Toward an Optimal Joint Recognition of the S<sub>1</sub>‘ Subsites of Endothelin Converting Enzyme-1 (ECE-1), Angiotensin Converting Enzyme (ACE), and Neutral Endopeptidase (NEP)

Inguimbert, Nicolas; Coric, Pascale; Poras, Hervé; Meudal, Hervé; Teffot, Franck; Fournié‐Zaluski, Marie‐Claude; Roques, Bernárd P.

doi:10.1021/jm0005454

Cited by 22 publications

(22 citation statements)

References 31 publications

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“…Diabetes attenuated the acute antihypertrophic effects of selective ACE and neutral endopeptidase inhibitors in isolated hearts, but the long-term antihypertensive and antihypertrophic effects of dual ACE/neutral endopeptidase inhibition are preserved in vivo (31). Given that neutral endopeptidase may also modulate local levels of angiotensin II and ET-1, future therapeutic strategies are likely to include the recently described compounds that inhibit all three key metallopeptidases, neutral endopeptidase, ACE, and endothelin-converting enzyme (40). These may be of particular benefit in diabetic patients, who often require multiple therapies to achieve adequate blood pressure control and thereby reduce cardiovascular risk.…”

Section: Discussionmentioning

confidence: 99%

B-Type Natriuretic Peptide Prevents Acute Hypertrophic Responses in the Diabetic Rat Heart

et al. 2003

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Stimulation of cardiomyocyte guanosine 3,5-cyclic monophosphate (cyclic GMP) via endothelial-derived nitric oxide (NO) is an important mechanism by which bradykinin and ACE inhibitors prevent hypertrophy. Endothelial NO dysfunction and cardiac hypertrophy are morbid features of diabetes not entirely prevented by ACE inhibitors. In cardiomyocyte/endothelial cell cocultures, bradykinin efficacy is abolished by highglucose-induced endothelial NO dysfunction. We now demonstrate that antihypertrophic actions of natriuretic peptides, which stimulate cyclic GMP independently of NO, are preserved in cardiomyocytes despite high-glucose-induced endothelial dysfunction. Further, streptozotocin-induced diabetes significantly impairs the effectiveness of acute antihypertrophic strategies in isolated rat hearts. In hearts from citrate-treated control rats, angiotensin II-stimulated [ 3 H]phenylalanine incorporation and atrial natriuretic peptide and ␤-myosin heavy chain mRNA expression were prevented by B-type natriuretic peptide (BNP), bradykinin, the ACE inhibitor ramiprilat, and the neutral endopeptidase inhibitor candoxatrilat. These antihypertrophic effects were accompanied by increased left ventricular cyclic GMP. In age-matched diabetic hearts, the antihypertrophic and cyclic GMP stimulatory actions of bradykinin, ramiprilat, and candoxatrilat were absent. However, the blunting of hypertrophic markers and accompanying increases in cyclic GMP stimulated by BNP were preserved in diabetes. Thus BNP, which increases cyclic GMP independently of NO, is an important approach to prevent growth in the diabetic myocardium, where endothelium-dependent mechanisms are compromised.

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Section: Discussionmentioning

confidence: 99%

B-Type Natriuretic Peptide Prevents Acute Hypertrophic Responses in the Diabetic Rat Heart

et al. 2003

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“…9B and Table 1). Compounds with nanomolar affinity toward NEP, ACE, and ECE-1 have previously been obtained through rational design (27,28), but their affinities toward ECE-2 have never been investigated. While testing such inhibitors using PL405, compounds 1 and 2 were found to bind ECE-2 with high affinity.…”

Section: Resultsmentioning

confidence: 99%

Identification of an Endothelin-converting Enzyme-2-specific Fluorigenic Substrate and Development of an in Vitro and ex Vivo Enzymatic Assay

Ouimet

Orng

Poras

et al. 2010

Journal of Biological Chemistry

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Endothelin-converting enzyme-2 (ECE-2) is a membranebound zinc-dependent metalloprotease that shares a high degree of sequence homology with ECE-1, but displays an acidic pH optimum characteristic of maturing enzymes acting late in the secretory pathway. Although ECE-2, like ECE-1, can cleave the big endothelin intermediate to produce the vasoconstrictive endothelin peptide, its true physiological function remains to be elucidated, a task that is hampered by the lack of specific tools to study and discriminate ECE-2 from ECE-1, i.e. specific substrates and/or specific inhibitors. To fill this gap, we searched for novel ECE-specific peptide substrates. To this end, peptides derived from the big endothelin intermediate were tested using ECE-1 and ECE-2, leading to the identification of an ECE-1-specific substrate. Moreover, screening of our proprietary fluorigenic peptide Fluofast libraries using ECE-1 and ECE-2 allowed the identification of Ac-SKG-Pya-F-W-Nop-GGK-NH 2 (PL405), as a specific and high affinity ECE-2 substrate. Indeed, ECE-2 cleaved PL405 at the Pya-F amide bond with a specificity constant (k cat /K m ) of 8.1 ؎ 0.9 ؋ 10 3 M ؊1 s ؊1 . Using this novel substrate, we also characterized the first potent (K i ‫؍‬ 7.7 ؎ 0.3 nM) and relatively selective ECE-2 inhibitor and developed a quantitative fluorigenic ECE-2 assay. The assay was used to study the ex vivo ECE-2 activity in wild type and ECE-2 knockout tissues and was found to truly reflect ECE-2 expression patterns. The PL405 assay is thus the first tool to study ECE-2 inhibition using high throughput screening or for ex vivo ECE-2 quantification.Endothelin-converting enzyme-2 (ECE-2) 2 is a zinc-dependant metalloprotease belonging to the M13 family, in which it shares the highest degree of sequence homology with ECE-1 (EC 3.4.24.71) (1, 2). The family is also constituted by neprilysin (NEP, EC 3.4.24.11) (3); damage-induced neuronal endopeptidase, also known as endothelin converting enzyme like (ECEL) (4, 5); the Kell blood group antigen (6); phosphate-regulating neutral endopeptidase on the X chromosome (7); and neprilysin 2 (NEP2) (8 -11). These peptidases are type II integral membrane glycoproteins composed of a small cytosolic tail followed by a transmembrane region and a large C-terminal core that is typically exposed at the cell surface. The active site of the enzyme is located within this last domain and is characterized by two highly conserved zinc-binding consensus motifs HEXXH and EXXXD (where X is any amino acid) (12). The structure of the human ECE-1 enzyme co-crystallized with phosphoramidon, a generic inhibitor of the family, was recently reported (13) showing the conserved "thermolysin-like" arrangement of its catalytic site (14, 15).The endothelin-converting enzymes were first characterized as cleaving the big endothelin inactive intermediate at its Trp 21 -Val 22 bond to produce the highly vasoconstrictive endothelin peptide (1, 2). It was thereafter found that ECE-1 can also hydrolyze smaller peptides in vitro such as big endothel...

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“…The family of zinc metallopeptidases involved in cardiovascular regulation includes angiotensin converting enzyme (ACE), endothelin converting enzyme (ECE-1), and neprilysin (NEP). ACE and ECE-1 are involved in the generation of the strong vasoconstrictors angiotensin II (Ang II) and endothelin I (ET-I) that are derived from the inactive precursors angiotensin I (Ang I) and endothelin, respectively [52], while ET-I is degraded by NEP. There is substantial interest in evolving compounds with synergistic therapeutic profiles in comparison to individual selective inhibitors [52].…”

Section: Zinc-metalloproteases As Drug Targets In Cardiovascular Diseasementioning

confidence: 99%

“…ACE and ECE-1 are involved in the generation of the strong vasoconstrictors angiotensin II (Ang II) and endothelin I (ET-I) that are derived from the inactive precursors angiotensin I (Ang I) and endothelin, respectively [52], while ET-I is degraded by NEP. There is substantial interest in evolving compounds with synergistic therapeutic profiles in comparison to individual selective inhibitors [52]. Since NEP achieves the degradation of ET-I, inhibition of the former results in increased plasma levels of ET-I, thereby limiting the application of the dual NEP/ACE inhibition approach.…”

Section: Zinc-metalloproteases As Drug Targets In Cardiovascular Diseasementioning

confidence: 99%

“…In addition to these, a small number of drugs that act by a dual mechanism (such as the calcium channel β 1 adrenergic blockers 1, calcium channel α 1 adrenergic blockers, 5HT 2A antagonists, endothelin and angiotensin receptor blockers, ECE and ACE inhibitors, and NEP and ECE inhibitors) increasingly present an alternative therapeutic approach [52].…”

Section: Zinc-metalloproteases As Drug Targets In Cardiovascular Diseasementioning

confidence: 99%

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Catalytic metallodrugs

Cowan

2008

Pure and Applied Chemistry

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Drug discovery remains a top priority in medical science. The phenomenon of drug resistance has heightened the need for both new classes of pharmaceutical, as well as novel modes of action. A new paradigm for drug activity is presented, which includes both recognition and subsequent irreversible inactivation of therapeutic targets. Application to both RNA and enzyme therapeutic targets has been demonstrated, while incorporation of both binding and catalytic centers provides a double-filter mechanism for improved target selectivity and lower dosing. In contrast to RNA targets that are subject to strand scission chemistry mediated by ribose H-atom abstraction, proteins appear to be inactivated through oxidative damage to amino acid side chains around the enzyme active site. Methods to monitor both intracellular delivery and activity against RNA targets have been developed based on plasmid expression of the green fluorescent protein (GFP). Herein, the activity of representative metallodrugs is described in the context of both in vitro and cellular assays, and the mechanism of action is discussed. Studies with scavengers of reactive oxygen species (ROS) confirmed hydrogen peroxide to be an obligatory diffusible intermediate, prior to formation of a Cu-bound hydroxyl radical species generated from Fenton-type chemistry.

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Toward an Optimal Joint Recognition of the S₁‘ Subsites of Endothelin Converting Enzyme-1 (ECE-1), Angiotensin Converting Enzyme (ACE), and Neutral Endopeptidase (NEP)

Cited by 22 publications

References 31 publications

B-Type Natriuretic Peptide Prevents Acute Hypertrophic Responses in the Diabetic Rat Heart

B-Type Natriuretic Peptide Prevents Acute Hypertrophic Responses in the Diabetic Rat Heart

Identification of an Endothelin-converting Enzyme-2-specific Fluorigenic Substrate and Development of an in Vitro and ex Vivo Enzymatic Assay

Catalytic metallodrugs

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Toward an Optimal Joint Recognition of the S1‘ Subsites of Endothelin Converting Enzyme-1 (ECE-1), Angiotensin Converting Enzyme (ACE), and Neutral Endopeptidase (NEP)

Cited by 22 publications

References 31 publications

B-Type Natriuretic Peptide Prevents Acute Hypertrophic Responses in the Diabetic Rat Heart

B-Type Natriuretic Peptide Prevents Acute Hypertrophic Responses in the Diabetic Rat Heart

Identification of an Endothelin-converting Enzyme-2-specific Fluorigenic Substrate and Development of an in Vitro and ex Vivo Enzymatic Assay

Catalytic metallodrugs

Contact Info

Product

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Toward an Optimal Joint Recognition of the S₁‘ Subsites of Endothelin Converting Enzyme-1 (ECE-1), Angiotensin Converting Enzyme (ACE), and Neutral Endopeptidase (NEP)