Folate-based thymidylate synthase inhibitors as anticancer drugs

Abstract

The enzyme, thymidylate synthase (TS) is considered an important target for the development of new anticancer agents. Moreover, the folate-binding site in TS is believed to offer better opportunities for the design of highly specific inhibitors than the pyrimidine (dUMP) binding site. This belief led to the design of N10-propargyl-5,8-dideazafolic acid (CB3717), a quinazoline-based drug which had antitumour activity in clinical studies. Occasional, but serious nephrotoxicity led to the withdrawal of CB3717 from further clinical study. More water-soluble and non-nephrotoxic analogues were developed with an interesting diversity in biochemical profile, particularly with respect to interactions with the reduced-folate cell membrane carrier (RFC) and folylpolyglutamate synthetase (FPGS). An example of a compound that uses both of these processes well is the quinazoline, ZD1694 (Tomudex), a drug which is about to complete phase III evaluation for colorectal cancer. High chain length polyglutamates are formed that are up to 70-fold more potent TS inhibitors than the parent drug (Ki tetraglutamate = 1 nM). Furthermore they are retained in cells/tissues for a prolonged period. A number of other novel folate-based TS inhibitors are currently in pre-clinical or clinical study. For example, LY231514 is a pyrrolopyrimidine analogue in phase I study and, although less potent as a TS inhibitor, has biochemical properties similar to ZD1694. Another compound in phase I study is the benzoquinazoline, BW1843U89 which has somewhat different properties. It is a very potent TS inhibitor (Ki = 0.09 nM) and an excellent substrate for the RFC (human) and FPGS, but polyglutamation proceeds to diglutamate only and is not accompanied by increased TS inhibition. Another highly water-soluble compound in pre-clinical development is ZD9331 which was specifically designed to use the RFC but not be a substrate for FPGS. Potent TS inhibition (Ki = 0.4 nM) was achieved through a rational programme of computerised molecular modelling of the active site of TS and a large database of structure-activity relationships. Two lipophilic compounds were designed to be devoid of interactions with either the RFC or FPGS. High resolutions crystal complexes of E. coli TS were central to obtaining potent TS inhibitors and both AG337 (Ki human recombinant TS = 16 nM) and AG331 (Ki = 12 nM) are in clinical studies. This portfolio of novel compounds therefore comprehensively addresses the potential of TS as a target for cancer chemotherapy.