Limits...
Discovery and characterization of novel inhibitors of the sodium-coupled citrate transporter (NaCT or SLC13A5).

Huard K, Brown J, Jones JC, Cabral S, Futatsugi K, Gorgoglione M, Lanba A, Vera NB, Zhu Y, Yan Q, Zhou Y, Vernochet C, Riccardi K, Wolford A, Pirman D, Niosi M, Aspnes G, Herr M, Genung NE, Magee TV, Uccello DP, Loria P, Di L, Gosset JR, Hepworth D, Rolph T, Pfefferkorn JA, Erion DM - Sci Rep (2015)

Bottom Line: NaCT transports citrate from the blood into the cell coupled to the transport of sodium ions.Binding and transport experiments indicate that 2 specifically binds NaCT in a competitive and stereosensitive manner, and is recognized as a substrate for transport by NaCT.The favorable pharmacokinetic properties of 2 permitted in vivo experiments to evaluate the effect of inhibiting hepatic citrate uptake on metabolic endpoints.

View Article: PubMed Central - PubMed

Affiliation: Worldwide Medicinal Chemistry, 610 Main street, Cambridge, MA 02139.

ABSTRACT
Citrate is a key regulatory metabolic intermediate as it facilitates the integration of the glycolysis and lipid synthesis pathways. Inhibition of hepatic extracellular citrate uptake, by blocking the sodium-coupled citrate transporter (NaCT or SLC13A5), has been suggested as a potential therapeutic approach to treat metabolic disorders. NaCT transports citrate from the blood into the cell coupled to the transport of sodium ions. The studies herein report the identification and characterization of a novel small dicarboxylate molecule (compound 2) capable of selectively and potently inhibiting citrate transport through NaCT, both in vitro and in vivo. Binding and transport experiments indicate that 2 specifically binds NaCT in a competitive and stereosensitive manner, and is recognized as a substrate for transport by NaCT. The favorable pharmacokinetic properties of 2 permitted in vivo experiments to evaluate the effect of inhibiting hepatic citrate uptake on metabolic endpoints.

No MeSH data available.


Related in: MedlinePlus

Structure of dicarboxylates 1–3 and in vitro profile of dicarboxylate 2 (PF-06649298).(A) Chemical structure of racemic dicarboxylate 1 and its enantiomers 2 and 3. (B) In vitro profile of dicarboxylate 2. Inhibition of citrate uptake was measured in mouse hepatocytes. IC50 is reported as a geometric mean of at least 3 replicates with pIC50 ± SD in parentheses. mwt = molecular weight. Papp = apparent passive permeability. Heps = hepatocytes. CLint = intrinsic clearance. fu = fraction unbound.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4664966&req=5

f1: Structure of dicarboxylates 1–3 and in vitro profile of dicarboxylate 2 (PF-06649298).(A) Chemical structure of racemic dicarboxylate 1 and its enantiomers 2 and 3. (B) In vitro profile of dicarboxylate 2. Inhibition of citrate uptake was measured in mouse hepatocytes. IC50 is reported as a geometric mean of at least 3 replicates with pIC50 ± SD in parentheses. mwt = molecular weight. Papp = apparent passive permeability. Heps = hepatocytes. CLint = intrinsic clearance. fu = fraction unbound.

Mentions: To identify NaCT inhibitors, a virtual search of Pfizer’s compound library was conducted based on structural similarities to the transporter’s preferred substrate citrate. 500 compounds were selected for testing in a HEK-293-derived stable cell line overexpressing SLC13A5 (HEKNaCT) to measure their effect on cellular citrate uptake. This effort led to the identification of racemic dicarboxylate 1 (Fig. 1A) which inhibited 50% of the citrate transport from the media into the cells at a concentration of 0.80 μM (Table 1). No cellular toxicity was observed under these conditions using a CellTiter-glo® assessment.


Discovery and characterization of novel inhibitors of the sodium-coupled citrate transporter (NaCT or SLC13A5).

Huard K, Brown J, Jones JC, Cabral S, Futatsugi K, Gorgoglione M, Lanba A, Vera NB, Zhu Y, Yan Q, Zhou Y, Vernochet C, Riccardi K, Wolford A, Pirman D, Niosi M, Aspnes G, Herr M, Genung NE, Magee TV, Uccello DP, Loria P, Di L, Gosset JR, Hepworth D, Rolph T, Pfefferkorn JA, Erion DM - Sci Rep (2015)

Structure of dicarboxylates 1–3 and in vitro profile of dicarboxylate 2 (PF-06649298).(A) Chemical structure of racemic dicarboxylate 1 and its enantiomers 2 and 3. (B) In vitro profile of dicarboxylate 2. Inhibition of citrate uptake was measured in mouse hepatocytes. IC50 is reported as a geometric mean of at least 3 replicates with pIC50 ± SD in parentheses. mwt = molecular weight. Papp = apparent passive permeability. Heps = hepatocytes. CLint = intrinsic clearance. fu = fraction unbound.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664966&req=5

f1: Structure of dicarboxylates 1–3 and in vitro profile of dicarboxylate 2 (PF-06649298).(A) Chemical structure of racemic dicarboxylate 1 and its enantiomers 2 and 3. (B) In vitro profile of dicarboxylate 2. Inhibition of citrate uptake was measured in mouse hepatocytes. IC50 is reported as a geometric mean of at least 3 replicates with pIC50 ± SD in parentheses. mwt = molecular weight. Papp = apparent passive permeability. Heps = hepatocytes. CLint = intrinsic clearance. fu = fraction unbound.
Mentions: To identify NaCT inhibitors, a virtual search of Pfizer’s compound library was conducted based on structural similarities to the transporter’s preferred substrate citrate. 500 compounds were selected for testing in a HEK-293-derived stable cell line overexpressing SLC13A5 (HEKNaCT) to measure their effect on cellular citrate uptake. This effort led to the identification of racemic dicarboxylate 1 (Fig. 1A) which inhibited 50% of the citrate transport from the media into the cells at a concentration of 0.80 μM (Table 1). No cellular toxicity was observed under these conditions using a CellTiter-glo® assessment.

Bottom Line: NaCT transports citrate from the blood into the cell coupled to the transport of sodium ions.Binding and transport experiments indicate that 2 specifically binds NaCT in a competitive and stereosensitive manner, and is recognized as a substrate for transport by NaCT.The favorable pharmacokinetic properties of 2 permitted in vivo experiments to evaluate the effect of inhibiting hepatic citrate uptake on metabolic endpoints.

View Article: PubMed Central - PubMed

Affiliation: Worldwide Medicinal Chemistry, 610 Main street, Cambridge, MA 02139.

ABSTRACT
Citrate is a key regulatory metabolic intermediate as it facilitates the integration of the glycolysis and lipid synthesis pathways. Inhibition of hepatic extracellular citrate uptake, by blocking the sodium-coupled citrate transporter (NaCT or SLC13A5), has been suggested as a potential therapeutic approach to treat metabolic disorders. NaCT transports citrate from the blood into the cell coupled to the transport of sodium ions. The studies herein report the identification and characterization of a novel small dicarboxylate molecule (compound 2) capable of selectively and potently inhibiting citrate transport through NaCT, both in vitro and in vivo. Binding and transport experiments indicate that 2 specifically binds NaCT in a competitive and stereosensitive manner, and is recognized as a substrate for transport by NaCT. The favorable pharmacokinetic properties of 2 permitted in vivo experiments to evaluate the effect of inhibiting hepatic citrate uptake on metabolic endpoints.

No MeSH data available.


Related in: MedlinePlus