TY - JOUR
T1 - Phenylquinoxalinone CFTR activator as potential prosecretory therapy for constipation
AU - Cil, Onur
AU - Phuan, Puay Wah
AU - Son, Jung Ho
AU - Zhu, Jie S.
AU - Ku, Colton K.
AU - Tabib, Niloufar Akhavan
AU - Teuthorn, Andrew P.
AU - Ferrera, Loretta
AU - Zachos, Nicholas C.
AU - Lin, Ruxian
AU - Galietta, Luis J.V.
AU - Donowitz, Mark
AU - Kurth, Mark J.
AU - Verkman, Alan S.
N1 - Funding Information:
This work was supported by grants DK099803 , DK72517 , DK101373 , DK35124 , EB00415 , EY13574 , DK26523 , DK61765 , DK089502 , TR000552 , TR000504 , and TR000504 from the National Institutes of Health (NIH) , a Research Development Program grant from the Cystic Fibrosis Foundation , a Catalyst grant from funds awarded to the UCSF CTSI ( UL1 TR000004 ), a Gates Foundation Grand Challenges Grant , and grants from Ministero della Salute (Ricerca Corrente: Cinque per mille).
Publisher Copyright:
© 2016 Elsevier Inc.
PY - 2017/4/1
Y1 - 2017/4/1
N2 - Constipation is a common condition for which current treatments can have limited efficacy. By high-throughput screening, we recently identified a phenylquinoxalinone activator of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel that stimulated intestinal fluid secretion and normalized stool output in a mouse model of opioid-induced constipation. Here, we report phenylquinoxalinone structure-activity analysis, mechanism of action, animal efficacy data in acute and chronic models of constipation, and functional data in ex vivo primary cultured human enterocytes. Structure-activity analysis was done on 175 phenylquinoxalinone analogs, including 15 synthesized compounds. The most potent compound, CFTRact-J027, activated CFTR with EC50∼ 200 nM, with patch-clamp analysis showing a linear CFTR current-voltage relationship with direct CFTR activation. CFTRact-J027 corrected reduced stool output and hydration in a mouse model of acute constipation produced by scopolamine and in a chronically constipated mouse strain (C3H/HeJ). Direct comparison with the approved prosecretory drugs lubiprostone and linaclotide showed substantially greater intestinal fluid secretion with CFTRact-J027, as well as greater efficacy in a constipation model. As evidence to support efficacy in human constipation, CFTRact-J027 increased transepithelial fluid transport in enteroids generated from normal human small intestine. Also, CFTRact-J027 was rapidly metabolized in vitro in human hepatic microsomes, suggesting minimal systemic exposure upon oral administration. These data establish structure-activity and mechanistic data for phenylquinoxalinone CFTR activators, and support their potential efficacy in human constipation.
AB - Constipation is a common condition for which current treatments can have limited efficacy. By high-throughput screening, we recently identified a phenylquinoxalinone activator of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel that stimulated intestinal fluid secretion and normalized stool output in a mouse model of opioid-induced constipation. Here, we report phenylquinoxalinone structure-activity analysis, mechanism of action, animal efficacy data in acute and chronic models of constipation, and functional data in ex vivo primary cultured human enterocytes. Structure-activity analysis was done on 175 phenylquinoxalinone analogs, including 15 synthesized compounds. The most potent compound, CFTRact-J027, activated CFTR with EC50∼ 200 nM, with patch-clamp analysis showing a linear CFTR current-voltage relationship with direct CFTR activation. CFTRact-J027 corrected reduced stool output and hydration in a mouse model of acute constipation produced by scopolamine and in a chronically constipated mouse strain (C3H/HeJ). Direct comparison with the approved prosecretory drugs lubiprostone and linaclotide showed substantially greater intestinal fluid secretion with CFTRact-J027, as well as greater efficacy in a constipation model. As evidence to support efficacy in human constipation, CFTRact-J027 increased transepithelial fluid transport in enteroids generated from normal human small intestine. Also, CFTRact-J027 was rapidly metabolized in vitro in human hepatic microsomes, suggesting minimal systemic exposure upon oral administration. These data establish structure-activity and mechanistic data for phenylquinoxalinone CFTR activators, and support their potential efficacy in human constipation.
UR - http://www.scopus.com/inward/record.url?scp=85006981607&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85006981607&partnerID=8YFLogxK
U2 - 10.1016/j.trsl.2016.10.003
DO - 10.1016/j.trsl.2016.10.003
M3 - Article
C2 - 27815136
AN - SCOPUS:85006981607
VL - 182
SP - 14-26.e4
JO - Translational Research
JF - Translational Research
SN - 1931-5244
ER -