9-羟基氮杂芴及其在凝血酶抑制剂中的应用。

9-hydroxyazafluorenes and their use in thrombin inhibitors.

作者信息

Stauffer Kenneth J, Williams Peter D, Selnick Harold G, Nantermet Philippe G, Newton Christina L, Homnick Carl F, Zrada Matthew M, Lewis S Dale, Lucas Bobby J, Krueger Julie A, Pietrak Beth L, Lyle Elizabeth A, Singh Rominder, Miller-Stein Cynthia, White Rebecca B, Wong Bradley, Wallace Audrey A, Sitko Gary R, Cook Jacquelyn J, Holahan Marie A, Stranieri-Michener Maria, Leonard Yvonne M, Lynch Joseph J, McMasters Daniel R, Yan Youwei

机构信息

Department of Medicinal Chemistry, Biological Chemistry, Pharmacology, Drug Metabolism, and Molecular Systems, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.

出版信息

J Med Chem. 2005 Apr 7;48(7):2282-93. doi: 10.1021/jm049423s.

DOI:10.1021/jm049423s

PMID:15801822

Abstract

Optimization of a previously reported thrombin inhibitor, 9-hydroxy-9-fluorenylcarbonyl-l-prolyl-trans-4-aminocyclohexylmethylamide (1), by replacing the aminocyclohexyl P1 group provided a new lead structure, 9-hydroxy-9-fluorenylcarbonyl-l-prolyl-2-aminomethyl-5-chlorobenzylamide (2), with improved potency (K(i) = 0.49 nM for human thrombin, 2x APTT = 0.37 microM in human plasma) and pharmacokinetic properties (F = 39%, iv T(1/2) = 13 h in dogs). An effective strategy for reducing plasma protein binding of 2 and improving efficacy in an in vivo thrombosis model in rats was to replace the lipophilic fluorenyl group in P3 with an azafluorenyl group. Systematic investigation of all possible azafluorenyl P3 isomers and azafluorenyl-N-oxide analogues of 2 led to the identification of an optimal compound, 3-aza-9-hydroxyfluoren-9(R)-ylcarbonyl-l-prolyl-2-aminomethyl-5-chlorobenzylamide (19b), with high potency (K(i) = 0.40 nM, 2x APTT = 0.18 microM), excellent pharmacokinetic properties (F = 55%, T(1/2) = 14 h in dogs), and complete efficacy in the in vivo thrombosis model in rats (inhibition of FeCl(3)-induced vessel occlusions in six of six rats receiving an intravenous infusion of 10 microg/kg/min of 19b). The stereochemistry of the azafluorenyl group in 19b was determined by X-ray crystallographic analysis of its N-oxide derivative (23b) bound in the active site of human thrombin.

摘要

通过取代氨环己基P1基团对先前报道的凝血酶抑制剂9-羟基-9-芴基羰基-L-脯氨酰-反式-4-氨基环己基甲酰胺（1）进行优化，得到了一种新的先导结构9-羟基-9-芴基羰基-L-脯氨酰-2-氨基甲基-5-氯苄酰胺（2），其活性（对人凝血酶的K(i)=0.49 nM，在人血浆中2倍活化部分凝血活酶时间（APTT）=0.37 microM）和药代动力学性质（犬体内的生物利用度F=39%，静脉注射半衰期T(1/2)=13小时）均得到改善。降低2的血浆蛋白结合率并提高其在大鼠体内血栓形成模型中的疗效的有效策略是用氮杂芴基取代P3中的亲脂性芴基。对2的所有可能的氮杂芴基P3异构体和氮杂芴基-N-氧化物类似物进行系统研究，得到了一种最佳化合物3-氮杂-9-羟基芴-9(R)-基羰基-L-脯氨酰-2-氨基甲基-5-氯苄酰胺（19b），其活性高（K(i)=0.40 nM，2倍APTT=0.18 microM），药代动力学性质优异（犬体内生物利用度F=55%，半衰期T(½)=14小时），并且在大鼠体内血栓形成模型中具有完全疗效（在接受10 microg/kg/min的19b静脉输注的6只大鼠中有6只抑制了FeCl(3)诱导的血管闭塞）。通过对结合于人凝血酶活性位点的其N-氧化物衍生物（23b）进行X射线晶体学分析，确定了19b中氮杂芴基的立体化学。