Cell lines

2 wild-type (wt) p53 (Lovo and LS174T) and 4 mutant (mt) p53 (WiDr, WiDr/F, HT29 and SW948) colon carcinoma cell lines

Preparation method

The solubility of this compound in DMSO is >154mg/mL. 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

50 and 100 nM; 24 and 48 hrs

Applications

Raltitrexed up-regulated p53 and p21 expression in wt p53 cells, but not in mt p53 cells. In the mt p53 cells HT29 and WiDr/F, the highest induction of thymidylate synthase (6 ~ 10 folds) was observed after Raltitrexed treatment. Moreover, Raltitrexed increased Bax expression up to 5 folds in wt p53 cells, but with only a very slight induction of Bax expression in mt p53 cells. In wt p53 cells, Raltitrexed treatment hardly changed Bcl-2 expression.

Animal models

C57BL/6J-ApcMin/+ mice

Dosage form

3 or 5 mg/kg; twice a week or five times a week

Applications

In C57BL/6J-ApcMin/+ mice, Raltitrexed (3 mg/kg; twice a week) increased the average tumor number in the small intestine by 4 folds. When the dose of Raltitrexed was elevated to 5 mg/kg, five times a week, it resulted in a 10-fold increase in the average tumor number. Under all administration schedules, Raltitrexed was well-tolerated with only few treatment-related deaths occurring. Raltitrexed-induced tumors commonly occurred in the duodenum and jejunum, with few in the ileum and none in the colon.

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.

Raltitrexed is an inhibitor of thymidylate synthase (Ki = 62 nM).[1] It inhibits the growth of L1210 mouse lymphocytic leukemia cells in vitro (IC50 = 8 nM) as well as reduces thymidylate synthase activity and completely eliminates tumors in a mouse L1210:ICR ascitic tumor model when administered at a dose of 4 mg/kg, effects that can be reversed by administration of thymidine. It also reduces tumor growth and the number of metastases in an HCT116-luc mouse xenograft model.[2] Formulations containing raltitrexed have been used in the treatment of advanced colorectal cancers.

Reference:
[1]. Jackman, A.L., Taylor, G.A., Gibson, W., et al. ICI D1694, a quinazoline antifolate thymidylate synthase inhibitor that is a potent inhibitor of L1210 tumor cell growth in vitro and in vivo: A new agent for clinical study. Cancer Res. 51(20), 5579-5586 (1991).
[2]. Qiu, C., Li, Y., Liang, X., et al. A study of peritoneal metastatic xenograft model of colorectal cancer in the treatment of hyperthermic intraperitoneal chemotherapy with Raltitrexed. Biomed Pharmacother. 92, 149-156 (2017).