In vitro activity: LY3104607 is a potent, selective and orally available G Protein-Coupled Receptor 40 (GPR40) agonist with potential antidiabetic application. It has optimized pharmacokinetic properties to support once daily oral treatment in patients with Type 2 Diabetes Mellitus. LY3104607 demonstrated functional potency and glucose-dependent insulin secretion (GDIS) in primary islets from rats. Potent, efficacious, and durable dose-dependent reductions in glucose levels were seen during glucose tolerance test (GTT) studies. Low clearance, volume of distribution, and high oral bioavailability were observed in all species. The combination of enhanced pharmacology and pharmacokinetic properties supported further development of this compound as a potential glucose-lowering drug candidate.

Kinase Assay: All in vitro assays including binding, calcium flux, β-arrestin agonist, and peroxisome proliferator-activated receptor (PPAR) α, δ, and γ assays were described previously in detail. The use of animals was in accordance with international guidelines (NIH 85-23) and was approved by the local animal ethics committee at Lilly Research Laboratories.

Cell Assay: Glucose-Dependent Insulin Secretion (GDIS) in Rat Islet. GDIS assays were performed in primary islets. Pancreatic islets of Langerhans were isolated from male SD (Sprague–Dawley) rats by collagenasedigestion and Histopaque density gradient separation. The islets were cultured overnight in RPMI-1640 medium with GlutaMAXn to facilitate recovery from the isolation process. Insulin secretion was determined using 90 min incubation in EBSS (Earle’s Balances Salt Solution) buffer in a 48-well plate. Islets were first preincubated in EBSS with 2.8 mM glucose for 60 min, then transferred to a 48-well plate (four islets/well) containing 150 μL of 2.8 mM glucose, and incubated with 150 μL of EBSS with 2.8 or 11.2 mM glucose in the presence or absence of test compounds for 90 min. The buffer was removed from the wells at the end of the incubation period and assayed for insulin levels using a rat insulin ELISA kit (Mercodia).