Takeo Tanaka

Endothelin ETB receptor antagonist, RES-701-1: effects on isolated blood vessels and small intestine.

RES-701-1 (cyclic (Gly1-Asp9)(Gly-Asn-Trp-His-Gly-Thr-Ala-Pro-Asp-Trp-Phe- Phe-Asn-Tyr-Tyr-Trp)), a peptide isolated from Streptomyces sp., has been reported to inhibit the endothelin ETB receptor. We examined the effects of this peptide on the blood vessels and the small intestine. In isolated rat aorta without endothelium, 10 microM RES-701-1 did not affect the resting tone, nor did it attenuate the contractions induced by endothelin-1, endothelin-3 or norepinephrine. In the aorta with endothelium, 3 microM RES-701-1 shifted the concentration-response curves for the contractile effects of endothelin-1 and endothelin-3 to the left. Removal of endothelium showed a similar effect to 3 microM RES-701-1. In the norepinephrine-stimulated aorta, endothelium-dependent relaxation induced by endothelin-3 was antagonized by 0.3-10 microM RES-701-1 in a concentration-dependent manner. In the guinea pig ileum stimulated by carbachol, endothelin-3 induced a transient relaxation followed by sustained relaxation. RES-101-1 (3 microM) selectively inhibited the transient relaxation. Since it has been shown that the contractile effects of endothelins in the aorta are mediated by the endothelin ETA receptor whereas the endothelium-dependent relaxation and the ileal relaxation are mediated by the endothelin ETB receptor, it is suggested that RES-701-1 is a selective antagonist against the endothelin ETB receptor.

Coupling of the endothelin ETA and ETB receptors to Ca2+ mobilization and Ca2+ sensitization in vascular smooth muscle

Effects of endothelins on cytosolic Ca2+ level ([Ca2+]i) and contraction were examined in the swine pulmonary artery and vein. In the artery, endothelin-1 and endothelin-3, but not sarafotoxin S6c and IRL 1620 (300 nM each), induced transient increase followed by sustained increase in [Ca2+]i and sustained contraction. These effects were inhibited by the ETA receptor antagonist, BQ-123. In the vein, endothelin-1 and endothelin-3 (300 nM each) induced sustained increase in [Ca2+]i and sustained contraction whereas sarafotoxin S6c and IRL 1620 (300 nM each) transiently increased both [Ca2+]i and contractile tension. The ETB receptor in the vein was desensitized by pretreatment with sarafotoxin S6c, abolishing the effects of sarafotoxin S6c and IRL 1620 without changing the effects of endothelin-1 and endothelin-3. In contrast, an ETB antagonist, RES-701-1, antagonized the effects of IRL 1620 without changing the effects of other stimulants. In both artery and vein, the maximum contraction induced by these stimulants was greater than that induced by KCl at a given [Ca2+]i. In the absence of external Ca2+, endothelin-1 and endothelin-3 induced transient increase in [Ca2+]i and slow sustained contraction in both artery and vein. In the vein, sarafotoxin S6c induced small sustained contraction without changing [Ca2+]i. In the permeabilized artery and vein, endothelin-1 augmented the contraction induced by Ca2+. These results suggest that the ETA receptors in the artery and vein are coupled to Ca2+ release (which does not seem to trigger contraction), Ca2+ influx and Ca2+ sensitization.(ABSTRACT TRUNCATED AT 250 WORDS)

HS-142-1, a Novel Non-peptide Antagonist for Atrial Natriuretic Peptide Receptor, Selectively Inhibits Particulate Guanylyl Cyclase and Lowers Cyclic GMP in LLC-PK1 Cells

HS-142-1, a novel atrial natriuretic peptide (ANP) antagonist isolated from the culture broth of Aureobasidium sp., selectively inhibits ANP-induced cyclic GMP accumulation in porcine kidney epithelial LLC-PK1 cells. At concentrations from 0.1 to 100 μg/ml (= 2.5 × 10(-8) - 2.5 × 10(-5) M, given the mean molecular weight is 4, 000), HS-142-1 prevents intracellular cyclic GMP accumulation initiated by 10(-8) M rat ANP in a dose-dependent manner, but not cyclic GMP accumulation produced by 10(-5) M sodium nitroprusside. HS-142-1 alone has no effects on the basal level of cyclic GMP seen in the absence of ANP. No change of intracellular cyclic AMP was observed upon the treatment of the cells with HS-142-1. Further, the selectivity of HS-142-1 for the guanylyl cyclase-linked receptor was confirmed by affinity labeling studies with bovine adrenocortical membranes. HS-142-1 specifically abolished the labeling of the guanylyl cyclase-linked 135-kDa band in a dose-dependent manner, but not the labeling of the 60-kDa band not coupled to the guanylyl cyclase. These results show that HS-142-1 selectively inhibits ANP-mediated accumulation of cyclic GMP in LLC-PK1 cells through interacting with guanylyl cyclase-linked receptors.

EI-1511-3, -5 and EI-1625-2, Novel Interleukin-1β Converting Enzyme Inhibitors Produced by Streptomyces sp. E-1511 and E-1625

EI-1511-3, -5 and EI-1625-2, novel interleukin-1 beta converting enzyme (ICE) inhibitors, were isolated from the culture broths of Streptomyces sp. E-1511 and E-1625. EI-1511-3, -5 and EI-1625-2 selectively inhibited the recombinant human ICE activity with IC50 values of 0.09, 0.38 and 0.2 microM, respectively. Taxonomy, fermentation of the producing strain and isolation of EI-1511-3, -5 and EI-1625-2 are described.

Hybrid peptides constructed from RES-701-1, an endothelin B receptor antagonist, and endothelin; binding selectivity for endothelin receptors and their pharmacological activity.

Hybrid peptides were constructed from endothelin B receptor (ET(B)) selective antagonist RES-701-1 (1) and endothelin (ET-1). They have N-terminal 10 amino acids derived from 1 and C-terminal 10 amino acids derived from ET-1. RES-701-1(1-10)-[Ala15]ET-1(12-21) and its analogues substituted or truncated at the residues derived from RES-701-1 had proved to possess high receptor binding activity selective for ETB as well as 1. Substitutions at the residues derived from ET-1 had produced some analogues that possessed high affinity not only for ETB but for ETA. Although all analogues had antagonistic effects on ETA, some analogues had proved to function as agonist on ETB confirmed by the changes in intracellular calcium concentrations of ET receptor-transfected COS-7 cells. We have found four types of ET receptor-binding peptides: (1) ETB-selective agonist with weak ETA antagonism (3, KT7421); (2) ETB-selective antagonist with weak ETA antagonism (29, KT7539); (3) ETB agonist with potent ETA antagonism (27, KT7538); and (4) non-selective ETA/ETB antagonist (26, KT7540).

RES-701-1/endothelin-1 hybrid peptide having a potent binding activity for type B receptor

Abstract N-Terminal cyclic peptide of RES-701-1, an endothelin (ET) type B receptor selective antagonist, was chemically combined to C-terminal peptide of ET family. Among a series of the hybrid peptide synthesized, RES-701-1(1–10)/[A15]ET-1(12–21) hybrid peptide possessed both a potent binding activity with an IC50 value of 0.24nM and a selectivity for type B receptor.

Effects of Endothelin B Antagonist RES−701−1 on Endothelin-induced Contractile Responses In-vivo and In-vitro in Guinea-pigs

The effects of an endothelin (ET)‐receptor B‐specific antagonist, RES−701−1, on ET‐induced contraction of guinea‐pig trachea and on ET‐induced bronchoconstriction in anaesthetized guinea‐pigs were investigated.

ANALOGS OF AN ENDOTHELIN ANTAGONIST RES-701-1 : SUBSTITUTIONS OF C-TERMINAL AMINO ACID

Abstract C-terminal substituted analogs of an ETB receptor specific antagonist RES-701-1 were prepared and evaluated of their receptor binding activities. C-terminal Trp deletion or substitutions for other aromatic amino acids, containing D-configurational ones, proved to be possible without reducing the receptor binding activity, which differs from the case of ET-1 previously reported on its Trp21 for its biological activities.

C-terminal modifications of an endothelin antagonist RES-701-1: Production of ETA/ETB dual selective analogs

RES-701-1 is an endothelin B receptor (ETB) selective peptidic antagonist, which has a novel cyclic structure of microbial origin. Modification at the C-terminal free carboxyl group of RES-701-1 by a methyl ester results in an ETA/ETB dual selective analog, which showed relatively high affinity for ETA receptor subtype, while retaining the affinity for ETB. The carboxyl-group-deleted analog with tryptamine as the C-terminal residue also showed relatively weak affinity for ETA; however, benzyl ester or amide analogs did not show remarkable affinity for ETA. It is suggested that the binding mode of RES-701-1 and its analogs is different from those of known ligands for ET receptors.