Ki16198

Inositolphosphate response

RH7777 cells expressing LPA1, LPA2, LPA3, LPA4, or LPA5 were cultured on collagen-coated 12-well dishes in the growth medium, and then the medium was changed to TCM199 containing 2 μCi/mL [3H]inositol and 0.1% (w/v) BSA (fraction V). After 24 hrs, the cells were washed 3 times with HEPES-buffered medium, which consisted of 20 mM Hepes (pH 7.4), 134 mM NaCl, 4.7 mM KCl, 1.2 mM KH2PO4, 1.2 mM MgSO4, 2 mM CaCl2, 2.5 mM NaHCO3, 5 mM glucose, and 0.1% (w/v) BSA, and incubated for 30 mins with the indicated concentrations of Ki16198 with or without 1 μM LPA in the presence of 10 mM LiCl in the same medium at final volume of 0.5 mL. The reaction was terminated by adding 1 N HCl (0.1 mL) and freezing the cells. The supernatant (acid extract in 0.5 mL) of the thawed cells was used for the separation of [3H]inositol phosphate fractions. The results were normalized to 105 dpm of the total radioactivity incorporated into the cellular inositol lipids. The radioactivity of the trichloroacetic acid (5%)-insoluble fraction was measured as the total radioactivity.

Cell lines

YAPC-PD cells

Preparation method

The solubility of this compound in DMSO is > 24.5 mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below - 20 °C for several months.

Reacting condition

10 μM; 24 hrs

Applications

In YAPC-PD cells, Ki16198 inhibited LPA-mediated migration and invasion. Moreover, Ki16198 inhibited LPA-induced expression of proMMP-9 protein and mRNA.

Animal models

Mice bearing YAPC-PD xenografts

Dosage form

2 mg/kg; p.o.

Applications

In mice bearing YAPC-PD xenografts, Ki16198 (2 mg/kg) substantially decreased the total metastatic node weight in the peritoneal cavity, as well as ascites formation by 50%.

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.

References:

[1]. Komachi M, Sato K, Tobo M, et al. Orally active lysophosphatidic acid receptor antagonist attenuates pancreatic cancer invasion and metastasis in vivo[J]. Cancer science, 2012, 103(6): 1099-1104.