AMPK Activator
BI-9774
BI-9774 is a well characterized, potent pan-AMPK activator. It acutely improved glucose tolerance, increased muscle glucose uptake and lactate and glycogen in rodents. BI-9774 also showed reduction in inotropism and heart rate in acute cardiovascular studies in anesthetized rats.
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Sensing and regulating the cellular energy status in response to environmental and/or nutritional stress is highly important and AMP-activated protein kinase (AMPK) is a major contributor for this task. Cellular energy depletion leads to the activation of AMPK thereby inhibiting ATP consuming and upregulating ATP generating pathways.
AMPK, a Ser/Thr protein kinase, is a heterotrimeric enzyme complex composed of a catalytic α-subunit (two isoforms) together with a β-scaffold subunit (two isoforms) and a γ-regulatory subunit (three isoforms):
Binding model of BI-9774 in an allosteric pocket of the kinase domain of AMPK (other parts of AMPK not shown).
An AMPK activator mimics the activation of the enzyme by a low energy status of the cell and changes metabolism from ATP consuming anabolic pathways to ATP-producing catabolic pathways. Therefore, an AMPK activator inhibits gluconeogenesis and fatty acid synthesis in liver and increases glucose uptake and fatty acid oxidation in skeletal muscle. The coordinated effects on glucose/fat metabolism should reduce plasma glucose and plasma lipids. Furthermore, an AMPK activator is expected to induce cardioprotective (e.g. ATP producing) pathways in cardiomyocytes.
BI-9774 displays an EC50 of 64 nM (Emax = 590 %) in an ADP-Glo kinase assay with human AMPK (α1β1γ1) and an EC50 of 8 nM (Emax = 168 nM %) in a cellular GLUT4 translocation assay.
| PROBE NAME | BI-9774 |
| MW [Da, free base]a | 541.0 |
| Human AMPK α1β1γ1 (EC50/Emax) b | 64 nM / 560 %CTL |
| Human AMPK α2β1γ1 (EC50/Emax) b | 88 nM / 580 %CTL |
| Human AMPK α2β2γ2 (EC50/Emax) b | 15 nM / 262 %CTL |
| Human AMPK α2β2γ3 (EC50/Emax) b | 24 nM / 396 %CTL |
| Human AMPK α1β1γ3 (EC50/Emax) b | 84 nM / 427 %CTL |
| Human AMPK α1β2γ2 (EC50/Emax) b | 41 nM / 148 %CTL |
| Human AMPK α1β2γ3 (EC50/Emax) b | 12 nM / 222 %CTL |
| GLUT4 transl. (L6 cells rat) (EC50/Emax) c | 8 nM / 168 %CTL |
aFor the salt form you will get, please refer to the label on the vial and fot the molecule weight of the salt, please refer to the FAQs
bAMPK Kinase Assay: Recombinant human AMPK (containing subunits alpha, beta and gamma) was obtained from a baculovirus expression system. The activity of the AMPK protein was determined using the ADP Glo® Luminescence Kinase test (Promega; V9103X). In this homogeneous test, the amount of ADP remaining after the kinase reaction is quantified by a luciferin-luciferase reaction using luminescence. The luminescence signal obtained correlates with the amount of ADP resulting from the kinase reaction and thus correlates with the activity of the protein kinase.
c Measurement of GLUT4myc translocation in L6 myoblasts: The amount of myc-tagged GLUT4 at the surface of intact cells is measured by an antibody-coupled colorimetric assay after the induced translocation of GLUT4 from an intracellular storage pool to the plasma membrane. A stimulator of GLUT4 translocation will give values above 100% CTL.
| PROBE NAME | BI-9774 |
| logD @ pH 11 | 0.6 |
| Solubility @ pH 6.8 [µg/mL] | 1 |
| Caco-2 permeability AB @ pH 7.4 [*10-6 cm/s] | 17 |
| Caco-2 efflux ratio | 4 |
| MDCK permeability PappAB @ 1µM [10-6 cm/s] | 0.05 |
| MDCK efflux ratio | 68 |
| Microsomal stability (human/mouse/rat) [% QH] | <23 / <23 / <22 |
| Hepatocyte stability (human/mouse/rat) [% QH] | 1 / 25 / 3 |
| Plasma Protein Binding (human/mouse/rat) [%] | 98.6 / 98.7 / 98.7 |
| hERG [inh. % @ 10 µM] | 9 |
| CYP 3A4 (IC50) [µM] | >50 |
| CYP 2C8 (IC50) [µM] | >50 |
| CYP 2C9 (IC50) [µM] | >50 |
| CYP 2C19 (IC50) [µM] | >50 |
| CYP 2D6 (IC50) [µM] | >50 |
| BI-9774 | MOUSE | RAT | MINIPIG |
| Clearance [% QH]a | 17 | 14 | 4b |
| Mean residence time after i.v. dose [h] | 1.5 | 3.8 | 5.8 |
| tmax [h] | 0.9 | 3.3 | 4.0 |
| Cmax [nM] | 50 | 74 | 111 |
| F [%] | 34 | 38 | 16 |
| Vss [L/kg] | 1.4 | 2.3 | 0.6 |
a i.v. dose: 0.54 mg/kg; p.o. dose: 5.4 mg/kg
b i.v. dose: 0.27 mg/kg; p.o. dose: 2.7 mg/kg
The AMPK activator BI-9774 showed dose dependent activation of BI-9774 with a maximal activation 10/20-fold. The activation was mainly driven by free concentration of BI-9774.
The molecule shows anti-diabetic efficacy in diabetic rodents. It also acutely improved glucose tolerance, increased muscle glucose uptake and lactate and glycogen in rodents. BI-9774 subchronically reduced blood glucose, HbA1c and body weight in ZDF rats.
An acute cardio protection in I/R rat model could be shown.
BI-9774 showed reduction in inotropism and heart rate in acute cardiovascular studies in anesthetized rats. Heart weight and heart glycogen increased in HanWistar rats treated for 2 glucose tolerance weeks with BI-9774.
Target engagement measurement in rat (HanWistar): PhosphoT172 AMPK in liver, skeletal muscle.
Dose dependent activation by BI-9774, max. activation 10/20-fold.
Acute efficacy in rat (HanWistar): Dose-dependent glucose lowering of BI-9774 was seen in non-diabetic rats.
Summary Antidiabetic profile:
| ACUTE GTT | SKELETAL MUSCLE | SUBCHRONIC STUDY | |
BI-9774 | oGTT↑ in normoglycemic rodents and mZDF rats | Acute glucose uptake↑, lactate↑, glycogen↑ | Blood glucose↓, moderate HbA1c↓ and BW↓ in ZDF rats (6 weeks) |
Summary Safety profile:
| RODENTS | ECG | CARDIAC FUNCTION | |
BI-9774 | HW= and H glycogen↑ in ZDF rats (6 weeks); HW↑ and H glycogen↑ in HW rats (2 weeks) | No WPW syndrome | HR↓ and EDP↑ in ZDF and HW rats |
Summary Cardiovascular safety:
Acute cardiovascular safety (anesthetized Wistar Rats), subchronic target related cardiovascular safety (anesthetized Wistar and ZDF rats).
*AP-arterial pressure, PP-pulsed pressure, PR, QRS, QT – EKG peaks
BI-9774 activates all human AMPK isoforms tested (see table above) as well as rat AMPK (α1β1γ1) with EC50 values below 100 nM. It is selective at 10µM on the SafetyScreen44TM and Invitrogen® panels. However, in the Invitrogen kinase panel, inhibits MYLK with an IC50 of 170 nM (Grempler et.al., 2018).
| SELECTIVITY DATA AVILABLE | BI-8925 |
SafetyScreen44™ with kind support of  | Yes |
| Invitrogen® | Yes |
| DiscoverX® | No |
| Dundee | No |
Download selectivity data:
BI-9774_selectivityData.xlsx
The X-ray crystal structure of target in complex with BI-9774 was not determined.
MK-8722 is another pan-AMPK activator available.
BI-9774 is a highly potent and well characterized AMPK activator which can be used for in vitro and in vivo studies.
2D structure formats available
Discovery of MK-8722: A Systemic, Direct Pan-Activator of AMP-Activated Protein Kinase
Feng D., Biftu T., Romero F. A., Kekec A., Dropinski J., Kassick A., Xu S., Kurtz M. M., Gollapudi A., Shao Q., Yang X., Lu K., Zhou G., Kemp D., Myers R. W., Guan H-P., Trujillo M. E., Li C., Weber A., Sebhat I. K.
ACS Medicinal Chemistry Letters 2018, 9(1), 39-44.
Systemic pan-AMPK activator MK-8722 improves glucose homeostasis but induces cardiac hypertrophy
Myers R. W., Guan H-P., Ehrhart J., Petrov A., Prahalada S., Tozzo E., Yang X., Kurtz M. M., Trujillo M., Trotter D. G., Feng D., Xu S., Eiermann G., Holahan M. A., Rubins D., Conarello S., Niu X., Souza S. C., Miller C.. Liu J., Lu K., Feng W., Li Y., Painter R. E., Milligan J. A., He H., Liu F., Ogawa A., Wisniewski D., Rohm R. J., Wang L., Bunzel M., Qian Y., Zhu W., Wang H., Bennet B., LaFranco Scheuch L., Fernandez G. E., Li C., Klimas M., Zhou G., van Heek M., Biftu T., Weber A., Kelley D. E., Thornberry N., Erion M. D., Kemp D. M., Sebhat I. K.
Science 2017, 357, 507–511.
When you plan a publication, please use the following acknowledgement:
BI-9774 was kindly provided by Boehringer Ingelheim via its open innovation platform opnMe, available at https://www.opnme.com.