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2 protocols using trifluoperazine dihydrochloride tfp

1

Synthesis and Glucuronidation of NAF Enantiomers

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R(+)-NAF (>99.5% ee) and S(-)-NAF (>99.5% ee) (Figure 1) were synthesized according to previous methods (Shivani et al., 2007 (link)). The internal standard (IS) Z10 (CN201110148322.7) was obtained from our laboratory. HPLC-grade methanol, acetonitrile, and acetic acid were obtained from Merck (Darmstadt, Germany). Uridine diphosphate glucuronic acid (UDPGA), alamethicin, Tris, magnesium chloride, methylum-belliferone (MU), 4-methylumbelliferone (4-MU), 4-methylum-belliferyl-β-D-glucuronide hydrate (4-MU-G), propofol, propofol glucuronide, zidovudine, zidovudine glucuronide, fluconazole, and niflumic acid, trifluoperazine dihydrochloride (TFP), and TFP-glucuronide (TFP-G) were purchased from Sigma–Aldrich (St. Louis, MO, United States). Recombinant human UGTs (UGT1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B15, and 2B17) and the commercially available microsomes from human liver (HLMs), intestine (HIMs), and kidney (HKMs), as well as those from rat liver (RLMs), intestine (RIMs), and kidney (RKMs), were purchased from BD Gentest (Woburn, MA, United States). Protein contents of the microsomes were used according to the manufacturers’ instructions. Ultrapure water was used in all experiments (Millipore, Billerica, MA, United States).
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2

Inhibition of DENV and ZIKV by TFP

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Trifluoperazine dihydrochloride (TFP) (Fig. 1a) was purchased from Sigma-Aldrich (USA). The stock solution was prepared in water at a concentration of 10 mM and sterilized by filtration.

Antiviral activity of TFP against DENV and ZIKV in Vero cells. (a) Chemical structure of trifluoperazine dihydrochloride. (b-f) Cells were infected with DENV-1 (b), DENV-2 (c), DENV-3 (d), DENV-4 (e), or ZIKV (f), and after 24 h of infection in the presence of different concentrations of TFP, extracellular virus yields were determined by plaque assay. Data are mean values from three independent experiments ± standard deviation (SD)

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