scholarly journals Endopeptidase 3.4.24.11 converts N-1-(R,S)carboxy-3-phenylpropyl-Ala-Ala-Phe-p-carboxyanilide into a potent inhibitor of angiotensin-converting enzyme

1993 ◽  
Vol 294 (3) ◽  
pp. 681-684 ◽  
Author(s):  
C H Williams ◽  
T Yamamoto ◽  
D M Walsh ◽  
D Allsop
Keyword(s):  
Angiotensin Converting Enzyme ◽  
Potent Inhibitor ◽  
Neutral Endopeptidase ◽  
Converting Enzyme ◽  
Inhibitory Effects ◽  
Rabbit Lung ◽  
Pig Kidney ◽  
Tissue Extracts ◽  
Ic50 Value

It was reported recently that N-1-(R,S)carboxy-3-phenylpropyl-Ala-Ala-Phe-p-carboxyanilide (CPP-A-A-F-pAB), an inhibitor of endopeptidase 3.4.24.15 (E-24.15), also inhibits angiotensin-converting enzyme (ACE) from rabbit lung. We have found that this compound is without effect on ACE purified from pig kidney, at a concentration some 1000-fold greater than the Ki reported for inhibition of the enzyme from lung. However, preincubation of CPP-A-A-F-pAB with neutral endopeptidase 3.4.24.11 (E-24.11) does result in potent inhibitory effects on ACE. We have shown this to be due to formation of a fragment, CPP-A-A, the structure of which is closely related to ACE inhibitors such as enalaprilat. CPP-A-A was found to be a potent inhibitor of pig ACE. Under the conditions used it had an IC50 value of 1.6 x 10(-8) M, compared with the value obtained for captopril of 7.5 x 10(-10) M. These results have important implications for studies of E-24.15 when using CPP-A-A-F-pAB in vivo or in crude tissue extracts where E-24.11 might also be present.

scholarly journals Development and characterization of novel potent and stable inhibitors of endopeptidase EC 3.4.24.15

2000 ◽  
Vol 345 (2) ◽  
pp. 351-356 ◽  
Author(s):  
Corie N. SHRIMPTON ◽  
Giovanni ABBENANTE ◽  
Rebecca A. LEW ◽  
A. Ian SMITH
Keyword(s):  
Potent Inhibitor ◽  
Solid Phase ◽  
Neutral Endopeptidase ◽  
Amide Bond ◽  
Converting Enzyme ◽  
Phase Synthesis ◽  
Tissue Extracts ◽  
Endothelin Converting Enzyme

Solid-phase synthesis was used to prepare a series of modifications to the selective and potent inhibitor of endopeptidase EC 3.4.24.15 (EP24.15), N-[1(R,S)-carboxy-3-phenylpropyl]-Ala-Ala-Tyr-p-aminobenzoate (cFP), which is degraded at the Ala-Tyr bond, thus severely limiting its utility in vivo. Reducing the amide bond between the Ala and Tyr decreased the potency of the inhibitor to 1/1000. However, the replacement of the second alanine residue immediately adjacent to the tyrosine with α-aminoisobutyric acid gave a compound (JA-2) that was equipotent with cFP, with a Ki of 23 nM. Like cFP, JA-2 inhibited the closely related endopeptidase EC 3.4.24.16 1/20 to 1/30 as potently as it did EP24.15, and did not inhibit the other thermolysin-like endopeptidases angiotensin-converting enzyme, endothelin-converting enzyme and neutral endopeptidase. The biological stability of JA-2 was investigated by incubation with a number of membrane and soluble sheep tissue extracts. In contrast with cFP, JA-2 remained intact after 48 h of incubation with all tissues examined. Further modifications to the JA-2 compound failed to improve the potency of this inhibitor. Hence JA-2 is a potent, EP24.15-preferential and biologically stable inhibitor, therefore providing a valuable tool for further assessing the biological functions of EP24.15.

Dual inhibition of angiotensin converting enzyme and neutral endopeptidase by omapatrilat in rat in vivo

2001 ◽  
Vol 44 (5) ◽  
pp. 411-418 ◽  
Author(s):  
Tom Bäcklund ◽  
Eeva Palojoki ◽  
Tina Grönholm ◽  
Anders Eriksson ◽  
Olli Vuolteenaho ◽  
...  
Keyword(s):  
Angiotensin Converting Enzyme ◽  
Neutral Endopeptidase ◽  
Converting Enzyme ◽  
Dual Inhibition

scholarly journals Novel activity of angiotensin-converting enzyme. Hydrolysis of cholecystokinin and gastrin analogues with release of the amidated C-terminal dipeptide

1989 ◽  
Vol 262 (1) ◽  
pp. 125-130 ◽  
Author(s):  
P Dubreuil ◽  
P Fulcrand ◽  
M Rodriguez ◽  
H Fulcrand ◽  
J Laur ◽  
...  
Keyword(s):  
Angiotensin Converting Enzyme ◽  
Chloride Ion ◽  
Peptide Bond ◽  
Major Product ◽  
Enzyme Hydrolysis ◽  
Ion Concentration ◽  
Converting Enzyme ◽  
Rabbit Lung ◽  
Hydrolysis Of

ACE (angiotensin-converting enzyme; peptidyl dipeptidase A; EC 3.4.15.1), cleaves C-terminal dipeptides from active peptides containing a free C-terminus. We investigated the hydrolysis of cholecystokinin-8 [CCK-8; Asp-Tyr(SO3H)-Met-Gly-Trp-Met-Asp-Phe-NH2] and of various gastrin analogues by purified rabbit lung ACE. Although these peptides are amidated at their C-terminal end, they were metabolized by ACE to several peptide fragments. These fragments were analysed by h.p.l.c., isolated and identified by comparison with synthetic fragments, and by amino acid analysis. The initial and major site of hydrolysis was the penultimate peptide bond, which generated a major product, the C-terminal amidated dipeptide Asp-Phe-NH2. As a secondary cleavage, ACE subsequently released di- or tri-peptides from the C-terminal end of the remaining N-terminal fragments. The cleavage of CCK-8 and gastrin analogues was inhibited by ACE inhibitors (Captopril and EDTA), but not by other enzyme inhibitors (phosphoramidon, thiorphan, bestatin etc.). Hydrolysis of [Leu15]gastrin-(14-17)-peptide [Boc (t-butoxycarbonyl)-Trp-Leu-Asp-Phe-NH2] in the presence of ACE was found to be dependent on the chloride-ion concentration. Km values for the hydrolysis of CCK-8, [Leu15]gastrin-(11-17)-peptide and Boc-[Leu15]gastrin-(14-17)-peptide at an NaCl concentration of 300 mM were respectively 115, 420 and 3280 microM, and the catalytic constants were about 33, 115 and 885 min-1. The kcat/Km for the reactions at 37 degrees C was approx. 0.28 microM-1.min-1, which is approx. 35 times less than that reported for the cleavage of angiotensin I. These results suggest that ACE might be involved in the metabolism in vivo of CCK and gastrin short fragments.

Airway vasodilation by bradykinin is mediated via B2 receptors and modulated by peptidase inhibitors

1994 ◽  
Vol 266 (2) ◽  
pp. L156-L162
Author(s):  
I. Yamawaki ◽  
P. Geppetti ◽  
C. Bertrand ◽  
B. Chan ◽  
J. A. Nadel
Keyword(s):  
Blood Flow ◽  
Left Ventricle ◽  
Angiotensin Converting Enzyme ◽  
Reference Sample ◽  
Neutral Endopeptidase ◽  
Converting Enzyme ◽  
B1 Receptor ◽  
Bradykinin B1 Receptor ◽  
B2 Receptors

We studied the effect of exogenous bradykinin on blood flow in the airway microcirculation of anesthetized F344 rats in vivo. We made three successive determinations of airway blood flow and cardiac output using a modification of the reference sample microsphere technique. Injection of bradykinin into the left ventricle increased airway blood flow in a dose-related manner. Pretreatment with the bradykinin B2 receptor antagonist, Hoe 140, completely abolished bradykinin-, but not histamine-induced vasodilation. A bradykinin B1 receptor agonist, [des-Arg9]bradykinin, did not affect airway blood flow. We also studied the effect of inhibitors of angiotensin-converting enzyme (captopril) and neutral endopeptidase (phosphoramidon) on bradykinin-induced vasodilation. Pretreatment with captopril, but not phosphoramidon, potentiated the bradykinin-induced vasodilation. However, the addition of phosphoramidon further potentiated the effect of captopril. We conclude that injection of bradykinin into the left ventricle produces a dose-related vasodilation in the airway microcirculation mediated via B2 receptors, an effect that is modulated primarily by angiotensin-converting enzyme and, to a lesser extent, by neutral endopeptidase.

Kinin B1 receptor antagonists with multi-enzymatic resistance properties

2002 ◽  
Vol 80 (4) ◽  
pp. 287-292 ◽  
Author(s):  
Witold Neugebauer ◽  
Paul A Blais ◽  
Stephanie Hallé ◽  
Catherine Filteau ◽  
Domenico Regoli ◽  
...  
Keyword(s):  
Angiotensin Converting Enzyme ◽  
Human Platelets ◽  
Converting Enzyme ◽  
Receptor Antagonists ◽  
Rabbit Lung ◽  
Duration Of Action ◽  
Prolonged Duration ◽  
Bradykinin Antagonists

The kinin B1 receptor has been implicated in a variety of pathological states; therefore, potent, selective, and specific antagonists with prolonged duration of action in vivo are needed. Using R-715 (AcLys[D-β-Nal7,Ile8] desArg9BK) as a template, new peptides containing α-MePhe in position 5, Oic in position 2, and AcOrn instead of AcLys at the N-terminal were prepared and tested for their antagonist potency, their selectivity, and their specificity for the kinin B1 receptor. In vitro metabolic stabilities toward aminopeptidase M (from human plasma), aminopeptidase P (from human platelets), and angiotensin-converting enzyme (purified from rabbit lung) were also investigated. The results of this study indicate that the three modifications applied separately are as well tolerated as they are when present conjointly in the template R-715. Indeed, pA2 values of R-715 (ranging from 8.40 to 8.5) do not differ significantly from the analogues R-954 and R-955 (both ranging from 8.4 to 8.6) when measured at kinin B1 receptors from rabbit aortas and human umbilical veins. Moreover, the chemical modifications utilized in the peptides R-954 and R-955 have provided resistance against aminopeptidases M and P, as well as the angiotensin-converting enzyme, unlike the early (e.g., Lys[Leu8]desArg9BK) and more recent (e.g., R-715, B-9858) generations of B1 receptor antagonists. Ongoing in vivo assays will validate the assumption that the analogues R-954 and R-955 have a prolonged duration of action.Key words: bradykinin, antagonists, B1 receptors, peptidases, metabolism.

In Vivo Pharmacology of Dual Neutral Endopeptidase/Angiotensin-Converting Enzyme Inhibitors

1996 ◽  
Vol 28 (5) ◽  
pp. 672-678 ◽  
Author(s):  
Andrea Ann Seymour ◽  
Magdi M. Asaad ◽  
Benoni E. Abboa-Offei ◽  
Patricia L. Smith ◽  
W. Lynn Rogers ◽  
...  
Keyword(s):  
Angiotensin Converting Enzyme ◽  
Enzyme Inhibitors ◽  
Angiotensin Converting Enzyme Inhibitors ◽  
Neutral Endopeptidase ◽  
Converting Enzyme ◽  
Converting Enzyme Inhibitors

Effect of T-kinin on microvascular permeability and its modulation by peptidases in rat airways

1995 ◽  
Vol 79 (4) ◽  
pp. 1129-1133 ◽  
Author(s):  
I. Yamawaki ◽  
J. Tamaoki ◽  
Y. Takeda ◽  
A. Chiyotani ◽  
N. Sakai ◽  
...  
Keyword(s):  
Angiotensin Converting Enzyme ◽  
Receptor Antagonist ◽  
Enzyme Inhibitor ◽  
Sensory Nerve ◽  
Neutral Endopeptidase ◽  
Rat Plasma ◽  
Plasma Extravasation ◽  
Dependent Manner ◽  
Converting Enzyme

T-kinin (Ile-Ser-bradykinin), the product of T-kininogen, has been found in rat plasma during systemic inflammation, but the effect of this kinin on airway inflammatory response is unknown. We examined the effect of T-kinin on vascular permeability in airways of anesthetized rats in vivo by using photometric measurement of the extravasated Evans blue. Intravenous injection of T-kinin (0.1–10 mumol/kg) increased dye extravasation in a dose-dependent manner, with 134% for trachea and 117% for bronchi by 1 mumol/kg. Pretreatment with bradykinin B2-receptor antagonist Hoe-140 (100 nmol/kg), but not the B1-receptor antagonist des-Arg9-Leu8-bradykinin (5 mg/kg), abolished plasma extravasation evoked by T-kinin (1 mumol/kg). NK1 tachykinin-receptor antagonist CP-99994 (4 mg/kg) did not affect T-kinin-induced vascular leakage. Pretreatment with captopril (2.5 mg/kg), angiotensin-converting enzyme inhibitor, potentiated T-kinin (100 nmol/kg)-induced plasma extravasation, whereas phosphoramidon (2.5 mg/kg), a neutral endopeptidase inhibitor, had no effect. We conclude that T-kinin produces airway vascular extravasation via stimulation of B2 receptors. The effect is modulated by endogenous angiotensin-converting enzyme and is not mediated via activation of sensory nerve.

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