phosphoramidon and amastatin

phosphoramidon has been researched along with amastatin* in 31 studies

Other Studies

31 other study(ies) available for phosphoramidon and amastatin

ArticleYear
Effect of three peptidase inhibitors on antinociceptive potential and toxicity with intracerebroventricular administration of dynorphin A (1-17) or (1-13) in the rat.
    Journal of anesthesia, 2015, Volume: 29, Issue:1

    The N- and C-terminal regions of dynorphin (Dyn) A (1-17) activate opioid and N-methyl-D-aspartate receptors, respectively. Earlier studies demonstrated that Dyn-converting enzyme cleaved Dyn A (1-17) mainly at the Arg(6)-Arg(7) bond, resulting in the production of N- and C-terminal region peptide fragments, and that this enzyme was not inhibited by a mixture of the three peptidase inhibitors (PIs) amastatin (A), captopril (C), and phosphoramidon (P). The purpose of the present study was to evaluate antinociceptive potential and toxicity with intracerebroventricular administration of Dyn A (1-17) or (1-13) under pretreatment with a mixture of A, C, and P and/or Dyn-converting enzyme inhibitor (p-hydroxymercuribenzoate).. Peptide fragments from Dyn A (1-17) following incubation with membrane preparation under pretreatment with a mixture of the three PIs was identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). Infusion of drugs and peptides into the third ventricle in rats was performed via indwelling cannulae. Induction of antinociception and toxicity by Dyn A (1-17), Dyn A (1-13), Dyn A (1-6), or Dyn A (7-17) were determined by the tail-flick test and induction of barrel rotation, respectively. The effects of the PIs on antinociception and toxicity were evaluated by a dose-response study and a comparison of differences among various combinations of Dyn A (1-17) or Dyn A (1-13) and the three PIs and p-hydroxymercuribenzoate.. MALDI-TOF-MS analysis identified Dyn A (1-6) and Dyn A (1-10) fragments as products following incubation of Dyn A (1-17) with membrane preparation of rat midbrain under pretreatment with a mixture of the three PIs. Pretreatment with a mixture of the three PIs produced an approximately 30-fold augmentation in antinociception induced by low-dose intracerebroventricular administration of Dyn A (1-17) or (1-13) in a μ-, δ- and κ-opioid receptor antagonist-reversible manner, but without signs of toxicity such as barrel rotation in the rat. Dyn A (1-17)-induced antinociception and toxicity was greater than that of Dyn A (1-6), Dyn A (1-13), or Dyn A (7-17) at the same dose. Dyn A (1-17)-induced antinociception and toxicity under pretreatment with various combinations of the three PIs and p-hydroxymercuribenzoate was greater than that with a mixture of the three PIs alone.. These findings suggest that administration of a mixture of the three PIs increases Dyn A (1-17)- or (1-13)-induced antinociception under physiological conditions without toxicity.

    Topics: Analgesics; Analgesics, Opioid; Animals; Brain Chemistry; Captopril; Dose-Response Relationship, Drug; Dynorphins; Glycopeptides; Injections, Intraventricular; Male; Pain Measurement; Peptides; Protease Inhibitors; Rats; Rats, Wistar; Receptors, Opioid

2015
Potentiation of [Met5]enkephalin-induced antinociception by mixture of three peptidase inhibitors in rat.
    Journal of anesthesia, 2014, Volume: 28, Issue:5

    Previous in vitro studies have shown that degradation of opioid peptides during incubation with cerebral membrane preparations is almost completely prevented by a mixture of three peptidase inhibitors (PIs), namely, amastatin, captopril, and phosphoramidon. In the present in vivo study, we evaluate the effects of intrathecal administration of these PIs on antinociception by [Met(5)]enkephalin (ME) or PIs themselves.. Drugs were administered into the thoracolumbar level of the spinal cord in the intrathecal space in rat. Induction of antinociception was measured by the tail immersion assay, with 55 °C as the nociceptive stimulus. Effects of PIs on antinociception were evaluated by dose-response study (ME, 1-20 nmol; PIs, 1-20 nmol each), by comparison of differences among two combinations of PIs (amastatin and captopril; captopril and phosphoramidon; amastatin and phosphoramidon) and three PIs (amastatin, captopril, and phosphoramidon), and by using opioid receptor selective antagonists.. Intrathecal administration of ME with these three PIs or PIs alone significantly and dose dependently increased antinociception in a μ- and δ-opioid receptor antagonist-reversible manner; moreover, the degree of antinociception with a combination of any two of these was less than that with all three, indicating that any residual single peptidase could inactivate significant amounts of ME.. The present data, together with those of earlier studies, clearly demonstrate that amastatin-, captopril-, and phosphoramidon-sensitive enzymes play an important role in inactivation of opioid peptides at the spinal level.

    Topics: Analgesics; Animals; Captopril; Drug Synergism; Enkephalin, Methionine; Glycopeptides; Male; Narcotic Antagonists; Peptides; Protease Inhibitors; Rats; Rats, Wistar

2014
Increase in antinociceptive effect of [leu5]enkephalin after intrathecal administration of mixture of three peptidase inhibitors.
    The Tokai journal of experimental and clinical medicine, 2013, Jul-20, Volume: 38, Issue:2

    Previous in vitro studies have shown that the degradation of [Leu5]enkephalin during incubation with cerebral membrane preparations is almost completely prevented by a mixture of three peptidase inhibitors such as amastatin, captopril, and phosphoramidon. The present in vivo study was performed to examine the antinociceptive effect of [Leu5]enkephalin administered intrathecally under pretreatment with these three peptidase inhibitors.. A tail-flick test was used to determine the nociceptive threshold after administration of [Leu5]enkephalin. The time-course profiles of the latency to flick the tail and the area under the time response curve were evaluated for the antinociceptive action of the drug.. The antinociceptive effect of [Leu5]enkephalin administered intrathecally on the tail-flick test was increased more than 100-fold under i.t. pretreatment with three peptidase inhibitors. The antinociceptive effect produced by [Leu5]enkephalin in rats pretreated with any single peptidase inhibitor increased antinociception compared to that with saline. The antinociceptive potency of [Leu5]enkephalin under pretreatment with any combination of two peptidase inhibitors was smaller than that in rats pretreated with three peptidase inhibitors together. These results indicate that any residual single peptidase inactivates significant amounts of [Leu5]enkephalin.. The present data, together with those of earlier studies, clearly demonstrate that amastatin-, captopril-, and phosphoramidon-sensitive enzymes play an important role in the inactivation of [Leu5]enkephalin administered intrathecally in rat.

    Topics: Analgesics; Animals; Captopril; Dose-Response Relationship, Drug; Drug Combinations; Drug Synergism; Enkephalins; Glycopeptides; Injections, Spinal; Male; Nociceptive Pain; Pain Threshold; Peptides; Protease Inhibitors; Rats; Rats, Wistar

2013
Antinociceptive effect of [Met5]enkephalin semicarbazide is not affected by dipeptidyl carboxypeptidase-I.
    Journal of peptide science : an official publication of the European Peptide Society, 2012, Volume: 18, Issue:2

    Dipeptidyl carboxypeptidase-I is an enzyme involved in the biological degradation of enkephalins. It has been suggested that C-terminal amidation of enkephalins enhances their resistance to dipeptidyl carboxypeptidase-I-mediated biodegradation. In this study, a novel [Met5]enkephalin amide (MEA) analogue [Met5]enkephalin (ME)-semicarbazide synthesized by another laboratory in our group was assessed for its antinociceptive effects compared with ME-ethylamide, MEA and ME, using tail flick test. To protect the administered drugs from biodegradation, rats were pretreated with peptidase inhibitors including amastatin, phosphoramidon and captopril. Then captopril (dipeptidyl carboxypeptidase-I inhibitor) was deleted from the peptidase inhibitors' combination for evaluating in vivo resistance of the synthetic drugs to dipeptidyl carboxypeptidase-I. According to the results, ME-semicarbazide and MEA were resistant enough to dipeptidyl carboxypeptidase-I to exert their strong antinociception following intrathecal administration even in the absence of captopril, whereas the antinociceptive effects produced by ME-ethylamide (10 nmol) were abolished in rats not pretreated with captopril, indicating that significant amounts of the ME-ethylamide were degraded by dipeptidyl carboxypeptidase-I. Replacement of the amide moiety of MEA with semicarbazide provides a new ME derivative, with high analgesic effects as well as more resistance to dipeptidyl carboxypeptidase-I-mediated biodegradation.

    Topics: Analgesics; Animals; Biotransformation; Captopril; Carboxypeptidases; Enkephalin, Methionine; Glycopeptides; Hydrolysis; Injections, Spinal; Male; Nociception; Peptides; Protease Inhibitors; Rats; Rats, Wistar; Semicarbazides

2012
Great increase in antinociceptive potency of [Leu5]enkephalin after peptidase inhibition.
    Journal of pharmacological sciences, 2008, Volume: 106, Issue:2

    Previous in vitro studies have shown that the degradation of [Leu(5)]enkephalin during incubation with cerebral membrane preparations is almost completely prevented by a mixture of three peptidase inhibitors: amastatin, captopril, and phosphoramidon. The present in vivo study shows that the inhibitory effect of [Leu(5)]enkephalin administered intra-third-ventricularly on the tail-flick response was increased more than 500-fold by the intra-third-ventricular pretreatment with the three peptidase inhibitors. The antinociceptive effect produced by the [Leu(5)]enkephalin in rats pretreated with any combination of two peptidase inhibitors was significantly smaller than that in rats pretreated with the three peptidase inhibitors, indicating that any residual single peptidase could inactivate significant amounts of the [Leu(5)]enkephalin. The present data, together with those obtained from previous studies, clearly demonstrate that amastatin-, captopril-, and phosphoramidon-sensitive enzymes play important roles in the inactivation of short endogenous opioid peptides, such as penta-, hepta-, and octa-peptides, administered intra-third-ventricularly to rats.

    Topics: Animals; Captopril; Drug Synergism; Enkephalin, Leucine; Glycopeptides; Guinea Pigs; Ileum; Injections, Intraventricular; Male; Mice; Mice, Inbred ICR; Muscle Contraction; Pain; Peptides; Protease Inhibitors; Rats; Rats, Wistar; Vas Deferens

2008
Functional evidence of des-Arg10-kallidin enzymatic inactivating pathway in isolated human umbilical vein.
    Naunyn-Schmiedeberg's archives of pharmacology, 2007, Volume: 375, Issue:3

    It has been known for many years that plasma and tissues contain a variety of enzymes capable of metabolizing kinins. The aim of the present study was to evaluate, by means of functional studies in a capacitance vessel such as the human umbilical vein (HUV), the possible role played by the metallopeptidases angiotensin-converting enzyme (ACE), neutral endopeptidase (NEP), and aminopeptidase M (APM) as an inactivating pathway of the B(1) receptor endogenous agonist des-Arg(10)-kallidin (DAKD). In HUV rings with and without endothelium, concentration-response curves (CRCs) to DAKD were determined after a 300-min incubation period, and enzymatic inhibitors were added to the organ baths 30 min before construction of the CRC. Presence of endothelial layer was confirmed by histological studies. There was a significant leftward shift observed in control HUV rings devoid of endothelium compared with intact tissues. Exposure to 1 microM captopril (ACE inhibitor) potentiated DAKD-elicited vasoconstrictor responses in HUV rings with endothelium while no such effect was observed in tissues devoid of endothelium. Application of 10 microM amastatin (APM inhibitor) induced a leftward shift of DAKD-elicited contractile responses in HUV with and without endothelium. On the other hand, 10 microM phosphoramidon (NEP inhibitor) showed no potentiating effect in HUV rings either with or without endothelium. However, under concurrent inhibition of ACE, NEP and APM, there was a higher potentiation of DAKD-elicited contractile responses compared with the effect observed with combined inhibition of ACE and APM. Moreover, when we evaluated contractile responses induced by Sar(0)-D-Phe(8)-des-Arg(9)-BK (a metabolically protected B(1) receptor agonist), no potentiating effect was observed under triple enzymatic inhibition. In conclusion, in the present study for the first time, we demonstrated in a capacitance vessel, HUV, that metallopeptidases ACE, NEP and APM represent a relevant functional inactivation pathway of DAKD.

    Topics: Angiotensin-Converting Enzyme Inhibitors; Bradykinin; Captopril; CD13 Antigens; Dose-Response Relationship, Drug; Endothelium, Vascular; Enzyme Inhibitors; Female; Glycopeptides; Humans; In Vitro Techniques; Kallidin; Metalloproteases; Neprilysin; Peptides; Protease Inhibitors; Receptor, Bradykinin B1; Umbilical Veins; Vasoconstriction

2007
The enhancing effects of peptidase inhibitors on the antinociceptive action of [Met5]enkephalin-Arg6-Phe7 in rats.
    Journal of pharmacological sciences, 2007, Volume: 105, Issue:1

    Previous in vitro studies have shown that the degradation of [Met(5)]enkephalin-Arg(6)-Phe(7) during incubation with cerebral membrane preparations is largely prevented by a mixture of three peptidase inhibitors: amastatin, captopril, and phosphoramidon. The present in vivo study shows that the inhibitory effect of [Met(5)]enkephalin-Arg(6)-Phe(7) administered intra-third-ventricularly on the tail-flick response was increased more than 1000-fold by the intra-third-ventricular pretreatment with three peptidase inhibitors. The antinociceptive effect produced by the [Met(5)]enkephalin-Arg(6)-Phe(7) in rats pretreated with any combination of two peptidase inhibitors was significantly smaller than that in rats pretreated with three peptidase inhibitors, indicating that any residual single peptidase could inactivate significant amounts of the [Met(5)]enkephalin-Arg(6)-Phe(7). The present data, together with those obtained from previous studies, clearly show that amastatin-, captopril-, and phosphoramidon-sensitive enzymes play important roles in the inactivation of endogenous opioid peptides, such as [Met(5)]enkephalin, [Met(5)]enkephalin-Arg(6)-Phe(7), [Met(5)]enkephalin-Arg(6)-Gly(7)-Leu(8), and dynorphin A (1-8), administered intra-third-ventricularly to rats.

    Topics: Analgesics; Animals; Captopril; Dose-Response Relationship, Drug; Drug Synergism; Enkephalin, Methionine; Glycopeptides; Injections, Intraventricular; Injections, Subcutaneous; Male; Naloxone; Narcotic Antagonists; Pain; Pain Measurement; Peptides; Protease Inhibitors; Rats; Rats, Wistar

2007
Peptidases prevent mu-opioid receptor internalization in dorsal horn neurons by endogenously released opioids.
    The Journal of neuroscience : the official journal of the Society for Neuroscience, 2003, Mar-01, Volume: 23, Issue:5

    To evaluate the effect of peptidases on mu-opioid receptor (MOR) activation by endogenous opioids, we measured MOR-1 internalization in rat spinal cord slices. A mixture of inhibitors of aminopeptidases (amastatin), dipeptidyl carboxypeptidase (captopril), and neutral endopeptidase (phosphoramidon) dramatically increased the potencies of Leu-enkephalin and dynorphin A to produce MOR-1 internalization, and also enhanced the effects of Met-enkephalin and alpha-neoendorphin, but not endomorphins or beta-endorphin. The omission of any one inhibitor abolished Leu-enkephalin-induced internalization, indicating that all three peptidases degraded enkephalins. Amastatin preserved dynorphin A-induced internalization, and phosphoramidon, but not captopril, increased this effect, indicating that the effect of dynorphin A was prevented by aminopeptidases and neutral endopeptidase. Veratridine (30 microm) or 50 mm KCl produced MOR-1 internalization in the presence of peptidase inhibitors, but little or no internalization in their absence. These effects were attributed to opioid release, because they were abolished by the selective MOR antagonist CTAP (D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH(2)) and were Ca(2+) dependent. The effect of veratridine was protected by phosphoramidon plus amastatin or captopril, but not by amastatin plus captopril or by phosphoramidon alone, indicating that released opioids are primarily cleaved by neutral endopeptidase, with a lesser involvement of aminopeptidases and dipeptidyl carboxypeptidase. Therefore, because the potencies of endomorphin-1 and endomorphin-2 to elicit internalization were unaffected by peptidase inhibitors, the opioids released by veratridine were not endomorphins. Confocal microscopy revealed that MOR-1-expressing neurons were in close proximity to terminals containing opioids with enkephalin-like sequences. These findings indicate that peptidases prevent the activation of extrasynaptic MOR-1 in dorsal horn neurons.

    Topics: Aminopeptidases; Animals; Anti-Bacterial Agents; Captopril; Endopeptidases; Enzyme Inhibitors; Glycopeptides; In Vitro Techniques; Lumbosacral Region; Narcotic Antagonists; Neprilysin; Opioid Peptides; Peptide Fragments; Peptide Hydrolases; Peptides; Posterior Horn Cells; Rats; Rats, Sprague-Dawley; Receptors, Opioid, mu; Somatostatin; Veratridine

2003
Effects of peptidase inhibitors on anti-nociceptive action of dynorphin-(1-8) in rats.
    Naunyn-Schmiedeberg's archives of pharmacology, 2000, Volume: 361, Issue:3

    Previous in vitro studies showed that the degradation of dynorphin-(1-8) [dyn-(1-8)] by cerebral membrane preparations is almost completely prevented by a mixture of three peptidase inhibitors (PIs), amastatin, captopril and phosphoramidon. In the present investigations, effects of the three PIs on the anti-nociception induced by the intra-third-ventricular (i.t.v.) administration of dyn-(1-8) were examined. The inhibitory effect of dyn-(1-8) on the tail-flick response was increased more than 100-fold by the i.t.v. pretreatment of rats with the three PIs. The inhibition produced by dyn-(1-8) in rats pretreated with any combination of two PIs was significantly smaller than that in rats pretreated with three PIs, indicating that any residual single peptidase could inactivate significant amounts of dyn-(1-8). The antagonistic effectiveness of naloxone, a relatively selective mu-opioid antagonist, indicates that dyn-(1-8)-induced inhibition of tail-flick response in rats pretreated with three PIs is mediated by mu-opioid receptors. Furthermore, mu-receptor-mediated inhibition induced by dyn-(1-8) was significantly greater than that produced by [Met5]-enkephalin in rats pretreated with three PIs. The data obtained in the present investigations together with those obtained in previous studies strongly indicate that dyn-(1-8) not only has well-known kappa-agonist activity but also has high mu-agonist activity.

    Topics: Analgesics, Opioid; Analysis of Variance; Angiotensin-Converting Enzyme Inhibitors; Animals; Anti-Bacterial Agents; Captopril; Drug Interactions; Dynorphins; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Glycopeptides; Injections, Intraventricular; Male; Naloxone; Narcotic Antagonists; Pain; Pain Measurement; Peptide Fragments; Peptides; Protease Inhibitors; Rats; Rats, Wistar; Receptors, Opioid, mu

2000
Identification of kallidin degrading enzymes in the isolated perfused rat heart.
    Japanese journal of pharmacology, 1999, Volume: 79, Issue:1

    Kallidin (KD) is an important vasoactive kinin whose physiological effects are strongly dependent on its degradation through local kininases. In the present study, we examined the spectrum of these enzymes and their contribution to KD degradation in isolated perfused rat hearts. By inhibiting angiotensin-converting enzyme (ACE), aminopeptidase M (APM) and neutral endopeptidase (NEP) with ramiprilat (0.25 microM), amastatin (40 microM) and phosphoramidon (1 microM), respectively, relative kininase activities were obtained. APM (44%) and ACE (35%) are the main KD degrading enzymes in rat heart; NEP (7%) plays a minor role. A participation of carboxypeptidase N (CPN) could not be found.

    Topics: Aminopeptidases; Angiotensin-Converting Enzyme Inhibitors; Animals; Anti-Bacterial Agents; Glycopeptides; In Vitro Techniques; Kallidin; Male; Myocardium; Neprilysin; Peptide Hydrolases; Peptides; Peptidyl-Dipeptidase A; Perfusion; Protease Inhibitors; Ramipril; Rats; Rats, Wistar

1999
Kallidin- and bradykinin-degrading pathways in human heart: degradation of kallidin by aminopeptidase M-like activity and bradykinin by neutral endopeptidase.
    Circulation, 1999, Apr-20, Volume: 99, Issue:15

    Since kinins kallidin (KD) and bradykinin (BK) appear to have cardioprotective effects ranging from improved hemodynamics to antiproliferative effects, inhibition of kinin-degrading enzymes should potentiate such effects. Indeed, it is believed that this mechanism is partly responsible for the beneficial effects of angiotensin-converting enzyme (ACE) inhibitors. In the heart, enzymes other than ACE may contribute to local degradation of kinins. The purpose of this study was to investigate which enzymes are responsible for the degradation of KD and BK in human heart tissue.. Cardiac membranes were prepared from the left ventricles of normal (n=5) and failing (n=10) hearts. The patients had end-stage congestive heart failure as the result of coronary heart disease or idiopathic dilated cardiomyopathy. Heart tissue was incubated with KD or BK in the presence or absence of enzyme inhibitors. We found no difference in the enzymes responsible for kinin metabolism or their activities between normal and failing hearts. Thus KD was mostly converted into BK by the aminopeptidase M-like activity. When BK was used as substrate, it was converted into an inactive metabolite BK-(1-7) mostly (80% to 90%) by the neutral endopeptidase (NEP) activity, with ACE unexpectedly playing only a minor role. The low enzymatic activity of ACE in the cardiac membranes, compared with that of NEP, was not due to chronic ACE inhibitor therapy, because the cardiac ACE activities of patients, whether receiving ACE inhibitors or not, and of normal subjects were all equal.. The present in vitro study shows that in human cardiac membranes, the most critical step in kinin metabolism, that is, inactivation of BK, appears to be mediated mostly by NEP. This observation suggests a role for NEP in the local control of BK concentration in heart tissue. Thus inhibition of cardiac NEP activity could be cardioprotective by elevating the local concentration of BK in the heart.

    Topics: Angiotensin-Converting Enzyme Inhibitors; Anti-Bacterial Agents; Bradykinin; Captopril; CD13 Antigens; Dipeptides; Female; Glycopeptides; Heart Failure; Humans; Kallidin; Leucine; Male; Membrane Proteins; Middle Aged; Myocardium; Neprilysin; Peptides; Peptidyl-Dipeptidase A; Protease Inhibitors; Substrate Specificity

1999
Effects of three peptidase inhibitors, amastatin, captopril and phosphoramidon, on the hydrolysis of [Met5]-enkephalin-Arg6-Phe7 and other opioid peptides.
    Naunyn-Schmiedeberg's archives of pharmacology, 1998, Volume: 357, Issue:3

    The contents of [Met5]-enkephalin-Arg6-Phe7 (met-enk-RF) and its six hydrolysis products: Y, YG, YGG, YGGF, YGGFM, and YGGFMR were estimated after incubating met-enk-RF with either a guinea-pig ileal or striatal membrane fraction for various times at 37 degrees C. After 45 min incubation with either ileal or striatal membranes, met-enk-RF was completely hydrolyzed, yielding Y as the major product. Incubation with either membrane preparation for 60 min in the presence of the aminopeptidase inhibitor amastatin hydrolyzed 90 or 92% of met-enk-RF, respectively, with YGG being the major product. If the dipeptidyl carboxypeptidase I inhibitor captopril is also included in the incubation, met-enk-RF hydrolysis decreases by about half for both membranes, with YGG remaining the major product. Inclusion of three peptidase inhibitors, amastatin, captopril, and phosphoramidon (inhibition of endopeptidase-24.11) further reduced met-enk-hydrolysis, with 87% or more remaining intact. This shows that met-enk-RF was mainly hydrolyzed by three enzymes, amastatin-sensitive aminopeptidase, captopril-sensitive dipeptidyl carboxypeptidase I and phosphoramidon-sensitive endopeptidase-24.11, in both ileal and striatal membranes. Additionally, estimations of [Leu5]-enkephalin (leu-enk), alpha- and beta-neoendorphins (alpha- and beta-neoends), and dynorphin B (dyn B) contents after incubating the individual peptides with striatal membrane for 60 min in the presence of the three peptidase inhibitors showed that 98, 32, 5, and 23%, respectively, remained intact. Our previous studies together with the data obtained here show that one group of endogenous opioid peptides: met-enk, leu-enk, met-enk-RF, met-enk-RGL, and dyn A-(1-8) are largely or almost exclusively hydrolyzed by the three enzymes, amastatin-sensitive aminopeptidase, captopril-sensitive dipeptidyl carboxypeptidase I, and phosphoramidon-sensitive endopeptidase-24.11, and indicate that an unidentified fourth enzyme(s) is involved in the hydrolysis of another group of peptides: alpha-neoend, beta-neoend, and dyn B.

    Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Anti-Bacterial Agents; Captopril; Enkephalins; Glycopeptides; Guinea Pigs; Hydrolysis; Ileum; Male; Membranes; Opioid Peptides; Peptides; Protease Inhibitors; Visual Cortex

1998
Protection against dynorphin-(1-8) hydrolysis in membrane preparations by the combination of amastatin, captopril and phosphoramidon.
    The Journal of pharmacology and experimental therapeutics, 1998, Volume: 286, Issue:2

    The amounts of dynorphin-(1-8) [dyn-(1-8)] and its seven hydrolysis products, Y, YG, YGG, YGGF, YGGFL, YGGFLR and YGGFLRR, were estimated after incubating dyn-(1-8) with a membrane fraction from either guinea-pig ileum or striatum for various times at 37 degrees C. The major hydrolysis products during the initial 5-min incubation were YGGFLR and Y, which indicates that dipeptidyl carboxypeptidase and aminopeptidase activities were mainly involved in the hydrolysis. After 60 min of incubation, dyn-(1-8) was completely hydrolyzed in both membrane preparations. When the ileal and the striatal preparations were incubated for 60 min in the presence of both captopril, a dipeptidyl carboxypeptidase inhibitor, and amastatin, an aminopeptidase inhibitor, 63.8 and 49.3% of dyn-(1-8), respectively, were hydrolyzed. The YGG fragment was the major hydrolysis product in both preparations. When the ileal and the striatal membrane fractions were incubated with dyn-(1-8) in the presence of three peptidase inhibitors, captopril, amastatin and phosphoramidon (an inhibitor of endopeptidase-24.11), approximately 95% of the opioid octapeptide remained intact in both cases. This shows that dyn-(1-8) was almost exclusively hydrolyzed by three enzymes, amastatin-sensitive aminopeptidase, captopril-sensitive dipeptidyl carboxypeptidase I and phosphoramidon-sensitive endopeptidase-24.11, in both ileal and striatal membranes. Additionally, the Ke (equilibrium dissociation constant) values of selective antagonists against dyn-(1-8) and its initial main hydrolysis product YGGFLR in two isolated preparations pretreated with the three peptidase inhibitors indicate that the latter acts on mu receptors in guinea pig ileum but delta receptors in mouse vas deferens and the former acts on kappa receptors in both preparations. It is indicated, therefore, that in the absence of peptidase inhibitors endogenously released dyn-(1-8) acts either through dyn-(1-8) itself on kappa receptors or through YGGFLR on mu or delta receptors depending on both the three peptidase activities and the three receptor type densities at the target synaptic membrane.

    Topics: Animals; Anti-Bacterial Agents; Biotransformation; Captopril; Chromatography, High Pressure Liquid; Dynorphins; Electrochemistry; Glycopeptides; Guinea Pigs; Hydrolysis; Hypothalamic Hormones; In Vitro Techniques; Kinetics; Male; Membranes; Mice; Mice, Inbred ICR; Peptide Fragments; Peptides; Protease Inhibitors; Receptors, Opioid, delta; Receptors, Opioid, kappa

1998
Effects of peptidase inhibitors on the enkephalin-induced anti-nociception in rats.
    Japanese journal of pharmacology, 1998, Volume: 78, Issue:4

    The intra-third-ventricular (i.t.v.) administration of [Met5]-enkephalin (enk) to rats pretreated i.t.v. with three peptidase inhibitors (PIs), amastatin, captopril and phosphoramidon, inhibited the tail-flick response. The enk-induced inhibition was augmented by increasing the doses of the three PIs, with the maximum inhibition being attained at the doses of 10 nmol each. The enk-induced inhibition in rats pretreated with any combination of two PIs, however, were markedly smaller than that in rats pretreated with all three PIs, indicating that three kinds of enzymes all played important roles in the inactivation of enk. The inhibitory effect of enk on the tail-flick response in rats pretreated with the three PIs at doses of 10 nmol each was approximately tenfold higher than that of morphine. The relative anti-nociceptive potencies of enk and morphine were similar to the relative inhibitory potencies obtained previously with the isolated guinea pig ileum pretreated with the three PIs, indicating that the hydrolysis of the i.t.v. administered enk was largely prevented by the three PIs. However, the magnitude of the enk-induced inhibition in rats pretreated s.c. with the three PIs indicated that the hydrolysis of enk injected i.t.v. was not largely prevented by the s.c. administration of three PIs at doses up to 10 micromol each/kg.

    Topics: Animals; Anti-Bacterial Agents; Area Under Curve; Captopril; Dose-Response Relationship, Drug; Drug Therapy, Combination; Enkephalin, Methionine; Glycopeptides; Injections, Intraventricular; Injections, Subcutaneous; Male; Naloxone; Naltrexone; Narcotic Antagonists; Nociceptors; Pain; Pain Measurement; Peptides; Protease Inhibitors; Rats; Rats, Wistar

1998
Almost complete protection from [Met5]-enkephalin-Arg6-Gly7-Leu8 (Met-enk-RGL) hydrolysis in membrane preparations by the combination of amastatin, captopril and phosphoramidon.
    The Journal of pharmacology and experimental therapeutics, 1997, Volume: 281, Issue:2

    The contents of [Met5]-enkephalin-Arg6-Gly7-Leu8 (met-enk-RGL) and its seven hydrolysis products-Y, YG, YGG, YGGF, YGGFM, YGGFMR, and YGGFMRG-were estimated after incubating met-enk-RGL with a membrane fraction from either guinea pig ileum or striatum for various times at 37 degrees C. After 15 min of incubation, met-enk-RGL was completely hydrolyzed in both the ileal and the striatal membrane preparations. The major hydrolysis products were YGGFMR, YGGF and Y, which indicates that dipeptidyl carboxypeptidase and aminopeptidase activities were mainly involved in the hydrolysis. Additionally, even when the ileal and the striatal preparations were incubated for 60 min in the presence of both captopril, a dipeptidyl carboxypeptidase inhibitor, and amastatin, an aminopeptidase inhibitor, 24% and 44% of enkephalin octapeptide, respectively, were hydrolyzed. The YGG fragment was the major hydrolysis product in both preparations. When the ileal and the striatal membrane fractions were incubated with met-enk-RGL in the presence of three peptidase inhibitors-captopril, amastatin, and phosphoramidon (an inhibitor of endopeptidase-24.11)-approximately 95% of the enkephalin octapeptide, remained intact in both cases. This shows that met-enk-RGL was almost exclusively hydrolyzed by three enzymes, amastatin-sensitive aminopeptidase, captopril-sensitive dipeptidyl carboxypeptidase I and phosphoramidon-sensitive endopeptidase-24.11, in both ileal and striatal membranes. We also reported the potencies of several opioids relative to that of met-enk-RGL in guinea pig ileum pretreated with the three peptidase inhibitors.

    Topics: Animals; Anti-Bacterial Agents; Captopril; Corpus Striatum; Enkephalin, Methionine; Glycopeptides; Guinea Pigs; Hydrolysis; Ileum; In Vitro Techniques; Kinetics; Male; Morphine; Peptides; Protease Inhibitors

1997
In vitro metabolism of an insect neuropeptide by neural membrane preparations from Lymantria dispar.
    Peptides, 1996, Volume: 17, Issue:2

    Neural membrane fractions, prepared from brain-subesophageal ganglion complexes of the adult lepidopteran Lymantria dispar, contain at least two peptidases capable of metabolizing locust adipokinetic hormone-I in vitro. The initial fragments, pGlu1-Leu2-Asn3 and Phe4-Thr5-Pro6-Asn7-Trp8-Gly9-Thr10, result from the action of an endopeptidase with properties similar to those reported for neutral metalloendopeptidase in Schistocerca gregaria and mammalian endopeptidase 24.11. The heptapeptide is further degraded by an aminopeptidase that exhibits kinetic properties similar to those described for aminopeptidase 3.4.11.2. These enzymes appear to be responsible for the first two steps in AKH catabolism in L. dispar.

    Topics: Amino Acid Sequence; Aminopeptidases; Animals; Anti-Bacterial Agents; Edetic Acid; Endopeptidases; Ganglia, Invertebrate; Glycopeptides; In Vitro Techniques; Insect Hormones; Lepidoptera; Leucine; Molecular Sequence Data; Peptides; Protease Inhibitors; Thiorphan

1996
In vitro metabolism of LVV-hemorphin-7 in human plasma studied by reversed-phase high-performance liquid chromatography and micro-electrospray mass spectrometry.
    Journal of chromatography. A, 1996, Aug-30, Volume: 743, Issue:1

    In these experiments we have studied the in vitro metabolism of LVV-hemorphin-7 in human plasma by using reversed-phase high-performance liquid chromatography (RP-HPLC) in combination with micro-electrospray mass spectrometry (micro-ES-MS). Tandem mass spectrometry (MS-MS) was performed in order to verify the structure of the peptide fragments found. Incubations were performed with and without different protease inhibitors. Results showed that LVV-hemorphin-7 was metabolized from the N-terminal end of the peptide, probably by an amastatin-sensitive exopeptidase.

    Topics: Amino Acid Sequence; Angiotensin-Converting Enzyme Inhibitors; Anti-Bacterial Agents; Captopril; Chromatography, High Pressure Liquid; Endorphins; Gas Chromatography-Mass Spectrometry; Glycopeptides; Hemoglobins; Humans; Molecular Sequence Data; Opioid Peptides; Peptide Fragments; Peptides; Plasma; Protease Inhibitors; Spectrophotometry, Ultraviolet; Time Factors

1996
Reversed-phase high-performance liquid chromatography combined with tandem mass spectrometry in studies of a substance P-converting enzyme from human cerebrospinal fluid.
    Journal of chromatography. A, 1996, Aug-30, Volume: 743, Issue:1

    The application of reversed-phase HPLC in combination with micro-electrospray mass spectrometry to study a substance P (SP)-hydrolysing endoprotease in human cerebrospinal fluid (hCSF) is reported. The enzyme was partially purified from the hCSF specimens by ion-exchange chromatography and molecular sieving. During the purification procedure the enzyme activity was monitored by measuring the formation of the SP-fragment 1-7 from SP by radioimmunoassay. Regarding its behaviour upon molecular sieve chromatography, the enzyme was suggested to be associated with an apparent molecular mass of around 100 x 10(3). In subsequent experiments using the partially purified endopeptidase, the hydrolysis of SP was demonstrated by HPLC. The reaction product mixture was resolved in several components including the N-terminal fragments 1-8, 1-7 and 1-6 and the C-terminal fragment 8-11. The identity of these fragments were confirmed by tandem mass spectrometry. It was concluded that the present SP-degrading enzyme is different from those previously identified and purified from hCSF. The applied techniques were proven to be highly efficient for the recovery and identification of the released peptide products.

    Topics: Animals; Anti-Bacterial Agents; Captopril; Chromatography, Agarose; Chromatography, Gel; Chromatography, High Pressure Liquid; Conductometry; Cross Reactions; Endopeptidases; Gas Chromatography-Mass Spectrometry; Glycopeptides; Humans; Hydrolysis; Immune Sera; Iodine Radioisotopes; Peptide Fragments; Peptides; Protease Inhibitors; Rabbits; Radioimmunoassay; Spectrophotometry, Ultraviolet; Substance P

1996
Metabolism of AF1 (KNEFIRF-NH2) in the nematode, Ascaris suum, by aminopeptidase, endopeptidase and deamidase enzymes.
    Molecular and biochemical parasitology, 1996, Volume: 75, Issue:2

    We have studied the metabolism and inactivation of AF1 (KNEFIRF-NH2) by membranes prepared from the locomotory muscle of Ascaris suum. FIRF-NH2 and KNEFIRF were identified as three primary degradation products, resulting from the action of an endopeptidase, aminopeptidase and a deamidase, respectively. The endopeptidase resembled mammalian neprilysin (NEP, endopeptidase 24.11) in that the enzyme activity was inhibited by phosphoramidon and thiorphan and that it cleaved AF1 on the amino side of phenylalanine. The aminopeptidase activity was inhibited by amastatin and bestatin but not by puromycin. The deamidation of AF1 was inhibited by phenylmethylsulfonyl fluoride, p-chloromercuricphenylsulfonate and mercuric chloride, indicating that the deamidase enzyme is a serine protease with a requirement for a free thiol group for activity. AF1 (1 microM) induces an increase in tension and an increase in the frequency and amplitude of spontaneous contractions of an A. suum muscle strip. None of the aforementioned AF1 metabolites (2-20 microM) retained biological activity in this bioassay, indicating that the endopeptidase, aminopeptidase and deamidase have the potential to terminate the action of AF1 on locomotory muscle of A. suum.

    Topics: Amidohydrolases; Amino Acid Sequence; Aminopeptidases; Animals; Anti-Bacterial Agents; Ascaris suum; Enzyme Inhibitors; Glycopeptides; Helminth Proteins; Hydrogen-Ion Concentration; Molecular Sequence Data; Muscle Proteins; Neprilysin; Neuropeptides; Peptide Fragments; Peptides; Protease Inhibitors; Serine Proteinase Inhibitors

1996
Metabolism of AF1 (KNEFIRF-NH2) in the nematode Ascaris suum.
    Biochemical Society transactions, 1994, Volume: 22, Issue:3

    Topics: Amino Acid Sequence; Animals; Anti-Bacterial Agents; Ascaris suum; Endopeptidases; Glycopeptides; Molecular Sequence Data; Muscles; Neuropeptides; Peptide Fragments; Peptides; Protease Inhibitors

1994
Tachykinin receptors in the rat isolated uterus.
    Regulatory peptides, 1993, Jul-02, Volume: 46, Issue:1-2

    Tachykinin receptors mediating uterotonic effects were examined in preparations from oestrogen-primed rats. In the absence of peptidase inhibitors [Lys5-MeLeu9-Nle10] NKA (4-10) was 14-fold more potent than neurokinin A (NKA), but the two peptides were equipotent in the presence of phosphoramidon alone and in combination with amastatin. The NK-2 receptor antagonist SR 48968 antagonised responses to the tachykinins. These findings indicate that an NK-2 receptor is present in the oestrogen-primed rat uterus and that endopeptidase 24.11 plays a major role to inactivate NKA in this tissue.

    Topics: Animals; Anti-Bacterial Agents; Benzamides; Carbachol; Estradiol; Female; Glycopeptides; Kinetics; Neurokinin A; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Peptides; Piperidines; Rats; Rats, Sprague-Dawley; Receptors, Neurokinin-1; Receptors, Neurokinin-2; Uterus

1993
Intracisternal neutral endopeptidase-24.11 inhibitors produce inhibition in gastric acid output: independence from opiate, bombesin, or neurotensin-mediated mechanisms.
    Regulatory peptides, 1993, Jul-23, Volume: 46, Issue:3

    Intracisternal (ic) injection of the neutral endopeptidase-24.11 inhibitor phosphoramidon (1-100 nmol) produced a dose-dependent inhibition of gastric acid secretion in 2-h pylorus-ligated rats. The response resulted from a reduction in acid concentration and volume. Likewise, ic injection of another neutral endopeptidase-24.11 inhibitor Zincov (200 nmol) produced a 63% inhibition in gastric acid output. In contrast, neither intravenous injection of phosphoramidon (100 nmol) nor ic injection of the aminopeptidase inhibitor amastatin (100 nmol) produced any change in gastric acid secretion. The inhibitory effect of ic phosphoramidon (10 nmol) was not reversed by a dose of naloxone sufficient to antagonize the acid inhibitory effects of ic [D-Ala2-D-met5]enkephalinamide (8.5 nmol). Moreover, phosphoramidon-induced inhibition of acid was not reduced by the centrally effective bombesin antagonist N-acetyl-GRP(20-26)-O-CH3 or by reserpine pretreatment at a dose effective to antagonize ic neurotensin-induced inhibition in acid secretion. These results suggest that an endogenous neutral endopeptidase-24.11 sensitive substrate may act in the brain to inhibit gastric acid output by mechanisms independent of CNS opiate, bombesin or neurotensin activity.

    Topics: Animals; Anti-Bacterial Agents; Drug Interactions; Gastric Acid; Gastrointestinal Hormones; Glycopeptides; Hydroxamic Acids; Injections, Intravenous; Injections, Intraventricular; Male; Naloxone; Neprilysin; Oligopeptides; Peptides; Rats; Rats, Sprague-Dawley; Reserpine

1993
Endogenous methionine enkephalin may play an anticonvulsant role in the seizure-susceptible El mouse.
    Neurochemical research, 1993, Volume: 18, Issue:12

    After the intracisternal injection of three protease inhibitors which prevent the degradation of methionine enkephalin (amastatin, Des-Pro2-bradykinin, and phosphoramidon) and a mixture of these protease inhibitors, we investigated the effect on convulsive seizures in the seizure-susceptible El mouse. We also measured the cerebral methionine enkephalin content by high-performance liquid chromatography coupled with radioimmunoassay. Protease inhibitors significantly decreased both the incidence of seizures and the seizure score in El mice in a dose-dependent manner. This anticonvulsant effect was reversed by naloxone (2 mg/kg, sc). The cerebral methionine enkephalin content increased significantly after the administration of protease inhibitors in comparison with saline injection. These findings suggest that it was not protease inhibitors but instead increase of endogenous methionine enkephalin that reduced the incidence of seizures and the seizure score in El mice. Together with our previous data, the present findings support our hypothesis that a deficit in anticonvulsant endogenous methionine enkephalin is involved in the pathogenesis of seizures in the El mouse.

    Topics: Animals; Anti-Bacterial Agents; Bradykinin; Brain; Chromatography, High Pressure Liquid; Enkephalin, Methionine; Glycopeptides; Mice; Mice, Mutant Strains; Naloxone; Peptides; Protease Inhibitors; Radioimmunoassay; Seizures

1993
Degradation of oxytocin by the human placenta: effect of selective inhibitors.
    Acta endocrinologica, 1992, Volume: 127, Issue:1

    The hydrolysis of oxytocin by human placental subcellular fractions was studied in the presence of selective inhibitors by measuring liberated amino acids by high performance liquid chromatography (HPLC). Oxytocin degradation by microsomal and lysosomal fractions was inhibited by bestatin, amastatin and puromycin. The IC50 values of these inhibitors on oxytocin degradation by both fractions were similar to those of these inhibitors on the human placental aminopeptidase M measured by L-Leu-p-nitroanilide as a substrate (LAP activity), which we reported previously. However, purified aminopeptidase M from human placental microsomal fractions could not liberate any amino acid from oxytocin. Since phosphoramidon (1 mumol/l), a putative metalloendopeptidase inhibitor, and N-benzylcarbonyl-valyl-prolinal (Z-Val-prolinal) (14 mumol/l), a selective inhibitor of post-proline endopeptidase, could not significantly influence the degradation of oxytocin by either subcellular fractions, neither enzyme seems to be actively involved in oxytocin degradation. These results strongly suggested the existence of oxytocinase(s) other than the above three enzymes in microsomal and/or lysosomal fractions of human placenta.

    Topics: Amino Acids; Aminopeptidases; Anti-Bacterial Agents; CD13 Antigens; Chromatography, High Pressure Liquid; Cystinyl Aminopeptidase; Female; Glycopeptides; Humans; Leucine; Leucyl Aminopeptidase; Lysosomes; Microsomes; Oligopeptides; Oxytocin; Peptides; Placenta; Pregnancy; Prolyl Oligopeptidases; Puromycin; Serine Endopeptidases; Subcellular Fractions

1992
Effects of the subcutaneous administration of enkephalins on tail-flick response and righting reflex of developing rats.
    Brain research. Developmental brain research, 1992, Oct-23, Volume: 69, Issue:2

    The s.c. administration of [Met5]-enkephalin to 10-day-old rats pretreated with the mixture of 3 peptidase inhibitors, amastatin, captopril and phosphoramidon, produced the inhibition of tail-flick response and loss of righting reflex. When infant rats were pretreated with the mixture of any combination of two peptidase inhibitors, however, the change in both the response and the reflex were not produced at all by enkephalin injection, indicating that 3 kinds of enzymes, amastatin-sensitive aminopeptidase(s), captopril-sensitive peptidyl dipeptidase A and phosphoramidon-sensitive endopeptidase 24.11, played an important role in the inactivation of enkephalin after its systemic administration. Additionally, the fact that the two enkephalin-induced effects were more effectively antagonized by naloxone, a relatively selective mu-opioid antagonist, than by naltrindole, a specific delta-antagonist, or by nor-binaltorphimine, a specific kappa-antagonist, showed that these two effects were produced by the interaction of enkephalin with mu receptors. Moreover the involvement of mu receptors in the production of these two effects was shown by the fact that the s.c. administration of [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin, a selective mu agonist, also produced these two effects which were more effectively antagonized by naloxone than by naltrindole or nor-binaltorphimine. Since the magnitude of the two effects induced by enkephalins in 15-day-old rats was significantly lower than that in 10-day-old rats, and the two enkephalin-induced effects were not produced at all in 20-day-old rats, a maturation-induced decrease in the permeability of the blood-brain barrier against opioid peptides was indicated.

    Topics: Aging; Animals; Anti-Bacterial Agents; Captopril; Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; Enkephalin, Methionine; Enkephalins; Female; Glycopeptides; Injections, Subcutaneous; Male; Oligopeptides; Pain; Peptides; Posture; Protease Inhibitors; Rats; Rats, Wistar; Reflex

1992
Inhibition of peptidases potentiates enkephalin-stimulated contraction of gastric muscle cells.
    The American journal of physiology, 1991, Volume: 261, Issue:3 Pt 1

    Cell surface peptidases degrade enkephalins and thereby restrict the number of molecules available to activate receptors. The effects of peptidase inhibitors on degradation of enkephalins and on enkephalin-stimulated contraction of gastric smooth muscle cells were examined. Muscle cells dispersed from the guinea pig stomach degraded [Tyr1-3H] [Leu5]enkephalin (41.6 +/- 9.0% degradation at 60 min incubation, mean +/- SD, n = 4 animals). Amastatin (10 microM, an aminopeptidase inhibitor) inhibited degradation by 72.1 +/- 1.5% The residual peptidase activity was inhibited by phosphoramidon (1 microM, an endopeptidase EC 3.4.24.11 inhibitor) by 58.0 +/- 11.0%. [Tyr1-125I] [Met5]enkephalin was similarly degraded. Phosphoramidon (1 microM) inhibited the degradation of the aminopeptidase-resistant peptide [Tyr1-3H] [D-Ala2]-[Leu5]enkephalin by greater than 95%. [Met5]enkephalin, incubated with cells for 30 s, stimulated contraction [50% maximal contraction (EC50) 120 +/- 50 nM, n = 6]. Pretreatment of cells with phosphoramidon alone, amastatin alone, or phosphoramidon plus amastatin, caused 20-fold (EC50 6.5 +/- 1.1 nM), 2-fold (EC50 63 +/- 23 nM), and 100-fold (EC50 1.1 +/- 0.3 nM) increase in potency of [Met5]enkephalin, respectively. The results show that endopeptidase EC 3.4.24.11 and aminopeptidases contribute to degradation of enkephalins by gastric muscle cells. The rapidity and magnitude of the potentiating effects of the inhibitors on enkephalin-stimulated contraction suggest a close physical relationship between the peptidases and the enkephalin receptors.

    Topics: Aminopeptidases; Animals; Anti-Bacterial Agents; Carbachol; CD13 Antigens; Enkephalin, Leucine; Enkephalin, Methionine; Glycopeptides; Guinea Pigs; In Vitro Techniques; Kinetics; Male; Muscle Contraction; Muscle, Smooth; Neprilysin; Oligopeptides; Peptides; Protease Inhibitors; Recombinant Proteins; Stomach

1991
Inactivation of dynorphin-(1-8) in isolated preparations by three peptidases.
    Japanese journal of pharmacology, 1988, Volume: 47, Issue:4

    Inactivation of dynorphin-(1-8) in three in vitro isolated preparations, guinea-pig ileum, mouse vas deferens and rabbit vas deferens, was estimated by employing the relatively specific inhibitors of enkephalin-hydrolyzing enzymes. All three enzyme inhibitors, amastatin, captopril and phosphoramidon, significantly enhanced the inhibitory potency of dynorphin-(1-8) in the three isolated preparations. The magnitude of the enhancement of the dynorphin potency by captopril was significantly higher than that by either amastatin or phosphoramidon in guinea-pig ileum; that by amastatin was significantly higher than that by either captopril or phosphoramidon in rabbit vas deferens; and that by amastatin was similar to that by captopril, but significantly higher than that by phosphoramidon in mouse vas deferens. The Ke values of three antagonists, naloxone, Mr 2266 and ICI 154129, against dynorphin-(1-8) in the presence of the three peptidase inhibitors indicated that dynorphin-(1-8) acted on kappa receptors in guinea-pig ileum and on both kappa and delta receptors in mouse vas deferens. Since amastatin, captopril and phosphoramidon produced the naloxone-reversible inhibition of contractions of guinea-pig ileum in the presence of dynorphin-(1-8), all three dynorphin-inactivating enzymes were indicated to be located very close to kappa receptors.

    Topics: Aminopeptidases; Animals; Anti-Bacterial Agents; Captopril; Dynorphins; Glycopeptides; Guinea Pigs; Hypothalamic Hormones; In Vitro Techniques; Male; Mice; Mice, Inbred ICR; Muscle Contraction; Muscle, Smooth; Neprilysin; Oligopeptides; Peptide Fragments; Peptides; Peptidyl-Dipeptidase A; Protease Inhibitors; Rabbits

1988
Estimation of relative importance of three enzymes in the inactivation of [Met5]-enkephalin and [Met5]-enkephalin-Arg6 in three isolated preparations by employing the inhibitor specific for each enzyme.
    Japanese journal of pharmacology, 1987, Volume: 44, Issue:3

    The relative importance of three enzymes, amastatin-sensitive aminopeptidase, captopril-sensitive peptidyl dipeptidase A and phosphoramidon-sensitive endopeptidase-24.11, to inactivate two opioid peptides, [Met5]-enkephalin and [Met5]-enkephalin-Arg6, was investigated in three in vitro isolated preparations, guinea-pig ileum, mouse vas deferens and rat vas deferens, by estimating the magnitude of the enhancement of the inhibitory potency of the opioid peptide by each peptidase inhibitor. Results showed that the relative importance of the three enzymes in the inactivation of the opioid peptide, whether it was [Met5]-enkephalin or [Met5]-enkephalin-Arg6, in guinea-pig ileum was significantly different from that in either mouse vas deferens or rat vas deferens. Additionally, the relative importance of the three enzymes in the preparation, whether it was guinea-pig ileum, mouse vas deferens or rat vas deferens, in the inactivation of [Met5]-enkephalin was significantly different from that of [Met5]-enkephalin-Arg6. The significance of the presence of plural inactivating-enzymes for opioid peptides was discussed.

    Topics: Aminopeptidases; Animals; Anti-Bacterial Agents; Captopril; Enkephalin, Methionine; Enkephalins; Glycopeptides; Guinea Pigs; Ileum; In Vitro Techniques; Male; Metalloendopeptidases; Mice; Mice, Inbred ICR; Neprilysin; Oligopeptides; Peptides; Peptidyl-Dipeptidase A; Rats; Rats, Inbred Strains; Vas Deferens

1987
Determination and characterization of succinyl tri-alanine p-nitroanilide hydrolyzing metalloendopeptidase in serum.
    Enzyme, 1987, Volume: 37, Issue:4

    Serum succinyl (Ala)3-p-nitroanilide hydrolyzing elastase-like activity which elevates in patients with obstructive jaundice, is due to the joint action of two enzymes: first, succinyl (Ala)3-p-nitroanilide is cleaved to succinyl (Ala)2 and Ala-p-nitroanilide by metalloendopeptidase, and then Ala-p-nitroanilide is cleaved to Ala and p-nitroaniline by aminopeptidase. We adopt a new assay method for serum endopeptidase activity using HPLC.

    Topics: Adult; Aminopeptidases; Anti-Bacterial Agents; Cholestasis; Chromatography, High Pressure Liquid; Diabetes Mellitus; Endopeptidases; Female; Glycopeptides; Humans; Immunodiffusion; Kidney; Leucine; Male; Metalloendopeptidases; Neoplasms; Oligopeptides; Peptides; Protease Inhibitors

1987
Inactivation of [Leu5]-enkephalin in three isolated preparations: relative importance of aminopeptidase, endopeptidase-24.11 and peptidyl dipeptidase A.
    NIDA research monograph, 1986, Volume: 75

    Enkephalin had been shown to be almost exclusively hydrolyzed by three peptidases in the previous studies. In the present investigation, the relative importance of three enzymes in the inactivation of [Leu5]-enkephalin was examined in three isolated preparations. Results showed that amastatin-sensitive aminopeptidase played the greatest role in both guinea-pig ileum and rat vas deferens while it played the similar role to either phosphoramdidon-sensitive endopeptidase-24.11 or captopril-sensitive peptidyl dipeptidase A in mouse vas deferens.

    Topics: Animals; Anti-Bacterial Agents; Captopril; Enkephalin, Leucine; Glycopeptides; Guinea Pigs; Hydrolysis; Ileum; Kinetics; Male; Metalloendopeptidases; Mice; Neprilysin; Oligopeptides; Peptides; Peptidyl-Dipeptidase A; Vas Deferens

1986
The enhancing effects of amastatin, phosphoramidon and captopril on the potency of [Met5]-enkephalin in rat vas deferens.
    Japanese journal of pharmacology, 1986, Volume: 42, Issue:1

    The enkephalin-inactivating enzymes in rat vas deferens were studied by using the relatively specific inhibitor of each enzyme. The results showed that the rat vas deferens, like the other three preparations, guinea-pig ileum, mouse vas deferens and striatal membranes of guinea-pig brain, which had been investigated previously, contained three distinct enkephalin-hydrolyzing peptidases. Additionally, the enkephalin-hydrolyzing aminopeptidase, endopeptidase-24.11 and peptidyl dipeptidase A in rat vas deferens were found to be inhibited maximally with 1 microM of amastatin, 1 microM of phosphoramidon and 1 microM of captopril, respectively. In contrast to these three enzymes, both L-tyrosyl-L-tyrosine-sensitive dipeptidyl aminopeptidase and D-phenylalanine-sensitive carboxypeptidase were suggested not to be involved significantly in the inactivation of exogenously given enkephalin in rat vas deferens. The characteristics of the enkephalin-degradative enzymes in rat vas deferens were discussed in terms of their similarities to and differences from those in the other preparations.

    Topics: Animals; Anti-Bacterial Agents; Captopril; Electric Stimulation; Enkephalin, Methionine; Glycopeptides; In Vitro Techniques; Male; Muscle, Smooth, Vascular; Naloxone; Oligopeptides; Peptides; Protease Inhibitors; Rats; Rats, Inbred Strains; Vas Deferens

1986