Indirubin-3'-monoxime (Indirubin-3'-oxime) is a potent inhibitor of GSK3β (IC50: 22 nM) and also inhibits CDKs ( (IC50s: 100/180/250 nM for Cdk5/p35, Cdk1/cyclin B, Cdk2/cyclin E).

CDK2-CyclinE:250 nM (cell free), CDK5-p25:100 nM (cell free), CDK1-CyclinB:180 nM (cell free), GSK-3β:22 nM (cell free)

Indirubins是强效的GSK-3 beta抑制剂（IC50：5-50 nM）。通过GSK-3β，细菌表达的重组人tau蛋白确实在体外被磷酸化，且这种磷酸化作用可被Indirubin-3'-monoxime 以剂量依赖的方式抑制，IC50值约为100 nM [1]。Indirubin-3'-monoxime可可逆性地在G2期阻滞异步HBL-100细胞。它抑制了共识CDK磷酸化位点以及在特定CDK1/cyclin B磷酸化位点上的核仁蛋白的磷酸化 [2]。在基于细胞和无细胞的实验中，Indirubin-3'-monoxime选择性地抑制了5-脂氧合酶（5-LO），这是LT生物合成中的关键酶，其IC50值处于低μM范围内 [3]。

经IMX处理的小鼠，在血浆中的葡萄糖、甘油三酯、胆固醇、胰岛素水平方面显示出显著下降，并在学习记忆能力上表现出改善；同时减轻了氧化应激和AChE活性。此外，IMX还能剂量依赖性地提高高脂饮食（HFD）喂养小鼠大脑中的胰岛素和BDNF水平[4]。

Kinase activities were assayed in Buffer A or C (unless otherwise stated), at 30?°C, at a final ATP concentration of 15 μM. Blank values were subtracted, and activities were calculated as picomoles of phosphate incorporated for a 10-min incubation. The activities are usually expressed in percentage of the maximal activity, i.e. in the absence of inhibitors. Controls were performed with appropriate dilutions of dimethyl sulfoxide. In a few cases, phosphorylation of the substrate was assessed by autoradiography after SDS-PAGE [1].

To determine the effects of aminopurvalanol and indirubin-3′-monoxime on tau phosphorylation, Sf9 cells infected with baculovirus expressing htau23 protein were treated 36 h post-infection (when cells have already expressed levels of tau sufficient for the outgrowth of cell processes) with 20 μM inhibitors for 3 h before being harvested [1].

Male mice (5–6 weeks old) were randomly assigned into five groups (n = 10). Group 1: received normal pellet diet (NPD); Group 2: received a HFD; Group 3–5 received HFD for 8 weeks followed by Indirubin-3'-monoxime (IMX) treatment (0.1, 0.2 and 0.4 mg/kg i.p, respectively) once daily for 1 week. IMX was dissolved in (2.5% v/v) DMSO in saline. The mice in NPD and HFD groups received an equivalent volume of vehicle (2.5% v/v DMSO in saline). The composition of HFD was similar as described by Srinivasan. Doses of IMX were selected based on the reports available in literature. Mice were kept under standard husbandry conditions (22 ± 1 C and 60% humidity) and maintained on a 12/12-h light/dark schedule with free access to food and water for 8 weeks. Body weight was recorded weekly throughout the experimental period [4].