Probing of primed and unprimed sites of calpains

Design, synthesis and evaluation of epoxysuccinyl-peptide derivatives as selective inhibitors

Levente E. Dókus, Dóra K. Menyhárd, Ágnes Tantos, F. Hudecz, Z. Bánóczi

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Calpains are intracellular cysteine proteases with important physiological functions. Up- or downregulation of their expression can be responsible for several diseases, therefore specific calpain inhibitors may be considered as promising candidates for drug discovery. In this paper we describe the synthesis and characterization of a new class of inhibitors derived from the analysis of amino acid preferences in primed and unprimed sites of calpains by incorporation of l- or d-epoxysuccinyl group (Eps). Amino acids for replacement were chosen by considering the substrate preference of calpain 1 and 2 enzymes. The compounds were characterized by RP-HPLC, amino acid analysis and ESI-MS. Selectivity of the compounds was studied by using calpain 1 and 2; and cathepsin B. We have identified five calpain specific inhibitors with different extent of selectivity. Two of these also exhibited isoform selectivity. Compound NH 2-Thr-Pro-Leu-(d-Eps)-Thr-Pro-Pro-Pro-Ser-NH2 proved to be a calpain 2 enzyme inhibitor with at least 11.8-fold selectivity, while compound NH2-Thr-Pro-Leu-(l-Eps)-Ser-Pro-Pro-Pro-Ser-NH2 possesses calpain 1 enzyme inhibition with at least 4-fold selectivity. The results of molecular modeling calculations suggest that the orientation of the bound inhibitor in the substrate binding cleft is markedly dependent on the stereochemistry of the epoxysuccinyl group.

Original languageEnglish
Pages (from-to)274-280
Number of pages7
JournalEuropean Journal of Medicinal Chemistry
Volume82
DOIs
Publication statusPublished - Jul 23 2014

Fingerprint

Calpain
Derivatives
Peptides
Amino Acids
Cathepsin B
Cysteine Proteases
Enzyme inhibition
Enzyme Inhibitors
Enzymes
Drug Discovery
Stereochemistry
Molecular modeling
Substrates
Protein Isoforms
Up-Regulation
Down-Regulation
High Pressure Liquid Chromatography

Keywords

  • Calpain
  • Enzyme inhibitor
  • Epoxysuccinic acid
  • Epoxysuccinyl-peptide

ASJC Scopus subject areas

  • Drug Discovery
  • Organic Chemistry
  • Pharmacology
  • Medicine(all)

Cite this

Probing of primed and unprimed sites of calpains : Design, synthesis and evaluation of epoxysuccinyl-peptide derivatives as selective inhibitors. / Dókus, Levente E.; Menyhárd, Dóra K.; Tantos, Ágnes; Hudecz, F.; Bánóczi, Z.

In: European Journal of Medicinal Chemistry, Vol. 82, 23.07.2014, p. 274-280.

Research output: Contribution to journalArticle

@article{8fdf1b035f6745a194db7d75d89ee859,
title = "Probing of primed and unprimed sites of calpains: Design, synthesis and evaluation of epoxysuccinyl-peptide derivatives as selective inhibitors",
abstract = "Calpains are intracellular cysteine proteases with important physiological functions. Up- or downregulation of their expression can be responsible for several diseases, therefore specific calpain inhibitors may be considered as promising candidates for drug discovery. In this paper we describe the synthesis and characterization of a new class of inhibitors derived from the analysis of amino acid preferences in primed and unprimed sites of calpains by incorporation of l- or d-epoxysuccinyl group (Eps). Amino acids for replacement were chosen by considering the substrate preference of calpain 1 and 2 enzymes. The compounds were characterized by RP-HPLC, amino acid analysis and ESI-MS. Selectivity of the compounds was studied by using calpain 1 and 2; and cathepsin B. We have identified five calpain specific inhibitors with different extent of selectivity. Two of these also exhibited isoform selectivity. Compound NH 2-Thr-Pro-Leu-(d-Eps)-Thr-Pro-Pro-Pro-Ser-NH2 proved to be a calpain 2 enzyme inhibitor with at least 11.8-fold selectivity, while compound NH2-Thr-Pro-Leu-(l-Eps)-Ser-Pro-Pro-Pro-Ser-NH2 possesses calpain 1 enzyme inhibition with at least 4-fold selectivity. The results of molecular modeling calculations suggest that the orientation of the bound inhibitor in the substrate binding cleft is markedly dependent on the stereochemistry of the epoxysuccinyl group.",
keywords = "Calpain, Enzyme inhibitor, Epoxysuccinic acid, Epoxysuccinyl-peptide",
author = "D{\'o}kus, {Levente E.} and Menyh{\'a}rd, {D{\'o}ra K.} and {\'A}gnes Tantos and F. Hudecz and Z. B{\'a}n{\'o}czi",
year = "2014",
month = "7",
day = "23",
doi = "10.1016/j.ejmech.2014.05.058",
language = "English",
volume = "82",
pages = "274--280",
journal = "European Journal of Medicinal Chemistry",
issn = "0223-5234",
publisher = "Elsevier Masson SAS",

}

TY - JOUR

T1 - Probing of primed and unprimed sites of calpains

T2 - Design, synthesis and evaluation of epoxysuccinyl-peptide derivatives as selective inhibitors

AU - Dókus, Levente E.

AU - Menyhárd, Dóra K.

AU - Tantos, Ágnes

AU - Hudecz, F.

AU - Bánóczi, Z.

PY - 2014/7/23

Y1 - 2014/7/23

N2 - Calpains are intracellular cysteine proteases with important physiological functions. Up- or downregulation of their expression can be responsible for several diseases, therefore specific calpain inhibitors may be considered as promising candidates for drug discovery. In this paper we describe the synthesis and characterization of a new class of inhibitors derived from the analysis of amino acid preferences in primed and unprimed sites of calpains by incorporation of l- or d-epoxysuccinyl group (Eps). Amino acids for replacement were chosen by considering the substrate preference of calpain 1 and 2 enzymes. The compounds were characterized by RP-HPLC, amino acid analysis and ESI-MS. Selectivity of the compounds was studied by using calpain 1 and 2; and cathepsin B. We have identified five calpain specific inhibitors with different extent of selectivity. Two of these also exhibited isoform selectivity. Compound NH 2-Thr-Pro-Leu-(d-Eps)-Thr-Pro-Pro-Pro-Ser-NH2 proved to be a calpain 2 enzyme inhibitor with at least 11.8-fold selectivity, while compound NH2-Thr-Pro-Leu-(l-Eps)-Ser-Pro-Pro-Pro-Ser-NH2 possesses calpain 1 enzyme inhibition with at least 4-fold selectivity. The results of molecular modeling calculations suggest that the orientation of the bound inhibitor in the substrate binding cleft is markedly dependent on the stereochemistry of the epoxysuccinyl group.

AB - Calpains are intracellular cysteine proteases with important physiological functions. Up- or downregulation of their expression can be responsible for several diseases, therefore specific calpain inhibitors may be considered as promising candidates for drug discovery. In this paper we describe the synthesis and characterization of a new class of inhibitors derived from the analysis of amino acid preferences in primed and unprimed sites of calpains by incorporation of l- or d-epoxysuccinyl group (Eps). Amino acids for replacement were chosen by considering the substrate preference of calpain 1 and 2 enzymes. The compounds were characterized by RP-HPLC, amino acid analysis and ESI-MS. Selectivity of the compounds was studied by using calpain 1 and 2; and cathepsin B. We have identified five calpain specific inhibitors with different extent of selectivity. Two of these also exhibited isoform selectivity. Compound NH 2-Thr-Pro-Leu-(d-Eps)-Thr-Pro-Pro-Pro-Ser-NH2 proved to be a calpain 2 enzyme inhibitor with at least 11.8-fold selectivity, while compound NH2-Thr-Pro-Leu-(l-Eps)-Ser-Pro-Pro-Pro-Ser-NH2 possesses calpain 1 enzyme inhibition with at least 4-fold selectivity. The results of molecular modeling calculations suggest that the orientation of the bound inhibitor in the substrate binding cleft is markedly dependent on the stereochemistry of the epoxysuccinyl group.

KW - Calpain

KW - Enzyme inhibitor

KW - Epoxysuccinic acid

KW - Epoxysuccinyl-peptide

UR - http://www.scopus.com/inward/record.url?scp=84902124324&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84902124324&partnerID=8YFLogxK

U2 - 10.1016/j.ejmech.2014.05.058

DO - 10.1016/j.ejmech.2014.05.058

M3 - Article

VL - 82

SP - 274

EP - 280

JO - European Journal of Medicinal Chemistry

JF - European Journal of Medicinal Chemistry

SN - 0223-5234

ER -