Nampt-IN-1 (LSN3154567)

Cell experiment:

Cell[1]The cell lines used as following: A2780 and KM-12, KMS-11 and MKN-74, OPM-2 and Kelly; and all other cancer cells. The identity of these cell lines is not tested or verified prior to use in this study. Cells are seeded in 96-well plates, cultured overnight, and treated with LSN3154567 (0.03 to 1,000 nM)±NA (10 μM) in duplicate at 37°C in 5% CO2 for 72 hours. Staurosporine (10 μM) is used as positive control. Cell viability is determined by a CytoTox-Glo Cytotoxicity assay kit[1].

Animal experiment:

Mice[1]Male CD-1 mice are dosed with LSN3154567 at 2 mg/kg intravenously in 20% Captisol (w/v), 25 mmol/L NaPO4, pH 2, q.s. formulation or 2 mg/kg orally in the same vehicle in a cross over design with 3-d wash period in between two arms. Blood samples are obtained through retro orbital at 0, 0.08 (IV only), 0.25, 0.75, 2, 4, 8, and 24 hours and frozen until analysis. For exposure analysis, blood samples (≈20 μL) are obtained through tail clipping at 0.5, 1, 2, 4, 8, and 24 hours. The samples are collected into EDTA-coated capillary tubes and spotted onto Whatman DMPK-C DBS collection cards.Rats and Dogs[1]Female rats or male and female dogs are dosed with LSN3154567 orally in the same vehicle. For NA rescue studies, NA is formulated in phosphate buffered saline (pH 7.4). Rats are given LSN3154567 orally once daily at 20, 40, or 80 mg/kg for two cycles of 4 days with 3 days between the first and second cycles. Dogs are also dosed orally at 1 or 2.5 mg/kg (without NA) or at 5 mg/kg (with NA; BID) for 4 days. Exposure is obtained and analyzed as described above.

References:

[1]. Zhao G, et al. Discovery of a Highly Selective NAMPT Inhibitor That Demonstrates Robust Efficacy and Improved Retinal Toxicity with Nicotinic Acid Coadministration. Mol Cancer Ther. 2017 Dec;16(12):2677-2688.