Hennequin L F, Thomas A P, Johnstone C, Stokes E S, Plé P A, Lohmann J J, Ogilvie D J, Dukes M, Wedge S R, Curwen J O, Kendrew J, Lambert-van der Brempt C
AstraZeneca, Zeneca Pharma, Centre de Recherches, Z.I. La Pompelle, B.P. 1050, Chemin de Vrilly, 51689 Reims Cedex 2, France.
J Med Chem. 1999 Dec 30;42(26):5369-89. doi: 10.1021/jm990345w.
A series of substituted 4-anilinoquinazolines and related compounds were synthesized as potential inhibitors of vascular endothelial growth factor (VEGF) receptor (Flt and KDR) tyrosine kinase activity. Enzyme screening indicated that a narrow structure-activity relationship (SAR) existed for the bicyclic ring system, with quinazolines, quinolines, and cinnolines having activity and with quinazolines and quinolines generally being preferred. Substitution of the aniline was investigated and clearly indicated that small lipophilic substituents such as halogens or methyl were preferred at the C-4' position. Small substituents such as hydrogen and fluorine are preferred at the C-2' position. Introduction of a hydroxyl group at the meta position of the aniline produced the most potent inhibitors of Flt and KDR tyrosine kinases activity with IC(50) values in the nanomolar range (e.g. 10, 12, 13, 16, and 18). Investigation of the quinazoline C-6 and C-7 positions indicates that a large range of substituents are tolerated at C-7, whereas variation at the C-6 is more restricted. At C-7, neutral, basic, and heteroaromatic side chains led to very potent compounds, as illustrated by the methoxyethoxy derivative 13 (IC(50) < 2 nM). Our inhibitors proved to be very selective inhibitors of Flt and KDR tyrosine kinase activity when compared to that associated with the FGF receptor (50- to 3800-fold). Observed enzyme profiles translated well with respect to potency and selectivity for inhibition of growth factor stimulated proliferation of human umbilical vein endothelial cells (HUVECs). Oral administration of selected compounds to mice produced total plasma levels 6 h after dosing of between 3 and 49 microM. In vivo efficacy was demonstrated in a rat uterine oedema assay where significant activity was achieved at 60 mg/kg with the meta hydroxy anilinoquinazoline 10. Inhibition of growth of human tumors in athymic mice has also been demonstrated: compound 34 inhibited the growth of established Calu-6 lung carcinoma xenograft by 75% (P < 0.001, one tailed t-test) following daily oral administration of 100 mg/kg for 21 days.
合成了一系列取代的4-苯胺基喹唑啉及其相关化合物，作为血管内皮生长因子（VEGF）受体（Flt和KDR）酪氨酸激酶活性的潜在抑制剂。酶筛选表明，双环系统存在较窄的构效关系（SAR），喹唑啉、喹啉和噌啉具有活性，其中喹唑啉和喹啉通常更受青睐。对苯胺的取代进行了研究，结果清楚地表明，在C-4'位优选小的亲脂性取代基，如卤素或甲基。在C-2'位优选氢和氟等小取代基。在苯胺的间位引入羟基产生了Flt和KDR酪氨酸激酶活性的最有效抑制剂，其IC(50)值在纳摩尔范围内（例如10、12、13、16和18）。对喹唑啉C-6和C-7位的研究表明，C-7位可耐受大范围的取代基，而C-6位的变化则更受限制。在C-7位，中性、碱性和杂芳族侧链产生了非常有效的化合物，甲氧基乙氧基衍生物13（IC(50) < 2 nM）就说明了这一点。与FGF受体相关的活性相比，我们的抑制剂被证明是Flt和KDR酪氨酸激酶活性的非常选择性的抑制剂（50至3800倍）。观察到的酶谱在抑制生长因子刺激的人脐静脉内皮细胞（HUVECs）增殖的效力和选择性方面表现良好。给小鼠口服选定的化合物后6小时，血浆总水平在3至49 microM之间。在大鼠子宫水肿试验中证明了体内疗效，其中间羟基苯胺基喹唑啉10在60 mg/kg时具有显著活性。在无胸腺小鼠中也证明了对人肿瘤生长的抑制作用：化合物34在每日口服100 mg/kg，持续21天后，抑制已建立的Calu-6肺癌异种移植瘤的生长达75%（P < 0.001，单尾t检验）。
