Niclosamide (Niclocide)(BAY2353) is used to treat most tapeworm infections by inhibiting DNA replication.

STAT3:0.7 μM

Niclosamide (< 5 μM) 在HeLa细胞中剂量依赖性抑制STAT3介导的荧光素酶报告活性，其IC50为0.25 μM。Niclosamide (< 2 μM) 在Du145细胞中剂量依赖性抑制STAT3的磷酸化。Niclosamide (1 μM) 抑制在Du145细胞中EGF诱导的STAT3核内转移。Niclosamide (< 2 μM) 在Du145细胞中剂量依赖性抑制STAT3下游基因的转录。Niclosamide (< 10 μM) 在Du145癌细胞中剂量依赖性诱导G0/G1停滞和凋亡。[1] Niclosamide能够在感染SARS-CoV的Vero E6细胞中以微摩尔浓度抑制SARS-CoV的复制。[2] Niclosamide (< 7.5 μM) 促进Frizzled1的内吞作用，下调Dishevelled-2蛋白，并在U2OS细胞中抑制Wnt3A刺激的beta-catenin稳定性和LEF/TCF报告活性。[3] Niclosamide在U2OS细胞中剂量和时间依赖性抑制TNF诱导的NF-κB报告活性。Niclosamide (125 nM) 抑制由p65、IKKα、IKKβ、IKKγ和TAK1在U2OS细胞中诱发的NF-κB激活。Niclosamide (< 500 nM) 完全阻断在HL-60细胞中TNFα诱导的NF-κB–DNA复合体的时间和剂量依赖性变化。Niclosamide (< 10 nM) 抑制U266细胞中固有的NF-κB激活。Niclosamide在HL-60、Molm13或AML原发细胞中剂量和时间依赖性抑制TNF诱导的IκBα降解和p65重新定位。Niclosamide (500 nM) 降低 TNF诱导的与HL-60细胞生存相关的NF-κB依赖性基因产品。Niclosamide剂量依赖性抑制AML细胞的生长并有效诱发凋亡，降低Mcl-1和XIAP水平，增加细胞内ROS水平。[4]

Niclosamide（每日每公斤40毫克，腹腔注射）抑制裸鼠体内荷HL-60异种移植瘤的AML细胞生长。[4]

Protein Kinase profiling assay: Assay for 22 different proteins kinases is carried out by ProQinase Gmbh. All of the protein kinases are expressed either in Sf9 insect cells or in E.coli as recombinant GST-fusion proteins or His-tagged proteins. Protein kinases are purified by affinity chromatography using either GSH-agarose or Ni_NTH-agarose. A radiometric protein kinase assay is used for measuring the kinase activity of the 22 protein kinases. Briefly, for each protein kinase, 50 μL reaction cocktail containing 60 mM HEPES-NaOH, 3 mM MgCl2, 3 mM MnCl2, 3 μM Na-orthovanadate, 1.2 mM DTT, 50 μg/mL PEG20000, 1 μM [γ-33P]-ATP, Niclosamide, adequate amount of enzyme and its substrate. The PKC-alpha assay additionally contain 1 mM Cacl2, 4 mM EDTA, 5 μg/mL phosphatidylserine and 1 μg/mL 1, 2-Dioleyl-glycerol. The reaction cocktails are incubated at 37 °C for 60 minutes and stop with 50 μL 2% (v/v) H3PO4. Incorporation of 33Pi is determined with a microplate scintillation counter. The activities and the IC50 values are calculated using Quattro Workflow V2.28.

Cells are plated in 96-well culture plates with cell density of 3-4 × 103 cells/well and treat with Niclosamide by adding 100 μL medium containing Niclosamide of various concentrations on the second day. After 72-hour's treatment, MTT is added to each well and incubated for additional 4-5 hours, and the absorbance is measured on a microplate reader at 570 nM. Cell growth inhibition is evaluated as the ratio of the absorbance of the sample to that of the control. The results are representative of at least 3 independent experiments. (Only for Reference)