Jackson D S, Fraser S A, Ni L M, Kam C M, Winkler U, Johnson D A, Froelich C J, Hudig D, Powers J C
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia 30332-0400, USA.
J Med Chem. 1998 Jun 18;41(13):2289-301. doi: 10.1021/jm970543s.
Thirty-six new amino acid and peptidyl diphenyl phosphonate esters were synthesized and evaluated to identify potent and selective inhibitors for four trypsin-like proteases: lymphocyte granzymes A and K, human mast cell tryptase, and pancreatic trypsin. Among five Cbz derivatives of Lys and Arg homologues, Z-(4-AmPhe)P(OPh)2 is the most potent inhibitor for granzyme A, and Z-LysP(OPh)2 is the best inhibitor for granzyme K, mast tryptase, and trypsin. The amidino P1 residue D,L-(4-AmPhGly)P(OPh)2 was utilized in a series of compounds with several different N-protecting groups and systematic substitutions at P2 in Cbz-AA derivatives and at P3 in Cbz-AA-Ala derivatives. Generally, these phosphonates inhibit granzyme A and trypsin more potently than granzyme K and tryptase. The P2 Thr and Ala dipeptide phosphonates, Cbz-AA-(4-AmPhGly)P(OPh)2, are the most potent inhibitors for granzyme A, and Cbz-Thr-(4-AmPhGly)P(OPh)2 (kobs/[I] = 2220 M-1 s-1) was quite specific with much lower inhibition rates for granzyme K and trypsin (kobs/[I] = 3 and 97 M-1 s-1, respectively) and no inhibition with tryptase. The most effective inhibitor of granzyme A was Ph-SO2-Gly-Pro-(4-AmPhGly)P(OPh)2 with a second-order rate constant of 3650 M-1 s-1. The most potent inhibitor for granzyme K was 3, 3-diphenylpropanoyl-Pro-(4-AmPhGly)P(OPh)2 with a kobs/[I] = 1830 M-1 s-1; all other phosphonates inhibited granzyme K weakly (kobs/[I] < 60 M-1 s-1). Human mast cell tryptase was inhibited slowly by these phosphonates with Cbz-LysP(OPh)2 as the best inhibitor (kobs/[I] = 89 M-1 s-1). The overall results suggest that scaffolds of Phe-Thr-(4-AmPhe) and Phe-Pro-Lys will be useful to create selective phosphonate inhibitors for granzymes A and K, respectively, and that P4 substituents offer opportunities to further enhance selectivity and reactivity.
合成并评估了36种新的氨基酸和肽基二苯基膦酸酯，以鉴定四种类胰蛋白酶的强效和选择性抑制剂：淋巴细胞颗粒酶A和K、人肥大细胞类胰蛋白酶和胰蛋白酶。在赖氨酸和精氨酸同系物的五种Cbz衍生物中，Z-(4-氨基苯丙氨酸)P(OPh)₂是颗粒酶A最有效的抑制剂，Z-赖氨酸P(OPh)₂是颗粒酶K、肥大细胞类胰蛋白酶和胰蛋白酶的最佳抑制剂。脒基P1残基D,L-(4-氨基苯甘氨酸)P(OPh)₂被用于一系列具有不同N-保护基的化合物中，并在Cbz-氨基酸衍生物的P2位和Cbz-氨基酸-丙氨酸衍生物的P3位进行系统取代。一般来说，这些膦酸酯对颗粒酶A和胰蛋白酶的抑制作用比对颗粒酶K和类胰蛋白酶更强。P2位苏氨酸和丙氨酸二肽膦酸酯Cbz-氨基酸-(4-氨基苯甘氨酸)P(OPh)₂是颗粒酶A最有效的抑制剂，Cbz-苏氨酸-(4-氨基苯甘氨酸)P(OPh)₂（kobs/[I]=2220 M⁻¹ s⁻¹）具有很高的特异性，对颗粒酶K和胰蛋白酶的抑制率低得多（分别为kobs/[I]=3和97 M⁻¹ s⁻¹），对类胰蛋白酶无抑制作用。颗粒酶A最有效的抑制剂是苯磺酰基-甘氨酸-脯氨酸-(4-氨基苯甘氨酸)P(OPh)₂，二级速率常数为3650 M⁻¹ s⁻¹。颗粒酶K最有效的抑制剂是3,3-二苯基丙酰基-脯氨酸-(4-氨基苯甘氨酸)P(OPh)₂，kobs/[I]=1830 M⁻¹ s⁻¹；所有其他膦酸酯对颗粒酶K的抑制作用较弱（kobs/[I]<60 M⁻¹ s⁻¹）。这些膦酸酯对人肥大细胞类胰蛋白酶的抑制作用缓慢，Cbz-赖氨酸P(OPh)₂是最佳抑制剂（kobs/[I]=89 M⁻¹ s⁻¹）。总体结果表明，苯丙氨酸-苏氨酸-(4-氨基苯丙氨酸)和苯丙氨酸-脯氨酸-赖氨酸支架分别有助于创建颗粒酶A和K的选择性膦酸酯抑制剂，并且P4取代基提供了进一步提高选择性和反应性的机会。
