BMS-5 (LIMKi 3) is a potent LIMK inhibitor with IC50s of 7 nM and 8 nM for LIMK1 and LIMK2, respectively.

LIMK2:8 nM, LIMK1:7 nM

BMS-5以剂量依赖的方式抑制Nf2ΔEx2小鼠施万细胞（MSCs）中的cofilin-Ser3磷酸化，其IC50约为2μM。同样以剂量依赖的方式，BMS-5降低Nf2ΔEx2 MSC的存活率，IC50为3.9μM，但在相应的BMS-5浓度下，对照组Nf2flox2/flox2 MSC的存活率并未显著降低。在10μM BMS-5时，Nf2ΔEx2 MSC的存活率为40%，而对照组的存活率为83%[2]。

BMS-5（20或200 μM/侧）在大鼠进行语境恐惧条件训练后立即双侧注入海马区。训练后48小时，通过测试大鼠的记忆巩固。事后分析显示，接受200 μM BMS-5处理的组比20 μM组和对照车辆组表现出更低的冻结水平（P）。

The protein kinase domains of human LIMK1 and LIMK2 are expressed as glutathione S-transferase fusion proteins using the Bac-to-Bac system in Sf9 cells. Compounds 1 to 6 (e.g., BMS-5) are assayed for inhibition of LIMK1 and LIMK2 protein kinase activity by radioactive phosphate incorporation into biotinylated full-length human destrin. Reactions are done with a concentration series of compound in 25 mM HEPES, 100 mM NaCl, 5 mM MgCl2, 5 mM MnCl2, 1 μM total ATP, 83 μg/mL biotinylated destrin, 167 ng/mL glutathione S-transferase-LIMK1, or 835 ng/mL glutathione S-transferase-LIMK2 in a total volume of 60 μL at room temperature for 30 min (LIMK1) or 60 min (LIMK2). Reactions are terminated by addition of 140 μL of 20% TCA/100 mM sodium pyrophosphate, and the precipitates are harvested onto GF/C unifilter plates. The radioactivity incorporated is determined using a TopCount after addition of 35 μL Microscint scintillation fluid[1]

Cell membrane asymmetry is measured. Nf2ΔEx2 MSCs plated in a 6-well format are incubated with 2 μM BMS-5 or DMSO vehicle for 24 hrs. Cell are harvested and assayed. Plasma membrane asymmetry is evaluated with the Violet ratiometric assay by flow cytometry[2] .

Rats [3]