMX NS004 UCCF-029 CFTRact-09 CFTRact-12

N R CFTR inhibitors
NBD1 NBD2

Cytoplasm C
NPPB glibenclamide CFTRinh-172

Current Opinion in Pharmacology

CFTR activators and inhibitors. (a) CFTR structure showing membrane-spanning domains (shaded), NBD1 and NBD2, and the regulatory domain (R).
(b) Chemical structures of selected CFTR activators and inhibitors.

mutants; however, the latter subject is beyond the scope of (NPPB) (Figure 1b) [15]. These compounds are non-
this review. specific Cl
channel blockers; patch-clamp and mutagen-
esis studies suggest voltage-dependent block of CFTR,
CFTR inhibitors resulting in CFTR pore occlusion. The sulfonylureas,
Two major classes of CFTR inhibitors are the arylami- originally used as bacteriostatic agents, were optimized
nobenzoates and sulphonylureas. The arylaminobenzo- for modulation of insulin release as oral hypoglycemic
ates, developed originally as blockers of kidney tubule agents in diabetes. Glibenclamide (Figure 1b) binds to
Cl
conductance [14], include diphenylamine-2-carbox- the ABC protein SUR, where it modulates Kir6.2 Kþ
ylate and 5-nitro-2-(3-phenylpropylamino)-benzoic acid channel activity and insulin release from pancreatic islet

Figure 2

(a) (b) (c)

Iodide
Cl– CFTR
Control CPT- 0.2 µM CFTR -172
inh
(no activators) cAMP 1
YFP Cl–
Activation by forskolin + IBMX + apigenin

forskolin Isc 2
5 min IBMX 5
YFP fluorescence

200
apigenin
Cl– µA

YFP Cl– Control 10 min

10 min 10 µM test
compound
100 glibenclamide
Inactive
YFP Cl– compounds 80

I– Isc% 60
I– 40
20 CFTRinh-172
YFP Cl– Cl–
Active 0
compounds 0.01 0.1 1 10 100 1000
2s
Fluorescence [CFTRinh-172] (µM)
Time
Current Opinion in Pharmacology

Identification of CFTR inhibitors by high-throughput screening. (a) Screening approach: CFTR is stimulated by multiple agonists in stably transfected
epithelial cells co-expressing human CFTR and a yellow fluorescent protein (YFP) having Cl
/I
-sensitive fluorescence. After addition of test
compound, I
influx is induced by adding an I
-containing solution. (b) Representative original fluorescence data from individual wells of a 96-well
plate showing controls (no activators, no test compound) and test wells. (c) Top panel: CFTRinh-172 inhibition of short-circuit current in
permeabilized Fisher rat thyroid cells expressing human CFTR after stimulation by 100 mM CPT-cAMP. Bottom panel: dose-inhibition data for
CFTRinh-172 and glibenclamide.

www.current-opinion.com Current Opinion in Pharmacology 2003, 3:594–599

596 Gastrointestinal

cells. At much higher concentrations, glibenclamide was
found to inhibit CFTR [16]. Glibenclamide and the
related sulfonylurea tolbutamide are likely to block
CFTR function by binding to the CFTR pore in its open
state [17]. Although arylaminobenzoates and sulphony-
lureas inhibit CFTR Cl
channel function, high concen-
trations are generally required (> 0.1 mM) where multiple
ion channels and transporters are affected. Disulfonic
stilbene inhibitors of anion transport also inhibit CFTR
but at very high concentrations (generally > 1 mM). A
component of boiled rice also appears to inhibit CFTR
Cl
secretion in intestinal cells [18], although its identity
remains unknown.
Recently, a collection of 50 000 drug-like molecules was
screened for CFTR inhibition. Fisher rat thyroid cells co-
expressing human CFTR and a green fluorescent protein-
based halide sensor [19] were stimulated by a CFTR-
activating cocktail and then subjected to an iodide gra-
dient (Figure 2a). Iodide addition produced a prompt
decrease in cell fluorescence after CFTR activation
(Figure 2b). Inhibitors (‘active compounds’) were identi-
fied from a reduction in the fluorescence slope. The best
inhibitor identified by screening and subsequent optim-
ization was the 2-thioxo-4-thiazolidinone compound
CFTRinh-172 (Figure 1b) [20]. CFTRinh-172 inhibited
CFTR function at submicromolar concentrations —
approximately 500 times better than glibenclamide when
studied under similar conditions (Figure 2c). Further anal-
ysis showed voltage-independent inhibition of CFTR
Cl
conductance with prolonged mean channel-closed time
but without change in unitary conductance. CFTRinh-172
did not affect other Cl
channels (calcium- and volume-
activated) or ABC transporters (e.g. multi-drug resistance

Figure 3

Intestinal
lumen Cholera STa Na+
toxin toxin CFTR inhibitor Rotavirus
Cl– Cl– H2O
? ClC Cl–
GM1 Guanylin – other Cl–
receptor receptor channels
CFTR CaCC

cGMP Ca2+
cAMP

Enterocyte
Entero-
2Cl–
chromaffin
2K+ Na+ K+ K+ Mast cells
cells
– Neutrophils

5-HT

3Na+ K+ Inflammatory
NKCC
5-HT VIP Na+K+ channels mediators
Enkephalin ATPase ACh
SP

Mu receptor Enteric neurons

Blood Entero-invasive bacteria
– Inflammatory bowel disease
Loperamide
Diphenoxylate
Current Opinion in Pharmacology

Intestinal secretory pathways. Cl
secretion involves Cl
uptake at the basolateral membrane of enterocytes by the NaþKþ2Cl
cotransporter (NKCC)
and its exit primarily via apical membrane CFTR. Other Cl
channels (such as CaCCs and ClCs) might also mediate Cl
secretion. Naþ and
water follow by a paracellular route. Secretion initiated by pathogens (cholera toxin, STa toxin and rotavirus) involves multiple pathways involving the
release of 5-hydroxytryptamine (5-HT) from enterochromaffin cells, neuronal signaling, vasoactive intestinal peptide (VIP), acetylcholine (ACh),
substance P (SP) and the release of inflammatory mediators from mast cells and neutrophils (e.g. prostaglandins and interleukins). Signals are
transduced by second messengers (cAMP, cGMP, Ca2þ) to activate membrane ion channels. Secretion can be blocked by CFTR inhibitors, inhibition
of neuronal signaling, 5-hydroxytryptamine receptor antagonists and anti-enkephalinases.

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 CFTR pharmacology and its role in intestinal fluid secretion Thiagarajah and Verkman 597

protein-1, SUR). Rodent pharmacology studies indicated
that CFTRinh-172 has low toxicity, a large volume of
distribution with slow elimination by renal glomerular
filtration, little metabolism, and enterohepatic circulation
with accumulation in bile and intestine [21].
Role of CFTR in intestinal secretion
Fluid secretion plays a key role in intestinal physiology.
Under normal conditions, the intestine carries out the
absorption of luminal fluid, electrolytes and nutrients. A
small amount of basal secretion facilitates hydration of the
intestinal mucosa and mixing of intestinal contents. Fluid
secretion is driven by active Cl
transport from the
basolateral to the apical side of enterocytes (Figure 3).
Cl
is taken up at the basolateral membrane via NaK2Cl
cotransporter, which is driven by Naþ and Cl
concentra-
tion gradients produced by the NaþKþ-ATPase and baso-
lateral Kþ channels. Cl
is electrochemically driven across
the cell apical membrane primarily through the CFTR, as
well as through Ca2þ-activated Cl
channels (CaCCs) and
other Cl
channels. Both Naþ and fluid follow Cl
para-
cellularly. As shown schematically in Figure 3, Cl
and
hence fluid secretion can be activated by different
mechanisms. For example, secretory neuronal pathways

Figure 4

(a) (b)
0.14 0.25
Saline
Cholera toxin-
CFTRinh–172 injected loops
0.12
0.20
Loop weight (g/cm)

Loop weight (g/cm)

50 60 70 80
0.10 % absorption at 30 min
0.15
0.08

0.10
0.06
Saline-injected loops
0.04 0.05

0 10 20 30 40 50 60 1 2 3 4 5 6
Time (min) Time (h)

(c) (d)
100
0.25

Cholera 80
0.20 toxin
Loop weight (g/cm)

Percent inhibition

CFTRinh–172 60
0.15
Saline 40

0.10 Cholera
20
toxin Measure

0.05 0

0 10 20 200 –12 –10 –8 –6 –4 –2 0 +2 +4 +6

CFTRinh–172 dose (µg) Time CFTRinh–172 given (h)

Current Opinion in Pharmacology

Antidiarrheal properties of a CFTR inhibitor. (a) Intestinal fluid absorption. Closed mouse ileal loops were injected with 200 ml saline and loop weight
measured at indicated times. Inset: shows % absorption at 30 minutes with and without CFTRinh-172. (b) Intestinal fluid secretion. Time-course
of cholera toxin-induced fluid secretion. Loops were injected with 1 mg cholera toxin. Dashed line shows control (saline-injected) loops.
(c) Dose-response for inhibition of fluid accumulation. Mice were given single doses of CFTRinh-172 by intraperitoneal injection and loop weight
was measured at six hours. Inset: intestinal loops six hours after cholera toxin or saline injection. Where indicated, the mouse was treated with
CFTRinh-172. (d) Persistence of CFTRinh-172 inhibition. Mice were injected with 20 mg CFTRinh-172 at indicated times before or after cholera
toxin administration.

www.current-opinion.com Current Opinion in Pharmacology 2003, 3:594–599

598 Gastrointestinal
involve the release of 5-hydroxytryptamine from enter-
ochromaffin cells, resulting in activation of cholinergic and
vasoactive intestinal peptide neurons and increased cAMP
and Ca2þ levels, causing subsequent Cl
channel activa-
tion. Inflammatory mediators, such as prostaglandins and
interleukins, also play an important role in initiating
Cl
secretion, and recent studies have begun to elucidate
their signaling pathways [22,23,24,25]; nucleotides
and purinergic signaling also stimulate Cl
secretion
through Ca2þ and cAMP signaling pathways [26,27].
Evidence suggests a pivotal role for CFTR in intestinal
Cl
and fluid secretion. Numerous in vitro studies have
shown that agonist-induced Cl
secretion in intestinal
sheets and cell-lines is blocked by glibenclamide and
NPPB [28,29]. CFTR knockout mice develop intestinal
obstruction because of defective intestinal Cl
and fluid
secretion and increased fluid absorption [30]. Increased
expression of alternative Cl
channels is found in CFTR
null mice that develop mild intestinal symptoms [31].
Cholera toxin, which causes massive fluid secretion in the
small intestine in normal mice, does not induce fluid
secretion in CFTR knockout mice [32].
CFTR inhibitors as anti-diarrheals
Secretory diarrhea is caused by intestinal infection by
various bacterial and viral pathogens. Bacterial enterotox-
ins, such as cholera toxin from V. cholerae and STa toxin
from E. coli, induce increases in the second messengers
cAMP and cGMP, respectively, resulting in activation of
CFTR Cl
conductance (Figure 3). Other pathogens,
such as entero-invasive bacteria, also stimulate Cl
secre-
tion, and recent studies on Salmonella and enteropatho-
genic E. coli provide evidence for the involvement of
inflammatory mediators and nucleotides in secretion
[25,27]. An important cause of secretory diarrhea is
the rotavirus viral pathogen, which is responsible for
20% of all diarrhea-related deaths in children under
the age of five [33]. Secretion in rotaviral diarrhea was
thought to involve inhibition of solute and fluid absorp-
tion. However, the extensive fluid loss during sympto-
matic infection suggests the involvement of active
secretory mechanisms. CaCCs, which provide an alter-
native apical exit pathway for Cl
(Figure 3), have been
postulated to play a role in some viral diarrheas [34].
Another diarrheal disorder, AIDS-related diarrhea, is
thought to have a multi-factorial etiology, including
opportunistic infection by pathogens such as enterotoxi-
genic bacteria (e.g. E. coli), parasites (e.g. cryptospori-
dium) and viruses (e.g. cytomegalovirus).
CFTR inhibition has been proposed as a therapy for some
forms of secretory diarrhea [35]. Indeed, previous in vitro
studies showed that a diarylsulfonylurea CFTR inhibitor
prevented Cl
secretion in porcine mucosa [36]. The
antidiarrheal efficacy of the thiazolidinone CFTRinh-
172 has recently been tested [20]. In a mouse closed-
Current Opinion in Pharmacology 2003, 3:594–599
ileal loop model, injected fluid was rapidly cleared and
remained unaffected by CFTRinh-172 administration
(Figure 4a), an important prerequisite for antidiarrheal
application of a CFTR inhibitor. Injection of cholera toxin
into loops produced fluid secretion over six hours after
a slow onset (Figure 4b). In a dose-response study, a
single intraperitoneal injection of CFTRinh-172 (just after
cholera toxin infusion) reduced fluid accumulation by
90%, with an IC50 of 5 mg CFTRinh-172 (Figure 4c).
Inhibition of fluid accumulation was seen when CFTRinh-
172 was administered three hours before or after cholera
toxin (Figure 4d), which is probably a consequence of
reduced fluid secretion with continued absorption. Orally
administered CFTRinh-172 was also effective in blocking
intestinal fluid secretion following oral cholera toxin in an
open-loop model. Finally, CFTRinh-172 inhibited fluid
secretion after E. coli STa toxin exposure, as well as cAMP-
and cGMP-stimulated Cl
currents in human intestine.
Conclusions
There is now good evidence for a central role of CFTR-
mediated Cl
secretion in enterotoxin-mediated intest-
inal fluid secretion in infectious diarrheas, including
cholera and traveler’s diarrhea. However, the relative
importance of CFTR, CaCCs and other Cl
channels
in different forms of diarrhea, such as viral diarrhea and
inflammatory bowel disease, remains unknown. CFTR is
thus an attractive target for development of inhibitors
with antidiarrheal efficacy in cholera and other disorders
of intestinal fluid secretion. CFTR inhibitors might also
be useful in creating cystic fibrosis animal models, and in
pharmacologically creating the cystic fibrosis phenotype
in excised human tissues.
Acknowledgements
This work is supported by the Cystic Fibrosis Foundation, and
NIH grants HL73856, HL59198, EB00415, EY13574 and DK35124.
Dr Thiagarajah was supported by a Cystic Fibrosis Foundation fellowship.
References and recommended reading
Papers of particular interest, published within the annual period of
review, have been highlighted as:
 of special interest
 of outstanding interest
1. Welsh MJ, Tsui LC, Boat TF, Beudet AL: Cystic Fibrosis.
In The Metabolic and Molecular Basis of Inherited Disease.
New York: McGraw-Hill Inc; 1995:3799-3876.
2. Sheppard DN, Welsh MJ: Structure and function of the CFTR
chloride channel. Physiol Rev 1999, 79:23-45.
3. Barrett KE, Keely SJ: Chloride secretion by the intestinal
epithelium: molecular basis and regulatory aspects. Annu Rev
Physiol 2000, 62:535-572.
4. Farthing MJ: Diarrhoea: a significant worldwide problem.
Int J Antimicrob Agents 2000, 14:65-69.
5. Farthing MJ: History and rationale of oral rehydration and recent
developments in formulating an optimal solution. Drugs 1988,
4:80-90.
6. Schultz BD, Singh AK, Devor DC, Bridges RJ: Pharmacology
of CFTR chloride channel activity. Physiol Rev 1999,
79:S109-S144.
www.current-opinion.com
 CFTR pharmacology and its role in intestinal fluid secretion Thiagarajah and Verkman 599
7. Illek B, Lizarzaburu ME, Lee V, Nantz MH, Kurth MJ, Fischer H:
Structural determinants for activation and block of CFTR-
mediated chloride currents by apigenin. Am J Physiol 2000,
279:C1838-C1846.
8. Dormer RL, Derand R, McNeilly CM, Mettey Y, Bulteau-Pignoux L,
Metaye T, Vierfond JM, Gray MA, Galietta LJ, Morris MR et al.:
Correction of delF508-CFTR activity with
benzo(c)quinolizinium compounds through facilitation of its
processing in cystic fibrosis airway cells. J Cell Sci 2001,
114:4073-4081.
9. Bulteau L, Derand R, Mettey Y, Metaye T, Morris MR, McNeilly CM,
Folli C, Galietta LJ, Zegarra-Moran O, Pereira MM et al.: Properties
of CFTR activated by the xanthine derivative X-33 in human
airway Calu-3 cells. Am J Physiol 2000, 279:C1925-C1937.
10. Gribkoff VK, Champigny G, Barbry P, Dworetzky SI, Meanwell NA,
Lazdunski M: The substituted benzimidazolone NS004 is an
opener of the cystic fibrosis chloride channel. J Biol Chem 1994,
269:10983-10986.
11. Galietta LJ, Springsteel MR, Eda M, Niedzinski EJ, By K,
Haddadin MJ, Kurth MJ, Nantz MH, Verkman AS: Novel CFTR
chloride channel activators identified by screening of
combinatorial libraries based on flavone and
benzoquinolizinium lead compounds. J Biol Chem 2001,
276:19723-19728.
12. Springsteel M, Galietta LJ, Ma T, By K, Berger TO, Yang H,
Dicus CW, Choung W, Quan C, Shelat AA et al.: Benzoflavone
activators of the cystic fibrosis transmembrane conductance
regulator: a pharmacophore model of flavone-CFTR binding.
Bioorgan Med Chem 2003, 11:4113-4120.
13. Ma T, Vetrivel L, Yang Y, Pedemonte N, Zegarra-Moran O,
 Galietta LJ, Verkman AS: High-affinity activators of CFTR
chloride conductance identified by high-throughput screening.
J Biol Chem 2002, 277:37235-37241.
Describes CFTR activators with submicromolar potency and novel struc-
tures discovered by screening 60 000 random compounds.
14. Di Stefano A, Wittner M, Schlatter E, Lang HJ, Englert H, Greger R:
Diphenylamine-2-carboxylate, a blocker of the ClS-conductive
pathway in ClS transporting epithelia. Pflugers Arch 1985,
405:95-100.
15. Wangemann P, Wittner M, Di Stefano A, Englert HC, Lang HJ,
Schlatter E, Greger R: ClS-channel blockers in the thick
ascending limb of the loop of Henle. Structure activity
relationship. Pflugers Arch 1986, 407:128-141.
16. Sheppard DN, Welsh MJ: Effect of ATP-sensitive KR channel
regulators on cystic fibrosis transmembrane conductance
regulator chloride currents. J Gen Physiol 1992, 100:573-591.
17. Zhou Z, Hu S, Hwang TC: Probing an open CFTR pore with
 organic anion blockers. J Gen Physiol 2002, 120:647-662.
Single-channel electrophysiological analysis of CFTR inhibition mech-
anisms.
18. Mathews CJ, MacLeod RJ, Zheng SX, Hanrahan JW, Bennett HP,
Hamilton JR: Characterization of the inhibitory effect of boiled
rice on intestinal chloride secretion in guinea-pig crypt cells.
Gastroenterology 1999, 116:1342-1347.
19. Jayaraman S, Haggie P, Wachter R, Remington SJ, Verkman AS:
Mechanism and cellular applications of a green
fluorescent protein-based halide sensor. J Biol Chem 2000,
275:6047-6050.
20. Ma T, Thiagarajah JR, Yang H, Sonawane ND, Folli C, Galietta LJ,
 Verkman AS: Thiazolidinone CFTR inhibitor identified by high-
throughput screening blocks cholera-toxin induced intestinal
fluid secretion. J Clin Invest 2002, 110:1651-1658.
Identification and characterization of a CFTR inhibitor with submicromolar
potency identified by random compound screening, and demonstration
of its antidiarrheal efficacy.
21. Nagatani R, Sonawane N, Song Y, Verkman AS:
Pharmacokinetics, tissue distribution and elimination
www.current-opinion.com
mechanism of a thiazolidinone CFTR inhibitor in rodents.
Pharm Res 2003, in press.
22. Buresi MC, Buret AG, Hollenberg MD, MacNaughton WK:
 Activation of proteinase-activated receptor 1 stimulates
epithelial chloride secretion through a unique MAP kinase- and
cyclo-oxygenase-dependent pathway. FASEB J 2002,
16:1515-1525.
Elucidates the mechanism by which thrombin, which is elevated in
inflammatory bowel diseases, causes intestinal Cl
secretion.
23. Uribe JM, McCole DF, Barrett KE: Interferon-gamma activates
EGF receptor and increases TGF-alpha in T84 cells:
implications for chloride secretion. Am J Physiol 2002,
283:G923-G931.
24. Oprins JC, van der Burg C, Meijer HP, Munnik T, Groot JA: Tumour
 necrosis factor alpha potentiates ion secretion induced by
histamine in a human intestinal epithelial cell line and in mouse
colon: involvement of the phospholipase D pathway. Gut 2002,
50:314-321.
Shows that an important mediator of inflammatory bowel disease, tumor
necrosis factor-a, potentiates histamine-stimulated Cl
secretion.
25. Resta-Lenert S, Barrett KE: Enteroinvasive bacteria alter barrier
 and transport properties of human intestinal epithelium: role of
iNOS and COX-2. Gastroenterology 2002, 122:1070-1087.
Shows that invasive diarrheal strains of bacteria induce Cl
secretion
through inflammatory mediators.
26. Kottgen M, Loffler T, Jacobi C, Nitschke R, Pavenstadt H,
 Schreiber R, Frische S, Nielsen S, Leipziger J: P2Y6 receptor
mediates colonic NaCl secretion via differential activation of
cAMP-mediated transport. J Clin Invest 2003, 111:371-379.
Elucidates the mechanism of intestinal CFTR-mediated Cl
secretion by
extracellular nucleotides.
27. Crane JK, Olson RA, Jones HM, Duffey ME: Release of ATP during
 host cell killing by enteropathogenic E. coli and its role as a
secretory mediator. Am J Physiol 2002, 283:G74-G86.
Provides evidence for the involvement of nucleotide release in Cl

secretion induced by invasive bacteria.
28. Ko WH, Law VW, Yip WC, Yue GG, Lau CW, Chen ZY, Huang Y:
Stimulation of chloride secretion by baicalein in isolated rat
distal colon. Am J Physiol 2002, 282:G508-G518.
29. Duszyk M, MacVinish L, Cuthbert AW: Phenanthrolines — a new
class of CFTR chloride channel openers. Br J Pharmacol 2001,
134:853-864.
30. Grubb BR, Gabriel SE: Intestinal physiology and pathology in
gene-targeted mouse models of cystic fibrosis. Am J Physiol
1997, 273:G258-G266.
31. Gyomorey K, Rozmahel R, Bear CE: Amelioration of intestinal
disease severity in cystic fibrosis mice is associated with
improved chloride secretory capacity. Pediatr Res 2000,
48:731-734.
32. Gabriel SE, Brigman KN, Koller BH, Boucher RC, Stutts MJ:
Cystic fibrosis heterozygote resistance to cholera toxin in the
cystic fibrosis mouse model. Science 1994, 266:107-109.
33. de Zoysa I, Feachem R: Interventions for the control of
diarrhoeal diseases among young children: rotavirus and
cholera immunization. Bull World Health Organ 1985, 63:569-583.
34. Morris AP, Scott JK, Ball JM, Zeng CQ, O’Neal WK, Estes MK:
NSP4 elicits age-dependent diarrhea and Ca2R-mediated IS
influx into intestinal crypts of CF mice. Am J Physiol 1999,
277:G431-G444.
35. Kunzelmann K, Mall M: Electrolyte transport in the mammalian
colon: mechanisms and implications for disease. Physiol Rev
2002, 82:245-248.
36. O’Donnell EK, Sedlacek RL, Singh AK, Schultz BD: Inhibition
of enterotoxin-induced porcine colonic secretion by
diarylsulfonylureas in vitro. Am J Physiol 2000,
279:G1104-G1112.
Current Opinion in Pharmacology 2003, 3:594–599