title: Structure-based design of lipophilic quinazoline inhibitors of thymidylate synthase.
abstract: To develop novel lipophilic thymidylate synthase (TS) inhibitors, the X-ray structure of Escherichia coli TS in ternary complex with FdUMP and the inhibitor 10-propargyl-5,8-dideazafolic acid (CB3717) was used as a basis for structure-based design. A total of 31 novel lipophilic TS inhibitors, lacking a glutamate residue, were synthesized; 26 of them had in common a N-((3,4-dihydro-2-methyl-6-quinazolinyl)methyl)-N-prop-2-ynylaniline+  ++ structure in which the aniline was appropriately substituted with simple lipophilic substituents either in position 3 or 4, or in both. Compounds were tested for their inhibition of E. coli TS and human TS and also for their inhibition of the growth in tissue culture of a murine leukemia, a human leukemia, and a thymidine kinase-deficient human adenocarcinoma. The crystal structures of five inhibitors complexed with E. coli TS were determined. Five main conclusions are drawn from this study. (i) A 3-substituent such as CF(3), iodo, or ethynyl enhances binding by up to 1 order of magnitude and in the case of CF(3) was proven to fill a nearby pocket in the enzyme. (ii) A simple strongly electron-withdrawing substituent such as NO(2) or CF(3)SO(2) in the 4-position enhances binding by 2 orders of magnitude; it is hypothesized that the transannular dipole so induced interacts favorably with the protein. (iii) Attempts to combine the enhancements of i and ii in the same molecule were generally unsuccessful (iv) A 4-C(6)H(5)SO(2) substituent provided both electron withdrawal and a van der Waal's interaction of the phenyl group with a hydrophobic surface at the mouth of the active site. The inhibition (K(is) = 12 nM) of human TS by this compound, 7n, showed that C(6)H(5)SO(2) provided virtually as much binding affinity as the CO-glutamate which it had replaced. (v) The series of compounds were poorly water soluble, and also the potent TS inhibition shown by several of them did not translate into good cytotoxicity. Compounds with large cyclic groups linked to position 4 by an SO or SO(2) group did, however, have IC(50)'s in the range 1-5 microM. Of these, 4-(N-((3,4-dihydro-2-methyl-6-quinazolinyl)methyl)-N-prop-2-ynylamino  )phenyl phenyl sulfone, 7n, had IC(50)'s of about 1 microM and was chosen for further elaboration.
authors.affiliation: Agouron Pharmaceuticals, Inc., San Diego, California 92121, USA.
authors.forename: T R
authors.initials: TR
authors.lastname: Jones
authors.forename: M D
authors.initials: MD
authors.lastname: Varney
authors.forename: S E
authors.initials: SE
authors.lastname: Webber
authors.forename: K K
authors.initials: KK
authors.lastname: Lewis
authors.forename: G P
authors.initials: GP
authors.lastname: Marzoni
authors.forename: C L
authors.initials: CL
authors.lastname: Palmer
authors.forename: V
authors.initials: V
authors.lastname: Kathardekar
authors.forename: K M
authors.initials: KM
authors.lastname: Welsh
authors.forename: S
authors.initials: S
authors.lastname: Webber
authors.forename: D A
authors.initials: DA
authors.lastname: Matthews
authors.forename: K
authors.initials: K
authors.lastname: Appelt
authors.forename: W W
authors.initials: WW
authors.lastname: Smith
authors.forename: C A
authors.initials: CA
authors.lastname: Janson
authors.forename: J E
authors.initials: JE
authors.lastname: Villafranca
authors.forename: R J
authors.initials: RJ
authors.lastname: Bacquet
authors.forename: E F
authors.initials: EF
authors.lastname: Howland
authors.forename: C L
authors.initials: CL
authors.lastname: Booth
authors.forename: S M
authors.initials: SM
authors.lastname: Herrmann
authors.forename: R W
authors.initials: RW
authors.lastname: Ward
authors.forename: J
authors.initials: J
authors.lastname: White
authors.forename: E W
authors.initials: EW
authors.lastname: Moomaw
authors.forename: C A
authors.initials: CA
authors.lastname: Bartlett
authors.forename: C A
authors.initials: CA
authors.lastname: Morse
mesh_terms: D000230:Adenocarcinoma; D000818:Animals; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000633:toxicity; D001665:Binding Sites; D002460:Cell Line; D018360:Crystallography, X-Ray; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology; D005493:Folic Acid Antagonists / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D007938:Leukemia; D007939:Leukemia L1210; D051379:Mice; D008958:Models, Molecular; D008968:Molecular Conformation; D015394:Molecular Structure; D011487:Protein Conformation; D011799:Quinazolines / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D005493:Folic Acid Antagonists; D011799:Quinazolines; D013940:Thymidylate Synthase