Wb 4101

Small-scale HuH-7^R-targeted quinazoline-derived small molecular compound screening was performed via IC₅₀ ranking based on MTT assays. HuH-7^R cells were seeded into 96-well plates and exposed to concentration gradients of prazosin (Sigma-Aldrich), cyproheptadine (Sigma-Aldrich), phentolamine (Sigma-Aldrich), doxazosin (Sigma-Aldrich), terazosin (Sigma-Aldrich), WB-4101 (Sigma-Aldrich), tamsulosin (Sigma-Aldrich), clozapine (Sigma-Aldrich), bunazosin (Angene Chemical), and ketanserin (Sigma-Aldrich) for 72 h. The IC₅₀ value of each compound was determined and ranked.

Noradrenaline-bitartrate(10 nM - 100 μM; I9278, Sigma), the α₁-AR antagonist prazosine (5 µM; P7791, Sigma), the α₂-AR antagonist yohimbine (5 µM; Y3125, Sigma), the α_2A-AR antagonist BRL 44408 (10 μM, C5776, Sigma), the α_2B-AR antagonist ARC 239, the α_1A-AR antagonist WB 4101, the α_1C, D-AR antagonist CEC (1 nM – 1 µM, Q102, Sigma) and the α_1A-AR agonist A61603 (100 nM; A5861, Sigma) were added to the normal aCSF.
To reduce glutamatergic and GABAergic synaptic input to the recorded neurons, 10 µM CNQX (6-cyano-7-nitroquinoxaline-2,3-dione, C127, Sigma-Aldrich), 50 µM DL-AP5 (DL-2-amino-5-phosphonopentanoic acid, BN0086, Biotrend), and 100 µM PTX (picrotoxin, P1675, Sigma Aldrich) was added to the extracellular saline.