Quantitative structure-activity relationship study of N-(3-oxo-3,4-dihydro-2H-benzo[1,4]thiazine-6-carbonyl)guanidines as potent Na/H exchange inhibitors.

We have previously reported that N-(4-isopropyl-2,2-dimethyl-3-oxo-3,4-dihydro-2H-benzo[1,4]oxazine-6-car bonyl)guanidine (4b) methanesulfonate salt (KB-R9032) is a potent and highly water-soluble Na/H exchange inhibitor. In a series of studies on Na/H exchange inhibitors, we designed and synthesized N-(3-oxo-3,4-dihydro-2H-benzo[1,4]thiazine-6-carbonyl)guanidines (5) as more potent inhibitors with high water-solubility. The design strategy for 5 was based on a quantitative structure-activity relationship (QSAR) study, involving the proportional relationship between the biological activity and hydrophobicity of the ring structure of compounds 4. As expected, compounds 5 showed more potent activity than 4. It was found by using the QSAR analysis that 5 were about five-fold more potent than 4. The increase in potency of compounds 5 well agreed with our previous QSAR analysis result. The most potent derivative was the methanesulfonate salt 5d of the 4-isopropyl derivative (IC50=0.0091 microM). And in addition to the in vitro study, 5d showed significant protective activity against a rat acute myocardial infraction model.

Synthesis and quantitative structure-activity relationships of N-(3-oxo-3,4-dihydro-2H-benzo[1,4]oxazine-6-carbonyl)guanidines as Na/H exchange inhibitors.

N-(3-Oxo-3,4-dihydro-2H-benzo[1,4]oxazine-6-carbonyl)guanidines 4 were prepared and tested for Na/H exchange inhibitory activities in order to clarify the structure-activity relationship (SAR). Quantitative SAR (QSAR) analysis of 6-carbonylguanidines 4 indicated that the length of the 4-substituent was parabolically related to activity and that the calculated optimum 4-substituents were propyl, ethyl and isopropyl groups. This SAR was similar to the SAR of the 2- and 4-substituents of 7-carbonylguanidine derivatives 3, although the position relative to the essential guanidinocarbonyl group was different. Larger 2-substituents, such as a phenyl group were unfavorable. The most potent derivative in this series was N-(4-isopropyl-2,2-dimethyl-3-oxo-3,4-dihydro- 2H-benzo[1,4]oxazine-6-carbonyl)guanidine 4 g, with an IC50 value of 0.12 microM. The methanesulfonate salt (KB-R9032) of 4g had excellent water-solubility and showed anti-arrhythmia activity against a rat acute myocardial infarction model. KB-R9032 was selected for further investigation as a therapy for ischemia-reperfusion induced injury.

Structural requirements for potential Na/H exchange inhibitors obtained from quantitative structure-activity relationships of monocyclic and bicyclic aroylguanidines.

The quantitative structure-activity relationship (QSAR) of N-(3-amino-6-chloro-5-ethylisopropylaminopyrazine-4-carbonyl) guanidine (EIPA) 1ac and its derivatives as Na/H exchange inhibitors was analyzed using th steric parameters and an indicator variable. The results indicated that bicyclic aroylguanidines might have Na/H exchange inhibitory activity. Therefore, various bicyclic aroylguanidines were synthesized and tested for Na/H exchange inhibitory activity. The QSAR study of the bicyclic aroylguanidines showed that hydrophobic bicyclic rings seemed to be preferable for potent activity. The hydrophobicity of the aroyl ring moiety was thought to be particularly important. Thus, the QSAR of EIPA and its derivatives was re-analyzed using hydrophobicity and steric parameters. The results indicated that high hydrophobicity of the pseudo-ring moiety and a substituent of appropriate length at the position corresponding to the 5-position of the naphthalene ring enhance the activity. As expected from the results, 5-bromo-2-naphthoylguanidine 3b and 5-methoxy-2-naphthoylguanidine 3c exhibited strong activity. These findings will be helpful to design new, potent Na/H exchange inhibitors.

Design, synthesis and quantitative structure-activity relationship study of N-(3-oxo-3,4-dihydro-2H-benzo[1,4]oxazine-7-carbonyl)guanidine derivatives as potent Na/H exchange inhibitors.

Inhibition of the Na/H exchanger is a promising approach for treating ischemia-reperfusion injury, but no clinical agent is yet available. Recently, we established the structural requirements for potent inhibitors of the Na/H exchanger. In the present work, we designed N-(3-oxo-3,4-dihydro-2H-benzo[1,4]oxazine-7-carbonyl)guanidine 3a as a new lead compound for potent inhibitors with good water-solubility, based on the previous information. During the structural optimization, care was taken to keep the hydrophobicity (clogP) in the range of about 1.5-2.0, which is considered optimum for good bioavailability. Various derivatives of 3a were synthesized and the quantitative structure-activity relationship (QSAR) was studied. The QSAR result indicated that the lengths of the substituents at the 2- and the 4-positions of the 2H-benzo[1,4]oxazine ring are parabolically related to activity. The most potent compounds were (R) and/or (S)-N-(2-ethyl-4-isopropyl(or ethyl)-3-oxo-3,4-dihydro-2H-benzo[1,4]oxazine-7-carbonyl)guanidines 3q-t with IC50 values of 0.036-0.073 microM. The water-solubility of the hydrochlorides and methanesulfonates is 3-5 mg/ml, which is sufficient for therapeutic use.

Physiological correlation between glycyrrhizin, glycyrrhizin-binding lipoxygenase and casein kinase II.

By means of glycyrrhizin (GL)-affinity column chromatography, a GL-binding lipoxygenase (gbLOX) was selectively purified from the partially purified soybean LOX-1 fraction. Polypeptide analysis of the purified gbLOX by SDS-PAGE detected two distinct polypeptides (p96 and p94), which were identical to LOX-3 as determined by their partial N-terminal amino acid sequences. Moreover, it was found that (i) phosphorylation of gpLOX by casein kinase II (CK-II) is significantly stimulated by 3 microM GL, but inhibited by 30 microM GL or 10 microM oGA; and (ii) gbLOX activity is enhanced when the enzyme is phosphorylated by CK-II in the presence of 3 microM GL. These results suggest that (i) CK-II is a kinase responsible for the activation of gbLOX through its specific phosphorylation; and (ii) GL is one of the regulatory substances for specific phosphorylation of gbLOX (LOX-3) by CK-II in plant cells.

Design and syntheses of a series of novel serotonin3 antagonists.

From a structural comparison study between serotonin and serotonin3 (5-HT3) antagonists using a two-dimensional grid template composed of regular hexagons, we deduced structural modification patterns from agonists to antagonists, and designed new 5-HT3 antagonist prototypes. Among them, 2-(4-methyl-1-piperazinyl)-1-butylbenzimidazole (6) was identified as a lead compound which has potent 5-HT3 antagonistic activity comparable to that of granisetron. Using a quantitative structure-activity relationships method, we optimized the structure of 6 and selected 6-amino-5-chloro-1-isopropyl-2-(4-methyl-1-piperazinyl)benzimidazole dimaleate (69, KB-6933), one of the most potent and long-acting 5-HT3 antagonists, as a candidate drug.

[Structure-activity relationship of flavonoids in suppressing rat liver lipid peroxidation].

145 flavonoids were studied for their inhibitory effects on the iron-induced lipid peroxidation in mitochondria obtained from rat livers. Of these compounds tested, 30, 57, 59, 67, 70, 72, 77, 102 and 110 (ED50 < or = 0.5 nmol/mg prot) showed distinctly more potent inhibitory activity than baicalein (ED50 < or = 5 nmol/mg prot) and 59 and 72 (ED50 < or = 0.05 nmol/mg prot) exhibited the most potent activity. In order to elucidate the relationships between substituents on the flavonoid skeleton and the biological activity, the quantitative structure-activity relationships (QSAR) were analyzed by the adaptive least-squares (ALS) method for 142 flavonoids. The analysis has shown that the presence of the 1,4- and 1,2-hydroquinone in A-ring and/or B-ring of flavonoids and hydrophobicity of the molecule are responsible for the in vitro inhibitory activity.

Structural modification patterns from agonists to antagonists and their application to drug design--a new serotonin (5-HT3) antagonist series.

Structural variations from agonists to their selective antagonists seemed to follow certain patterns. To analyze the variation patterns in the structural modification processes in past examples as well as to utilize the "common" variation patterns as possible principles to design new selective antagonistic drugs, the structures of agonists and their antagonists were superimposed on a two-dimensional grid template composed of regular hexagons and the topological similarities and dissimilarities of substructural elements between agonists and antagonists were examined. Between several pairs of neurotransmitter amines and their "selective" antagonists, similar patterns were disclosed in their structural modification processes. The generalized structural modification patterns were successfully applied as guiding principles to design and identify a new prototype structure of the 5-HT3 antagonist. The prototype structure was optimized by use of QSAR procedures leading to a compound which shows a potent antiemetic activity as well as a powerful gastrointestinal-motility modulation.

Effects of baicalein and alpha-tocopherol on lipid peroxidation, free radical scavenging activity and 12-O-tetradecanoylphorbol acetate-induced ear edema.

The effects of baicalein, a flavonoid, and alpha-tocopherol (vitamin E) on lipid peroxidation in rat forebrain homogenates, on free radical scavenging action against diphenyl-p-picrylhydrazyl (DPPH), and on 12-O-tetradecanoylphorbol acetate (TPA)-induced ear edema in mice were studied. Baicalein inhibited lipid peroxidation in forebrain homogenates, DPPH-induced free radical and TPA-induced ear edema as potently as did quercetin and nordihydroguaiaretic acid (NDGA), a lipoxygenase inhibitor, and more potently than BW755C, a mixed cyclooxygenase and lipoxygenase inhibitor. Lipid peroxidation in forebrain homogenates, DPPH-induced free radical and TPA-induced ear edema were also inhibited by alpha-tocopherol. Flavone showed no reaction. These results suggest that lipid peroxidation may play an important role in the pathogenesis of TPA-induced ear edema in mice.

Potential nootropic agents, 4-alkoxy-2-(1-piperazinyl)quinazoline derivatives.

A series of 4-alkoxy-2-(1-piperazinyl)quinazoline derivatives was synthesized and evaluated for its ability to reverse a scopolamine-induced learned impairment in a one-trial passive avoidance task (antiamnestic activity). 2-(4-Allyl-1-piperazinyl)-4-pentyloxyquinazoline (4) showed more potent antiamnestic activity than such reference compounds as aniracetam, idebenone and bifemelane at a wide dose range (1-30 mg/kg]. Compound 4 also exhibited potent anticonvulsive and antihypoxic activities, and was selected as the most promising nootropic candidate agent.

Catecholamine responses to histamine infusion in man.

To evaluate the effects of histamine-induced hypotension on plasma catecholamine levels, eight normal men, aged 20 to 40 years, were infused with incremental doses of histamine starting at 0.2 microgram/kg/min at a 30 degree tilt position with monitoring of blood pressure (BP) and heart rate. Histamine dosage was increased every 5 minutes by 0.1 to 0.2 microgram/kg/min until mean BP fell greater than 15 mm Hg or a dosage of 1.6 micrograms/kg/min was reached. Plasma catecholamine samples were taken between the fourth and fifth minute of each histamine dosage. Identical measurements were made during nitroglycerin-induced hypotension in these subjects. Histamine produced threefold greater increases in heart rate and plasma norepinephrine (NE) levels than did nitroglycerin for comparable decreases in BP. Although NE levels increased twofold to fivefold from baseline with histamine infusion, epinephrine levels increased minimally at the highest doses or not at all. Our data demonstrate that histamine selectively releases NE from adrenergic nerve terminals without significant adrenal catecholamine release. We suggest that neural NE release plays an important role in the cardiac effects of histamine.

Selective effect of KB-2796, a new calcium entry blocker, on cerebral circulation: a comparative study of the effects of calcium entry blockers on cerebral and peripheral arterial blood flows.

The effects of a new calcium entry blocker, KB-2796, on cerebral and peripheral circulation were compared with those of four other calcium entry blockers (flunarizine, cinnarizine, nicardipine, and diltiazem) and also papaverine in anesthetized dogs. KB-2796 increased the vertebral blood flow to the same extent as the other drugs. KB-2796 and the other drugs increased the coronary blood flow, although the effect of KB-2796 was obviously weaker than that of the other drugs. The dose of KB-2796 producing a 30% increase in coronary blood flow was about 20 times higher than that producing an equivalent increase in vertebral blood flow. KB-2796 and the other calcium entry blockers caused a moderate increase in both femoral and common carotid blood flow. The renal artery showed a biphasic response to all drugs, which consisted of an initial decrease and a subsequent increase in the flow. From these results, it is suggested that KB-2796 is a new type of calcium entry blocker that selectively affects the cerebral circulation. It is also concluded that calcium entry blockers have a different spectrum of vasodilatory action on coronary and cerebral vascular beds and that a selective coronary vasodilatory action is not a common feature among all calcium entry blockers.

Novel 4-substituted 2-piperazinylquinazolines as potent anticonvulsive and antihypoxic agents.

Several types of quinazoline derivatives were prepared and examined for anticonvulsive and antihypoxic activities. Many compounds showed potent anticonvulsive activity, and their anticonvulsive profile is similar to that of phenytoin. The analysis of quantitative structure-activity relationships indicated that the anticonvulsive activity was parabolically related to the lipophilicity of the compounds. Most of the 4-alkoxyquinazolines showed potent anticonvulsive and antihypoxic activities. It is confirmed that there is a good correlation between the potencies of these activities.

Retrograde thrombosis of feeding arteries after removal of arteriovenous malformations.

Five cases of retrograde thrombosis of former feeding arteries after removal of an arteriovenous malformation (AVM) are reported. The clinical features of these patients were studied and compared to those of 71 patients without this complication. The following characteristics were found to correlate with retrograde thrombosis: 1) advancing age of the patient; 2) large AVM size; and 3) markedly dilated and elongated feeders. It is suggested that the slow flow in the former feeding arteries that was observed immediately after AVM removal and pathological changes in these vessels due to long-standing hemodynamic stresses contributed to the development of retrograde thrombosis. Neurological manifestations related to retrograde thrombosis were noted in three of the five cases. Although infrequent, this complication should be considered as a serious possibility following removal of an AVM.

Novel 4-phenoxy-2-(1-piperazinyl)quinazolines as potent anticonvulsive and antihypoxic agents.

A series of 4-phenoxy-2-(1-piperazinyl)quinazolines was synthesized and examined for anticonvulsive and antihypoxic activities. Many of the compounds exhibited potent anticonvulsive activity comparable to that of carbamazepine or phenytoin. Among them, 4-phenoxy-2-(4-propyl-1-piperazinyl)quinazoline (5w) was selected as the most promising candidate antiepileptic drug with few side effects. It seemed that potent anticonvulsive activity was a prerequisite for potent antihypoxic activity.

Indomethacin enhances histamine-induced pulmonary hemodynamic changes.

To determine the role of prostaglandins in porcine pulmonary hemodynamic changes caused by histamine, we compared responses to intravenous histamine with and without pre-treatment with the cyclo-oxygenase inhibitor, indomethacin. In anesthetized pigs, pulmonary artery pressure (Ppa), pulmonary arterial wedge pressure (Ppaw), left ventricular end diastolic pressure (Plved) and cardiac output (Q) were measured repeatedly for 30 minutes, following a 1 ml intrajugular injection of histamine 0.6 microM/kg (n = 6), the identical histamine dose after pre-treatment with indomethacin 5 mg/kg (n = 7), or normal saline (n = 5). Pulmonary arterial resistance (Ra) and pulmonary venous resistance (Rv) were calculated as (Ppa-Ppaw)/Q and (Ppaw-Plved)/Q respectively. Indomethacin pre-treatment caused 2-fold greater increases in Ra and Rv with histamine and more prolonged changes. We conclude that inhibition of a vasodilatory prostaglandin released from pulmonary endothelial cells results in unopposed pulmonary vasoconstriction, thereby augmenting pulmonary resistance changes due to histamine.

Benzylpiperazine derivatives. XI. Synthesis of compounds related to the metabolites of 1-[bis(4-fluorophenyl)methyl]-4-(2,3,4-trimethoxybenzyl)piperazine dihydrochloride (KB-2796).

The metabolites of 1-[bis(4-fluorophenyl)methyl]-4-(2,3,4-trimethoxybenzyl)piperazine dihydrochloride (KB-2796, 1), a cerebral vasodilator, and related compounds were synthesized to confirm the proposed structures. The structures of the metabolites (3-5) in rats were identified by means of synthesis of the authentic compounds.

Bioisosteric transformation of H1-antihistaminic benzimidazole derivatives.

With the aim of obtaining new H1-antihistaminic agents, transformations of previously reported antihistaminic benzimidazoles were performed on the basis of the concept of bioisosterism. Among the compounds prepared, imidazo[4,5-b]pyridine (8) and 4(3H)-quinazolinone (11) exhibited significant H1-antihistaminic activity.

Syntheses of the metabolites of 1-(2-ethoxyethyl)-2-(hexahydro-4-methyl-1H-1,4-diazepin-1-yl)-1H benzimidazole difumarate (KG-2413) and related compounds.

The metabolites of 1-(2-ethoxyethyl)-2-(hexahydro-4-methyl-1H-1,4-diazepin-1-yl)-1H-b enzimidazole difumarate (KG-2413), which has a potent H1-antihistaminic activity, were predicted on the basis of metabolic studies of related compounds and were synthesized to aid in identification of the actual metabolites and for examination of their antihistaminic activity. Among the twelve compounds prepared, nine compounds were actually found as the metabolites of KG-2413 in rat urine. The antihistaminic activities of these metabolites were found to be lower than that of KG-2413.