Rational design of thioamide peptides as selective inhibitors of cysteine protease cathepsin L.
Rational design of thioamide peptides as selective inhibitors of cysteine protease cathepsin L.
复制标题
硫酰胺肽作为半胱氨酸蛋白酶组织蛋白酶L.的选择性抑制剂的合理设计。
DOI：
10.1039/d1sc00785h
复制
发表时间：
2021-08-18
期刊：
影响因子：
8.4
通讯作者：
Petersson EJ
中科院分区：
文献类型：
--
作者：
Phan HAT;Giannakoulias SG;Barrett TM;Liu C;Petersson EJ
组织蛋白酶L（Cts L）是一种广泛表达的内体半胱氨酸蛋白酶，其异常水平与许多疾病如癌症和糖尿病有关。值得注意的是，由于最近发现Cts L在SARS-CoV-2进入宿主细胞中发挥关键作用，因此它已被确定为治疗COVID-19的潜在靶点。然而，目前还没有临床批准的特异性Cts L抑制剂，因为获得针对许多高度同源的组织蛋白酶家族半胱氨酸蛋白酶的特异性通常具有挑战性。基于肽的试剂通常是有前途的蛋白酶抑制剂，因为它们提供高选择性和效力，但不幸的是在体内会发生降解。硫代酰胺取代（肽骨架中的单原子O-至-S修饰）已被证明可改善肽的蛋白水解稳定性，从而解决该问题。利用这种方法，我们在本文中证明，良好的肽基底物可以通过肽骨架中的单个硫代酰胺取代转化为亚微摩尔的Cts L抑制剂。我们已经设计并扫描了几种硫代酰胺稳定的肽支架，其中一种肽RS 1A被所有五种组织蛋白酶（Cts L、Cts V、Cts K、Cts S和Cts B）稳定以抵抗蛋白水解，同时抑制Cts L，对其他组织蛋白酶具有>25倍的特异性。我们进一步表明，这种稳定的RS 1A肽可以抑制人肝癌裂解物中的Cts L（IC 50 = 19 μM）。我们的研究表明，可以合理地设计一个稳定的，具体的肽基蛋白酶抑制剂的战略布局的硫代酰胺，并重申了这种单原子修饰的工具箱中的肽为基础的合理的药物设计的地方。侧链和骨架修饰对组织蛋白酶（Cts）L、V、K、S和B活性的影响的信息用于设计对所有Cts惰性并选择性抑制Cts L的硫代酰胺肽。
Aberrant levels of cathepsin L (Cts L), a ubiquitously expressed endosomal cysteine protease, have been implicated in many diseases such as cancer and diabetes. Significantly, Cts L has been identified as a potential target for the treatment of COVID-19 due to its recently unveiled critical role in SARS-CoV-2 entry into the host cells. However, there are currently no clinically approved specific inhibitors of Cts L, as it is often challenging to obtain specificity against the many highly homologous cathepsin family cysteine proteases. Peptide-based agents are often promising protease inhibitors as they offer high selectivity and potency, but unfortunately are subject to degradation in vivo. Thioamide substitution, a single-atom O-to-S modification in the peptide backbone, has been shown to improve the proteolytic stability of peptides addressing this issue. Utilizing this approach, we demonstrate herein that good peptidyl substrates can be converted into sub-micromolar inhibitors of Cts L by a single thioamide substitution in the peptide backbone. We have designed and scanned several thioamide stabilized peptide scaffolds, in which one peptide, RS1A, was stabilized against proteolysis by all five cathepsins (Cts L, Cts V, Cts K, Cts S, and Cts B) while inhibiting Cts L with >25-fold specificity against the other cathepsins. We further showed that this stabilized RS1A peptide could inhibit Cts L in human liver carcinoma lysates (IC50 = 19 μM). Our study demonstrates that one can rationally design a stabilized, specific peptidyl protease inhibitor by strategic placement of a thioamide and reaffirms the place of this single-atom modification in the toolbox of peptide-based rational drug design. Information on the effects of sidechain and backbone modification on the activity of cathepsin (Cts) L, V, K, S, and B was used to design a thioamide peptide that is inert to all Cts and selectively inhibits Cts L.
登录
查看更多内容
影响因子：
3.9
作者：
通讯作者：
--
影响因子：
15
作者：
CAMPBELL, P;NASHED, NT
通讯作者：
NASHED, NT
影响因子：
120.1
作者：
Drag, Marcin;Salvesen, Guy S.
通讯作者：
Salvesen, Guy S.
影响因子：
15
作者：
Goldberg JM;Chen X;Meinhardt N;Greenbaum DC;Petersson EJ
通讯作者：
Petersson EJ
影响因子：
3.7
作者：
Dennemaerker, Julia;Lohmueller, Tobias;Reinheckel, Thomas
通讯作者：
Reinheckel, Thomas