PF-06409577 is an effective, orally active, and specific allosteric activator of AMPK (EC50: 7 nM, for α1β1γ1).

AMPK α1β1γ1:7 nM (EC50)

在人类和大鼠中，PF-06409577对α1β1γ1亚型具有相似的效力。PF-06409577在一系列受体、离子通道、PDEs和激酶上显示出最小的非目标效应。PF-06409577在贴片钳测量中不抑制hERG（100 μM），并且不是主要人类细胞色素P450亚型的微粒体活性抑制剂（IC50>100 μM）[1]。

在大鼠、猴子和狗的研究中，PF-06409577展现了中等程度的血浆清除率，并且分布良好，达到稳态分布体积。经口服0.5%甲基纤维素悬浮液的结晶性PF-06409577后，PF-06409577在狗、大鼠和猴子体内快速吸收。相应的口服生物利用度在大鼠、猴子和狗中分别为15%、59%和100%。PF-06409577处理（300 mg/kg）可观察到整个肾组织中相对于总AMPK（tAMPK）的pAMPK水平呈剂量依赖性增加，最大响应为3.8倍。口服PF-06409577（每天10、30和100 mg/kg）在肥胖ZSF1动物中导致蛋白尿呈剂量依赖性减少，与车辆对照组相比，60天治疗后24小时尿白蛋白损失减少超过2倍[2]。

PF-06409577 is prepared in DMSO. PF-06409577 is incubated with fully phosphorylated AMPK in assay buffer at room temperature for 15 min followed by addition of PP2a and another incubation for 60 min at room temperature. The phosphatase treatment is quenched and the kinase assay initiated with the addition of okadaic acid (50 nM final), 50 nM Cy-5 SAMS peptide and ATP equal to Km for each isoform. Reactions are incubated for an additional 60 min and the kinase reaction is quenched with the addition of 10 mM EDTA and 2 nM Eu-pACC antibody in detection Buffer. Kinase activity is monitored by excitation at 320 nM and measuring emission at 665 and 615 nM, respectively[2].

PF-06409577 is prepared in 0.5% methylcellulose[2].Daily administration of 0.5% methylcellulose (p.o.), PF-06409577 at 10, 30, or 100 mg/kg (p.o.), PF- 249 at 3, 10, or 30 mg/kg (p.o.), or ramipril in drinking water (1 mg/kg/day) is initiated and continued for 68 days. Urine is collected for 24-hours and volume recorded from all lean and obese rats after 14, 28, 42, and 60 days of dosing. On Day 63 all rats are administered a final dose after 16-hour overnight fasting. One hour following the final dose, blood glucose is measured by glucometer and a 100 μL tail vein blood sample collected and processed for determination of insulin levels and total protein. Each rat is then anesthetized with isoflurane. The right kidney is collected and immediately freeze-clamped and transferred to liquid nitrogen storage; the left kidney is fixed in 10% formalin. Rats are then euthanized by exsanguination from the vena cava[2].