Cell lines

Non–small cell lung cancer (NSCLC) PC-9/ER, PC-9/ERC15, PC-9/ERC16 cell lines

Preparation method

Soluble in DMSO. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

1 μM, 72 h

Applications

Treatment with 1 μM ALW-II-41-27 inhibited cell proliferation and increased apoptosis in erlotinib-resistant NSCLC cell lines. Apoptosis induced by ALW-II-41-27 was accompanied by the increase of cleavage of caspase-3 and PARP as well as decreased expression of antiapoptotic proteins BCL-xL and MCL-1.

Animal models

6-week-old athymic nude mice

Dosage form

Intraperitoneal injection, 15, 30 mg/kg, twice daily

Application

Administration of ALW-II-41-27 to tumor-bearing mice significantly inhibited H358 tumor growth. Histological analysis showed that tumors treated with ALW-II-41-27 had a significant increase of apoptosis compared with tumors treated with NG-25 or vehicle alone, similar to genetic ablation of EPHA2.

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.

ALW-II-41-27 is a potent inhibitor of EPH family kinases, with an IC50 value of 11 nM to EPHA2 [1] [2].

EPH family proteins are key regulators of both disease and normal development. EPH receptors are involved in many intracellular signaling pathways such as PI3K/AKT/mTOR, RAS/RAF/MAPK, FAK, SRC, ABL, and RHO/RAC/CDC42 [2].

In H358 cells, treatment with ALW-II-41-27 at a concentration of 1 μM within 15 minutes impaired the tyrosine phosphorylation of the EPHA2 receptor and continued to inhibit the tyrosine phosphorylation through 6 hours. ALW-II-41-27 also dose-dependently inhibited the EPHA2 phosphorylation induced by ligand. When the EPHA2 was depleted by RNAi in NSCLC cell lines, cells were much less sensitive to ALW-II-41-27.

It was suggested that EPHA2 plays an oncogenic role according to results in lung cancers. In mice bearing non–small cell lung cancers (NSCLCs), intraperitoneal injection with ALW-II-41-27 at a dose of 15 mg/kg twice daily for 14 days significantly resulted in an inhibition of the growth of H358 tumors. ALW-II-41-27 significantly increased the apoptosis of tumors compared with the vehicle alone or NG-25. This was similar to the effect of the genetic ablation of EPHA2. Compared with treatments with vehicle alone or NG-25, treatment with ALW-II-41-27 did not result in significant differences in the vessel density or proliferation of tumors [2].

References:
[1]. Marialuisa Moccia, Qingsong Liu, Teresa Guida, et al. Identification of Novel Small Molecule Inhibitors of Oncogenic RET Kinase. PLOS ONE, 2015, 10(6):e0128364.
[2]. Katherine R. Amato, Shan Wang, Andrew K. Hastings, et al. Genetic and pharmacologic inhibition of EPHA2 promotes apoptosis in NSCLC. Journal of Clinical Investigation, 2014, 124(5):2037-2049.