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W Neugebauer

Publications and source records attributed to W Neugebauer.

At least 19 recordsLinked to original sources
PubMed1999

Agonist-dependent AT(4) receptor internalization in bovine aortic endothelial cells.

Recent studies have characterized a specific binding site for the C-terminal 3-8 fragment of angiotensin II (Ang IV). In the present study we looked at the internalization process of this receptor on bovine aortic endothelial cells (BAEC). Under normal culture conditions, BAEC efficiently internalized (125)I-Ang IV as assessed by acid-resistant binding. Internalization of (125)I-Ang IV was considerably decreased after pretreatment of cells with hyperosmolar sucrose or after pretreatment of BAEC with inhibitors of endosomal acidification such as monensin or NH(4)Cl. About 50% of internalized (125)I-Ang IV recycled back to the extracellular medium during a 2 h incubation at 37 degrees C. (125)I-Ang IV remained mostly intact during the whole process of internalization and recycling as assessed by thin layer chromatography. As expected, internalization of (125)I-Ang IV was completely abolished by divalinal-Ang IV, a known AT(4) receptor antagonist. Interestingly, (125)I-divalinal-Ang IV did not internalize into BAEC. These results suggest that AT(4) receptor undergoes an agonist-dependent internalization and recycling process commonly observed upon activation of functional receptors.

Angiotensin II↗
PubMed1999

[Occupational risk factors for bladder carcinoma. A case control study].

Aim of this case-control study, performed on 412 male bladder cancer cases and 414 controls with benign prostatic hyperplasia in a former area of coal, iron and steel industries in Germany, was to identify occupations with an increased bladder cancer risk. In bladder cancer cases, smokers were overrepresented (58.3%) compared to controls (35.2%). The percentage of patients who had stopped smoking for at least 10 years did not differ in cases (10.2%) and controls (9.7%). Significantly elevated smoking-adjusted bladder cancer odds ratios (MH) were observed in painters and lacquers (MH 2.24, 95% CI 1.07-5.13), chemistry-related occupations (MH 2.44, 95% CI 1.05-5.67), coke plant workers (MH 2.89, 95% CI 1.16-7.16) and hard coal miners (MH 2.33, 95% CI 1.52-3.58). Significantly decreased smoking-adjusted bladder cancer odds ratios (MH) were observed in businessmen (MH 0.64, 95% CI 0.45-0.92) and office personnel (MH 0.58, 95% CI 0.41-0.81). In these two groups a relevant exposure to occupational bladder carcinogens is not likely.

Aged↗
PubMed1999

Kinin B1 receptor antagonists containing alpha-methyl-L-phenylalanine: in vitro and in vivo antagonistic activities.

-To protect from metabolism and to improve potency of the AcLys-[D-betaNal7,Ile8]desArg9-bradykinin (BK) (R 715), we prepared and tested 3 analogues containing alpha-methyl-L-Phe ([alphaMe]Phe) in position 5: these are the AcLys-[(alphaMe)Phe5,D-betaNal7, Ile8]desArg9BK (R 892), Lys-Lys-[(alphaMe)Phe5,D-betaNal7, Ile8]desArg9BK (R 913), and AcLys-Lys-[(alphaMe)Phe5,D-betaNal7, Ile8]desArg9BK (R 914). The new compounds were tested against the contractile effect induced by desArg9BK on 2 B1 receptor bioassays, the human umbilical vein, and the rabbit aorta. Their antagonistic activities were compared with those of the early prototypes (Lys-[Leu8]desArg9BK and [Leu8]desArg9BK) and with other recently described peptide antagonists. The 3 (alphaMe)Phe analogues showed high antagonistic potencies (pA2) at both the human (8.8, 7.7, and 8. 7, respectively) and rabbit (8.6, 7.8, and 8.6, respectively) B1 receptors. No antagonistic effects (pA2<5) were observed on the B2 receptors that mediate the contractile effects of BK on the human umbilical vein, the rabbit jugular vein, and the guinea pig ileum. Moreover, these new B1 antagonists were found to be resistant to in vitro degradation by purified angiotensin-converting enzyme from rabbit lung. The Nalpha-acetylated forms, R 892 and R 914, were resistant to aminopeptidases from human plasma. In vivo antagonistic potencies (ID50) of B1 receptor antagonists were evaluated in anesthetized lipopolysaccharide-treated (for B1 receptor) and nontreated (for B2 receptor) rabbits against the hypotensive effects of exogenous desArg9BK and BK. R 892 efficiently inhibited (ID50 2.8 nmol/kg IV) hypotension induced by desArg9BK without affecting that evoked by BK (ID50 >600 nmol/kg IV). Conversely, the peptide antagonists Lys-Lys-[Hyp3,Igl5,D-Igl7,Oic8]desArg9BK (B 9858) and DArg-[Hyp3,Thi5,D-Tic7,Oic8] desArg9BK (S 0765) showed dual B1/B2 receptor antagonism in vitro and in vivo. It is concluded that R 892 and congeners provide selective, highly potent, and metabolically stable B1 kinin receptor antagonists that can be useful for the assessment of the physiological and pathological roles of kinin B1 receptors.

Animals↗
PubMed1998

Adenovirus endopeptidase and papain are inhibited by the same agents.

Adenoviruses encode a cysteine protease (AVP) which carries out highly specific cleavages on at least seven viral proteins and two cellular proteins. Virus infectivity is dependent on this function. The three-dimensional positions of the amino acids involved in catalysis display a striking similarity to those of papain, suggesting a similar catalytic mechanism. This similarity has prompted us to compare the effect of papain inhibitors on the two enzymes. AVP and papain activity was tested on a fluorescent peptide substrate as well as on metabolically labeled adenovirus (Ad2) precursor proteins. Hep2 cells infected with Ad2 were exposed to inhibitors and assayed for, (a) infectious virus, (b) in situ Ad2 protease activity, (c) physical particle production and their polypeptide composition. We found that in both substrate systems AVP was sensitive to the papain inhibitors benzamidoacetonitrile, acetamidoacetonitrile and N-methoxyphenylalanine glycylnitrile, and that the degree of sensitivity was influenced by the substrate. Unlike papain, AVP was relatively insensitive to E64. In ex vivo tests, Hep2 cells infected with Ad2 were exposed to inhibitors and assayed for, (a) infectious virus, (b) in situ Ad2 protease activity, (c) physical particle production and their polypeptide composition. A 4-fold reduction in virus titer was obtained when the inhibitors were added between 17 and 25 h after infection. Processing of precursor proteins was also inhibited yet the production of physical particles was only reduced 2-fold. These experiments show that papain inhibitors are also capable of inhibiting the adenovirus protease both in vitro and ex vivo, thus forging a possible link between structural similarity and functionality.

Acetonitriles↗
PubMed1997

Angiotensin analogues palmitoylated in positions 1 and 4.

Lipidated angiotensin II (Ang) agonists and antagonists were synthesized and evaluated for their biological activities for eventual use an antimyoproliferative agents. Solid phase peptide synthesis was used for the assembly of the peptides with the Fmoc protection scheme. N-Acetyl-Ser1 Ang was palmitoylated on the serine hydroxyl function. The nonpalmitoylated analogue retained one-third of Ang's affinity toward the AT1 receptor on bovine adrenal cortex membranes, and the palmitoylated analogue was essentially inactive. Upon enzymatic lipolysis or mild saponification of the palmitoylated peptide, biological activity was restored. An analogous compound of Ang, N-acetyl-Ser1,beta-D-naphthylalanine8 ([NAcSer1,D-Nal8]Ang), was a pure antagonist on rabbit aorta but with lower affinity. Its O-palmitoylated form was inactive as well but was easily converted to the nonlipidated active form by lipolysis or saponification. Direct palmitoylation of [sarcosine1]Ang with palmitoyl chloride was obtained on the free phenolic hydroxyl of Tyr4 on solid phase on an otherwise fully protected peptide. This lipopeptide was fully active, was comparable to [Sar1]Ang, and exhibited strongly prolonged activity. Lipolysis and saponification under mild conditions yielded standard [Sar1]Ang. The corresponding [Sar1,D-Nal8]Ang was a potent and very long-lasting antagonist (pA2 = 8.1), and its analogous palmitoyl phenyl ester in position 4 was active in its palmitoylated form (antagonist) and, again, returned to the nonlipidated form upon saponification or lipolysis. [Sar1,Tyr4(O-octadecyl)]Ang, an analogue to Tyr-palmitoylated [Sar1]Ang with an octadecyl phenyl ether in position 4, was also prepared. Surprisingly, the ether compound was inactive. Premature hydrolysis of the palmitoyl phenyl ester peptide was excluded by HPLC analysis, and the activity of the ester peptide is attributed to a putative hydrogen bond that may be critical for biological activity. The discovery of potent biologically active lipidated antagonists of Ang gives access to potential antimyoproliferative agents with numerous application possibilities.

Adrenal Cortex↗
PubMed1997

Bioactivities and secondary structures of constrained analogues of human parathyroid hormone: cyclic lactams of the receptor binding region.

In a search for analogues of human parathyroid hormone (hPTH) with improved activities and bioavailabilities, we have prepared the following three lactam analogues of hPTH-(1-31)-NH2 (1) or [Leu27]hPTH-(1-31)-NH2 (2): [Leu27]cyclo(Glu22-Lys26)-hPTH-(1-31)-NH2 (3), [Leu27]cyclo(Lys26-Asp30)-hPTH-(1-31)-NH2 (4), and cyclo(Lys27-Asp30)-hPTH-(1-31)-NH2 (5). Analogues 1, 2, and 5 had seven or eight residues of alpha-helix, as estimated from their circular dichroism (CD) spectra, in contrast to 12 residues in cyclic analogues 3 and 4. Thus, lactams 3 and 4 stabilized a helix previously shown to exist within residues 17-29. The adenylyl cyclase activity (EC50), measured in rat osteosarcoma 17/2 cells, of 5 (40.3 +/- 2.3 nM) was half that of its linear form 1 (19.9 +/- 3.9 nM). The linear Leu27 mutant 2 was twice as active (11.5 +/- 5.2) as analogue 1, and lactam analogue 3 was 6-fold more active (3.3 +/- 0.3 nM). Lactam analogue 4 had less activity (16.9 +/- 3.3 nM) than 2, its linear form. Peptides hPTH-(1-30)-NH2 (6), [Leu27]hPTH-(1-30)-NH2 (7), and [Leu27]cyclo(Glu22-Lys26)-hPTH-(1-30)-NH2 (8) all had AC-stimulating activities similar to that of 1. When injected intravenously, with a dose of 0.8 nmol/100 g of analogue in acid saline, hypotensive effects paralleled their adenylyl cyclase activities. They behaved quite differently when applied subcutaneously. Analogues 1, 5, and 6, the weakest, showed about half the drop in blood pressure observed with 3 and 4, the most active. In contrast, the time required to reach a maximum drop in blood pressure of 4-8, after subcutaneous administration, was 2-4 times that of the other analogues. Thus, the bioavailabilities of the lactam analogues, unlike their adenylyl cyclase-stimulating activities, were highly dependent on the presence or conformation of Val31.

Adenylyl Cyclases↗
PubMed1997

Pharmacological heterogeneity of neurotensin receptors: an in vitro study.

Neurotensin (NT), a linear tridecapeptide, has been shown to exert a variety of biological effects in the periphery and in the central nervous system. The aim of the present study was to characterize the NT receptors mediating the contractions of two isolated organs, the rat stomach strip and the guinea pig ileum. More than 20 compounds, peptides, nonpeptides, or pseudopeptides, were tested for their agonistic and antagonistic effects against NT and a series of potent analogs or fragments. Receptors were characterized using the two classical criteria suggested by Schild, the order of potency of agonists and the affinity of antagonists. The results shown in this study indicate that the contractions of the guinea pig ileum in response to NT are mediated by acetylcholine and prostaglandins because they are blocked by atropine and indomethacin. The contractions induced by NT in the rat stomach are not influenced by atropine, indomethacin, methysergide, and diphenhydramine and may result from the direct activation of smooth muscle receptors. Differences in the order of potency of agonists were also found between the two preparations: in the rat stomach strip, the order of potency was AcNT(8-13) > Arg-NT(8-13) > Lys-NT(8-13) > NT = NT(8-13) and in the guinea pig ileum was Arg-NT(8-13) > AcNT(8-13) > NT = NT(8-13) > Lys-NT(8-13). The nonpeptide antagonist SR 48692 was shown to possess higher apparent affinity for the rat stomach functional sites (pA2 8.0) than for those of the guinea pig ileum (pA2 6.45). The results presented in this paper suggest that two different pharmacological entities may subserve the myotropic effect of NT and some analogs and fragments in the gastrointestinal tract of the guinea pig and the rat.

Amino Acid Sequence↗
PubMed1997

Antagonists for kinin B1 and B2 receptors in the mouse.

Contractile responses to B1 and B2 receptor agonists have been demonstrated in the mouse stomach; the mouse urinary bladder responds only to B2 receptor agonists. These tissues were used in this study to investigate the antagonistic effect of four B2 receptor antagonists, namely, DArg[Hyp3,DPhe7,Leu8]BK (BK, bradykinin), HOE-140, WIN 64338, and FR-173657 (B2 receptor antagonists), as well as three B1 kinin receptor antagonists; [Leu8]desArg9BK, Lys[Leu8]desArg9BK, and AcLys[D beta Nal7,Ile8]desArg9BK, were investigated. Results shown indicate that DArg[Hyp3,DPhe7,Leu8]BK is a partial agonist, while HOE-140 and FR-173657 are pure antagonists, devoid of direct myotropic effects, and quite selective for the B2 receptor. WIN 64338 was essentially inactive on both B1 and B2 receptors. The myotropic effect of DArg[Hyp3,DPhe7,Leu8]BK is blocked by HOE-140. Similarly, Lys[Leu8]desArg9BK and [Leu8]desArg9BK are B1 receptor partial agonists whose activities are blocked by AcLys[D beta Nal7,Ile8]desArg9BK (code name R 715), a fairly pure B1 receptor antagonist. Both HOE-140 and FR-173657 are long-acting, slowly reversible compounds that exert a noncompetitive type of antagonism, while R 715 is rapidly reversible and, thus, possibly competitive. Data presented in this paper provide a pharmacological characterization of B1 and B2 receptor antagonists in the mouse and underline the positive features of FR-173657 as a potent and selective B2 receptor antagonist, as well as the potency and purity of R 715 as a B1 receptor antagonist in the mouse.

Animals↗
PubMed1997

Pharmacological profiles of the human and rabbit B1 receptors.

Twenty-two peptides related to kinins were used (i) to examine some chemical features required for the human and rabbit B1 receptor activation or blockade and (ii) to establish the existence of a correlation between the pharmacological spectrum of the B1 receptor obtained on the rabbit aorta (rbA) and the human umbilical vein (hUV). The apparent affinities of these peptides were measured in vitro using classical bioassays and are expressed in terms of pD2 (for agonists) or pA2 values (for antagonists). Selectivity for the B1 receptor was demonstrated by testing the peptides against the effect of bradykinin (BK) on the hUV and the rabbit jugular vein (rbJV), two preparations containing B2 receptor-mediating vasoconstriction. The results show that (i) lysyl-peptide agonists and antagonists demonstrate higher affinities than nonlysyl compounds on human and rabbit B1 receptors, (ii) peptides containing hydrophobic D-residues (e.g., Tic, beta Nal, Hyp(trans-propyl), Igl) in position 7 are suitable for B1 receptor antagonism, and (iii) the additive substitution of an Oic residue in position 8 leads to nonselective kinin receptor antagonists. Moreover, a high (r = 0.92) and positive (regression slope = 0.99 +/- 0.09) correlation between the affinities measured for the kinin analogues in two B1 receptor bioassay systems has been revealed. Based on the similarity of pharmacological profiles observed in the rabbit and human B1 receptors, we suggest that the B1 receptor domain in which peptide agonists and antagonists interact may be similar in these two species.

Amino Acid Sequence↗
PubMed1997

Pharmacological characterization of nociceptin receptor: an in vitro study.

The newly discovered neuropeptide nociceptin (NC) (alias orphanin FQ) was tested for its potential direct effects as well as for its ability to modify the electrically evoked contractions in several isolated organs suspended in vitro. NC was inactive both as stimulant and as inhibitor of smooth muscle in several preparations, whereas it inhibited the contractions induced by electrical field stimulation in the mouse vas deferens and guinea pig ileum. NC showed the same potency (IC50 = 10 nM) in the two preparations. However, it was significantly more effective in the mouse vas deferens (maximal effect -80%) than in the guinea pig ileum (maximal effect -50%). The inhibitory effect exerted by NC in the two preparations was not affected by naloxone or more selective opioid receptor antagonists. Moreover, by truncation of C-terminal sequences, NC fragments were designed. These fragments were subsequently tested in the mouse vas deferens and in the guinea pig ileum: NC(1-13)-NH2 was the smallest peptide maintaining the same efficacy and potency as the natural peptide. Finally, NC-NH2 and its fragments NC(1-13)-NH2 and NC(1-9)-NH2 were modified by substituting the phenylalanine 1 residue with a tyrosine. These peptides were tested in the guinea pig ileum, where they behaved as mixed NC-opioid receptor agonists ([Tyr1]NC-NH2 and [Tyr1]NC(1-13)-NH2) or as pure opioid receptor agonists ([Tyr1]NC(1-9)-NH2. In conclusion, the present paper demonstrated that the electrically stimulated mouse vas deferens and guinea pig ileum can be used as a sensitive bioassay for studying the pharmacology of NC and related compounds.

Amino Acid Sequence↗
PubMed1997

The use of confocal microscopy in the investigation of cell structure and function in the heart, vascular endothelium and smooth muscle cells.

In recent years, fluorescence microscopy imaging has become an important tool for studying cell structure and function. This non invasive technique permits characterization, localisation and qualitative quantification of free ions, messengers, pH, voltage and a pleiad of other molecules constituting living cells. In this paper, we present results using various commercially available fluorescent probes as well as some developed in our laboratory and discuss the advantages and limitations of these probes in confocal microscopy studies of the cardiovascular system.

Animals↗
PubMed1996

Peptide synthesis on chitin.

The use of chitin as a support for solid-phase peptide synthesis is described and illustrated by synthesis of four peptides, varying in length from 10 to 29 residues. Syntheses were performed in a continuous-flow peptide synthesizer, using Fmoc chemistry. A cleavable linker, p-[(R,S)-alpha-[1-(9H-fluoren-9-yl)-methoxyformamido]-2,4-di methoxybenzyl]- phenoxyacetic acid, was attached to chitosan at the desired substitution level, and the complex acetylated to yield a linker substituted chitin. The effects of temperature, solvents and degree of linker substitution on the syntheses were studied. Acyl carrier peptide (ACP) synthesis studies indicated that temperature was the single most important parameter. Increasing the temperature of the synthesis from 20 to 55 degrees C resulted in an enormous improvement of this synthesis, with about 90% of the crude product being the correct peptide. Denaturing solvents, such as DMSO, could be used without significant effect on the flow properties of the support. The synthesis of one peptide was mainly improved by lowering the degree of substitution from 0.3 to 0.1 mmol/g, suggesting peptide aggregation was a problem in this case. The results of three syntheses on chitin were comparable with those obtained with a commonly used commercial support. This work shows that, under appropriate conditions, chitin can be utilized directly as a support for peptide synthesis.

Amino Acid Sequence↗
PubMed1996

Structure-activity studies of B1 receptor-related peptides. Antagonists.

We tested several peptides related to des-Arg9-bradykinin as stimulants or inhibitors of B1 (rabbit aorta, human umbilical vein) and B2 (rabbit jugular vein, guinea pig ileum, human umbilical vein) receptors. We also incubated the compounds with purified angiotensin-converting enzyme from rabbit lung to test their resistance to degradation. We evaluated apparent affinities (in terms of the affinity constant pA2) of compounds and their potential residual agonistic activities (alpha E). Bradykinin and des-Arg9-bradykinin were used as agonists for the B2 and B1 receptors, respectively. Degradation of peptides by the angiotensin-converting enzyme was prevented in the presence of a D-residue in position 7 of des-Arg9-bradykinin. Replacement of Pro7 with D-Tic combined with Leu, Ile, Ala, or D-Tic in position 8 led to weak B1 receptor antagonists, some of which had strong residual agonistic activities on the B2 receptor preparations. The use of D-beta Nal in position 7, combined with Ile in position 8 and AcLys at the N-terminal (eg, AcLys[D-beta Nal7, Ile8]des-Arg9-bradykinin) gave the most active B1 receptor antagonist (pA2 of 8.5 on rabbit aorta and human umbilical vein), which is also partially resistant to enzymatic degradation. Extension of the N-terminal end by Sar-Tyr-epsilon Ahx (used for labeling purposes) and even cold-labeling of Tyr with iodine were compatible with high, selective, and specific antagonism of the B1 receptors. We compared some compounds with some already known B1 receptor antagonists to underline the novelty of new peptidic compounds.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1995

Solution structure and adenylyl cyclase stimulating activities of C-terminal truncated human parathyroid hormone analogues.

Analogues of human parathyroid hormone (hPTH) truncated at the C-terminal end have been studied for adenylyl cyclase (AC) activity and for solution conformation by circular dichroism (CD) spectroscopy. Analogues of hPTH-(1-34)-NH2, containing the first 28-31 residues, had only a slightly diminished ability to stimulate AC in rat osteosarcoma (ROS) cells as compared to that of the parent analogue. CD data on hPTH-(16-34)-NH2 and C-terminal deletion mutants of hPTH-(1-34)-NH2 supported the presence of a partially stable alpha-helix over residues 17-28. A carboxyl-terminal mutant, hPTH-(1-30)-OH, showed both reduced helix and greatly reduced AC-stimulating activity as compared to the corresponding amide analogue. In contrast, both of these analogues, in the presence of palmitoyloleoylphosphatidylserine (POPS) vesicles, showed an equal stabilization of alpha-helix. All other analogues showed at least some enhancement of alpha-helix in the presence of POPS. However, both in neutral, aqueous buffer and in POPS, the relative amount of alpha-helix decreased greatly as the peptide was shortened below the 1-28 sequence. These data provide additional support for an amphiphilic alpha-helix over residues 21-28 being the conformation for receptor binding of hPTH for stimulation of AC activity. Modeling human parathyroid hormone-related peptide as an alpha-helix over this same region, and comparison to hPTH, suggests that both may bind via the hydrophobic face to the receptor.

Adenylyl Cyclases↗
PubMed1995

Kinin B1 and B2 receptors in the mouse.

A systematic study has been performed in various segments of the intestine and in the urinary bladder of the mouse to identify tissues that respond to kinins and possess B1 and (or) B2 receptors. The stomach was found to contain B1 and B2 functional sites that show pharmacological profiles compatible with B1 and B2 receptors, whereas the urinary bladder possesses only B2 sites. Myotropic responses mediated by B1 receptors show slow onset and reversibility compared with responses evoked by the activation of B2 receptors. The order of potency of agonists is bradykinin (BK) > or = [Hyp3]BK > [Aib7]BK on the B2 of both the stomach and urinary bladder, while desArg9-BK is inactive. The order of potency of agonists on the B1 receptor is [Lys]desArg9BK < or = desArg9BK, while BK and the other B2 agonists are inactive. B2 antagonists of the first generation, such as DArg[Hyp3,DPhe7]BK, act as partial agonists and show residual agonistic activities higher than 0.5, while HOE-140 shows high affinity and very little residual agonistic activity; WIN 64338 is almost inactive. On the B1 receptor, classical antagonists, such as [Leu8]desArg9BK and Lys[Leu8]desArg9BK, act as partial agonists. A modification of their structures has led to a new compound (R-715) that shows fairly high affinity (pA2 7.0) and little residual agonistic effect. This compound has been used for B1 receptor characterization in the stomach. Residual agonistic activities of both B2 and B1 antagonists appear to be mediated by B2 and B1 receptors, respectively. Data presented in this paper provide the pharmacological basis for sensitive and selective preparations to be used for studying B1 and B2 receptors in the mouse.

Animals↗
PubMed1995

[Phe22]-big endothelin-1[19-37]: a new and potent inhibitor of the endothelin-converting enzyme.

To develop a new family of endothelin-converting enzyme (ECE) inhibitors, we assessed the inhibitory properties of an N- and C-terminal-truncated analogue of big endothelin (ET)-1[1-38], [Phe22]-big-ET-1 [19-37], on the renal vasoconstrictor properties of big ET-1 and ET-1. We had previously shown that a 60-min infusion of phosphoramidon (100 microM) potently inhibits big-ET-1 but not ET-1-induced vasoconstriction in the rabbit perfused kidney. A 1-min preinfusion of the analogue (100 microM) was sufficient to markedly blunt the vasoconstrictor actions of big ET-1 (250 pmol) (control 19.3 +/- 1.0 mm Hg; in presence of [Phe22]-big ET-1[19-37] 4.8 +/- 1.6 mm Hg; n = 6, p < 0.001). A 10- and 60-min preinfusion of the substrate analogue at concentrations of 50 and 5 microM, respectively, reduced the response of big ET-1 to 9.3 +/- 2.3 mm Hg and 3.6 +/- 1.8 mm Hg (n = 3, p < 0.01 compared to control). [Phe22]-big ET-1[19-37] was inactive against ET-1-induced vasoconstriction (10 pmol) and was devoided of intrinsic vasoactivity. All experiments were performed on kidneys pretreated with SQ-28603 (1 microM), a neutral endopeptidase inhibitor. Our results suggest that [Phe22]-big ET-1[19-37] may be a useful lead molecule for the development of more selective and enzyme-resistant inhibitors of the membrane-bound ECE.

Animals↗