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六氟亮氨酸和三氟异亮氨酸对蛋白酶消化的位置依赖性影响

Position-dependent impact of hexafluoroleucine and trifluoroisoleucine on protease digestion.

作者信息

Huhmann Susanne, Stegemann Anne-Katrin, Folmert Kristin, Klemczak Damian, Moschner Johann, Kube Michelle, Koksch Beate

机构信息

Department of Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 3, 14195 Berlin, Germany.

出版信息

Beilstein J Org Chem. 2017 Dec 22;13:2869-2882. doi: 10.3762/bjoc.13.279. eCollection 2017.

DOI:10.3762/bjoc.13.279
PMID:29564015
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5753150/
Abstract

Rapid digestion by proteases limits the application of peptides as therapeutics. One strategy to increase the proteolytic stability of peptides is the modification with fluorinated amino acids. This study presents a systematic investigation of the effects of fluorinated leucine and isoleucine derivatives on the proteolytic stability of a peptide that was designed to comprise substrate specificities of different proteases. Therefore, leucine, isoleucine, and their side-chain fluorinated variants were site-specifically incorporated at different positions of this peptide resulting in a library of 13 distinct peptides. The stability of these peptides towards proteolysis by α-chymotrypsin, pepsin, proteinase K, and elastase was studied, and this process was followed by an FL-RP-HPLC assay in combination with mass spectrometry. In a few cases, we observed an exceptional increase in proteolytic stability upon introduction of the fluorine substituents. The opposite phenomenon was observed in other cases, and this may be explained by specific interactions of fluorinated residues with the respective enzyme binding sites. Noteworthy is that 5,5,5-trifluoroisoleucine is able to significantly protect peptides from proteolysis by all enzymes included in this study when positioned N-terminal to the cleavage site. These results provide valuable information for the application of fluorinated amino acids in the design of proteolytically stable peptide-based pharmaceuticals.

摘要

蛋白酶的快速消化作用限制了肽作为治疗剂的应用。提高肽的蛋白水解稳定性的一种策略是用氟化氨基酸进行修饰。本研究系统地研究了氟化亮氨酸和异亮氨酸衍生物对一种设计为包含不同蛋白酶底物特异性的肽的蛋白水解稳定性的影响。因此，亮氨酸、异亮氨酸及其侧链氟化变体被位点特异性地掺入该肽的不同位置，从而产生了一个包含13种不同肽的文库。研究了这些肽对α-胰凝乳蛋白酶、胃蛋白酶、蛋白酶K和弹性蛋白酶蛋白水解的稳定性，并通过FL-RP-HPLC分析结合质谱对这一过程进行了跟踪。在少数情况下，我们观察到引入氟取代基后蛋白水解稳定性异常增加。在其他情况下观察到相反的现象，这可能是由于氟化残基与各自酶结合位点的特异性相互作用所致。值得注意的是，当5,5,5-三氟异亮氨酸位于切割位点的N端时，能够显著保护肽不被本研究中包括的所有酶进行蛋白水解。这些结果为氟化氨基酸在蛋白水解稳定的肽基药物设计中的应用提供了有价值的信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/1b8ce45480cf/Beilstein_J_Org_Chem-13-2869-g011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/c83d6fa7e383/Beilstein_J_Org_Chem-13-2869-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/cf9a5b567c5f/Beilstein_J_Org_Chem-13-2869-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/4c3f5b61b9db/Beilstein_J_Org_Chem-13-2869-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/45fef1cf0d7a/Beilstein_J_Org_Chem-13-2869-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/f5574e8fb0c8/Beilstein_J_Org_Chem-13-2869-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/7033fea592ca/Beilstein_J_Org_Chem-13-2869-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/c5224d467d8a/Beilstein_J_Org_Chem-13-2869-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/17ba02f164b4/Beilstein_J_Org_Chem-13-2869-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/ff44d92578bb/Beilstein_J_Org_Chem-13-2869-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/1b8ce45480cf/Beilstein_J_Org_Chem-13-2869-g011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/c83d6fa7e383/Beilstein_J_Org_Chem-13-2869-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/cf9a5b567c5f/Beilstein_J_Org_Chem-13-2869-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/4c3f5b61b9db/Beilstein_J_Org_Chem-13-2869-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/45fef1cf0d7a/Beilstein_J_Org_Chem-13-2869-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/f5574e8fb0c8/Beilstein_J_Org_Chem-13-2869-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/7033fea592ca/Beilstein_J_Org_Chem-13-2869-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/c5224d467d8a/Beilstein_J_Org_Chem-13-2869-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/17ba02f164b4/Beilstein_J_Org_Chem-13-2869-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/ff44d92578bb/Beilstein_J_Org_Chem-13-2869-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f1e/5753150/1b8ce45480cf/Beilstein_J_Org_Chem-13-2869-g011.jpg

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2
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Acc Chem Res. 2017 Sep 19;50(9):2093-2103. doi: 10.1021/acs.accounts.7b00226. Epub 2017 Aug 12.
3
Oral Administration of Peptide-Based Drugs: Beyond Lipinski's Rule.基于肽的药物的口服给药：超越Lipinski规则
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Environ Sci Technol. 2023 Jul 4;57(26):9762-9772. doi: 10.1021/acs.est.3c01240. Epub 2023 Jun 21.
4
Hexafluoroisobutylation of enolates through a tandem elimination/allylic shift/hydrofluorination reaction.通过串联消除/烯丙基迁移/氢氟化反应实现烯醇盐的六氟异丁基化。
Chem Sci. 2022 Jul 20;13(33):9507-9514. doi: 10.1039/d2sc02871a. eCollection 2022 Aug 24.
5
Fluorine-induced polarity increases inhibitory activity of BPTI towards chymotrypsin.氟诱导的极性增加了BPTI对胰凝乳蛋白酶的抑制活性。
RSC Chem Biol. 2022 May 19;3(6):773-782. doi: 10.1039/d2cb00018k. eCollection 2022 Jun 8.
6
Fluorinated peptide biomaterials.氟化肽生物材料。
Pept Sci (Hoboken). 2021 Mar;113(2). doi: 10.1002/pep2.24184. Epub 2020 Jul 28.
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4
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6
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9
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