N-(4-iodophenyl)-N′-(2-chloroethyl)urea as a microtubule disrupter: in vitro and in vivo profiling of antitumoral activity on CT-26 murine colon carcinoma cell line cultured and grafted to mice

Abstract

The antitumoral profile of the microtubule disrupter N -(4-iodophenyl)- N ′-(2-chloroethyl)urea (ICEU) was characterised in vitro and in vivo using the CT-26 colon carcinoma cell line, on the basis of the drug uptake by the cells, the modifications of cell cycle, and β -tubulin and lipid membrane profiles. N -(4-iodophenyl)- N ′-(2-chloroethyl)urea exhibited a rapid and dose-dependent uptake by CT-26 cells suggesting its passive diffusion through the membranes. Intraperitoneally injected ICEU biodistributed into the grafted CT-26 tumour, resulting thus in a significant tumour growth inhibition (TGI). N -(4-iodophenyl)- N ′-(2-chloroethyl)urea was also observed to accumulate within colon tissue. Tumour growth inhibition was associated with a slight increase in the number of G2 tetraploid tumour cells in vivo , whereas G2 blockage was more obvious in vitro. The phenotype of β -tubulin alkylation that was clearly demonstrated in vitro was undetectable in vivo . Nuclear magnetic resonance analysis showed that cells blocked in G2 phase underwent apoptosis, as confirmed by an increase in the methylene group resonance of mobile lipids, parallel to sub-G1 accumulation of the cells. In vivo , a decrease of the signals of both the phospholipid precursors and the products of membrane degradation occurred concomitantly with TGI. This multi-analysis established, at least partly, the ICEU activity profile, in vitro and in vivo , providing additional data in favour of ICEU as a tubulin-interacting drug accumulating within the intestinal tract. This may provide a starting point for researches for future efficacious tubulin-interacting drugs for the treatment of colorectal cancers.

N-4-iodophenyl-N′-2-chloroethylurea, a novel potential anticancer agent with colon-specific accumulation: radioiodination and comparative in vivo biodistribution profiles

“…Moreover, metabolization rate was generally slower in the tumor than in blood. These data are in accordance with our previous results [17][18]. The antitumor activity of itadministered 4-ICEU was reproduced in ip administered, with high tolerance and efficacy (around 60% tumor growth inhibition and 45% life-span increase).…”

“…We recently focused our efforts on the iodinated bioisostere N-4-iodophenyl-N′-(2-chloroethyl)urea (4-ICEU; Fig. 1) which exhibits not only improved antiproliferative and antiangiogenic activity in various cancer cell lines but also increased antitumoral activity and life-span in murine colon carcinoma [14,17,18]. We hypothesized that replacing the tert-butyl group with the bioisosteric iodine atom should increase in vivo stability.…”

Nuclear Magnetic Resonance (NMR) Study of the Effect of Cisplatin on the Metabolic Profile of MG-63 Osteosarcoma Cells

“…Several possible origins have been suggested for the lipid signals visible in the NMR spectrum, namely lipid droplets in the cytosol, or membrane lipids in the form of glycerophospholipids 15,16 or triglycerides microdomains embedded in the plasma membrane. 28 In several types of tumor cells, the enhancement of these lipid signals, as well as of the unsaturion degree, has been consistently noted and correlated to cells undergoing apoptosis, this effect having been observed for instance in MG-63 cells undergoing radiation-induced apoptosis. 11 A lengthening of the fatty acid CH 2 chain has also been associated with apoptosis in cervical carcinomas.…”

“…13 In addition, many different anticancer drugs have been investigated through NMR analysis of other types of cultured cells, mostly in the form of cell suspensions [14][15][16][17] or extracts. [18][19][20][21][22] High resolution magic angle spinning (HRMAS) NMR spectroscopy, which is well established in tissue analysis, 23,24 has been shown to enable well-resolved spectra to be obtained for intact or lysed cells 25 and to provide valuable information on both lipid and small metabolite changes upon treatment with anticancer drugs of endometrial, 26 melanoma, 27 colon carcinoma, 28 hepatocarcinoma and osteosarcoma 13 cells. In the present study, 1 H HRMAS NMR spectroscopy was used to assess the changes in the intracellular metabolic profile of MG-63 human OS cells induced by the chemotherapy agent cisplatin (cis-diamminedichloroplatinum(II) or CDDP) at different times of exposure.…”

Analytical Approaches toward Successful Human Cell Metabolome Studies by NMR Spectroscopy

“…High resolution magic angle spinning (HRMAS) 1 H NMR spectroscopy has, during the last 10 years, allowed significantly improved resolution to be achieved for tissues [19][20][21] and cell samples. [22][23][24][25][26][27][28][29][30] HRMAS has been successfully employed for the metabolic profiling of different types of tissues [19][20][21] and has been increasingly applied to identify metabolic changes in mammalian cells, e.g., associated with cell type, 22 differentiation, 23 genetic modification, 24,25 and response to drug exposure. [26][27][28] As for biological tissues, most studies on cells have used freezing and storage at -80°C to preserve the cells prior to NMR analysis, generally working under the assumption that cell membrane integrity is retained.…”