Cariporide

Manufactured by Sanofi

Sourced in Germany

Cariporide is a lab equipment product. It is used for in vitro research purposes. The core function of Cariporide is to inhibit the sodium-hydrogen exchanger isoform 1 (NHE1).

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Lab products found in correlation

5 protocols using cariporide

Antibody Preparation and Experimental Reagents

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The following antibodies were purchased from Cell Signaling: Ezrin/Radixin/Moesin (#3142), PARP (#9542) and cleaved PARP [Asp214] (#5625). NHE1 (#sc-136239), MCT4 (#sc-50329) and ZO-1 (#sc-10804) were from Santa Cruz Biotechnology, and MCT1 (#AB3538P) from Millipore. E-cadherin (#610181) and p150 [Glued] (#610473) were from BD Transduction Laboratories and CAIX (M75, [28 (link)]) was from BioScience, Slovakia. The NBCn1 antibody was a kind gift from Jeppe Praetorius, Aarhus University, Denmark, and the polyclonal NHE1 antibody (Xb-17) was a kind gift from Mark Musch, University of Chicago, IL. Antibody against the S703-phosphorylated NHE1 was from DSTT, University of Dundee, Ireland. β-actin (#A5441), anti-Mouse (#A1293) and anti-Rabbit (#A3937) alkaline phosphatase-conjugated secondary antibodies were from Sigma-Aldrich. anti-Mouse (#P0447) and anti-Rabbit (#P0448) Horseradish Peroxidase (HRP)-conjugated secondary antibodies were from Dako. Cariporide and S0859 were kind gifts from Sanofi-Aventis. AR-C155858 was acquired from AdooQ BioScience, Irvine, CA, USA. The Hydroxyprobe-1 kit including Pimonidazole and anti-pimonidazole mouse monoclonal antibody was purchased from Hydroxyprobe, Inc., Burlington, MA, USA.

Andersen A.P., Flinck M., Oernbo E.K., Pedersen N.B., Viuff B.M, & Pedersen S.F. (2016). Roles of acid-extruding ion transporters in regulation of breast cancer cell growth in a 3-dimensional microenvironment. Molecular Cancer, 15, 45.

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Inhibition of Ion Transporters

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Cariporide (NHE inhibitor) and S0859 (NBC inhibitor) were kindly provided by Sanofi Aventis (Germany). Dimethylamiloride (DMA; NHE inhibitor) was purchased from Sigma (Poole, UK).

Villafuerte F.C., Swietach P., Youm J.B., Ford K., Cardenas R., Supuran C.T., Cobden P.M., Rohling M, & Vaughan-Jones R.D. (2014). Facilitation by intracellular carbonic anhydrase of Na+–HCO3− co-transport but not Na+/H+ exchange activity in the mammalian ventricular myocyte. The Journal of Physiology, 592(Pt 5), 991-1007.

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Western Blot and Immunofluorescence Antibodies

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A table containing all employed antibodies as well as their cellular target, supplier and catalogue number is provided as Supplementary Table S2.
Cariporide was a gift from Sanofi-Aventis and 5-(N-ethyl-N-isopropyl) amiloride (EIPA) (#E3111) was from Thermo-Fisher. β-actin antibody (#A5441), tamoxifen (#T5648), cisplatin (#P4394), 5-fluorouracil (5-FU) (#F6627), 5-(N,N-Dimethyl) amiloride (DMA) (#A4562), 5-(N,N-Hexamethylene) amiloride (HMA) (#A9561) and amiloride (#A7410) were from Sigma-Aldrich, and doxorubicin (#120629) and eniporide (#HY-106150B) from Abcam and MedChemExpress, respectively. Akt (Akti, #124018) and MEK (U0126, #662005) inhibitors were from Calbiochem. Horseradish Peroxidase (HRP)-conjugated anti-mouse- (#P0447) and anti-rabbit (#P0448) antibodies were from Dako, and Alexa Fluor anti-mouse (#A11019) and anti-rabbit (#A11070) fluorophore-conjugated secondary antibodies were from Invitrogen.

Rolver M.G., Elingaard-Larsen L.O., Andersen A.P., Counillon L, & Pedersen S.F. (2020). Pyrazine ring-based Na+/H+ exchanger (NHE) inhibitors potently inhibit cancer cell growth in 3D culture, independent of NHE1. Scientific Reports, 10, 5800.

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EMPA Concentration Optimization for In Vitro Studies

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EMPA was obtained independently from Boehringer Ingelheim² (link)^,²⁰ (link)^,²³ (link) and from MedChemExpress (MCE),⁹ (link)^,¹⁰ (link)^,²⁴ (link)^,²⁵ (link) and referred to herein as EMPA and mEMPA, respectively. Dapagliflozin (DAPA) and canagliflozin (CANA) were obtained from MCE. Cariporide, a selective NHE1 inhibitor, was obtained from Sanofi-Aventis. The structure and physical characteristics of EMPA were verified using high-pressure liquid chromatography-mass spectrometry (HPLC-MS), 1D/2D NMR and circular dichroism (CD) spectroscopy. Therapeutically relevant plasma concentrations of EMPA have been reported to be in the range ∼200–700 nmol/L with plasma protein binding of ∼90%.²⁶ (link) To ensure we adequately cover or exceeded both the concentrations used in previously studies in vitro and the therapeutically relevant concentrations, we have used concentrations ranging from 1 to 30 µM in DMSO (0.01% v/v, or 0.03% v/v in the case of 30 µM EMPA).

Chung Y.J., Park K.C., Tokar S., Eykyn T.R., Fuller W., Pavlovic D., Swietach P, & Shattock M.J. (2020). Off-target effects of sodium-glucose co-transporter 2 blockers: empagliflozin does not inhibit Na+/H+ exchanger-1 or lower [Na+]i in the heart. Cardiovascular Research, 117(14), 2794-2806.

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Measurement of NHE1 H+ Efflux Kinetics

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All chemicals were purchased from MilliporeSigma (UK) unless indicated otherwise. A superfusion system controlled the immediate environment of cells. Solutions were delivered at 37°C to a custom-made Perspex superfusion chamber with a coverslip glass bottom (for experiments on adult myocytes) or an Ibidi μ-slide (for experiments on cultured cells). Normal Tyrode contained (in mM) 135 NaCl, 4.5 KCl, 1 CaCl₂, 1 MgCl₂, 11 glucose, 20 HEPES at pH 7.4. In ammonium-containing solutions, NaCl was iso-osmotically replaced with NH₄Cl. Calibration of cytoplasmic dye fluorescence as a function of pH_i was obtained using the K⁺/H⁺ ionophore nigericin (22 (link)). H⁺-efflux on NHE1 was calculated from the time-course of pH_i recovery in Hepes-buffered superfusates at pH 7.4, following a displacement of pH_i to an acidic level attained by a 4–6 min prepulse with 15 or 20 mM ammonium chloride. In some experiments 30 μM cariporide (a selective NHE inhibitor, Sanofi Aventis) was included in superfusates.

Kandilci H.B., Richards M.A., Fournier M., Şimşek G., Chung Y.J., Lakhal-Littleton S, & Swietach P. (2021). Cardiomyocyte Na+/H+ Exchanger-1 Activity Is Reduced in Hypoxia. Frontiers in Cardiovascular Medicine, 7, 617038.

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