Summary
Since the mitogenic action of EGF is mediated by ligand-induced autophosphorylation of the EGF receptor (EGFR), and EGFR is commonly overexpressed in solid human tumours, inhibitors of receptor tyrosine kinase activity (RTK) could prove to be effective antitumour agents. Screening of a compound library using an EGF-RTK enzyme prepared from human tumour derived A431 cells identified a series of potent (IC50<1µM) enzyme inhibitors. These inhibitors are quinazolines bearing a variety of substituted anilines at the 4-position. The most potent 4-anilinoquinazolines (IC50 ≅ 20nM) have small non-polar meta substituents on the aniline ring, and are competitive with ATP and non-competitive with substrate. The growth inhibitory activity of these agents was assessed in vitro using KB cells (human oral squamous tumour) grown in the absence or presence of EGF. A selected compound, 4-(3-chloroanilino)quinazoline (CAQ), inhibited EGF-stimulated growth in a concentration dependent manner and complete blockade was observed at concentrations (1–10 µM) which had no effect on basal growth. Selectivity of growth inhibition by CAQ was further exemplified in IGF1-stimulated KB cells where no effect was detected at concentrations which completely blocked EGF-stimulated growth. Similarly, CAQ blocked TGFα-stimulated growth in MCF-7 human breast cancer cells without affecting insulin-stimulated growth. These studies define a novel class of EGF-RTK inhibitors which are also potent and selective inhibitors of EGF-stimulated human tumour cell growthin vitro.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Sainsbury JRC, Malcolm AJ, Appleton DR, Farndon JR, Harris AL: Presence of epidermal growth factor receptor as an indicator of poor prognosis in patients with breast cancer. J Clin Pathol 38:1225–1228, 1985.
Gullick WJ: Prevalence of aberrant expression of the epidermal growth factor receptor in human cancers. Br Med Bull 47:87–98, 1991.
Rajkumar T, Gullick WJ: The Type I growth factor receptors in human breast cancer. Breast Cancer Res Treat 29:3–9, 1994.
Yarden Y, Ullrich A: Growth factor receptor tyrosine kinases. Ann Rev Biochem 57:443–478, 1998.
Schlessinger J: How receptor tyrosine kinases activate Ras. Trends Biochem Sci 18:273–275, 1993.
Ennis BW, Lippman ME, Dickson RB: The EGF receptor system as a target for antitumor therapy. Cancer Investigation 9:553–562, 1991.
Workman P: The potential for molecular oncology to define new drug targets.In Kerr DJ, Workman P (eds) New Molecular Targets for Cancer Chemotherapy. CRC Press, London, 1994, pp 1–29.
Burke TR Jr: Protein-tyrosine kinase inhibitors. Drugs of the Future 17:119–131, 1992.
Hanks SK: Eukaryotic protein kinases. Current Opin Struct Biol 1:369–383, 1991.
Yaish P, Gazit A, Gilon C, Levitzki A: Blocking of EGF-dependent cell proliferation by EGF receptor kinase inhibitors. Science 242:933–935, 1988.
Ward WHJ, Cook PN, Slater AM, Davies DH, Holdgate GA, Green LR: Epidermal growth factor receptor tyrosine kinase. Investigation of catalytic mechanism, structure-based searching and discovery of a potent inhibitor. Biochem Pharmacol 48:659–666, 1994.
Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 56:55–63, 1983.
Posner I, Engel M, Levitzki A: Kinetic model of the epidermal growth factor (EGF)-receptor tyrosine kinase and a possible mechanism of its activation by EGF. J Biol Chem 267:20638–20647, 1992.
Levitzki A: Tyrphostins — potential antiproliferative agents and novel molecular tools. Biochem Pharmacol 40:913–918, 1990.
Thompson AM, Rewcastle GW, Tercel M, Dobrusin EM, Fry DW, Kraker AJ, Denny WA: Tyrosine kinase inhibitors. 1. Structure-activity relationships for inhibition of epidermal growth factor receptor tyrosine kinase activity by 2,4-dihydro-2-thioxo-1H-indole-3-alkanoic acids and 2,2'-dithiobis(1H-indole-3-alkanoic acids). J Med Chem 36:2459–2469, 1993.
Buchdunger E, Trinks U, Mett H, Regenass U, Muller M, Meyer T, McGlynn E, Pinna LA, Traxler P, Lydon NB: 4,5-dianilinophthalimide: a protein-tyrosine kinase inhibitor with selectivity for the epidermal growth factor receptor signal transduction pathway and potentin vivo antitumor activity. Proc Natl Acad Sci USA 91:2334–2338, 1994.
Fry DW, Kraker AJ, McMichael A, Ambroso LA, Nelson JM, Leopold WR, Connors RW, Bridges AJ: A specific inhibitor of epidermal growth factor receptor tyrosine kinase. Science 265:1093–1095, 1994.
Osherov N, Gazit A, Gilon C, Levitzki A: Selective inhibition of the epidermal growth factor and HER2/Neu receptors by tyrphostins. J Biol Chem 268: 11134–11142, 1993.
Vignon F, Bouton M-M, Rochefort H: Antiestrogens inhibit the mitogenic effect of growth factors on breast cancer cells in the total absence of estrogens. Biochem Biophys Res Commun 146:1502–1508, 1987.
Koga M, Sutherland RL: Epidermal growth factor partially reverses the inhibitory effects of antioestrogens on T47D human breast cancer cell growth. Biochem Biophys Res Commun 146:739–747, 1987.
Wakeling AE, Newboult E, Peters SW: Effects of antioestrogens on the proliferation of MCF-7 human breast cancer cells. J Molec Endocr 2:225–234, 1989.
Ignar-Trowbridge DM, Nelson KG, Bidwell MC, Curtis SW, Washburn TF, McLachlan JA, Korach KS: Coupling of dual signalling pathways: Epidermal growth factor action involves the estrogen receptor. Proc Natl Acad Sci USA 89:4658–4662, 1992.
Newton CJ, Buric R, Trapp T, Brockmeier S, Pagotto U, Stalla GK: The unliganded estrogen receptor (ER) transduces growth factor signals. J Steroid Biochem Mol Biol 48:481–486, 1994.
Reddy KB, Mangold GL, Tandon AK, Yoneda T, Mundy GR, Zilberstein A, Osborne CK: Inhibition of breast cancer cell growthin vitro by a tyrosine kinase inhibitor. Cancer Res 52:3636–3641, 1992.
Yoneda T, Lyall RM, Alsina MM, Persons PE, Spada AP, Levitzki A, Zilberstein A, Mundy GR: The antiproliferative effects of tyrosine kinase inhibitors tyrphostins on a human squamous cell carcinomain vitro and in nude mice. Cancer Res 51:4430–4435, 1991.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wakeling, A.E., Barker, A.J., Davies, D.H. et al. Specific inhibition of epidermal growth factor receptor tyrosine kinase by 4-anilinoquinazolines. Breast Cancer Res Tr 38, 67–73 (1996). https://doi.org/10.1007/BF01803785
Issue Date:
DOI: https://doi.org/10.1007/BF01803785