Ca influx activity of CHO cells expressing human GPR40 (FLIPR assay)

CHO cells stably expressing human GPR40 were plated and incubated overnight in 5% CO2 at 37 ℃. Then, cells were incubated in loading buffer (recording medium containing 2.5 μg/mL fluorescent calcium indicator Fluo 4-AM, 2.5 mM probenecid and 0.1% fatty acid-free BSA) for 60 mins at 37 ?C. Various concentrations of TAK-875 were added into the cells and increase of the intracellular Ca2+ concentration after addition was monitored by FLIPR Tsystem for 90 seconds. EC50 value of TAK-875 was obtained with Prism 5 software.

Animal models

A rat model of diabetes

Dosage form

0.3 ~ 3 mg/kg, p.o.

Applications

In a rat model of diabetes, single dose of TAK-875 at 0.3 ~ 3 mg/kg reduced the blood glucose excursion and increased insulin secretion during an oral glucose tolerance test.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

TAK-875 is a potent, selective, and oral GPR40 agonist. GPR40 is one of the G protein-coupled receptors predominantly expressed in pancreatic β-cells, mediating enhancement of glucose-stimulated insulin secretion by free fatty acids.

In vitro: TAK-875 exhibited potent agonist activity and high binding affinity to the human receptor. In addition, TAK-875 showed excellent agonist potency selectivity for GPR40 receptor over other members of the FFA receptor family (for which EC50>10 μM) [1].

In vivo: TAK-875 showed potent plasma glucose-lowering action and insulinotropic action during an oral glucose tolerance test in female Wistar fatty rats with impaired glucose tolerance [2].

Clinical trial: TAK-875 acts as a glucose-dependent insulinotropic agent with low hypoglycemic risk. Its P K is suitable for once-daily oral administration.

References:
[1] Negoro N, Sasaki S, Mikami S, Ito M, Suzuki M, Tsujihata Y, Ito R, Harada A, Takeuchi K, Suzuki N, Miyazaki J, Santou T, Odani T, Kanzaki N, Funami M, Tanaka T1, Kogame A, Matsunaga S, Yasuma T, Momose Y.  Discovery of TAK-875: A Potent, Selective, and Orally Bioavailable GPR40 Agonist. ACS Med Chem Lett. 2010 Jun 18;1(6):290-4.
[2] Leifke E, Naik H, Wu J, Viswanathan P, Demanno D, Kipnes M, Vakilynejad M.  A multiple-ascending-dose study to evaluate safety, pharmacokinetics, and pharmacodynamics of a novel GPR40 agonist, TAK-875, in subjects with type 2 diabetes. Clin Pharmacol Ther. 2012 Jul;92(1):29-39.