Cell lines

DLD-1, HT29, SW620 and HCT116 CRC cell lines and the non-tumourigenic CCD-841 cell line

Preparation Method

6000 cells/well were seeded into 96- well flat-bottom plates and incubated for 24 h under standard conditions. MN58b or RSM-932A (TCD-717) were added at different concentrations from stocks solutions.

Reaction Conditions

2, 3, 4 μM, 24h

Applications

RSM-932A (TCD-717; 2-4 μM; for 24 hours) promotes cell death of colon cancer cells. ChoKα specific inhibitor RSM-932A (TCD-717), has demonstrated a potent antitumoral activity both in vitro and in vivo against several tumor-derived cell line xenografts including CRC-derived cell lines.

Animal models

Six-week-old nude mice

Preparation Method

Mice were s.c. inoculated at lowers flanks with 1 × 106 of HT-29 cells suspended in 100μL of serum-free DMEM media mixed (1:1) with Matrigel. Measurable subcutaneous tumors (~0.15-0.2 cm3) were randomized to the treatment and control group. Tumor growth was monitored three times per week.

Dosage form

Effective dose of RSM-932A is 7.5 mg/kg, RSM-932A was dissolved at 50 mM 100% DMSO and administrated by i.v.. The successive dilutions were done fresh every day of treatment in sterile 5% dextrose water.

Applications

This could be used to perform growth inhibition studies at the specified effective dose. No significant organ damage was observed apart from hepatotoxicity that was found in all treatments except for the less aggressive treatment with RSM-932A that showed no liver toxicity while retaining a potent antitumoral activity.

RSM-932A is a ChoKα inhibitor with potent in vitro antiproliferative activity against a large variety of tumor-derived cell lines and in vivo antitumoral activity against human xenographs in mice. RSM-932A also shows high efficacy with low toxicity at the effective doses.^[1]

In vitro experiments indicated that RSM-932A has a potent antiproliferative activity against most tumor-derived cell lines tested, including those derived from breast, lung, colon, bladder, liver, ovary, bone, cervix, kidney, pancreas, melanoma and brain tumors, with IC50 (1.15 ± 0.14 μM) in the low, single-digit micromolar range, consistent with a cytotoxic effect.

Table 1.Panel of human cancer cell lines tested with RSM-932A for 72 hours and determination of IC₅₀, GI₅₀, TGI, and LC₅₀parameters

In vivo experiments demonstrated that RSM-932A shows no detectable toxicity in mice at highly effective doses with 77% tumor growth inhibition.^[1]

References:
[1]. Lacal JC, et al. Preclinical characterization of RSM-932A, a novel anticancer drug targeting the human choline kinase alpha, an enzyme involved in increased lipid metabolism of cancer cells. Mol Cancer Ther. 2015 Jan;14(1):31-9.