Nanomolar CFTR Inhibition by Pore-Occluding Divalent Polyethylene Glycol-Malonic Acid Hydrazides

N. D. Sonawane, Dan Zhao, Olga Zegarra-Moran, Luis J V Galietta, A. S. Verkman

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel have potential application as antisecretory therapy in cholera. We synthesized mono- and divalent CFTR inhibitors consisting of a malonic acid hydrazide (MalH) coupled via a disulfonic stilbene linker to polyethylene glycols (PEGs; 0.2-100 kDa). IC50 values for CFTR inhibition were 10-15 μM for the monovalent MalH-PEGs, but substantially lower for divalent MalH-PEG-MalH compounds, decreasing from 1.5 to 0.3 μM with increasing PEG size and showing positive cooperativity. Whole-cell patch-clamp showed voltage-dependent CFTR block with inward rectification. Outside-out patch-clamp showed shortened single-channel openings, indicating CFTR pore block from the extracellular side. Luminally added MalH-PEG-MalH blocked by >90% cholera toxin-induced fluid secretion in mouse intestinal loops (IC50 ∼10 pmol/loop), and greatly reduced mortality in a suckling mouse cholera model. These conjugates may provide safe, inexpensive antisecretory therapy.

Original languageEnglish
Pages (from-to)718-728
Number of pages11
JournalChemistry and Biology
Volume15
Issue number7
DOIs
Publication statusPublished - Jul 21 2008

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Polyethylene glycols
Cholera
Clamping devices
Inhibitory Concentration 50
Fluids and Secretions
Stilbenes
Chloride Channels
Cholera Toxin
malonic acid
Fluids
Mortality
Electric potential
Therapeutics

Keywords

  • CELLBIO
  • CHEMBIO
  • SIGNALING

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Nanomolar CFTR Inhibition by Pore-Occluding Divalent Polyethylene Glycol-Malonic Acid Hydrazides. / Sonawane, N. D.; Zhao, Dan; Zegarra-Moran, Olga; Galietta, Luis J V; Verkman, A. S.

In: Chemistry and Biology, Vol. 15, No. 7, 21.07.2008, p. 718-728.

Research output: Contribution to journalArticle

Sonawane, N. D. ; Zhao, Dan ; Zegarra-Moran, Olga ; Galietta, Luis J V ; Verkman, A. S. / Nanomolar CFTR Inhibition by Pore-Occluding Divalent Polyethylene Glycol-Malonic Acid Hydrazides. In: Chemistry and Biology. 2008 ; Vol. 15, No. 7. pp. 718-728.
@article{e4123862f2a74a46a1c125707cbf8bdd,
title = "Nanomolar CFTR Inhibition by Pore-Occluding Divalent Polyethylene Glycol-Malonic Acid Hydrazides",
abstract = "Inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel have potential application as antisecretory therapy in cholera. We synthesized mono- and divalent CFTR inhibitors consisting of a malonic acid hydrazide (MalH) coupled via a disulfonic stilbene linker to polyethylene glycols (PEGs; 0.2-100 kDa). IC50 values for CFTR inhibition were 10-15 μM for the monovalent MalH-PEGs, but substantially lower for divalent MalH-PEG-MalH compounds, decreasing from 1.5 to 0.3 μM with increasing PEG size and showing positive cooperativity. Whole-cell patch-clamp showed voltage-dependent CFTR block with inward rectification. Outside-out patch-clamp showed shortened single-channel openings, indicating CFTR pore block from the extracellular side. Luminally added MalH-PEG-MalH blocked by >90{\%} cholera toxin-induced fluid secretion in mouse intestinal loops (IC50 ∼10 pmol/loop), and greatly reduced mortality in a suckling mouse cholera model. These conjugates may provide safe, inexpensive antisecretory therapy.",
keywords = "CELLBIO, CHEMBIO, SIGNALING",
author = "Sonawane, {N. D.} and Dan Zhao and Olga Zegarra-Moran and Galietta, {Luis J V} and Verkman, {A. S.}",
year = "2008",
month = "7",
day = "21",
doi = "10.1016/j.chembiol.2008.05.015",
language = "English",
volume = "15",
pages = "718--728",
journal = "Chemistry and Biology",
issn = "1074-5521",
publisher = "Cell Press",
number = "7",

}

TY - JOUR

T1 - Nanomolar CFTR Inhibition by Pore-Occluding Divalent Polyethylene Glycol-Malonic Acid Hydrazides

AU - Sonawane, N. D.

AU - Zhao, Dan

AU - Zegarra-Moran, Olga

AU - Galietta, Luis J V

AU - Verkman, A. S.

PY - 2008/7/21

Y1 - 2008/7/21

N2 - Inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel have potential application as antisecretory therapy in cholera. We synthesized mono- and divalent CFTR inhibitors consisting of a malonic acid hydrazide (MalH) coupled via a disulfonic stilbene linker to polyethylene glycols (PEGs; 0.2-100 kDa). IC50 values for CFTR inhibition were 10-15 μM for the monovalent MalH-PEGs, but substantially lower for divalent MalH-PEG-MalH compounds, decreasing from 1.5 to 0.3 μM with increasing PEG size and showing positive cooperativity. Whole-cell patch-clamp showed voltage-dependent CFTR block with inward rectification. Outside-out patch-clamp showed shortened single-channel openings, indicating CFTR pore block from the extracellular side. Luminally added MalH-PEG-MalH blocked by >90% cholera toxin-induced fluid secretion in mouse intestinal loops (IC50 ∼10 pmol/loop), and greatly reduced mortality in a suckling mouse cholera model. These conjugates may provide safe, inexpensive antisecretory therapy.

AB - Inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel have potential application as antisecretory therapy in cholera. We synthesized mono- and divalent CFTR inhibitors consisting of a malonic acid hydrazide (MalH) coupled via a disulfonic stilbene linker to polyethylene glycols (PEGs; 0.2-100 kDa). IC50 values for CFTR inhibition were 10-15 μM for the monovalent MalH-PEGs, but substantially lower for divalent MalH-PEG-MalH compounds, decreasing from 1.5 to 0.3 μM with increasing PEG size and showing positive cooperativity. Whole-cell patch-clamp showed voltage-dependent CFTR block with inward rectification. Outside-out patch-clamp showed shortened single-channel openings, indicating CFTR pore block from the extracellular side. Luminally added MalH-PEG-MalH blocked by >90% cholera toxin-induced fluid secretion in mouse intestinal loops (IC50 ∼10 pmol/loop), and greatly reduced mortality in a suckling mouse cholera model. These conjugates may provide safe, inexpensive antisecretory therapy.

KW - CELLBIO

KW - CHEMBIO

KW - SIGNALING

UR - http://www.scopus.com/inward/record.url?scp=47049088617&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=47049088617&partnerID=8YFLogxK

U2 - 10.1016/j.chembiol.2008.05.015

DO - 10.1016/j.chembiol.2008.05.015

M3 - Article

VL - 15

SP - 718

EP - 728

JO - Chemistry and Biology

JF - Chemistry and Biology

SN - 1074-5521

IS - 7

ER -