SAR-100842

Cell lines

Preparation Method

SAR-100842 were dissolved into 100% DMSO to 10 mM stock and further diluted in serum free media.cells were serum starved and incubated in vehicle or varying concentrations of drug for 24 hours. Lower wells contained DMEM with or without attractants (1-5% FBS or 5-10 µM LPA). 4T1-Luc2 or MDA-MB-231T cells were added to the upper wells at a concentration of 2×105 cells/mL in serum-free DMEM containing vehicle or SAR-100842 and incubated for 4 hours in a humidified chamber at 37⊿in 5% CO2. The top chamber was removed and the cells that had migrated to the bottom of the membrane were stained using Diff Quick Staining kit. Using an inverted brightfield microscope with a 10× objective, the number of cells that had migrated through the membrane was counted in three fields in the center of each filter.

Reaction Conditions

0.5-50µM for 24, 48, 72 hours

Applications

50 µM SAR-100842 reduced the migration of MDA-MD-231T cells through a collagen membrane by 1.92-fold and 3.15-fold to FBS and LPA chemoattractants, respectively. In 4T1-Luc2 cells 50 µM SAR-100842 reduced migration by 10.8-fold and 13.6-fold to FBS and LPA, respectively.

Animal models

6-8 week old athymic nu/nu mice

Preparation Method

Mice received 30 mg/kg SAR-100842 by oral gavage twice daily for the duration of the experiment. 6-8 week old athymic nu/nu mice received either 1.0 × 106 SKVO3 cells or 3.5 × 106 OVCAR5 cells in an intraperitoneal injection. On day two post cell injection, mice were randomized to three groups. Group one began vehicle on day 2, group two began 30 mg/kg SAR-100842 twice daily on day two for the duration of the experiment, and group three began 30 mg/kg SAR-100842 twice daily on day 10 for the duration of the experiment. On day 70 post cell injection all mice were euthanized and necropsied. Liver, abdominal lymph nodes or masses, omentum, peritoneum and any other organ suspected of harboring tumor were collected, fixed in 10% NBF and prepared for histological analysis.

Dosage form

30 mg/kg twice daily, oral

Applications

In the SKOV3 model treated early with SAR-100842, Only a weak reduction in diaphragm, kidney and lymph node metastases was evident ; none of these reductions were maintained in the group that started treatment on day 10 post-injection. For the OVCAR5 model, metastases were observed at day 70 post-injection on the omentum, liver, diaphragm, pancreas and peritoneum. Of these, SAR-100842 apparently reduced diaphragm tumor deposits, both surface and invasive, but was without significant effect in other locations.

References:

[1]: Brooks D, Zimmer A, Wakefield L, et al. Limited fibrosis accompanies triple-negative breast cancer metastasis in multiple model systems and is not a preventive target[J]. Oncotarget, 2018, 9(34): 23462.