5-(2-(4-Methoxyphenyl)ethyl)-2-amino-3-methylcarboxylate
thiophene (TR560) is the prototype drug of a recently discovered novel class of
tumor-selective compounds that preferentially inhibit the proliferation of specific
tumor cell types (e.g.
leukemia/
lymphoma). Here, we further increased
tumor selectivity by simplification of the molecule through replacing the 4-methoxyphenyl moiety by an alkyl chain. Several 2-amino-3-methylcarboxylate
thiophene derivatives containing at C-5 an alkyl group consisting of at least 6 (hexyl) to 9 (nonyl)
carbon units showed pronounced anti-proliferative activity in the mid-nanomolar range with 500- to 1000-fold
tumor cell selectivity. The compounds preferentially inhibited the proliferation of T-
lymphoma CEM and Molt/4, prostate PC-3, kidney Caki-1 and
hepatoma Huh-7
tumor cells, but were virtually inactive against other tumor cell lines including B-
lymphoma Raji and cervix
carcinoma HeLa cells. The novel prototype drug 3j (containing a 5-heptyl chain) elicited a cytotoxic, rather than
cytostatic activity, already after 4 h of exposure. The unusual
tumor selectivity could not be explained by a differential uptake (or efflux) of the drug by sensitive versus resistant
tumor cells. Exposure of a fluorescent derivative of 3j revealed pronounced uptake of the drug in the cytoplasm, no visible appearance in the nucleus, and a predominant localization in the endoplasmic reticulum. These observations may be helpful to narrow down the intracellular localization and identification of the molecular target of the 5-substituted
thiophene derivatives.