Zinc is retained in the mouth after use of a toothpaste containing 0.5% zinc citrate. More than one third of the dose was found to be retained after normal brushing. Elevated zinc levels were also found in plaque. Saliva zinc levels were significantly above background for at least 2 h after brushing. In-vitro experiments demonstrated that zinc can bind to the pellicle-coated tooth surface and can subsequently adsorb into saliva. Plaque can calcify to form calculus containing appreciable levels of hydroxyapatite. Zinc adsorbs to hydroxyapatite inhibiting crystal growth. Levels of zinc in plaque were found to be considerably higher than those taken up by hydroxyapatite in an in-vitro test of crystal growth inhibition indicating the potential of zinc to inhibit calculus formation.
Alpha-2 adrenoceptor agonists have been used in association with local anaesthetic to increase the duration of spinal anaesthesia. Intrathecal administration of clonidine prolonged motor blockade induced by local anaesthetic. Since the affinity of dexmedetomidine (DEX) to alpha-2 adrenoceptors is eight-times greater than clonidine, it is expected that DEX could be advantageous in clinical anaesthesia. We investigated the duration of motor nerve block induced by spinal injection of 0.5% levobupivacaine (LVB) associated with intrathecal or intraperitoneal administration of DEX. Seventy-two guinea-pigs were randomly divided in 12 groups, which were all treated with intrathecal injection of 50 microL of LVB. DEX was injected intrathecally with LVB in 6 groups or injected intraperitoneally after LVB in another 6 groups. Intrathecal DEX (0.1, 0.2 and 0.4 microg) increased the LVB-induced motor anaesthesia from 48 (41-66) min to 84.5 (52-91) min (P < 0.05), 101.5 (83-115) min (P < 0.05) and 105 (97-114) min (P < 0.05), respectively. Similarly, intraperitoneal DEX (20 and 40 microg kg(-1)) increased the motor blockade from 48.5 (33-59) min to 88 (71-114) min (P < 0.05) and 114.5 (103-156) min (P < 0.05), respectively. Pre-treatment with yohimbine reduced the duration of motor block from 101.5 (83-115) to 76.5 (68-86) min (P < 0.05) or from 114.5 (103-156) to 90 (83-93) min (P < 0.05) when DEX was administered by the intrathecal or intraperitoneal routes. Motor block induced by spinal injection of LVB was prolonged by intrathecal and systemic administration of DEX, which was partially dependent on activation of alpha-2 adrenoceptors.
D−002 is an anti-ulcerogenic product, isolated from beeswax, which consists of a well-defined mixture of higher primary aliphatic alcohols. It is highly effective against ethanol-induced ulcers. This study was designed to determine if D-002 shows cytoprotective properties on gastric mucosa in ethanol-induced ulcers. The involvement of endogenous prostaglandins in the protective effect of D−002 was also investigated.
When a subulcerogenic dose of indomethacin (10 mg kg−1) was injected simultaneously with oral administration of ethanol, oral pre-treatment with D-002 (5−100 mg kg−1) partially inhibited the gastric protection. D-002 (5 and 25 mg kg−1) administered to normal rats significantly increased the soluble mucus content and also prevented its reduction in rats with ethanol-induced ulcers. In addition, D-002 administered at 5 and 25 mg kg−1 prevented the increase of vascular permeability induced by ethanol (60%) and reduced the concentration of thromboxane B2 (TXB2) in gastric mucosa of rats with ethanol-induced ulcers.
These results support the hypothesis that the anti-ulcerogenic properties of D-002 could be related to a cytoprotective mechanism.
D-002 is a natural mixture of higher aliphatic primary alcohols, isolated and purified from beeswax which has anti-inflammatory properties, reduces leukotrienes (LTB4) and thromboxane B2 (TXB2) in exudated carrageenan-induced pleurisy, and has anti-ulcer activity in different experimental models. This study was conducted to determine the effect of D-002 on the pre-ulcerative phase of carrageenan-induced colonic ulceration in guinea-pigs.
Animals were randomly distributed among a negative control, a positive control group treated with the vehicle Tween 20 in H2O and two experimental groups receiving D-002 at 25 and 50 mg kg−1. All treated animals received degraded carrageenan for three days for induction of colonic ulceration. Significant reductions in wet weight, wall thickness, counts of infiltrating polymorphonuclear neutrophils and of macrophages, and histological index were observed in colonic mucosa of D-002-treated animals compared with controls.
It is concluded that D-002 has a protective effect on the pre-ulcerative phase of carrageenan-induced colonic ulceration in the guinea-pig.
D-003 is a mixture of long-chain fatty acids purified from sugarcane wax that inhibits both cholesterol synthesis prior to mevalonate formation, and lipid peroxidation. D-003 has been shown to prevent bone loss and bone resorption in ovariectomized rats, and significantly improves bone resorption markers in postmenopausal women with reduced bone mineral density. As hormone-replacement therapy, D-003 displays cholesterol-lowering and anti-resorptive effects. We have studied its potential oestrogenic activity in-vivo using the uterotrophic assay. Rats were randomly distributed into five groups: a sham-operated group and four groups of ovariectomized rats, one treated with vehicle, one with D-003 (50 mg kg(-1)), one with oestradiol benzoate (30 microg kg(-1)) and one with D-003 (50 mg kg(-1)) plus oestradiol benzoate (30 microg kg(-1)). Treatments were administered for 14 days. Ovariectomy decreased the values of relative uterus weight, epithelium cell height and endometrial thickness compared with sham-operated rats, and these effects were all significantly reduced with oestradiol benzoate, but not with D-003. Concurrent administration of D-003 and oestradiol benzoate had statistically similar effects on all variables as oestradiol benzoate alone. In conclusions, D-003 orally given at 50 mg kg(-1), a dose that prevents bone loss and bone resorption in ovariectomized rats, did not display oestrogenic/anti-oestrogenic activity in-vivo, as assessed in the uterotrophic assay.
The activity of RS-61756-007 (methyl-9-oxo, 15 alpha-hydroxy, 16-phenoxy, 17,18,19,20-tetranor prosta 4,5, 13(E) trienoate, 4,5,6(R), 8(R)) has been assessed at prostanoid receptors (DP, EP1, EP2, FP, IP and TP) in-vitro. The activity profile of RS-61756-007 resembles that of U46619, in that agonism was observed at TP and to a lesser extent at FP receptors, but there was no activity at the remaining subtypes. The actions of both RS-61756-007 and U46619 were antagonized in a similar manner by the TP antagonist SQ 29,548. We conclude that RS-61756-007 is a highly potent TP agonist.
Enprostil is a prostaglandin E2 analogue characterized as a racemic mixture of four stereoisomers. Enprostil and a single isomer, RS-86505-007, were evaluated for their effects on the permeability of actively and passively transported compounds in segments of small intestine from rabbits and monkeys. Consistent with human in-vivo studies, which have demonstrated decreases in absorption of D-xylose, both compounds inhibited D-glucose transport. The passively transported compounds mannitol and progesterone were also less permeable in this model in the presence of enprostil or RS-86505-007. In contrast to the concentration-dependent inhibition displayed by ouabain, RS-86505-007 had no effect on purified Na+K(+)-ATPase. It is suggested that an effect of a general nature, possibly an increase in the barrier properties at the intestinal surface, may explain the transport inhibition. Of two other enprostil isomers, RS-86812-007 inhibited D-glucose transport in rabbit small intestine, while RS-86505-008 had no effect. The prostaglandin E1 analogue misoprostol was ineffective in monkey and poorly effective in rabbit. This suggests that the inhibition of D-glucose transport by enprostil and its active stereoisomers is mediated through some structurally specific receptor interaction.
7-Hydroxy-staurosporine (UCN-01) is now under development as a novel anticancer drug. In clinical studies, different infusion schedules are being investigated in the USA and Japan. To examine the effect of different infusion schedules on the pharmacokinetics and cardiohaemodynamics of UCN-01, dogs were treated with UCN-01 as either a 3-h or a 24-h constant intravenous infusion. Blood pressure and heart rate, together with UCN-01 concentrations during and after infusion, were monitored. To analyse the relationship between the pharmacokinetics and cardiohaemodynamics of UCN-01, the plasma concentration of UCN-01 at the end of infusion (Cend), the area under the plasma concentration versus time curves (AUC0-∞) and the mean residence time (MRT) were used. As indices of cardiohaemodynamic changes, the area under decreasing systolic blood pressure and increasing heart rate versus time curves (dAUCpressure and AUCheart rate) were calculated by the trapezoidal method.
For the 3-h (0.22 and 0.65 mgkg−1) and 24-h infusion (0.81 to 6.48 mgkg−1), systolic and diastolic blood pressures fell after or during infusions, accompanied by a dose-dependent increase in heart rate for both infusions. During both infusion schedules, the plasma concentrations of UCN-01 gradually increased and Cend showed a dose-proportional increase. After that, UCN-01 was eliminated bi-exponentially with an elimination half-life of 5.14 ± 1.12 to 8.32 ± 1.80 h. The total clearance (CLtotal) ranged from 0.383 to 0.666 ± 0.149L h−1kg−1. There was no significant difference in these parameters among the doses in each infusion schedule, indicating that UCN-01 has a linear pharmacokinetic profile over the dose range examined for each infusion, and there were also no significant differences between the 3-h and 24-h infusion except for MRT. The pharmacokinetic parameters of Cend, AUC0-∞ and slope0-3h exhibited a degree of correlation with the AUCheart rate in the 3-h infusion and correlated significantly with the dAUCpressure in the 24-h infusion. The MRT did not correlate with cardiohaemodynamic changes during either infusion.
In conclusion, the pharmacokinetic profile of UCN-01 after the shorter infusion is similar to that after the longer one. However, a longer dosing period of UCN-01 increased the residence time in comparison with the shorter infusion. This may be due to the effect on the circulatory function in dogs.
Lu 3–010 [3,3-dimethyl-1-(3-methylaminopropyl)-1-phenylphthalan], administered intraperitoneally, blocks the uptake of [3H]noradrenaline into the mouse and rat heart and has an activity 5 times greater than imipramine and comparable to that of desipramine. Lu 3–010 inhibits basal gastric acid secretion in the rat and is 4 and 2 times more potent than imipramine and desipramine, respectively. The drug is about 9 times more potent than desipramine in preventing the pentagastrin-induced stimulation of gastric acid secretion in the rat. Lu 3–010 reduces the incidence of stress-induced gastric lesions in the rat, exhibiting an ED50 ± s.e. of 6.3 ± 1.4 mg/kg, and at 10 mg/kg, ulcer development in the 17-hour Shay test is reduced by 50%.
The effects of the thymoleptic drug, LU 3–010, on adrenergic transmitter release and reuptake and on effector cell function in rat isolated portal veins have been investigated. The contractile response to exogenous noradrenaline was potentiated by LU 3–010 and its concentration-effect curves after LU 3–010 10−7 g/ml were displaced to the left to a similar extent as had been found in chronically denervated preparations, indicating efficient blockade of the membrane pump mechanism. LU 3–010 10−7g/ml slightly potentiated the neurogenic response to electrical field stimulation, whereas 10−6 g/ml or more reduced the neural effector response in a concentration- dependent manner. The output of radioactive material per nerve impulse after incubation with 3H-NA was also studied. At 10−7 and 3 × 10−7 g/ml of LU 3–010 no significant change in transmitter overflow was observed. At 10−6 g/ml there was a decrease in the “fractional release”, suggesting that LU 3–010 concentrations of 10−6 g/ml or higher reduce transmitter release. Evidence is also given for a direct inhibitory effect of LU 3–010 on the smooth muscle.
The pharmacological activity of 3−((4−(2−methoxyphenyl)piperazin−1−yl)methyl)−2,3−dihydroimidazo(1,2−c)quinazolin−5(6H)-one (DC−015), a newly synthesized quinazoline derivative, was determined in rat isolated thoracic aorta and pressor responses were determined in spontaneously hypertensive rats (SHR).
Experimental results indicated that DC−015 is an α1-adrenoceptor-blocking agent in rat thoracic aorta as revealed by its competitive antagonism of phenylephrine-induced vasocontraction (pA2 = 10ṁ54 ± 0ṁ55). These effects still persisted in denuded aorta. It was as potent as prazosin (pA2 = 10ṁ04 ± 0ṁ63). At higher concentrations (1ṁ0 μM), DC−015 also expressed 5−hydroxytryptamine (5−HT) receptor competitive antagonism, but this 5−HT blocking effect was not found in the prazosin-administration group. [3H]Inositol monophosphate formation stimulated by phenylephrine (30 μM) in rat thoracic aorta was diminished by DC−015 (3 and 10 nM) and prazosin (10 nM); whereas the cAMP content of rat thoracic aorta was not altered by DC−015 and prazosin. Furthermore, intravenous administration of DC−015 and prazosin (both at 0ṁ01, 0ṁ05 and 0ṁ1 mg kg11) induced a dose-dependent reduction of mean arterial pressure which reached a maximal effect at 5 min after injection and persisted over 2 h in SHR. A higher dose of DC−015 (0ṁ1 mg kg−1, i.v.) did not cause any significant changes in heart rate, whereas, the same dose of prazosin (0ṁ1 mg kg−1, i.v.) produced a decrease which seems to parallel the time course of the hypotensive response.
We can conclude that the DC−015 is a potent, highly selective α1-adrenoceptor antagonist in vascular smooth muscle.
Rigosertib (ON 01910.Na, Estybon) is a novel, anticancer agent undergoing phase 3 clinical trials for a lead indication against myelodysplastic syndromes (MDS). In this research, the permeability of rigosertib was evaluated using the in-situ perfused rat intestine (IPRI) model to support development of an oral formulation for rigosertib for treating cancer patients.
Experiments (n = 6 per group) were conducted using male Sprague-Dawley rats. Studies evaluated permeability across various intestinal segments and assessed the dose-linearity of absorption over the entire intestinal length. Drug concentrations in the portal and jugular vein were collected to correlate permeability parameters with presystemic and systemic exposure.
Rigosertib permeability was highest in the jejunum, although parameter estimates indicated that rigosertib was a medium permeability compound. The compound displayed nonlinear absorption in the IPRI model, suggesting a saturable transport process. Transport inhibition studies using Caco-2 cells demonstrated that rigosertib was a P-glycoprotein (P-gp) substrate. Absolute bioavailability of rigosertib (10 and 20 mg/kg, 1-h infusion) in rats was estimated to be 10-15%. However, the fraction absorbed in humans predicted from IPRI data (52%) was consistent with published clinical data for rigosertib (35% oral bioavailability).
The results of this research indicated that rigosertib is a promising candidate for oral delivery. Further studies are needed to evaluate the potential impact of P-gp and other intestinal transporters on the oral absorption of this promising anticancer agent.
Prostaglandin E2 (PGE2) produced by cyclooxygenase (COX) is a potent pro-inflammatory mediator. We have recently discovered CJ-023,423, a highly selective antagonist of EP4 receptors, one of the PGE2 receptors. This agent is suitable for exploring the effects of blocking EP4 receptors following oral administration in rats. In this study, CJ-023,423 was used in rats with adjuvant-induced arthritis (AIA) to investigate the role of the EP4 receptor in chronic inflammation and bone destruction. These effects were compared with those of rofecoxib, a selective COX-2 inhibitor. CJ-023,423 had significant inhibitory effects on paw swelling, inflammatory biomarkers, synovial inflammation and bone destruction in AIA rats. In particular, the inhibitory effect on paw swelling in AIA rats was comparable to that of rofecoxib. These results suggest that PGE2 acting via the EP4 receptor is involved in the development of chronic inflammation and bone destruction, particularly with respect to oedema in AIA rats. This is the first study to confirm the in-vivo effects of EP4 receptor blockade on inflammation and bone destruction in AIA rats with a small-molecule compound.
The in-vivo activities of eight carbamate prodrugs of the D2-agonist N-0437 were determined by examining the effects of the prodrugs, after their oral administration in rats with unilateral 6-OHDA lesions of the striatum. The resulting contralateral turning was used as an index of the activity of the compounds. A comparison of the area under the curve of the time-effect curves of the prodrugs, revealed a significantly improved duration of action compared with N-0437 during the period 11-15 h after administration, for the propylcarbamate and the dimethoxyphenylcarbamate derivatives. The 2,4-dimethylphenylcarbamate showed a significantly enhanced turning behaviour over the whole 15 h time interval in comparison with N-0437. Three of the nine carbamates were virtually unhydrolysed in rat serum at 37 degrees C, while the other test compounds were hydrolysed relatively slowly, with t1/2 values ranging from 1.5-6 h. The test compounds differed greatly in partition coefficients, which were estimated by RP-HPLC (1-12 times more lipophilic than N-0437). The potential cholinesterase inhibiting properties of the carbamate prodrugs were assessed by a simple in-vitro incubation assay, which showed that only two carbamates were very weak cholinesterase inhibitors.
The anti-anginal effect of CP-060S, a new cardioprotective agent that prevents myocardial Na+-, Ca2+-overload and has Ca2+-channel blocking activity, was evaluated in a rat model of arginine8-vasopressin (AVP)-induced cardiac ischaemia. Infusion of AVP (0.2 IU kg(-1)) depressed the electrocardiogram (ECG) ST segment, an index of myocardial ischaemia. Vehicle, CP-060S and diltiazem were given orally 1, 2, 4, 8, 12 and 24 h before the administration of AVP. CP-060S, at 3 mg kg(-1) and 10 mg kg(-1), suppressed AVP-induced ST-segment depression for 2 h and 12 h, respectively. In contrast, diltiazem, at 10 and 30 mg kg(-1), suppressed AVP-induced ST-segment depression for only 1 h. The persistent suppression of the AVP-induced ST-segment depression by CP-060S correlated with the time course of changes in its plasma concentration. The minimum effective concentration of CP-060S was estimated to be 30 ng mL(-1) (approximately 50 nM), consistent with its vasorelaxant potency in rat isolated aortic strips (concentration producing 50% relaxation of KCl contraction, IC50 = 32.6+/-8.3 nM). Intravenously administered CP-060S, at 300 microg kg(-1) and diltiazem at 500 microg kg(-1) showed similar haemodynamic changes, whereas CP-060S, at 300 microg kg(-1), significantly suppressed AVP-induced ST-segment depression and diltiazem, at 500 microg kg(-1), had no effect on AVP-induced ST-segment depression. In summary, orally administered CP-060S exerted a long-lasting anti-anginal effect proportionate to the time course of changes in its plasma concentration in a rat model of AVP-induced ischaemia.
Resorcinol derivatives are known to inhibit melanin synthesis. In this study, resorcinol derivatives were synthesized and screened for their activity on melanogenesis. KI-063 (a tyrosinase inhibitor) was examined for its effects on melanogenesis using a spontaneously immortalized mouse melanocyte cell line (Mel-Ab). In a cell-free system, KI-063 directly inhibited tyrosinase, the rate-limiting melanogenic enzyme. Moreover, in a cell system, it inhibited melanin synthesis in a concentration-dependent manner. In addition, KI-063 inhibited the activity of cellular tyrosinase. Thus, this study examined the effects of a combination of KI-063 with terrein, an agent that down-regulates microphthalmia-associated transcription factor. The data suggest that KI-063 has an additive effect in combination with terrein. Thus, the suppression of tyrosinase activity by KI-063 and the inhibition of tyrosinase production by terrein appear to be an optimal combination for skin whitening.
The pharmacological activity of CL-065 (trans-3-acetamido-2, 2-dimethyl-4-hydroxy-3, 4-dihydro-2H-1-benzopyran-6-carbonitrite) was investigated in anaesthetized spontaneously hypertensive rats (SHR) and isolated thoracic aorta of Sprague-Dawley rats.
The intravenous administration of CL-065 (0.1–2.0 mg kg−1) to anaesthetized SHR induced a dose-dependent reduction of mean arterial pressure (MAP) with maximum effect approximately 5 min after injection and which persisted for over 3 h. CL-065 also induced a reflex tachycardia which seemed to parallel the time course of the hypotensive effect. The hypotensive effect of CL-065 was blocked by pretreatment with glibenclamide (5 mg kg−1, i.v.), a specific ATP-sensitive potassium (KATP) channel blocker. Moreover, CL-065 (0.01–10 μM) resulted in dose-dependent vasodilatory effects on phenylephrine (0.3 μM)-induced vasoconstriction in isolated thoracic aorta. The vasorelaxation elicited by CL-065 was antagonized competitively by pretreatment with glibenclamide (0.1–1.0 μM; pA2 = 6.90 ± 0.09; slope = 1.03 ± 0.18). Similarly, the other two KATP-channel openers cromakalim (1.0 nM–1.0 μM) and nicorandil (0.1–30 μM) also induced vasorelaxation in thoracic aorta. The EC50 of cromakalim, CL-065 and nicorandil (i.e. the doses having half the maximum effect) were approximately 0.083, 0.17, and 4.5 μM, respectively, for phenylephrine (0.3 μM)-induced vasoconstriction in isolated thoracic aorta. Moreover, increased extracellular potassium levels (20–60 mM) resulted in concentration-dependent attenuation of the vasodilator effect of CL-065.
In conclusion, CL-065 induces a depressor effect via activation of KATP channels.
The promoting action of the calcium chelating compound EDTA on intestinal drug absorption is supposed to be based on Ca2+ depletion, inducing widening of tight junctions. The aim of the present study was to evaluate the effects of the calcium-binding agent 3-amino-1-hydroxypropylidene-1,1-diphosphonate disodium salt (APD) on rectal cefoxitin absorption in rats. The extent of rectal cefoxitin absorption was enhanced by 0.5 to 6% w/v of APD, on rectal infusion as well as on bolus delivery, the latter regimen tending to result in lower bioavailabilities. A maximal cefoxitin bioavailability of 85 +/- 10% was achieved by infusion with 4% w/v of APD, compared with 14 +/- 12% without APD.
The effect of 1,1-dimethyl-4-phenylpiperazinium (dmpp) on the response to sympathetic nerve stimulation of rat mesenteric arteries perfused with Tyrode solution at a constant flow has been studied. dmpp (0·3 μg/ml) infused for 3 min enhanced the vasoconstriction caused by stimulation. Infusion of the same concentration for 16–40 min greatly reduced the response to nerve stimulation but did not affect the vasoconstrictor response to injected noradrenaline. The blockade of the response to nerve stimulation produced by dmpp was overcome either by adding (+)-amphetamine to the perfusion fluid or by raising the calcium concentration. Neither effect of dmpp was altered by the infusion of atropine. These effects of dmpp were similar to those seen when acetylcholine was added to the perfusion fluid except that the effects of acetylcholine were diminished or abolished by a concentration of atropine much higher than that of acetylcholine. It is concluded that the receptors at the adrenergic nerve terminals are partly muscarinic and partly nicotinic.
Replacement of the 1-adamantyl group with a 1,1′-biadamantyl group in the N-adamant-1-yl-N'-arylsulphonylureas leads to compounds having the same or a doubled hypoglycaemic activity. The new biadamantyl derivatives exert a delayed onset of action compared with that of the corresponding adamantane derivatives. The most promising, 3,3′-di(N'-p-toluenesulphonylureido)-1,1′-biadamantyl (Compd No. 1) and 3,3′-di(N'-p-methoxybenzenesulphonylureido)-1,1′-biadamantyl (Compd No. 3) are only slightly toxic and appear not to have other pharmacological activity.
A series of 2-aralkyl-4H-pyridothiadiazine 1,1-dioxides and 3-aralkylamino-2-aryl-2H-pyrido[4,3-e]-1,2,4-thiadiazine 1,1-dioxides structurally related to quinazolinone CCK receptor antagonists were synthesized and evaluated as CCK-A and CCK-B receptor ligands.
The compounds were effective as cholecystokinin-ligands in the micromolar range of concentration, c.f. the cholecystokinin receptor antagonists asperlicin, lorglumide or benzotript, and were thus less potent than the best quinazolinones previously reported.
Although the compounds were unsuitable for drug use, the work contributed to our understanding of the chemistry of unusual 2,3-disubstituted pyridothiadiazinedioxides.
7-Chloro-3-pyridyl(alkyl)amino-4H-1,2,4-benzothiadiazine 1,1-dioxides and 3-alkylamino-7-chloro-4H-1,2,4-benzothiadiazine 1,1-dioxides containing one or more heteroatoms on the side chain in the 3 position have been synthesized in an attempt to discover new potent KATP-channel openers. The compounds were tested as putative pancreatic B-cells KATP channel openers by measuring their inhibitory activity on the insulin releasing process. The influence on the biological activity of the nature of the side chain in the 3 position is discussed.
The Kampo medicine, Ninjin-yoei-to, scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals in a dose-dependent fashion as did ascorbic acid and alpha-tocopherol. Ninjin-yoei-to, which is composed of 12 herbs, had a potent DPPH radical scavenging ability. We investigated the transition of the materials that scavenge DPPH radicals in plasma after oral administration of Ninjin-yoei-to to rats. When 1.0 g kg(-1) Ninjin-yoei-to was administered, the DPPH radical scavenging ability increased at 30 min and biphasic peaks were observed at 2 h and at 10 h. From the response-time profile, kinetic parameters including values for K(a) (absorption rate constant), t(max) (peak concentration time), t(1/2) (half-life) and MRT (mean residence time) of the radical scavenging ability in plasma could be calculated for DPPH radicals. K(a) values were 0.53 +/- 0.03 and 0.36 +/- 0.07 h, t(max) values were 2.1 +/- 1.04 and 8.56 +/- 2.69 h, t(1/2) values were 1.60 +/- 0.12 and 3.39 +/- 1.72 h, and MRT values were 4.14 +/- 1.59 and 8.18 +/- 2.55 h, respectively. These parameters calculated from the antioxidation dynamics were considered to offer a very meaningful procedure for examining the effects of Ninjin-yoei-to.
The topical anti-inflammatory activity of a series of N-pyridinyl(methyl)1,2-dihydro-4-hydroxy-2-oxoquinoline-3-carboxamides, analogues of roquinimex, has been evaluated by measuring their inhibitory effect in the phorbol myristate acetate (PMA)-induced mouse ear swelling test, used as a screening test.
All the eight carboxamides tested (9–16) exhibited significant inhibitory activity at 0.4 and 0.2 mm kg−1. The most potent compound, the 6-bromo derivative 12, induced a 73% inhibition at 0.2 mm kg−1. Pharmacomodulation was carried out by heterocycle opening and molecular simplification leading to pentafluorobenzoylacetamide 17, pentafluorocinnamamides 18 and 19, and pentafluorobenzaldimines 20 and 21. All the five compounds exerted a reduction in swelling (49–63% at 0.2 mm kg−1) comparable with ibuprofen (56%). Anti-inflammatory activity of the most efficient compounds was evaluated by carrageenan-induced rat paw oedema inhibition. The pentafluorobenzaldimine 20 showed the highest activity with an inhibition percentage of 85% at 0.2 mm kg−1.
1,2-Diethyl-3-hydroxypyridin-4-one (CP94) is an orally active iron chelator with potential for use in photodynamictherapy. This investigation reports the formation and characterization of two isomeric glucuronides of CP94 in rat liver homogenate incubates. To assign the glucuronidation sites in the CP94 molecule, two O-methylated derivatives of CP94 have been synthesized. By comparing the spectral characteristics of the CP94 3-O- and 4-O-methyl derivatives with CP94 and the CP94 glucuronides formed during incubation, evidence was obtained which enabled the assignment of these two isomeric glucuronides to the 3-O-glucuronide and 4-O-glucuronide of CP94. It was found that the 3-O-glucuronide was the dominant CP94 metabolite under in-vitro conditions. In an attempt to understand the potential influence of structural variation on the glucuronidation of CP94 analogues, the 1-and 2-monoethyl derivatives of CP94 were investigated. The 2-monoethyl derivative of CP94 yielded only the 3-O-glucuronide in rat liver homogenate incubate, while no glucuronide was formed from the 1-monoethyl derivative. In addition, no glucuronide from the 3-O-methyl or 4-O-methyl derivatives of CP94 could be detected. The relevance of these findings to the development of new 3-hydroxypyridin-4-one iron chelators is discussed.
Nitric oxide (NO) possesses antitumour activity. It induces differentiation and apoptosis in acute myeloid leukaemia (AML) cells. The NO prodrug O(2) -(2,4-dinitrophenyl)1-((4-ethoxycarbonyl)piperazin-1-yl)diazen-1-ium-1,2-diolate, or JS-K, has potent antileukaemic activity. JS-K is also active in vitro and in vivo against multiple myeloma, prostate cancer, non-small-cell lung cancer, glioma and liver cancer. Using the Pluronic P123 polymer, we have developed a micelle formulation for JS-K to increase its solubility and stability. The goal of the current study was to investigate the cellular distribution of JS-K in AML cells.
We investigated the intracellular distribution of JS-K (free drug) and JS-K formulated in P123 micelles (P123/JS-K) using HL-60 AML cells. We also studied the S-glutathionylating effects of JS-K on proteins in the cytoplasmic and nuclear cellular fractions.
Both free JS-K and P123/JS-K accumulate primarily in the nucleus. Both free JS-K and P123/JS-K induced S-glutathionylation of nuclear proteins, although the effect produced was more pronounced with P123/JS-K. Minimal S-glutathionylation of cytoplasmic proteins was observed.
We conclude that a micelle formulation of JS-K increases its accumulation in the nucleus. Post-translational protein modification through S-glutathionylation may contribute to JS-K's antileukaemic properties.
In the search for antipsychotic agents that are not associated with extrapyramidal side effects, efforts have been focused on finding selective D4-receptor antagonists and investigating their pharmacology. Our laboratory has developed a synthesis program for new pyrroloquinoxalines with therapeutic potential. We have described the synthesis of some new pyrroloquinoxalines with substituted arylpiperazino or aryltetrahydropyrido chain at position 3 of the quinoxaline ring (2-(4-phenylpiperazin-1-ylmethyl)-4-phenylpyr-rolo[1,2-a]quinoxalinium oxalate (3a), 2-[4-(2-methoxyphenyl)piperazin-1-ylmethyl]-4-phenylpyrrolo[1,2-a]quinoxalinium oxalate (3b), 2-[4-(3-trinuoromethylphenyl)piperazin-1-ylmethyl]-4-phenylpyrrolo[l,2-a]quinoxalinium oxalate (3c), 2-[4-(4-chlorophenyl)piperazin-1-ylmethyl]-4-phenylpyrrolo[1,2-a]quinoxalinium oxalate (3d), 2-(4-pyridin-2-ylpiper-azin-1-ylmethyl)-4-phenylpyrrolo[1,2-a]quinoxalinium oxalate (3e), and 2-(4-phenyl-1,2,3,6-tetrahydropyridin-1-ylmethyl)-4-phenylpyrrolo[1,2-a]quinoxalinium oxalate (3f)).
A preliminary pharmacological study of these products was conducted using climbing behaviour induced by apomorphine (2.5 mg kg−1, s.c.) in mice. The derivatives were administered intraperitoneally 30 min before apomorphine. Haloperidol, chlorpromazine and clozapine were used as references.
Among this series, 3b, 3c and 3f revealed a central dopamine antagonist activity. The most active derivative was 3b, which exhibited a profile relatively close to clozapine.
The solvent-water distribution coefficients of 17 alpha-methyltestosterone have been determined in selected alkanes, alkanols and alkane-alkanol blends. The oral availability of tritium from solutions of [3H]methyltestosterone in these solvents and blends was studied in rats by measuring radioactivity in plasma, faeces and urine. Disappearance of radioactivity from the gut and biliary recirculation of 3H were also examined. Variation of the solvent led to marked changes in the absorption of radioactivity, suggesting a possible method of varying bioavailability from soft gelatin capsules. Neither distribution coefficients nor tritium concentrations changed significantly from one homologue to another, but there was a marked difference when the alkane series was compared with the alkanol series. Blends of octane and octanol, together with the pure solvents, provided a suitable spread of distribution coefficients and tritium concentrations, but no simple relationship could be established between the two properties.
It has been known that reactive oxygen and nitrogen species such as nitric oxide (NO), superoxide radical (*O2-) and their byproduct peroxynitrite (ONOO-) induce cellular and tissue injury, ultimately resulting in several human diseases. In this study, we examined scavenging effects of 3-methyl-1,2-cyclopentanedione (MCP) from coffee extract on the reactivity of those toxic molecules. MCP significantly inhibited both the oxidation of 2,7-dichlorodihydrofluorescein diacetate (H2DCFDA) by reactive oxygen species (ROS) (mainly *O2-) from kidney homogenate (41% at 100 microM) and the generation of fluorescent 4,5-diaminofluorescein (DAF-2) by NO from sodium nitroprusside (IC50 (concn producing 50% inhibition), 63.8 microM). More potently, however, MCP suppressed the oxidation of dihydrorhodamine 123 (DHR 123) to fluorescent rhodamine 123 mediated by authentic ONOO- with an IC50 value of 3.3 microM. The neutralizing effect of the reactivity of ONOO- by MCP was due to electron donation, not nitration of the compound. Additionally, MCP also decreased ONOO- formation of nitrotyrosine adducts of glutathione (GSH) reductase, and consequently protected the enzyme activity of GSH reductase against decreasing by ONOO-, indicating that MCP may prevent ONOO- -induced damage of GSH reductase. Furthermore, MCP only weakly suppressed NO production, which is one of the upstream sources of ONOO- in-vivo, suggesting that NO production may be not a pharmacological target for MCP. Taken together, our results suggest that MCP may be regarded as a selective regulator of ONOO- -mediated diseases via direct scavenging activity of ONOO-.
Colon cancer is a major cause of morbidity and mortality in developed and developing countries. Diet and dietary constituents play a major role in the aetiology of colon cancer. We have investigated the effect of an aqueous extract of oregano (Origanum vulgare. L.) on lipid peroxidation and anti-oxidant status in 1,2-dimethylhydrazine (DMH)-induced rat colon carcinogenesis. We aimed to identify the important antioxidants present in Indian oregano using RP-HPLC. DMH (20 mgkg(-1)) was administered subcutaneously once a week for the first four weeks and then discontinued. Oregano was supplemented every day orally at a dose of 20, 40 or 60 mgkg(-1) to different groups of rats for 15 weeks. After this time the rats were killed and the colons were examined visually and evaluated biochemically. The levels of lipid peroxidation products, such as thiobarbituric acid reactive substances and conjugated dienes were significantly higher in the liver whereas in caecum and colon the levels were lower in DMH-treated animals as compared with control rats. The levels of the anti-oxidants superoxide dismutase, catalase, reduced glutathione, glutathione reductase, glutathione peroxidase and glutathione-S-transferase were decreased in DMH-treated rats, but were significantly reversed on oregano supplementation. Oregano supplementation (40 mgkg(-1)) had a modulatory role on tissue lipid peroxidation and antioxidant profile in colon cancer-bearing rats, which suggested a possible anti-cancer property of oregano.
Over the last 30 years, desferrioxamine has been the only iron chelator in clinical use. This chelator is expensive and must be given by injection. A new class of chelators, namely 1-alkyl-2-methyl-3-hydroxypyrid-4-ones, have been shown to be orally effective. Using 1,2 dimethyl-3-hydroxy-pyrid-4-one (DMHP), we have carried out a study to clarify the mechanism of intestinal absorption of this new class of drug, using an in-situ system of the intestine from rabbit. The major site of DMHP absorption is in the intestine and is linear with increasing drug concentration. DMHP absorption per unit length of jejunum and ileum is similar; however, due to the larger surface area of jejunum, the absorption by ileum segment is more effective per unit surface. L-Proline, L-tryptophan (amino acids), 2-deoxyglucose, and sodium iodoacetate (metabolic inhibitors) have no effect on DMHP absorption, but L-phenylalanine, an amino acid with a 6-member carbon ring, significantly inhibits the DMHP absorption from the intestinal segment. We conclude that the mechanism of DMHP absorption in the intestine is mainly by simple passive diffusion based on the linear relationship found between drug concentration and absorption. However, the inhibitive effect of L-phenylalanine suggests that the co-existence of a facilitated uptake cannot be ruled out.
We have studied the effects of some hexahydroimidazo[1,2-c]pyrimidine derivatives (HIPs) on leucocyte functions in-vitro and we have assayed the anti-inflammatory activity of these compounds in two models of inflammation. All HIPs inhibited the human neutrophil degranulation process and superoxide generation at concentrations in the microM range. In mouse peritoneal macrophages stimulated with lipopolysaccharide, HIP-4 and HIP-5 inhibited nitrite production without affecting prostaglandin E2 (PGE2) accumulation. HIP-4 was also active in the zymosan-injected mouse air pouch model (at 100 nmol/pouch), with significant reductions in leucocyte migration and PGE2 and leukotriene B4 levels in the air pouch exudate. To confirm the anti-inflammatory effects of this compound, we tested HIP-4 orally (10-40 mg kg(-1)) on carrageenan mouse-paw oedema where it exerted a dose-dependent inhibition of paw swelling with significant reductions of myeloperoxidase and elastase activity and PGE2 levels in paw homogenates. This study demonstrates that some HIPs inhibit leucocyte functions and one of these derivatives (HIP-4) shows anti-inflammatory activity when administered by the oral route, which can be related to inhibition of leucocyte migration.
1,2,3-Benzotriazin-4-one derivatives are extensively used as herbicides, insecticides and nematicides. Incorporation of these compounds into the human food chain is a cause for concern, since their toxicity to man is well documented.
This work describes the development of a data-base of classical 1,2,3-benzotriazin-4-one fragmentation patterns and consequent sequential neutral mass losses which allows the prediction of the presence of a triazinone derived compound.
Positive and negative ion tandem mass spectrometry techniques were employed since they allow rapid and sensitive structural characterization of drugs and their metabolites in biological fluids through the unambiguous monitoring of daughter ions from pre-selected primary ions.
MX, (N-[3-(3-(1-piperidinylmethyl)phenoxy)propyl]-hydroxyacetamide 2-hydroxypropane-1,2,3-tricar-boxylate bismuth (3+) complex) is a novel salt of the active metabolite of H2-antagonist roxatidine with a complex of bismuth with citric acid.
In a model of ethanol-induced ulcers in male Wistar rats, both roxatidine and the bismuth salt reduced the number and the total length of lesions. Comparison of roxatidine and MX, at equimolar doses of 160 μmol kg−1 showed a more potent cytoprotective effect of MX1. The potency of anti-secretory and antiacidic effects of MX1 was more than twice that of roxatidine on histamine-stimulated secretion in female Wistar pylorus-ligated rats. Microbiological tests with the reference bismuth preparation De-Nol showed prominent anti-Helicobacter properties of MX1 in-vitro. Both test compounds had similar range of MICs to Helicobacter pylori, from 4 to 64 μg bismuth mL−1.
The cytoprotective, antisecretory, anti-acidic and anti-Helicobacter properties of the new agent MX1 warrant further more extensive pharmacological and clinical trials.
A variety of dopamine derivatives and analogues were investigated to assess their potential to act as catechol-O-methyltransferase (COMT) substrates using purified, homogeneous pig liver enzyme. This enabled accurate kinetic constants to be determined as opposed to previous in-vivo studies (Rollema et al 1980; Horn et al 1981; Costall et al 1982; Feenstra et al 1983). 2-Amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (A-6,7-DTN) proved to be a far better substrate (Km = 0.082 mM; Vmax = 300 mu mg-1 protein) than its 5,6-dihydroxy isomer (Km = 2.60 mM; Vmax = 113.9 mu mg-1 protein). This result supports evidence suggesting that differences in brain concentration of these isomers are due to their differential susceptibility to O-methylation by COMT (Rollema et al 1980). A similar result was obtained with a series of NN-di-n-alkyl substituted ADTN derivatives: the same pattern of preferential O-methylation of A-6,7-DTN derivatives over the corresponding A-5,6-DTN isomers was observed. However, increasing the length of the alkyl chain increased the susceptibility of both isomers to metabolism by COMT as shown by a decline in Km. An homologous series of NN-di-n-alkylated dopamines showed a similar trend implying that more hydrophobic compounds are better COMT substrates.
M-7, 1 and 3 mg kg-1 s.c., elicits an antihypertensive response and bradycardia in conscious spontaneously hypertensive rats (SHR) and causes inhibition of stimulation-evoked pressor response and tachycardia in pithed SHR. Metoclopramide (30 mg kg-1 i.p.), but not piperoxan (5 mg kg-1 i.p.), abolished the antihypertensive effect and inhibition of stimulation-evoked pressor responses produced by M-7 (1 mg kg-1 s.c.) in SHR. Conversely, piperoxan, but not metoclopramide, reduces the bradycardia and inhibition of stimulation-evoked tachycardia produced by M-7. Metoclopramide (30 mg kg-1 i.p.) did not affect the cardiovascular responses elicited by intracerebroventricular administration of either clonidine (1 microgram) of M-7 (3 micrograms). These results suggest that the antihypertensive effect of M-7 may be mediated by stimulation of presynaptic dopamine receptors on sympathetic nervous to the vasculature and is independent of the bradycardia, which is probably due to stimulation of presynaptic alpha 2-adrenoceptors on cardiac sympathetic nerve endings.
Some acute pharmacological effects have been examined of racemic ADT 16 (1,2,3,5,6,11b-hexahydrobenzothieno[3,2-g]indolizine hydrochloride), on peripheral and central responses mediated by 5-HT and adrenergic systems in the rat. In-vitro, ADT 16 (10-1000 nM), similarly to mianserin, antagonized the inhibitory responses to B-HT 920 of the electrically-stimulated rat isolated prostatic vas deferens. High concentrations of ADT 16 (10 microM), also resembled those of mianserin by potentiating twitch responses to electrical stimulation of the tissue. Contractile responses to phenylephrine of rat isolated epididymal vas deferens were antagonized by ADT 16 (0.3-1 microM). In the rat stomach fundus strip, ADT 16 (1-3 microM) antagonized contractions due to 5-HT. ADT 16 (0.1-1 microM) had no effect on responses to acetylcholine of the guinea-pig isolated ileum. In-vivo, in spinalized, decerebrated rats, fenfluramine- or clonidine-induced facilitation of flexor reflex activity of the anterior tibialis muscle was attenuated by ADT 16 (3 and 10 mg kg-1, i.v., and 3 mg kg-1, i.v. respectively). In the anaesthetized rat, L-3,4-dihydroxyphenylalanine (L-dopa)- or L-5-hydroxytryptophan (L-5-HTP)-induced increases in the frequency of spontaneous twitches of the anterior digastricus muscle were attenuated by ADT 16 (1 and 3 mg kg-1, i.v.; n = 4). It is concluded that ADT 16, similarly to mianserin, is a novel peripherally and centrally active antagonist of 5-HT and adrenergic responses in the rat.