PREPARATION AND ASSAY OF NEW SERINE PROTEASE INHIBITORS

硼酸是凝血酶和其他化合物的过渡态类似物 丝氨酸水解酶。七十年代的S，笔者等人研究简单 作为胰凝乳酶抑制剂的烷基和芳基硼酸& 枯草杆菌/1.3。在80年代的S，酰基硼氨基酸，如乙酰基- 硼苯丙氨酸，其中硼酸基取代了羧基 由Matteson等人制备，并对其进行了研究 由Lienard&Philipp/4.5领导的小组使用糜蛋白酶和枯草杆菌酶。 最近，已经制备了多肽硼酸，它显示了 被认为是潜在的治疗方法所需的特异性。群组 在Kettner/6和Katzenellenbogen/7的领导下，制备了寡肽硼酸 弹性蛋白酶和胰凝乳酶所特有的酸。Kettner和同事们 杜邦对弹性酶抑制剂/6进行了广泛的研究，与 肺气肿。凯特纳的GROUP/8和这个GROUP/9.10已经准备了具体的 凝血酶抑制剂。 本文作者和同事研制的凝血酶抑制物为Z-D- Phe-前甲氧基丙基硼甘氨酸。这是一种强有力的、高度具体的 凝血酶抑制剂。与其他化合物不同的是，它能抑制血栓溶解 纤溶酶或纤溶酶原激活剂等酶仅微弱。不像 含硼精氨酸的多肽，对血液没有致死作用 压力。 过渡态类似物硼多肽抑制剂是一种特异性的探针 如过渡态所见的酶结合部位。我们之前的工作是 集中在N-酰基上含有寡肽的硼多肽 边上。未来在这一领域的工作将集中在硼肽抑制剂上。 在抑制剂的C-末端含有寡肽。 他们将研究酶结合部位的不同区域，还 可能会导致产生生物医学上有趣的抑制剂。在建议的 项目中，我们将使用化学合成的多肽库 使用Guysen/11的方法，这种方法将 促进更针对凝血酶的硼多肽的产生。 这种方法成功地将过渡状态模拟组结合在一起 而多肽库的产生，可能会导致其他酶的产生 在酶过渡态研究中有兴趣的抑制剂 有约束力的。

Boronic acids are transition state analog inhibitors of thrombin and other serine hydrolases. In the 70's, this author and others studied simple alkyl and aryl boronic acids as inhibitors of chymotrypsin & subtilison/1.3. In the 80's, acylboroamino acids, such as acetyl- borophenylalanine, where the boronic acid group replaces the carboxyl group of an amino acid, were prepared by Matteson et al., and studied using chymotrypsin and subtilisin by groups led by Lienhard & Philipp/4.5. More recently, peptide boronic acids have been prepared that show the specificity required for consideration as potential therapeutics. Groups led by Kettner/6 and Katzenellenbogen/7 have prepared oligopeptide boronic acids specific to elastase and chymotrypsin. Kettner and coworkers at DuPont did extensive work on the elastase inhibitors/6 in connection with emphysema. Kettner's group/8 and this group/9.10 have prepared specific inhibitors of thrombin. The thrombin inhibitor prepared by this author and coworkers/9.10 is Z-D- Phe-Pro-methoxypropylboroglycine. This is a potent and highly specific inhibitor of thrombin. Unlike other compounds, it inhibits thrombolytic enzymes such as plasmin or plasminogen activators only weakly. Unlike boroarginine-containing peptides, it has no lethal effect on blood pressure. Transition state analog boropeptide inhibitors are specific probes of enzyme binding sites as seen for transition states. Our previous work has concentrated on boropeptides that contain the oligopeptide on the N-acyl side. Future work in this area will concentrate on boropeptide inhibitors that contain the oligopeptide on the C-terminal side of the inhibitor. These will study a different area of the enzyme binding site and also perhaps lead to biomedically interesting inhibitors. In the proposed project, we will use chemically synthesized peptide libraries of boropeptides, using the approach of Guysen/11. This approach will facilitate the production of boropeptides more specific to thrombin. Success in this approach, which combines transition state analog groups and the generation of peptide libraries, may lead to other enzyme inhibitors interesting for their use in studying enzyme-transition state binding.