Vinblastine sulfate (Vincaleukoblastine sulfate salt) can inhibit the formation of microtubule, it also inhibits nAChR(IC50=8.9 uM).

nAChR:8.9 μM

Vinblastine的平均终末半衰期为14.3小时。在新鲜分离的大鼠肝细胞中孵育时，VLB迅速且强烈地渗透进入细胞内，这一过程很可能通过被动扩散机制实现，随后紧密地与细胞结合[3]。Vinblastine通过抑制由肾上腺髓质素诱导的血管生成反应，同时对于有丝分裂滑移也表现出积极效应，导致带有细胞分裂阻滞的单核细胞中出现微核[4]。在产生约50%的细胞死亡和细胞静止或更低浓度下，根据RPD、RICC和RCC的计算，vinblastine显著增加了微核化的单核细胞数量[2]。

Vinblastine是一种广泛使用的抗癌化合物，但具有不希望的副作用[6]。VBL与RAP的低剂量组合对人类HCC在体内展现出令人满意的抗血管生成效果[4]。临床相关剂量的Vinblastine在体内抑制CEM细胞中tubulin的棕榈酰化作用（对tubulin去棕榈酰化的影响）[5]。

Cell based receptor autophosphorylation assays: Autophosphorylation of PDGFR family kinase assays are cell-based enzyme-linked immunosorbent (ELISA) assays using CHO cells expressing wild-type PDGFRβ, chimeric protein PDGFRβ/c-Kit, and PDGFRβ/Flt3 which contain the extracellular and transmembrane domains of PDGFRβ and the cytoplasmic domain of c-Kit, and Flt-3. Cells are grown to confluency in 96-well microtiter plates under standard tissue culture conditions, followed by serum starvation for 16 hours. Briefly, quiescent cells are incubated at 37 °C with increasing concentrations of Tandutinib for 30 minutes followed by the addition of 8 nM PDGF-BB for 10 minutes. Cells are lysed in 100 mM Tris, pH 7.5, 750 mM NaCl, 0.5% Triton X-100, 10 mM sodium pyrophosphate, 50 mM NaF, 10 μg/mL aprotinin, 10 μg/mL leupeptin, 1 mM phenylmethylsulfonyl fluoride, 1 mM sodium vanadate, and the lysate is cleared by centrifugation at 15,000 g for 5 minutes. Clarified lysates are transferred into a second microtiter plate in which the wells are previously coated with 500 ng/well of 1B5B11 anti-PDGFRβ mAb and then incubated for 2 hours at room temperature. After washing three times with binding buffer (0.3% gelatin, 25 mM HEPES, pH 7.5, 100 mM NaCl, 0.01% Tween 20), 250 ng/mL of rabbit polyclonal anti-phosphotyrosine antibody is added and plates are incubated at 37 °C for 60 minutes. Subsequently, each well is washed three times with binding buffer and incubated with 1 μg/mL of horseradish peroxidase-conjugated anti-rabbit antibody at 37 °C for 60 minutes. Wells are washed prior to adding 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), and the rate of substrate formation is monitored at 650 nm.

Six-well treatment plates are set up that contained 5 × 104 cells/mL in each well, suspended in 3 mL culture medium, and these are treated with vinblastine for 3 h followed by 21 h growth. (Only for Reference)