PREPARATION AND ASSAY OF NEW SERINE PROTEASE INHIBITORS

硼酸是凝血酶和其他凝血酶抑制剂的过渡态类似物。 丝氨酸水解酶 在70年代，作者和其他人研究了简单的 作为胰凝乳蛋白酶抑制剂的烷基和芳基硼酸 subtilison/1.3. 在80年代，酰基硼氨基酸，如乙酰基- 硼苯丙氨酸，其中硼酸基团取代羧基 一组氨基酸，由Matteson等人制备，和研究 由Lienhard & Philipp/4.5领导的小组使用糜蛋白酶和枯草杆菌蛋白酶。 最近，已经制备了肽硼酸，其显示出 考虑作为潜在治疗剂所需的特异性。 组 由Kettner/6和Katzenellenbogen/7领导制备了寡肽硼酸 对弹性蛋白酶和胰凝乳蛋白酶具有特异性的酸。 Kettner和同事们在 杜邦公司在弹性蛋白酶抑制剂/6方面做了大量的工作， 肺气肿 Kettner的小组/8和这个小组/9.10已经准备了具体的 凝血酶抑制剂。 作者和同事制备的凝血酶抑制剂/9.10是Z-D- Phe-Pro-甲氧基丙基硼甘氨酸。 这是一个有效的和高度具体的 凝血酶抑制剂。 与其他化合物不同， 例如纤溶酶或纤溶酶原激活剂的酶的活性很弱。 不像 含有硼氢化钠的肽，它对血液没有致命作用 压力 过渡态类似物硼肽抑制剂是 酶结合位点，如过渡态所见。 我们之前的工作 集中于在N-酰基上含有寡肽的硼肽 的方面想 今后的工作将集中在硼肽抑制剂 在抑制剂的C末端侧含有寡肽。 这些将研究酶结合位点的不同区域， 也许会导致生物医学上有趣的抑制剂。 拟议 项目，我们将使用化学合成的肽库， 硼肽，使用Guysen/11的方法。 这种方法将 促进对凝血酶更具特异性的硼肽的产生。 这种方法的成功，它结合了过渡态类似物组 和肽库的产生，可能会导致其他酶 用于研究酶过渡态的有意义的抑制剂 约束力

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.