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Concern over environmental pollution generated by chemical fungicides has led to the introduction of alternative pest management strategies to chemical fungicide application. One of those strategies is the induction of plant defense response by an abiotic elicitor. In the present study, field-grown grapevines were subjected to electrical stimulation using a solar panel from two weeks before flowering to harvest in the 2016 and 2020 growing seasons. In both years, electrical stimulation decreased the incidence of gray mold and/or ripe rot on bunches and downy mildew on leaves of the field-grown grapevine. Transcription of a gene encoding β-1,3-glucanase but not class IV chitinase in leaves of potted grapevine seedlings was upregulated 20 days after electrical stimulation, suggesting that electrical stimulation acts as an abiotic elicitor of plant defense response to fungal diseases. The gene expression of PR1 but not PDF1.2 was upregulated in Arabidopsis plants subjected to electrical stimulation. On the other hand, PR1 gene expression was not induced in salicylic acid (SA)-insensitive Arabidopsis mutant npr1-5 subjected to electrical stimulation. Taken together, electrical stimulation is responsible for plant defense response through the SA-dependent defense pathway. These findings would help us develop a novel and innovative practical technique that uses electrical stimulation in integrated pest management.

Objective: To evaluate the clinical usefulness of effective renal plasma flow (ERPF) measured using preoperative mercaptoacetyltriglycine-3 (MAG3) renogram for the prediction of chronic renal insufficiency after nephrectomy. Methods: A total of 47 patients underwent preoperative MAG3 renal scintigraphy and subsequent unilateral nephrectomy. Correlations between the 5-year postoperative estimated glomerular filtration rate (eGFR) and the preoperative ERPF of the contralateral kidney (cERPF), ERPF of the diseased kidney (dERPF), total ERPF (tERPF), cERPF to dERPF ratio, serum creatinine (sCr) level, eGFR, as well as the influence of preoperative comorbidities (diabetes, hypertension) on the postoperative eGFR, were evaluated with both univariate and multivariate analyses. Results: Multiple linear regression analysis showed that preoperative cERPF significantly correlated with postoperative eGFR. However, a much stronger correlation was observed between the preoperative and postoperative eGFR. Multiple logistic regression analysis showed that only preoperative eGFR was a significant predicator of the development of advanced-stage chronic kidney disease (CKD). Conclusions: Preoperative MAG3 renogram is not superior to eGFR measurement as a prognostic indicator of long-term renal function after unilateral nephrectomy.

The in vitro metabolism of amrubicin by rat and human liver microsomes and cytosol was examined. The main metabolic routes in both species were reductive deglycosylation and carbonyl group reduction in the side-chain. In vitro metabolism of amrubicinol by rat and human liver microsomes and cytosol was also examined and the main metabolic route of this active metabolite was reductive deglycosylation. Metabolism of amrubicin in human liver microsomes was inhibited by TlCl(3) and that in human liver cytosol was inhibited by dicumarol and quercetin. Generation of amrubicinol was inhibited only by quercetin. The results indicate that metabolism of amrubicin is mediated by NADPH-cytochrome P450 reductase, NADPH:quinone oxidoreductase and carbonyl reductase. In addition, generation of amrubicinol is mediated by carbonyl reductase. Metabolism of amrubicinol in human liver microsomes was inhibited by TlCl(3) and that in human liver cytosol was inhibited by dicumarol. The results indicate that metabolism of amrubicinol is mediated by NADPH-cytochrome P450 reductase and NADPH:quinone oxidoreductase. To investigate the influence of cisplatin on the metabolism of amrubicin and amrubicinol, human liver microsomes and cytosol were pre-incubated with cisplatin. This did not change the rates of amrubicin and amrubicinol metabolism in either human liver microsomes or cytosol.

Several basic metal oxide catalysts were tested for the synthesis of 1,3-disubstituted urea from ethylene carbonate (EC) and amine. Among the catalysts used, CaO has been found to be an excellent recyclable catalyst for the reaction. It has been suggested that strongly basic property of CaO results in its high activity. Disubstituted ureas are obtained from propylamine and butylamine with high yields at 100 °C. Slightly higher reaction temperatures are necessary for obtaining good yields from amines having larger molecular weights and urea is not produced from dibutylamine as a secondary amine. Propylene carbonate can be used instead of EC for the reaction. A reaction mechanism was proposed, which involves reaction between EC and amine giving a carbamate followed by catalytic reaction between the carbamate and amine, yielding 1,3-disubstituted urea. It is suggested that the latter reaction is the rate-determining step. On the basis of this reaction mechanism, the synthesis of unsymmetric urea was also examined. 2-Hydroxyethyl butylcarbamate is selectively produced from EC with butylamine in the absence of the catalyst at a low temperature and reacts with benzylamine producing 1-butyl-3-benzylurea along with symmetric dialkyl ureas.

26,26,26,27,27,27-Hexafluoro-1α,25(OH) 2 vitamin D 3 , a hexafluorinated analog of 1α,25(OH) 2 vitamin D 3 , has been reported to be several times more potent than the parent compound with respect to some vitamin D actions. The reason for enhanced biological activity in the bones, kidneys, and small intestine appears to be related to F 6 -1α,25(OH) 2 vitamin D 3 metabolism to ST-232 (26,26,26,27,27,27-hexafluoro-1α,23 S ,25-trihydroxyvitamin D 3 ), a bioactive 23 S -hydroxylated form that is resistant to further metabolism. We compared the disposition and metabolism of [1β- 3 H]F 6 -1α,25(OH) 2 vitamin D 3 and [1β- 3 H]1α,25(OH) 2 vitamin D 3 in parathyroid glands of rats intravenously administered with labeled compounds at a dose of 10 μg/kg. In the [1β- 3 H]F 6 -1α,25(OH) 2 vitamin D 3 -dosed group, radioactivity was highly detected in the kidneys, parathyroid glands, and the small intestine. The radioactivity in the parathyroid glands remained high until 48 h postdosing, with values of 2.5, 8.4, and 14.6 times higher at 6, 24, and 48 h postdosing than after dosing with [1β- 3 H] 1α,25(OH) 2 vitamin D 3 . In the group given [1β- 3 H]F 6 -1α,25(OH) 2 vitamin D 3 , the unchanged compound was mainly detected with a small amount of ST-232 at 6 h postdosing. At the 24- and 48-h time points, over half of the radioactivity was observed as ST-232, and additionally, ST-233, the 23-oxo form, accounted for a small amount at the 48-h time point. The present study demonstrated local retention of [1β- 3 H]F 6 -1α,25(OH) 2 vitamin D 3 and the bioactive metabolite ST-232 in parathyroid glands after intravenous administration. The findings may indicate one of the reasons for the higher potency of F 6 -1α,25(OH) 2 vitamin D 3 than 1α,25(OH) 2 vitamin D 3 in parathyroid.

In vitro metabolism studies were conducted to assess drug-drug interactions between perospirone, an antipsychotic agent, and concomitantly administered drugs--biperiden, flunitrazepam, haloperidol, and diazepam--using human liver microsomes. The metabolism of perospirone in the presence of 100 microg/ml drugs was decreased to 45-73% of that in their absence, whereas no effects were observed with any of the drugs at 1 microg/ml or lower. The effects of perospirone on the metabolism of concomitantly administered drugs were also assessed, and no inhibitory effect was observed. Thus, the metabolism of perospirone and concomitantly administered drugs did not demonstrate any marked mutual inhibition in the human liver microsomes. On the other hand, the perospirone metabolism was markedly reduced by ketoconazole indicating a major role for CYP 3A4. Based on the inhibition constant (Ki) for perospirone metabolism and the plasma unbound concentration of ketoconazole, in vivo perospirone clearance was estimated to be reduced to 64-90% of the control level. Thus careful attention should be paid to the possibility of increase in unchanged perospirone concentration when perospirone is co-administered with drugs that are known as CYP3A4 inhibitors, including macrolide antibiotics and other imidazole antifungals.

(S)-9-chloro-5-[p-aminomethyl-o-(carboxymethoxy)phenylcarbamoylmethyl]-6,7-dihydro-1 H,5 H-pyrido[1,2,3-de]quinoxaline-2,3-dione hydrochloride trihydrate (SM-18400) was given intravenously to rats and dogs and its pharmacokinetics was investigated. By LC/MS/MS analysis, the major metabolite in the rat serum was identified as N-acetylated SM-18400 (SM-NAc). In rats, AUC ratio of SM-NAc to SM-18400 was approximately 50%. However, 71% of the dose was excreted as unchanged SM-18400 and only 9.8% as SM-NAc in the urine and bile, indicating that the contribution of N-acetylation clearance (CL(NAc)) to the total clearance (CL(tot)) is limited to 10-30% in rats. No SM-NAc or other metabolites were detected in the dog serum, urine or bile. The in vitro intrinsic clearance (CL(int), ml/min/mg cytosolic protein) of N-acetyltransferase (NAT) activities of dog liver cytosol towards SM-18400 and hepatic N-acetylation clearance (CL(NAc), ml/min/kg body weight) estimated by well-stirred model were both only 5% of the respective rat value, well reflecting the relative in vivo CL(NAc)/CL(tot) ratios. CL(int) values for human liver cytosol samples (n = 4) and estimated CL(NAc) were all less than 18% and 7% of the rat, respectively. Based on these results, we concluded that the CL(NAc)/CL(tot) of human would be small enough to avoid major inter-individual variance in SM-18400 pharmacokinetics due to N-acetylation polymorphism. In addition, even a human liver cytosol sample lacking polymorphic NAT2 activity as determined by sulfamethazine (SMZ) N-acetylation analysis, proved capable of acetylating SM-18400, suggesting that NAT2 is not the major enzyme responsible for N-acetylation of SM-18400 in human. This fact would also reduce the risk of N-acetylation polymorphism playing a role in clinical use of this drug.

Following the administration of [Carbonyl-14C]perospirone ([CO-14C]perospirone) and [isothiazolyl-3-14C]perospirone ([TH-14C]perospirone) to male-pigmented rats at a single oral dose of 10 mg/kg, serum radioactivity reached maxima of 499 ng eq./g and 446 ng eq./g, respectively, at 1 hour, then decreased rapidly to below 2 ng eq./g at 168 hours, and radioactivity in eyeballs reached maxima of 576 ng eq./g and 3449 ng eq./g, respectively, at 24 hours, then decreasing slowly to become 120 ng eq./g and 314 ng eq./g at 26 weeks. Apparent elimination half-lives of radioactivity in eyeballs were 197 and 126 days for [CO-14C]perospirone and [TH-14C]perospirone, respectively. With radioluminograms, radioactivity was detected specifically in the uvea, reaching maxima at 168 hours for [CO-14C]perospirone and at 24 hours for [TH-14C]perospirone. In addition to unchanged compound, many metabolites were found in serum and eyeballs, but those were also detectable in SD albino rats and cynomolgus monkeys. Among radioactive components in eyeballs after administration of [CO-14C]perospirone, ratios of ID-15036 and MX11 followed by unchanged compound and MX9 are high. After administration of [TH-14C]perospirone, ID-11614 accounted for nearly half of the radioactivity. Values decreased in line with radioactivity in eyeballs over time and 26 weeks only a few polar metabolites could be detected.

Vanadium pentoxide is used as a catalyst and a ferrovanadium alloy ingredient in automotive steels and in jet engines and airframes. In addition, vanadium is found in fuel oils. Thus, occupational exposures to vanadium pentoxide and trioxide may occur during the cleaning of oil-fired ship boilers, and from oil-fired power station boilers. Occupational exposure to vanadium pentoxide induces green tongue, asthmatic symptoms and albuminuria with cast. Urinary citrate is freely filtered at the glomerulus, and its reabsorption in the proximal tubule is the major determinant of the rate of renal excretion. In this study, we exposed rat renal brush border membrane vesicles (BBMV) to vanadium pentoxide and examined their citrate uptake characteristics. The preincubation of BBMV with 1 mM V2O5 for 8 hours significantly inhibited citrate uptake compared with that of BBMV without V2O5, preincubation. These findings indicate that the preincubation of BBMV with vanadium pentoxide results in a time-dependent inhibition of citrate uptake by BBMV. These findings might contribute to nephrotoxicity in vanadium exposure.

1. The disposition of SM-11355, an anticancer platinum complex for hepatocellular carcinoma, was investigated in dog by measuring platinum (Pt) and radioactivity levels following intrahepatic arterial administration of (14)C-SM-11355 suspended in Lipiodol, an oily lymphographic agent. Plasma and excretion profiles were monitored in six animals, with tissue distribution studied after 1 day, 4 and 13 weeks (n = 2/time point). 2. SM-11355 was released very slowly into the systemic circulation from Lipiodol, resulting in very low levels of Pt compounds in plasma, urine, faeces and organs. Plasma levels of Pt and radioactivity declined with apparent half-lives of 5-7 weeks. Excretion continued even at 3 months after the administration with proportions excreted for Pt and radioactivity up to 30-60% in urine and 8-10% in faeces. 3. The Pt and radioactivity in the liver accounted for 80-100% of the dose at 1 day and for 20-50% at 13 weeks after the administration, predominately as intact SM-11355. The concentrations were highest in the left lobe of the liver, the administration site, but levels in the remainder of the liver were also markedly higher than those in plasma and other tissues. 4. The results strongly support the concept that SM-11355 targets the liver with highly selectivity and sustained release of Pt compounds.

Bland-White-Garland syndrome (BWG) is a rare disorder that includes abnormalities of the coronary arteries that cause severe myocardial ischemia or infarction in infancy. In this case report, we describe an infant with BWG evaluated by dual single photon emission computed tomography using thallium-201 and I-123 beta-methyl-p-iodophenyl-pentadecanoic acid before and after surgery.

The metabolism of amrubicin hydrochloride (SM-5887), a novel antitumor agent, was investigated in rats and dogs after intravenous administration of 14C-labeled SM-5887. 1. Nine metabolites in rat urine, ten metabolites in rat bile, six metabolites in dog urine and eleven metabolites in dog bile were observed after intravenous administration of 14C-SM-5887. Major radioactive components in rat urine were amrubicinol (SM-5887-13-OH) and the unchanged SM-5887, and those in dog urine were M-2 and Met B. Major radioactive components in bile of rats and dogs were polar metabolites, such as M-1 and M-2. 2. In rat tissues, ma jor radioactive components were nonpolar or less polar metabolites, such as Met B and SM-5887-13-OH, and the unchanged SM-5887. 3. The structures of the nonpolar or less polar metabolites (SM-5887-13-OH and Met A ?? Met D) were determined by comparing them to the authentic samples using HPLC and mass spectrometry. The identified metabolites were an alcoholic derivative (SM-5887-13-OH), deoxyaglycones (Met A and Met B) formed by reductive glycosidic cleavage, an aglycone (Met B) formed by glycosidic cleavage and a deaminated derivative (Met D). Two polar metabolites (M-4 and M-6) were also identified as the conjugates of SM-5887-13-OH with glucuronic acid. The other polar metabolites (M-1 ?? M-3, M-5 and M-7), which were assumed to be conjugates based on their high polarities, could not be identified due to their low stabilities.

Pharmacokinetics on plasma and blood cells concentration, tissue distribution and excretion of amrubicin hydrochloride (SM-5887), a novel antitumor agent, were investigated after a single intravenous administration of 14C-labeled SM-5887 at the doses of 0.1, 1 and 10 mg/kg to rats. 1. The radioactivity levels in plasma and blood cells at these doses decreased multi-exponentially, and the levels of radioactivity in blood cells were 1/2 ?? 1/3 of those present in plasma. The time course profiles were almost parallel among these doses (0.1 ?? 10 mg/kg), and the linear relationship in the area under the concentration time-curve and administered doses was observed. 2. Plasma and blood cell concentrations of the unchanged SM-5887 at these doses decreased more rapidly than those of total radioactivity, and the levels in blood cells were about a half of those in plasma. The α, β and γ-phase half-lives of SM-5887 concentration in plasma were 1.8 min, 29 min and 1.9 hr, respectively. 3. The concentrations of amrubicinol (SM-5887-13-OH), the major bioactive metabolite, in plasma and blood cells increased rapidly, and then decreased slowly. In blood cells, the level of SM-5887-13-OH was three times as high as that in plasma. Other aglycone metabolites without the sugar moiety were also detected in plasma and blood cells. 4. The radioactivity level was relatively high in such tissues as bone marrow, intestinal wall, skin, adrenal, spleen, lung, Harderian gland, submaxillary gland, kidney and liver at 1 ?? 4 hr after administration. Then the radioactivity disappeared gradually from most tissues in a similar profile as that in plasma, except a few tissues such as testis, submaxillary gland, thymus and hair. 5. Within 168 hr after administration, 13% and 76% of the dosed radioactivity was excreted into urine and feces, respectively. Within 72 hr after administration, 58% of the dosed radioactivity was excreted into bile. After intraduodenal injection of the bile to the bile-duct cannulated rats, 31% of the injected radioactivity was reabsorbed. 6. Little difference in pharmacokinetic profile was observed between male and female rats. 7. In vitro protein binding of SM-5887 and SM-5887-13-OH in mammalian plasma and HSA were 92 ?? 97% and 78 ?? 93%, respectively. In vivo plasma protein binding of SM-5887 and SM-5887-13-OH after intravenous administration of SM-5887 to rats were 93 ?? 96% and 82 ?? 89%, respectively.

Metabolism of 4-[1-(2-fluoro-4-biphenylyl)ethyl]-2-methylaminothiazole (SM-8849), a novel immunomodulatory agent, in rats was investigated. By co-chromatography with authentic samples, desmethylated (SM-8800) p-hydroxylated (SL-5512) and desmethylated-p-hydroxylated SM-8849 (SL-5515) were detected in the bile. Thermospray mass spectrometry (TSP-MS) analysis of five metabolites isolated from the bile revealed molecular ions of both conjugates (glucuronides and a sulfate) and their aglycones. Aglycone structures were determined by comparison of their product spectra with those of authentic standards. Further analyses of conjugation sites were carried out by 1H-NMR including differential NOE. As a result, the sulfate of SL-5515 (5515-S), the N-glucuronides of SL-5512 and SM-8849 (5512-NG and 8849-NG, respectively), the glucuronide of SL-5512 (5512-G) and the O-glucuronide of 4-hydroxy-3-methoxy-SM-8849 (CatOMe-OG) were identified. In addition, N-methylthiouras was identified in urine by LC/MS/MS.

(+)-Methyl [2-[(2R,3aS,4R,5R,6aS)-octahydro-5-hydroxy-4-[(E)-(3S,5S)-3-hydroxy-5-methyl-1-[3- 14 C]nonenyl]-2-pentalenyl]ethoxy]-acetate ([nonenyl-3- 14 C]SM-10902) (1) was labeled with carbon-14 for use in mammalian metabolic studies. The synthesis was achieved in 7 steps from [ 14 C]carbon dioxide: including : 1) Grignard reaction, 2) esterification, 3) condensation with dimethyl methylphosphonate, 4) Wittig-Horner reaction, 5) separation of isomers by HPLC. Stereoselective reduction of the protected ketone with sodium borohydride in the presence of cerium (III) chloride and subsequent desilylation produced 1. The overall yield was 11.1% from Ba[ 14 C]CO 3

(2S,3R)-2-[3-(4-Fluorophenyl)]propylamino-3-(3,4-dihydroxyphenyl)-3-hydroxypropionic acid pyrrolidine amide hydrobromide (SM-11044) was labeled with carbon-14 for use in mammalian metabolic studies. The synthesis was achieved according to the scheme shown in Fig. 5. Grignard reaction of 3,4-methylenedioxy-phenylmagnesium bromide with [14C]carbon dioxide liberated from barium [14C]carbonate ((UNDERLINE)6(/UNDERLINE)) gave the acid ((UNDERLINE)5(/UNDERLINE)). Reduction of (UNDERLINE)5(/UNDERLINE) with lithium aluminum hydride followed by oxidation of the resulting benzyl alcohol ((UNDERLINE)11(/UNDERLINE)) with chromium oxide-pyridine complex afforded the aldehyde ((UNDERLINE)3(/UNDERLINE)). Condensation of (UNDERLINE)3(/UNDERLINE) with the optically active (R)-oxazolidinone ((UNDERLINE)10(/UNDERLINE)) yielded the alcohol ((UNDERLINE)12(/UNDERLINE)). Catalytic hydrogenation of (UNDERLINE)12(/UNDERLINE) and subsequent hydrolysis produced the optically active β-hydroxy-α-amino acid ((UNDERLINE)2(/UNDERLINE)), which was treated with N-carbomethoxyphthalimide to give the hydroxy acid ((UNDERLINE)13(/UNDERLINE)). Reaction of (UNDERLINE)13(/UNDERLINE) with pivaloyl chloride gave the corresponding mixed anhydride, which was allowed to react with pyrrolidine to provide the amide ((UNDERLINE)14(/UNDERLINE)). Deprotection of (UNDERLINE)14(/UNDERLINE) with hydrazine hydrate afforded the hydroxyamide (15). Condensation of (UNDERLINE)15(/UNDERLINE) with the aldehyde ((UNDERLINE)16(/UNDERLINE)) and subsequent reduction of the resulting imine with sodium cyanoborohydride produced the fluoride ((UNDERLINE)17(/UNDERLINE)). Cleavage of the methylenedioxy group of (UNDERLINE)17(/UNDERLINE) gave the free base of (UNDERLINE)1(/UNDERLINE), which was treated with hydrobromic acid to afford (UNDERLINE)1(/UNDERLINE). The overall yield was 11.1% from (UNDERLINE)6(/UNDERLINE).

The effect of unilateral nephrectomy on the growth of bladder tumors was investigated in male F344 rats treated with 0.05% N-butyl-N-(4-hydroxybutyl)nitrosamine(BBN) for 6 weeks by means of macroscopical and microscopical examinations. Unilateral nephrectomy 4 weeks after the initiation of BBN administration significantly increased both the number and the volume of visible tumors per rat as compared with the sham-operated group. Histological examination also revealed that the numbers of papillonodular hyperplasia and cancer per basement membrane were significantly increased by unilateral nephrectomy. The results imply a growth-enhancing effect of unilateral nephrectomy on carcinogenesis of the rat bladder.

The serum concentrations of 3 labeled ST-630 [(+)-(5Z, 7E)-26, 26, 26, 27, 27, 27-hexafluoro-9, 10-secocholesta-5, 7, 10(19)-triene-1α, 3β, 25-triol], a new fluorine-substituted analog of 1α, 25-dihydroxyvitamin D3, were determined in male rats after single oral or intravenous administration at a dose of 0.3, 1.0 or 3 μg/kg. The distribution, metabolism and excretion of 3H labeled ST-630 were also studied in male rats after single intravenous administration at a dose of 0.3 μg/kg. 1. The serum levels of radioactivity reached a maximum (Cmax 614 pg eq./ml) at 2 hr after oral dosing at a dose of 1 μg/kg, and then declined with a half life (t1/2) of 1.6 day (24-168 hr after dosing), the AUC[0-168 hr] was 9.48 ng eq.·.hr/ml. The serum concentrations of ST-630 reached a Cmaxs (620 pg/ml) at 2 hr after oral dosing and then declined with a t1/2 of 20 hr (24-72 hr after dosing), the AUC[0-72 hr] was 6.64 ng·hr/ml. 2. Cmax and AUC of the serum levels of radioactivity and ST-630, after oral dosing showed a good linearity to dose ranging from 0.3 to 3 μg/kg. 3. Cmax and AUC of radioactivi ty and ST-630 after oral dosing to fasting rats were about half of those observed in non-fasting rats. 4. After intravenous dosing at a dose of 0.3 μg/kg, the serum levels of radioactivity declined with a t1/2 of 1.4 day (24-168 hr after dosing), the AUC[0-168 hr] was 15.3 ng eq.·hr/ml, and the serum concentrations of ST-630 declined with a t1/22 of 25 hr (24-120 hr after dosing), and the AUC[0-120 hr] was 13.9 ng·hr/ml. The bioavailability was estimated to be about 17%. 5. The tissue distribution pattern of rad ioactivity after intravenous administration was not significantly different from that after oral dosing, except that the level of the small intestine was lower than that after the oral dosing. 6. The main component of radioactivity in the kidney at 24 hr after intravenous dosing was ST-232, a bioactive metabolite. However, in the small intestine at 24 hr after intravenous dosing, ST-630 was mainly detected. 7. Within 168 hr after intravenous dosing at 0.3 μg/kg, 8.5% and 71.2% of dosed radioactivity were excreted in the urine and feces, respectively.

The absorption, distribution and excretion of 3H labeled ST-630 [(+)-(5Z, 7E)-26, 26, 26, 27, 27, 27-hexafluoro-9, 10-secocholesta-5, 7, 10(19)-triene-1α, 3β, 25-triol], a new fluorine-substituted analog of 1α, 25-dihydroxyvitamin D3, were studied after single oral administration at a dose of 1 or 10 μg/kg in male and female rats. 1. The serum levels of radioactivity reached a maximum (Cmax) at about 2 hr after dosing at a dose of 1 μg/kg to male and female rats, and then declined with a half life (t1/2) of 2.8 day (24 ?? 168 hr after dosing) or 2.0 day (24 ?? 168 hr after dosing) in male and female rats, respectively. The serum concentrations of ST-630 reached a Cmax at 2 hr after dosing and then declined with a t1/2 of 1.0 day (male, 24 ?? 72 hr after dosing) or 0.9 day (female, 24-48 hr after dosing). Most of the radioactivity at 2 hr after dosing was found to be ST-630, and, the ratio of ST-630 to the radioactivity was decreased with time. 2. High levels of radioactivity were detected in the small intestine at 2 ?? 24 hr and in the kidney at 24 hr after dosing at a dose of 1 μg/kg to male and female rats. The autoradiograms of the small intestine showed that the radioactivity was distributed selectively at the intestinal mucosa besides the intestinal contents at 2 ?? 24 hr after dosing with 10 μg/kg to male rats. 3. The ratios of non-volatile radioactive component excluding tritiated water to total radioactivity (dry/wet) in many tissues were decreased with time. However little decrease of dry/wet in the liver, kidney and fat were observed at 168 hr after dosing. 4. Excretion of radioactivities in the urine, feces and expired air were 4.8%, 86.1% and 1.2% respectively for male rats, and 5.0%, 87.9% and 0.8% respectively for female rats within 168 hr after dosing of 1 μg/kg. 5. The t1/2 and AUC of radioactivity in male rats were about 1.4 times higher than those in female rats. However there were no marked differences in pharmacokinetics of ST-630 between male and female rats. No significant sex-related differences in the distribution and excretion of radioactivity were observed. 6. In bile-duct cannulated rats, 16% of dosed radioactivity was excreted into the bile for 72 hr after dosing at 1 μg/kg, and then approximately 40% of radioactivity presented in the bile was reabsorbed within 48 hr.

The effects of oral administration of nordihydroguaiaretic acid (NDGA), an antioxidant and inhibitor of arachidonic acid metabolism, on rat bladder carcinogenesis were examined. Six‐week‐old male Fischer 344 rats were given drinking watar containing 0.05% N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine for 4 weeks. Following this 4‐week period, diet containing 5% sodium saccharin (SS) with or without 0.1% NDGA supplement was given to the rats for 36 weeks. The incidences of papillary or nodular (PN) hyperplasia and of papilloma in the group treated with SS plus NDGA were significantly lower than those in the group treated with SS alone. The number of PN hyperplasic foci per 10 cm of basement membrane in rats treated with SS plus NDGA was also lower than that in the group treated with SS alone. These results suggest that NDGA has an anti‐tumor‐promoting effect on rat bladder carcinogenesis.

9-Amino-8-fluoro-2, 4-methano-1,2,3,4-tetrahydroacridine citrate (SM-10888), a novel cholinesterase inhibitor, was labeled with carbon-14 at C9 of the tetrahydroacridine ring for use in metabolic studies. Carbonation of 2,6-difluorophenyllithium (3) with [14C]carbon dioxide gave the acid (4). Chlorination of 4 followed by treatment of the resulting acid chloride with ammonia afforded the amide (5). Dehydration of 5 with thionyl chloride and subsequent displacement reaction with ammonia gave the aminobenzonitrile (7). Condensation of 7 with the ketone (8) in the presence of anhydrous zinc chloride yielded the aminoacridine (9), which was treated with citric acid to afford [9-14C]SM-10888 (1). The overall yield of 1 was 37% from 2, and the specific activity was 1.35 GBq/mmol.

N-[4-[4-(2-Pyrimidinyl)-1-piperazinyl]butyl]bicyclo-[2.2.1]heptane-2,3-di-exo-carboxyimide dihydrogen citrate (tandospirone), a novel anxiolytic agent, was labeled with carbon-14 individually at the imido carbonyl group and the pyrimidinyl ring. The synthesis of carbonyl-labeled tandospirone was achieved according to the scheme shown in Fig. 3. Diels-Alder reaction of maleic anhydride (2) with cyclopentadiene (3) afforded the endo anhydride (4), which was transformed into the imide (5) by treating with ammonia water. Thermal isomerization of the endo imide (5) and subsequent chromatographic separation gave the pure exo compound (6). Catalytic hydrogenation of 6 followed by alkylation with 1,4-dibromobutane (8) yielded the bromide (9a). Condensation of 9a with N-(2-pyrimidinyl)piperazine (10a) followed by treatment of the resulting disubstituted piperazine (11a) with citric acid afforded [carbonyl-14C]tandospirone (1a). The overall yield of 1a was 22% from 2. The similar method was applied to the synthesis of pyrimidinyl-labeled tandospirone as shown in Fig. 4. Condensation of 2-chloro[2-14C]pyrimidine (12) with anhydrous piperazine gave the pyrimidinylpiperazine (10b). N-alkylation of 10b followed by treatment with citric acid afforded [pyrimidinyl-2-14C]tandospirone (1b). The overall yield of 1b was 68% from 12.

(2R, 5S) - (+) -cis-3, 5-Dimethyl-2- (3-pyridyl) thiazolidin-4-one hydrochloride (SM 12502), a new PAF antagonist, was labeled with carbon-14 at C 2 of the thiazolidinone ring for use in metabolic studies. The synthesis was achieved according to the scheme illustrated in Fig, 3. 14C-Carbonation of 3-pyridyllithium (3) followed by chlorination and esterification gave mehyl [carbonyl-14C] nicotinate (6) . Reduction of 6 with diisobutylaluminium hydride produced a mixture of the alcohol (7) and the aldehyde (8) . Oxidation of the mixture with manganese dioxide gave 8 as a sole product. Condensation of 8 with methylamine and subsequent cyclocondensation of the imine (9) with (S) -2-mercaptopropionic acid (10) yielded a cis and trans mixture of the thiazolidinone (11) . Isolation of the desired stereoisomer by chiralphase HPLC and subsequent treatment with hydrochloric acid afforded 1. The overall yield of 1 was 22% from 2.

(1R,5S,6S)-2- [(3S,5S)-5-Dimethylaminocarbonylpyrrolidin-3-ylthio]-6- [(1R)-1-hydroxyethyl]-1-methylcarbapen-2-em-3-carboxylic acid trihydrate (SM-7338), a novel 1 β-methyl carbapenem antibiotic, was labeled with carbon-14 at the C3 position of the carbapenem nucleus for use in metabolic studies. The synthesis was achieved according to the scheme illustrated in Fig. 3. Selective esterification of [2-14C]malonic acid (2) with 4-nitrobenzyl bromide (3) gave its monoester (4). Condensation of the magnesium salt of 4 with the imidazolide derived from the azetidinone carboxylic acid (5) provided the β-keto ester (6), which was desililated with hydrochloric acid to give the alcohol (7). Treatment of 7 with carboxybenzenesulfonyl azide followed by decomposition of the resulting diazo intermediate (8) in the presence of a rhodium catalyst produced the bicyclic keto ester (9). Reaction of 9 with diphenyl chlorophosphate in the presence of N,N-diisopropylethylamine and subsequent displacement reaction of the vinyl phosphate (10) with mercaptopyrrolidine (11) gave bis-protected SM-7338 (12). Catalytic hydrogenolysis of 12 afforded [carbapenem-3-14C]-SM-7338 (1). The overall yield of 1 was 5.8% from 2.

Two stereoisomers of empenthrin, (E) -1-ethynyl-2-methyl-2-pentenyl (1 R) -cis, trans-chrysanthemates, were regioselectively labeled with carbon-14 for use in metabolic studies on the characteristic ethynyl alcohol moiety. Action of ethyl [14C] formate (2) on ethylidenetri-phenylphosphorane gave the 14C-formylated phosphorane (3) . Wittig reaction of 3 with propanal stereoselectively yielded the E isomer of 2-methyl-2- [1-14C] pentenal (4), which was derivatized to a crystalline imidazolidine (5) for purification. Regeneration of 4 from purified 5 followed by reaction with ethynylmagensium bromide gave (E) -1-ethynyl-2-methyl-2- [1-14C] -pentenol (6), the a-ethynyl alcohol of 14C-labeled empenthrin, in 36% overall radiochemical yield from 2. Esterification of 6 with (1R) -cis- and (1R) -traps-chrysanthemoyl chlorides afforded the corresponding optically active [pentenyl-1-14C] empenthrins (1a and 1b) in good yields after chromatographic purification.

In vitro studies were carried out to identify the major contribution of CYP2C8, CYP2D6 and CYP3A4 to the metabolism of perospirone (cis-N-[4-[4-(1,2-benzisothiazol-3-yl)-1-piperazinyl]butyl]cyclohexane-1,2-dicarboximide monohydrochloride dehydrate), a novel antipsychotic agent, using human liver microsomes and expressed P450 isoforms. Quinidine (a specific inhibitor of CYP2D6) did not markedly affect the metabolism of perospirone, whereas quercetin (an inhibitor of CYP2C8) and ketoconazole (an inhibitor of CYP3A4) caused a decrease in the metabolism with human liver microsomes in a concentration dependent fashion. With 10 microM quercetin, the metabolism of perospirone was inhibited by 60.0% and with 1 microM ketoconazole almost complete inhibition was apparent. Anti-CYP2C8 and anti-CYP2D6 antisera did not exert marked effects, whereas anti-CYP3A4 antiserum caused almost complete inhibition. With expressed P450s, K(m) and V(max) values were 1.09 microM and 1.93 pmol/min/pmol P450 for CYP2C8, 1.38 microM and 5.73 pmol/min/pmol P450 for CYP2D6, and 0.245 microM and 61.3 pmol/min/pmol P450 for CYP3A4, respectively. These results indicated that the metabolism of perospirone in human liver was mainly catalysed by CYP3A4, and to a lesser extent CYP2C8 and CYP2D6 were responsible because kinetic data (K(m) and V(max)) of CYP2C8 and CYP2D6 suggested catalytic potential.

In vitro metabolism of perospirone was examined with rat, monkey and human liver S9, human liver microsomes and yeast microsomes expressing human P450, using 14C labeled perospirone. With rat liver S9, the major metabolites were MX9 and ID-11614, produced by cleavage at the butylene chain. However, some butylene non-cleavage and hydration of the cyclohexane ring were found, although limited in extent. Unknown metabolites accounted for about 10% of the total. After incubation for 10 minutes with monkey liver S9, the major metabolites were ID-15036 and MX11, hydrated in the cyclohexane ring. After incubation for 60 minutes, ID-15001, i.e. the butylene chain cleavage type increased. Unknown metabolites accounted for about 20%. After incubation for 10 minutes with human liver S9, the major metabolite was ID-15036, hydrated in the cyclohexane ring. In addition, MX11 and many unknown metabolites were evident. After incubation for 60 minutes, the butylene chain cleavage type and unknown metabolites increased. Individual differences were found in the metabolic reaction rate. With human liver microsomes. MX11, ID-15001 and unknown metabolites were again the major metabolites. With yeast microsomes expressing human P450 subtypes, CYP1A1, 2C8, 2D6, 3A4 were responsible for the metabolism in particular, and CYP3A4 contributes greatly. Therefore it is unlikely that genetic polymorphism will arise a present a problem with regard to the clinical drug. The results demonstrated that the main metabolic pathway in human liver S9 and liver microsomes involve oxidation at cyclohexane, oxidative cleavage of the butylene side chain and S-oxidation. The same was the case in rat and monkey S9, but species differences were found in the proportions of the metabolites produced.

The pharmacological profile of N-(6-(2-(5-bromopyrimidine-2-yloxy)ethoxy)-5-(4-methylphenyl)pyrimidin-4-yl)-4-(2-hydroxy-1,1-dimethylethyl) benzensulfonamide sodium salt sesquihydrate (TA-0201), a new antagonist of endothelin receptors, was examined, using human recombinant and prostate endothelin receptors. In binding experiments with [125I]endothelin-1, TA-0201 showed extremely high affinity for recombinant endothelin ET(A) receptors (pK(i)=10.7), as compared with that for recombinant endothelin ET(B) receptors (pK(i)=7.8). Endothelin ET(A) and ET(B) receptors coexisted in human prostate with different proportions (endothelin ET(A) receptor: approximately 70%), which were distinguished by TA-0201 in the same manner as with recombinant receptors. Human prostate strips contracted in response to endothelin-1 and sorafotoxin S6c, but the maximum contraction induced by endothelin-1 was approximately three times greater than that induced by sarafotoxin S6c. The response to endothelin-1, but not to sarafotoxin S6c, was inhibited by TA-0201 and cyclo(D-Asp-Pro-D-Val-Leu-D-Trp) (BQ123) (endothelin ET(A) receptor antagonist) but not by BQ788 (endothelin ET(B) receptor antagonist). These results suggest that TA-0201 is a highly selective endothelin ET(A) receptor antagonist and will be useful for understanding the physiological and pathological roles of the endothelin ET(A) receptor in human prostate and other tissues.

Since amphiphilic drugs are known to interact with biomembranes, we investigated local vessel damage and thrombosis which might be brought about by intravenous dosing using chlorpromazine (CPZ) as a representative compound. CPZ-induced hemolysis was suppressed by an increase in sucrose concentration in the medium, characterizing this hemolysis to be colloid-osmotic lysis, which includes the enhancement of membrane phospholipid fluidity and consequent small pore formation in the membranes. This was supported by the observation that hemolysis by filipin, not featuring the stage of small pore formation, was not affected by sucrose. [14C]Glucose-entrapping liposomes were degraded by CPZ, and this degradation was enhanced by an increase in the intravesicle glucose concentration. These results indicated that the compound could induce colloid-osmotic lysis in erythrocytes and artificial membrane vesicles. CPZ also injured cultured porcine aortic endothelial cells (PAEC), as evidenced by lactate dehydrogenase (LDH) leakage. This injury was also suppressed by increase in sucrose concentration in the medium, suggesting that colloid-osmotic lysis again occurred. When rats were intravenously injected with CPZ, local endothelial cell (EC) injury and associated thrombus formation were observed, suggesting that CPZ's action was also evident in vivo. To our knowledge, this is the first finding which suggests that an intravenously dosed amphiphilic drug can injure local ECs based on a colloid-osmotic lysis mechanism leading to thrombosis.

1. 26,26,26,27,27,27-F6,-1,25(OH)2 vitamin D3, Falecalcitriol, the hexafluorinated analogue of 1,25(OH)2 vitamin D3, has been reported to be several times more potent than the parent compound regarding some vitamin D actions. The reason for enhanced biological activity appears related to F6-1,25(OH)2 vitamin D3 metabolism to F6-1,23S,25(OH)3 vitamin D3, a bioactive 23S-hydroxylated form which is resistant to further metabolism. 2. In the present in vivo studies, the repeated oral administration of [3H]F6-1,25(OH)2 vitamin D3 to rat resulted in a significant reduction of the radioactivity and the F6-1,25(OH)2 vitamin D3 concentrations in serum, especially at the 2 h maximum point after each dosing. Additionally, F6-1,23S,25(OH)3 vitamin D3 in the serum and small intestine was increased by the prior administration of F6-1,25(OH)2 vitamin D3. 3. Further in vitro investigation showed [3H]F6-1,25(OH)2 vitamin D3 to be metabolized to F6-1,23S,25(OH)3 vitamin D3 by kidney and small intestine homogenates of rat, the reaction being increased by the prior administration of F6-1,25(OH)2 vitamin D3. Moreover, this latter treatment was associated with a marked increase of CYP24 mRNA in the small intestine within 4 h after dosing. 4. The results indicate that in vivo metabolism of F6-1,25(OH)2 vitamin D3 to F6-1,23S,25(OH)3 vitamin D3 is catalysed by CYP24, the enzyme being induced by prior substrate exposure.

26,26,26,27,27,27-Hexafluo-1,25(OH) 2 vitamin D 3 , the hexafluorinated analog of 1,25(OH) 2 vitamin D 3 , has been reported to be several times more potent than the parent compound regarding some vitamin D actions. The reason for enhanced biologic activity in the kidneys and small intestine appears to be related to F 6 -1,25(OH) 2 vitamin D 3 metabolism to ST-232, 26,26,26,27,27,27-hexafluoro-1α,23S,25-trihydroxyvitamin D 3 , a bioactive 23S-hydroxylated form that is resistant to further metabolism. Since F 6 -1,25(OH) 2 vitamin D 3 is considered to prevent osteoporotic decrease in bone mass by suppressing bone turnover, we here compared the distribution and metabolism of [1β- 3 H]F 6 -1,25(OH) 2 vitamin D 3 and [1β- 3 H]1,25(OH) 2 vitamin D 3 in bones of rats by autoradiography and radio-HPLC. In the dosed groups, radioactivity was detected locally in the metaphysis, the modeling site in bones. As compared with the [1β- 3 H]1,25(OH) 2 vitamin D 3 case, [1β- 3 H]F 6 -1,25(OH) 2 vitamin D 3 was significantly retained in this site, and moreover, it mainly persisted as unchanged compound and ST-232. These findings indicate that the reason for the higher potency of F 6 -1,25(OH) 2 vitamin D 3 than 1,25(OH) 2 vitamin D 3 in bones are linked with increased distribution and reduced metabolism.

We previously reported the expression of hepatocyte growth factor (HGF), a ligand for c-MET, at the level of mRNA and protein in human prostate tissues. The present study was carried out to evaluate the relationship between c-MET expression and cancer cell proliferation or effect of cancer therapy. The expression both in mRNA and protein levels of c-MET proto-oncogene was determined semi-quantitatively by reverse transcription-competitive polymerase chain reaction (RT-competitive PCR) and Western blot analysis for prostate tissues from 32 Japanese male subjects. In addition, tissue localization of c-MET translation product was examined by immunohistochemistry for corresponding specimens. Although there was significantly higher c-MET protein expression in malignant (prostate cancer treated with/without neoadjuvant endocrine therapy) than in non-malignant prostate tissues (normal prostate and benign prostate hyperplasia; BPH), unexpectedly, c-MET mRNA showed high expression in the non-malignant group. Thus, there was no parallelism between mRNA and protein expressions of c-MET. Endocrine therapy did not alter c-MET mRNA and protein expressions in human prostate cancer. Immunohistochemical localization and expression of c-MET protein was found to be intense in cancer cells and weak in epithelia of normal and hyperplastic prostates. Unconcerted expression of mRNA and protein of c-MET, the reason of which is uninterpretable, is supposed to be one of characteristics of human prostate cancer.

Both of the components of prallethrin SF [(4S)-1-oxo-2-propargyl-3-methylcyclopent-2-en-4-yl (1R)-cis,trans-chrysanthemate], a novel potent insecticidal pyrethroid, were labelled with carbon-14 at the cyclopentenyl-2 position for use in metabolic studies. Carbonation of 2-lithio-5-methylfuran (4) with carbon-14 dioxide (3) gave 5-methyl-2-[carboxyl-14C]furoic acid (5). Reduction of 5 with lithium aluminium hydride followed by oxidation with manganese dioxide afforded 5-methyl[carbonyl-14C]-furfural (7). Modified Grignard reaction of 7 with propargyl bromide and magnesium in the presence of mercuric chloride yielded 1-(5-methyl-2-furyl)[1-14C]but-3-yn-1-ol (8). Molecular rearrangement of 8 followed by isomerization afforded 2-propargyl-3-methyl[2-14C]cyclopent-2-en-4-ol-1-one (9) in 30% yield from barium [14C]carbonate (2). Direct optical-resolution of 9 by preparative HPLC with a chiral-phase column gave (+)-(4S) and (-)-(4R) isomers (10 and 11) quantitatively. Condensation of 10 with (1R)-cis- or (1R)-trans-chrysanthemoyl chloride (12a or 12b) yielded (4S),(1R)-cis- or (4S),(1R)-trans-[cyclopentenyl-2-14C]prallethrin (1a or 1b). Both overall radiochemical yields of 1a and 1b were about 9% from 2.

A novel fungicide, (-)(E)-1-(2,4-dichlorophenyl)-4, 4-dimethyl-2(1,2,4-triazol-1-yl)-1-penten-3-ol (S-3308 L) and its three optically active stereoisomers were labeled with carbon-14 at the triazole ring for use in the metabolic and environmental fate studies. 1,2,4-Triazole-14C (4) prepared from formamide-14C (3) was treated with bromopinacolone to give triazolylpinacolone-14C (5), which was condensed with 2,4-dichlorobenzaldehyde giving (Z)-ketone-14C (6). (Z)-Ketone-14C (6) was photoisomerized to (E)-ketone- 14C (7), which was reduced to racemic (Z)-S-3308- 14C (1) in 20% yield from 3. Reduction of 6 gave racemic (Z))S-3308-14C (2) in 23% field from 3. Optical resolution of both racemates 1 and 2 by a HPLC method with a chiral column gave (-)(E)-, (+)(E)-, (-)(Z)- and (+)(Z)-1-(2,4-dichlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-[14C]-1-yl)-1-penten-3-ols (1a, 1b, 2a and 2b) in essentially Quantitative yields.

(S)-α-Cyano-3-phenoxybenzyl (S)-2-(4-chlorophenyl)-3-methylbutyrate (fenvalerate Aα), an optically active insecticide, was labelled with carbon-14 individually at the benzyl ring (1a) and the chlorophenyl ring (1b) for use in metabolic studies. Ullmann reaction of 3-methylphenol-14C6 (2) with bromobenzene followed by oxygen oxidation gave 3-phenoxybenzoic-14C6 acid (4). Reduction of 4 followed by potassium dichromate oxidation of the alcohol (5) gave 3-phenoxybenzaldehyde-14C6 (6). Condensation of 6 with (S)-2-(4-chlorophenyl)-3-methylbutanoyl chloride and sodium cyanide afforded fenvalerate A-(benzyl-14C6), which on optical resolution gave 1a in 29% yield from 2. Isopropylation of 4-chlorophenyl-14C6-acetonitrile (9) followed by hydrolysis gave 2-(4-chlorophenyl-14C6)-3-methylbutyric acid which on optical resolution gave the (S)-acid (12). Esterification of the acid with α-cyano-3-phenoxybenzyl bromide yielded fenvalerate A-(phenyl-14C6), which was resolved to give 1b in 9% yield from 9.

O- (2,6-Dichloro-4-methylphenyl) O,O-dimethyl phosphorothioate (5-3349) (1), a fungicide in soil, was labelled with carbon-14 individually at the aryl methyl and the phenyl ring for use in metabolic studies. The synthetic procedures are illustrated in Fig. 1. 4-Cyano-14C-phenol was quantitatively converted to 4-methyl-14C-Phenol (3a) by catalytic hydrogenation using 10% Pd-C as a catalyst. Dichlorination of 3a with chlorine gave 2,6-di-chloro-4-methyl-14C-phenol (4a). Condensation of 4a with O,O-dimethyl phosphorochloridothioate gave S-3349-(aryl methyl-14C) (1a) in the overall yield of 22% from potassium cyanide-14C. By the similar procedures, 4-methylphenol-14C6 (3b) was converted to S-3349-(Phenyl-14C6) (1b) in 70% yield.

The four 14C-labelled stereoisomers of both 3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropanecarboxylic acid and 3-(2,2-dibromoethenyl)-2,2-dimethylcyclopropanecarboxylic acid, the improtant acid components of various potent insecticidal pyrethroids, were prepared efficiently from [1-14C]chrysanthemic acid (5) for use in comparative metabolic studies. α-Bromo-β-naphthyl d,l-cis, trans-[1-14C]chrysanthemate (6) was oxidized with osmium tetroxide-sodium periodate to give the formyl ester (7) in 78% yield. Wittig reaction of 7 with dichloromethylenetris(dimethylamino)phosphorane at −75 - −70 °C afforded the dichlorovinylated ester (8) quantitatively. Optical resolution of 8 by preparative HPLC using a chiral-phase column led to the four stereoisomers (8a - 8d) with optical purities more than 99%. Hydrolyses of the esters (8a - 8d) yielded the corresponding optically active 3-(2,2-dichloroethenyl)-2,2-dimethyl [1-14C]cyclopropanecarboxylic acids (1a - 1d) quantitatively. Similarly, Wittig reaction of the formyl ester (7) with dibromomethylenetris(dimethylamino)phosphorane, followed by optical resolution and hydrolysis, gave four optically active 3-(2,2-dibromoethenyl)-2,2-dimethyl [1-14C]cyclopropanecarboxylic acids (2a - 2d) in considerably high yields.

Optically active cyphenothrins, a-cyano-3-phenoxybenzyl (1R) -cis- and (1R) -trans-chrysan-themates were labelled with 14C at the carbonyl carbon for use in metabolic studies. Ethyl glycinate-1-14C hydrochloride (6), prepared from potassium cyanide-14C (2) in the three steps, was condensed with 2, 5-dimethyl-2, 4-hexadiene via the diazo intermediate to give ethyl cis, trans-chrysanthemate-14C (7) . Hydrolysis of the ester 7 followed by geometrical separation gave cis-and trans-chrysanthemic-14C acid (8a and 8b) . Each geometrical isomer (8a or 8b), was optically resolved to afford (1R) -cis- or (1R) -trans-chrysanthemic-14C acid (8c or 8d) . Esterification of 8c and 8d with a-cyano-3-phenoxybenzyl bromide (9) gave (1R) -cis-and (1R) -trans-eyphenothrin- (carbonyl-14C) (la and 1b) in the yields of 4 and 6% from 2, respectively.

Farnesylacetic acid was efficiently labelled with 14C at the 5-position and gefarnate, a potent ulcer inhibitor, was prepared from it in radioactive form for use in metabolic studies. Condensation of [carbonyl-14C]acetyl chloride (5) with t-butyl 2-ethoxymagnesiomalonate (6) followed by acid-catalyzed deprotection and decarboxylation gave ethyl 3-oxo[3-14C]butanoate (8). Alkylation of the keto ester (8) with geranyl bromide (9) afforded the unsaturated keto ester (10), which was hydrolyzed and decarboxylated to give geranyl[2-14C]acetone (11). Grignard reaction of 11 with cyclopropylmagnesium bromide followed by treatment with hydrobromic acid yielded [4-14C]homofarnesyl bromide (13). Cyanation of 13 with potassium cyanide and subsequent hydrolysis gave [5-14C]farnesylacetic acid (1) in 6.1% yield from barium [14C]carbonate (3). Chlorination of 1 followed by esterification with geraniol afforded [5-14C]gefarnate (2) in 88% yield.

Bromobutide, a novel herbicide, was labeled with carbon-14 independently at the carbonyl group and the phenyl ring for use in metabolic studies. 14C-Carbonation of neopentylmagnesium chloride (3) gave 3,3-dimethyl[1-14C]butyric acid (4a) quantitatively. Chlorination of 4a with thionyl chloride followed by alpha-bromination with bromine yielded 2-bromo-3,3-dimethyl[1-14C]-butyryl halide (5a), which was subsequently condensed with alpha, alpha-dimethylbenzylamine (6a) to afford [carbonyl-14C]bromobutide (1a). The overall yield of 1a was 76% from barium [14C]-carbonate (2). Similarly, condensation of alpha, alpha-dimethyl[phenyl-14C]benzylamine (6b), which was prepared from alpha-methyl[phenyl-U-14C]styrene (7) in three steps, with 2-bromo-3,3-dimethylbutyryl halide (5b) gave [phenyl-14C]bromobutide (1b) in 67% yield after purification. The specific activities of 1a and 1b were 1.38 and 0.781 GBq/mmol (37.2 and 21.1 mCi/mmol), respectively.

A method for measuring specific activities of 14C-labelled compounds by gas chromatography-mass spectrometry-computer system (GC-MS-CPU) was developed. This method was proved to provide practically precise and accurate specific activities of various 14C-labelled compounds with such merits as requirement of small amount of samples, being applicable to volatile compounds, and convenience. The C. V. percent obtained for tested compounds was within 3.9 and the reliable sensitivity should be over 37 MBq/mM (1 mCi/mM) . This method was also useful for obtaining information on the labelling pattern and the synthetic procedures applied.

Effects of retinoids and inhibitors of arachidonic acid metabolism on tumor-promoter-induced soft agar colony formation of mouse epidermal cells and rat bladder cells were evaluated. Topical application of retinoic acid, an anti-tumor-promoter, to female SENCAR mouse skin inhibited 12-O-tetradecanoylphorbol-13-acetate-induced soft agar colony formation of mouse epidermal cells, an event proposed to be essential for tumor promotion. Effects of dietary retinyl acetate, nordihydroguaiaretic acid (NDGA) and quinacrine hydrochloride on colony formation of rat bladder cells were then examined. Male Fischer 344 rats were given 0.05% N-butyl-N-(4-hydroxybutyl)nitrosamine for 3 weeks, followed immediately by the administration for 9 weeks of 5% sodium saccharin supplemented with or without 0.05% retinyl acetate, 0.1% NDGA or 0.01% quinacrine hydrochloride. Saccharin-induced colony growth was significantly inhibited by the administration of retinyl acetate or NDGA, suggesting that these two agents have anti-tumor-promoting effects on rat bladder carcinogenesis. Thus, the colony-forming assay might be useful for early detection of anti-tumor-promoters of skin and bladder.

Several ionic liquids were applied as catalysts for the synthesis of cyclic urethanes from amino alcohols and pressurized CO2 in the presence of alkali metal compounds as promoters. A comparative study was made for the catalytic performance using different ionic liquids, substrates, promoters, and pressures. The optimum catalytic system was BMIM-Br promoted by K2CO3, which, for 1-amino-2-propanol, produced cyclic urethane in 40% yield with a smaller yield of substituted cyclic urea and no oligomeric byproducts. For other amino alcohols, cyclic urethanes, cyclic ureas, and/or undesired byproducts were produced in different yields depending on the substrates used. Possible reaction mechanisms are proposed.