Yoshiyuki Inada

CV-3988 - a specific antagonist of platelet activating factor (PAF).

CV-3988, rac-3-(N-n-octadecylcarbamoyloxy)-2-methoxypropyl 2-thiazolioethyl phosphate was shown to be a specific inhibitor of platelet activating factor (PAF). This compound in concentrations of 3 x 10(-6) to 3 x 10(-5)M inhibited aggregation of rabbit platelets induced by PAF (3 x 10(-8)M), while it had no effect on the aggregation induced by arachidonic acid, ADP, collagen or A-23187. CV-3988 alone even at a concentration of 10(-3)M had no effect on platelet aggregation. The inhibitory action of CV-3988 on the PAF-induced aggregation was independent of the formation of micelles. The PAF (0.1 to 1.0 micrograms/kg, i.v.)-induced hypotension in anesthetized rats was also inhibited dose-dependently by the i.v. administration of CV-3988 (1 and 10 mg/kg), while the hypotensive actions induced by the i.v. administration of acetylcholine (1 micrograms/kg), arachidonic acid (1 mg/kg), bradykinin (10 micrograms/kg), isoproterenol (1 microgram/kg) and histamine (100 micrograms/kg) were not altered by CV-3988 (10 mg/kg, i.v.). All these findings indicate that CV-3988 specifically inhibits the action of PAF in vitro and in vivo. This is the first report of a PAF antagonist which can specifically inhibit the PAF-induced hypotension as well as the PAF-induced platelet aggregation.

Protective effects of candesartan cilexetil (TCV-116) against stroke, kidney dysfunction and cardiac hypertrophy in stroke-prone spontaneously hypertensive rats.

The effects of chronic treatment with an angiotensin II receptor antagonist, candesartan cilexetil (TCV-116, 0.1, 1, 10 mg/kg), and an angiotensin converting enzyme inhibitor, enalapril maleate (enalapril, 10 mg/kg), on the development of end-organ damage were examined in stroke-prone spontaneously hypertensive rats (SHRSP). The control SHRSP developed severe hypertension with stroke signs and increased urinary protein excretion. TCV-116 (0.1 mg/kg) reduced the stroke incidence and urinary protein excretion without affecting the blood pressure. TCV-116 (1 and 10 mg/kg) and enalapril reduced blood pressure, the stroke incidence, the urinary indices and left ventricular weight. Circulating renin-angiotensin system (RAS) and renal renin mRNA expression were significantly accelerated or tended to be accelerated in the control SHRSP with end-organ damages. A low dose of TCV-116 tended to reduce the RAS indices in plasma by improving the damages, whereas a high dose (10 mg/kg) increased them by the reflexes with blocking RAS. The present results indicate that chronic All blockade reduces the increase in blood pressure, end-organ damages and RAS related to the damages in SHRSP.

Enhanced thromboxane A2 biosynthesis in the kidney of spontaneously hypertensive rats during development of hypertension.

Arachidonic acid (AA)-induced pressor response and production of thromboxane YXB2, the stable metabolite of TXA2, prostaglandin (PG)-like substance (PLS) and 6-keto-PGF1 alpha the stable metabolite of prostacyclin (PGIs), were studied using isolated, perfused kidneys of 6- and 18-week old spontaneously hypertensive rats (SHR), Wistar-Kyoto rats (WKY), two-kidney, one clip hypertensive rats (RHR) and DOCA/salt hypertensive rats (DOCA/salt HR). The AA-induced pressor response and release of TXB2 were highest in the 6-week old SHR, whereas, the release of PLS and 6-keto-PGF 1 alpha was marked in the 18-wek old SHR and the established hypertensive stages of both RHR and DOCA/salt HR. In the kidneys of SHR and WKy, exogenous TXA2 induced a severe vasoconstriction and there was a positive correlation between the AA-induced pressor response and the release of TXB2 or PLS. Thus, the initiation of hypertension in SHR may follow an accelerated synthesis of TXA2 against PGI2 in response to stimuli which induce a release of AA.

Angiotensin-II-stimulated release of thromboxane a2 and prostacyclin (PGI2 ..) in isolated, perfused kidneys of spontaneously hypertensive rats

Abstract Angiotensin-II(A-II)-stimulated release of rabbit aorta-contracting substance (RCS), prostaglandin-like substance (PLS) and dog coronary-relaxing substance (DRS) was studied in isolated, perfused kidneys of spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). When the perfusion pressure was set at 60–70 mmHg, the dose-related pressor response of the kidney to A-II did not differ significantly between the strains, or between the ages of 6, 12 and 18 weeks. The A-II-induced release of RCS from the kidney was most remarkable in 6-week SHR, slight in 12- and 18-week SHR, but negligible for all ages of WKY. The DRS release was remarkable in 12- and 18-week SHR, but was hardly detectable in 6-week SHR and all ages of WKY. The PLS release was much more remarkable in 18-week SHR than in 6- and 12-week SHR and all ages of WKY. The RCS was confirmed to be thromboxane A2 (TXA2) by: (1) its half-life of 38 sec at 37°, (2) a concomitant release of radio-immunoassayable TXB2: (a stable metabolite of TXA2) into the perfusate and (3) specific inhibition of its release by L-8027 (a TXA2 synthetase inhibitor). The DRS was considered to be prostacyclin (PGI2) by: (1) its half-life of 5.2 min at 37° and (2) its anti-platelet aggregation activity. It is speculated that an enhanced TXA2 synthesis in the 6-week SHR kidney may be involved in increasing the renal vascular resistance and in initiating the process of hypertension.

A new class of angiotensin II receptor antagonists with a novel acidic bioisostere

Abstract The synthesis and angiotensin II (AII) antagonistic activity of 2-substituted benzimidazole-7-carboxylic acids ( 8 ) bearing a novel acidic bioisostere, a 5-oxo-1,2,4-oxadiazole ring, are described. These compounds had in vitro and in vivo AII receptor antagonistic activity comparable to that of tetrazole analogs. The representative compound, TAK-536 ( 8e ) is a potent, orally active and specific AII receptor antagonist.

Delay of the initiation of hypertension in spontaneously hypertensive rats by CV-4151, a specific thromboxane A2 synthetase inhibitor

When CV-4151, a specific thromboxane (TX) A2 synthetase inhibitor, was given orally to 4 week old (4w) spontaneously hypertensive rats (SHR) daily for 3 weeks, the initiation of hypertension was delayed by about one week. The agent increased urinary excretion of water, sodium and creatinine, reduced that of TXA2 (as TXB2), increased that of PGI2 (as 6-keto-PGF1 alpha) and enhanced urinary PGI2/TXA2. In 4w Wistar-Kyoto rats (WKY) and 18w SHR was established hypertension, the agent had little effect on blood pressure and renal function. In isolated, perfused kidneys of 6w SHR, CV-4151 markedly inhibited both arachidonic acid-induced pressor action and production of TXA2. TXA2 synthetase activity in renal cortical microsomes of 5w SHR was approximately 1.5 times higher than that in age-matched WKY. CV-4151 inhibited TXA2 synthetase activity of medullary and cortical microsomes more effectively in 5w SHR than in age-matched WKY. Thus, in young SHR, the TXA2 synthetase inhibitor seemed to improve renal function by altering the balance of renal TXA2 and PGI2 biosynthesis and subsequently caused a delay in the initiation of hypertension. The present findings lend support to the idea that an imbalance in the renal TXA2-PGI2 biosynthesis may be involved in the initiation of hypertension in SHR.

Effect of an angiotensin II receptor antagonist, CV-11974, and its prodrug, TCV-116, on production of aldosterone

In dispersed rabbit adrenocortical glomerulosa cells, a non-peptide angiotensin II (AT1) receptor antagonist, CV-11974 (10(-10)-10(-5) M), competitively inhibited angiotensin II- or angiotensin III-stimulated aldosterone production, whereas PD123177, an angiotensin AT2 receptor antagonist, did not. CV-11974 inhibited aldosterone production induced by 4 mM K+ but not by 12 mM K+. CV-11974 had no effect on adrenocorticotropic hormone-stimulated aldosterone or corticosterone production, but inhibited angiotensin II-stimulated corticosterone production. In the rat, TCV-116, the prodrug of CV-11974, (0.1 and 1 mg/kg, p.o.) markedly reduced the elevation of both plasma aldosterone concentration and blood pressure induced by i.v. infusion of angiotensin II. In spontaneously hypertensive rats, TCV-116 at daily p.o. doses of 0.1, 1 and 10 mg/kg for 2 weeks caused a dose-dependent reduction of blood pressure and plasma aldosterone concentration without affecting plasma corticosterone. Thus, TCV-116 inhibited the induction of aldosterone production by not only exogenous but also endogenous angiotensin II.

MAPPING OF ANGIOTENSIN II RECEPTOR SUBTYPES IN PERIPHERAL TISSUES OF SPONTANEOUSLY HYPERTENSIVE RATS BY IN VITRO AUTORADIOGRAPHY

1. The regulation of angiotensin II (AII) receptor subtypes was studied in peripheral tissues of 20 week old male spontaneously hypertensive rats (SHR) and age‐matched normotensive Wistar‐Kyoto (WKY) rats.

Anti-hypertensive effect of oral controlled-release microspheres containing an ACE inhibitor (Delapril hydrochloride) in rats

Abstract— An oral controlled‐release drug delivery system based on microspheres of polyglycerol esters of fatty acids (PGEFs), was applied to an anti‐hypertensive, delapril hydrochloride. The in‐vitro release profile was controlled by selecting a PGEF with an appropriate hydrophilic‐lipophilic balance value for the matrix. The microspheres from which 80% of the drug was released in 6 h were orally administered to rats. The plasma concentration of the active metabolite was sustained after administration of the microspheres in comparison with administration of a solution. The in‐vivo release profile was in good agreement with the in‐vitro release profile. When the microspheres were administered, the pharmacological effect of delapril hydrochloride on the angiotensin I‐induced pressor response was also sustained showing consistency with the plasma concentration‐time curve.

Renal effects of pinane-thromboxane A2 and indomethacin in saline volume-expanded spontaneously hypertensive rats

In 6-week old spontaneously hypertensive rats (SHR), indomethacin (5 mg/kg i.v.) and pinane-thromboxane A2 (PTA2) (50 micrograms/kg per h i.v.) a thromboxane A2 (TXA2) antagonist as well as a TXA2 synthetase inhibitor, resulted in natriuresis accompanied by an increase in p-aminohippuric acid and inulin clearances. Indomethacin acted as an antidiuretic in 6 week Wistar-Kyoto rats (WKY). PTA2 did not alter renal functions in either 6 week WKY and 18 week SHR. Basal urinary excretion of TXB2 (UTXB2V) was greater in 6 week SHR than in 6 week WKY and 18 week SHR, and that of 6-keto-PGF1 alpha (U6-keto-PGF1 alpha V) did not differ among these strains. Thus, U6-keto-PGF1 alpha V/UTXB2V was lower in the 6 week SHR. Basal urinary excretion of PGE (UPGEV) was much greater in 18 week SHR than in the 2 other groups. In the 6 week SHR, PTA2 decreased UTXB2V and increased U6-keto-PGF1 alpha V without affecting UPGEV, and indomethacin reduced UTXB2V more markedly than did U6-keto-PGF1 alpha V and UPGEV. Thus, both PTA2 and indomethacin increased U6-keto-PGF1 alpha V/UTXB2V in the 6 week SHR. These findings indicate that a disequilibrium in the biosynthesis of vasoconstrictive TXA2 and of vasodilator PGI2 may be involved in water and sodium retention in SHR during the developmental phase of hypertension.