Pharmacodynamic study of procaterol hydrochloride dry powder inhaler: evaluation of pharmacodynamic equivalence between procaterol hydrochloride dry powder inhaler and procaterol hydrochloride metered-dose inhaler in asthma patients in a randomized, double-dummy, double-blind crossover manner.

Therapeutic equivalence between procaterol hydrochloride dry powder inhaler (Meptin DPI) and procaterol hydrochloride metered-dose inhaler (Meptin MDI), the currently marketed formulation, was assessed in 16 patients with bronchial asthma. The study was conducted in a randomized, double-dummy, double-blind crossover manner, using forced expiratory volume in the first second (FEV1) as an index of bronchodilatory effect. In Period I, the patients received 20 mcg of either Meptin DPI or Meptin MDI, and then crossed over in Period II after a washout interval of 3--28 days. Pharmacodynamic equivalence was accessed using AUC (FEV1)/h and peak FEV1 as indices, and the data were analyzed by analysis of variance (ANOVA). Factors used for the analysis were the treatment group and/or carryover effect, patients within each group, period, and treatment. The 90% confidence intervals for the differences between the two treatments were --0.0995 to --0.0204 (L) for mean AUC (FEV1)/h and --0.102 to --0.022 (L) for mean peak FEV1, both within the acceptance criteria of --0.15 to 0.15 (L). Meptin DPI was therefore assessed as being equivalent to the current Meptin MDI.

A new model for pancreatitis.

Pancreatitis induced by ligation of the pancreatic duct produces morphologic similarities to human pancreatitis. This model is easily performed in big animals, but it is very difficult to perform pancreatic duct ligation in small animals. Many experimental studies of pharmaceutical treatments for pancreatitis used pancreatic duct-ligation models, but it is also difficult to evaluate the efficacy of the drugs used, because the animals used are of different species with individual differences. To overcome these problems, we ligated the main pancreatic duct of the splenic lobe by a 5.0 absorbable suture by using a surgical microscope and left the gastroduodenal lobe intact in the same rats. This model produced damaged pancreatic tissue in one part and normal pancreatic tissue in another part of the pancreas in the same animals, biochemically and histologically. We evaluated the effect of a new protease inhibitor (ONO-3404) on this preliminary model and found this new protease inhibitor demonstrated a hypertrophic effect on the damaged pancreatic tissue and the normal pancreatic tissue in the same animals. This model is also useful to study pharmaceutic treatment for pancreatic insufficiency and to study chemically induced pancreatic carcinogenesis in the damaged pancreatic tissue and the normal pancreatic tissue in the same animals.

Inhibitory effect of green tea extract on the process of pancreatic carcinogenesis induced by N-nitrosobis-(2-oxypropyl)amine (BOP) and on tumor promotion after transplantation of N-nitrosobis-(2-hydroxypropyl)amine (BHP)-induced pancreatic cancer in Syrian hamsters.

Epidemiologic studies have shown a lower risk of gastrointestinal cancer in green tea drinkers. In the present study, the inhibitory effect of green tea extract (GTE) on the process of pancreatic carcinogenesis induced by N-nitrosobis-(2-oxypropyl)amine (BOP) and on tumor promotion after transplantation of N-nitrosobis-(2-hydroxypropyl)amine (BHP)-induced pancreatic cancer were investigated in hamsters. In the first experiment, shortly after the initiation of pancreatic carcinogenesis by BOP, the animals in the GTE group were given GTE (0.5 mg/L) in their drinking water and the control group was given tap water. All animals were sacrificed 24 weeks later. There were no significant differences in body weight, water intake, or food consumption between the two groups during the experiments. GTE consumption was approximately 1.25 mg/day/100 g body weight during this experiment. Seven of the 13 hamsters (54%) in the control group were found to have pancreatic tumors, versus six of the 18 hamsters (33%) in the GTE group. The average number of tumors in the control group was 1.0/hamster, compared with 0.5/hamster in the GTE group. The overall incidence of macroscopic pancreatic tumors in the GTE group was about half that in the control group. The incidence of pancreatic cancer was 54% (12/13) in the control group and 44% (8/18) in the GTE group. The number of pancreatic cancers, including invasive carcinoma and carcinoma in situ, in the GTE group was 0.88/hamster, significantly lower than in the control group (1.68/hamster) (p < 0.05). The incidence of atypical ductal hyperplasia, which is thought to be an early pancreatic cancer, was also significantly lower in the GTE group than in the control group (1.50/hamster vs. 4.65/hamster) (p < 0.05). In the second experiment, 1-mm3 pieces of BHP-induced pancreatic cancer were transplanted into the back of hamsters. The control group (N = 16) was maintained on the basal diet and tap water throughout the experiment, and the GTE group (N = 16) was also maintained on the basal diet and tap water for the first 3 weeks after transplantation, when successful transplantation was confirmed and, thereafter, given tap water containing GTE (0.5 mg/L) for an additional 12 weeks. Tumor growth was similar in both groups until 11 weeks after transplantation, but inhibition of tumor growth became apparent after 11 weeks in the GTE group. At 13 weeks, the average tumor volume in the GTE group was 1.01 +/- 0.11 x 104 mm3, significantly smaller than that in the control group (1.98 +/- 0.37 x 104 mm3) (p < 0.05). The results demonstrated that GTE has an inhibitory effect on the process of pancreatic carcinogenesis and on tumor promotion of transplanted pancreatic cancer. These results suggest that GTE may come to serve as a chemopreventive and chemotherapeutic agent for pancreatic cancer.

Effects of a new synthetic lipid A on endogenous tumor necrosis factor production and antitumor activity against human pancreatic cancer cells.

The effects of a synthetic lipid A on tumor necrosis factor (TNF) production and antitumor activity against human pancreatic cancer cells were investigated. Lipid A (10 mg/kg) was injected into normal rats and mice and serum TNF levels were measured. Lipid A-induced inhibition of Molt 4 and MIA paca-2 cells in culture were measured by counting viable cells. Activity of lipid A against transplanted human pancreatic cancer cells (MIA paca-2, Panc-1) was examined by determining tumor volume, necrosis, and survival rate after intraperitoneal injections of lipid A (10 and 20 mg/kg) over 4 weeks. Serum TNF levels increased 80-fold in rats and 100-fold in mice after intravenous lipid A injection. Although specific tumor growth inhibition by lipid A was not observed in vitro, tumor growth was significantly inhibited, and the survival rate was improved in pancreatic cancer cell-transplanted nude mice treated with lipid A compared with controls. Synthetic lipid A induces TNF production and has antitumor activity against transplanted pancreatic cancer cells. Further studies of this lipid A as an agent for pancreatic cancer are warranted.

Inhibitory effect of sarcophytol A on pancreatic carcinogenesis after initiation by N-nitrosobis(2-oxypropyl)amine in Syrian hamsters.

Sarcophytol A (SaA), a cembrane-type diterpene, inhibits pancreatic carcinogenesis induced by N-nitrosobis(2-oxypropyl)amine (BOP) in hamsters. The experimental groups received two injections of BOP at 70 mg/kg dose, followed 2 weeks later by a 20 mg/kg dose injection, and were fed a basal diet or 0.01 and 0.05% SaA diets starting 1 week after the second injection of BOP. Control groups were injected with normal saline and fed the basal diet or the 0.05% SaA diet. All animals were killed 30 weeks after the start of the experiments. Seventeen BOP-treated hamsters fed the basal diet developed pancreatic tumors (77.3%) while only 12 of 21 hamsters fed the 0.01% SaA diet (57.1%) and 12 of 23 hamsters fed the 0.05% SaA diet (52.2%) developed pancreatic tumors. Pancreatic lesions included ductal hyperplasia, atypical ductal hyperplasia, and carcinoma in situ. Microscopic invasive carcinoma induced by BOP and the incidence of larger pancreatic tumors in hamsters were significantly higher in hamsters fed the basal diet than in hamsters fed the SaA diet (p < 0.05). The proliferating cell nuclear antigen (PCNA) labeling index of pancreatic carcinoma in BOP-treated hamsters fed the basal diet was 41.2 +/- 13.4%, whereas BOP-treated hamsters fed 0.01 and 0.05% SaA diets yielded PCNA indexes of 26.8 +/- 8.3 and 28.4 +/- 7.0%, respectively. k-ras mutation was detected in 40% of cancers in both groups. No pancreatic tumors developed in saline-treated groups, and no differences in body weights or histological findings in their organs, including the pancreas, were observed in either group. These findings suggest that SaA not only inhibits BOP-induced pancreatic carcinogenesis in hamsters, but also provides antipromotion and antiprogression effects on these tumors, even when SaA commences 1 week after the initiation of pancreatic carcinogenesis.

Effects of bradykinin receptor antagonist on the release of beta-endorphin and bradykinin and on hemodynamic changes in a canine model of experimental acute pancreatitis.

Bradykinin and beta-endorphin increases during acute pancreatitis are thought to contribute to the development of hypotension and myocardial depression in acute pancreatitis. beta-Endorphin release is mediated by trypsin-like enzymes and bradykinin from the pituitary gland. This study was undertaken to investigate the effect of a long-acting bradykinin receptor antagonist on bradykinin and beta-endorphin release and on hemodynamic changes during acute pancreatitis. Pancreatitis was induced by the injection of autologous bile mixed with trypsin into the main pancreatic duct after ligation of the accessory duct. Serum bradykinin and plasma beta-endorphin levels and cardiovascular function were measured. Twelve dogs (control group) were given 10 ml/kg/h of lactate Ringer's solution intravenously beginning 1 h before the induction of pancreatitis and continuing throughout the experiments. Six dogs received an intravenous infusion of 0.6 mg/kg/h of a new bradykinin receptor antagonist, HOE 140, D-Arg-[Hyp3, Thi5, D-Tic, Oic8]-bradykinin, in lactate Ringer's solution soon after the induction of pancreatitis. Six of twelve dogs in the control group, and none of the six dogs in the bradykinin receptor antagonist group, died during the experiments. Serum bradykinin levels in both groups increased until 1 h after the induction of pancreatitis, but thereafter the levels in the bradykinin receptor antagonist group decreased gradually until 5 h after induction, and levels were significantly lower than those in the control group (p < 0.05). Plasma beta-endorphin levels in the control group increased significantly, to 291.8 pg/ml (+/- 6.6 SEM) 5 h after the induction of pancreatitis, from the mean levels of 47.8 pg/ml before the induction of pancreatitis, while the mean beta-endorphin level in the bradykinin receptor antagonist group did not increase after the induction of pancreatitis. Infusion of the bradykinin receptor antagonist improved survival rates, hypotension, myocardial depression, and plasma lactate, suggesting that the bradykinin receptor antagonist inhibited the release of bradykinin and beta-endorphin, which contributed to the clinical improvement.

Effect of a new synthetic protease inhibitor on beta-endorphin release during acute pancreatitis in dogs.

The effect on endogenous beta-endorphins of a new synthetic protease inhibitor was studied in acute pancreatitis. Pancreatitis was induced by the injection of autologous bile mixed with trypsin into the main pancreatic duct after ligation of the accessory duct. Plasma beta-endorphin concentrations and cardiovascular function were measured. Ten dogs (control group) were given 10 ml/kg/h of lactate Ringer's solution intravenously beginning 1 h before the induction of pancreatitis and continuing throughout the experiments. Six dogs received an intravenous infusion of 3 mg/kg/h of a new synthetic protease inhibitor, E-3123 (4-(2-succinimidoethylthio)4-geranidinobenzoate methanesuLfonate), in lactate Ringer's solution soon after the induction of pancreatitis. Plasma beta-endorphin concentrations in the control group increased significantly. However, plasma beta-endorphin levels in the protease inhibitor group did not increase as in the control group. The protease inhibitor infusion improved hypotension, myocardial depression, and plasma lactate, suggesting that the inhibitory effect of the protease inhibitor on beta-endorphin release contributed to the improvement.

Sarcophytol A: a new chemotherapeutic and chemopreventive agent for pancreatic cancer.

Sarcophytol A (SaA), a cembrane-type diterpene isolated from the marine soft coral Sarcophyton glaucum showed anticancer and cancer preventive effects in two different experiments. Growth of transplanted human pancreatic cancer cells (MIA paca-2, 1 x 10(7)) in nude mice (BALB/C 4W female) (Experiment 1) and pancreatic carcinogenesis induced by N-nitrobis-(2-hydroxypropyl) amine (BHP, 500 mg/kg) in Syrian golden hamsters (7W female) (Experiment 2) were inhibited by feeding the animals a diet containing 0.01% SaA. In Experiment 1, on day 29 after transplantation, tumor volume was significantly less in the SaA group than in the control group (1,759 + 310 mm3 vs. 2,364 + 467 mm3) (p < 0.05). In Experiment 2, the incidence of pancreatic tumors in the SaA group was 42.8% and that in the control group was 90.0% at 25-27 weeks. Thus, pancreatic carcinoma developed more slowly in the SaA group than in the control group. In addition, the incidence of atypical ductal hyperplasia and carcinoma in situ was lower in the SaA group. These results indicate that oral SaA administration is an effective vehicle for inhibition of certain types of cancer in hamsters.

Effect of a new cholecystokinin receptor antagonist (KSG 504) on the early stage of the healing process in acute pancreatitis induced in rats by the closed duodenal loop technique.

Creation of a closed duodenal loop produced edematous acute pancreatitis within 6 h and hemorrhagic acute pancreatitis within 12 h in male Sprague-Dawley rats. The pancreatitis thus established tended to improve after releasing the loop. We investigated the effect of a new cholecystokinin receptor antagonist, KSG 504, on the healing process in edematous and hemorrhagic acute pancreatitis after releasing the loop. Serum amylase and lipase levels in the control group decreased gradually after releasing the loop, but the reductions were not significant. In both the group treated with KSG 504 intravenously and the group treated subcutaneously, serum amylase and lipase levels decreased markedly upon release of the loop in edematous acute pancreatitis. Furthermore, the histologic changes in edematous acute pancreatitis improved more rapidly than in the control group. However, no such biochemical or histologic evidence of improvement was observed in hemorrhagic acute pancreatitis. The new cholecystokinin receptor antagonist, KSG 504, displayed a therapeutic effect in edematous acute pancreatitis but not in hemorrhagic acute pancreatitis. These findings suggest that endogenous cholecystokinin release induced by the closed duodenal loop may have a contributory role in the development of edematous acute pancreatitis but not of hemorrhagic acute pancreatitis.

Effect of capsaicin on neurites of cultured dorsal root ganglia and isolated neurons of chick embryos.

The effects of capsaicin on neurites of cultured chick lumbar dorsal root ganglia and their isolated single neurons were investigated. Capsaicin-treated neurites showed a disappearance of their tips, dumpling-like structures, and retardation as a result of degeneration. However, some of the retarded neurites showed a reextension. By electron microscopy the degeneration appeared similar to that caused by beta,beta'-iminodipropionitrile with respect to segregation of the cytoskeleton. This finding provides support for the idea that capsaicin inhibits axonal transport.

Quantitative estimation of the effects of capsaicin on the mouse primary sensory neurons.

Studies were made on the quantitative effects of capsaicin on the L4 dorsal roots and their ganglia (DRG) in mice. Capsaicin at a dose of 50 mg/kg was injected s.c. 1-3 times into neonatal mice, while other mice received only vehicle as controls. After capsaicin treatment unmyelinated fibers decreased by 41.3-75.3% compared to controls. After 2 or 3 capsaicin injections there was a greater decrease in unmyelinated fibers than after a single treatment. Capsaicin had most effect on Schwann cell subunit size of more than 13 axons. Reduction of myelinated fibers ranged from 6.1 to 11.6% (mean: 7.9) in 4 mice. A beta and A delta fibers of 4-5 microns diameter were significantly reduced, whereas A delta fibers of less than 2.0 microns diameter were slightly increased. Small DRG neurons (50-500 microns 2 area) decreased by 50.6-77.4% (mean: 69.6) compared to controls. In addition, capsaicin caused a decrease of 14.2-51.6% (mean: 35.2, n = 4) in large-sized neurons (500-1950 microns 2 area). Thus, capsaicin widely affects small and certain types of large neurons as well as unmyelinated and certain myelinated fibers.

Taste-modifying triterpene glycosides from Staurogyne merguensis.

Five new oleanane-type triterpene glycosides named strogins 1-5 were isolated from leaves of Staurogyne merguensis. Their structures were determined on the basis of chemical and spectral evidence. After strogins 1, 2 and 4 were held in mouth, water elicited a sweet taste. On the other hand, strogins 3 and 5 had no activity. The structure-activity relationship is discussed.

Role of endogenous and exogenous cholecystokinin in experimental acute pancreatitis induced in rats by the duodenal loop technique.

The role of endogenous cholecystokinin (CCK) release and exogenous CCK-8 administration in the development and progression of acute pancreatitis and in the early recovery phase of acute pancreatitis were investigated in rats with closed duodenal loop (CDL)-induced pancreatitis. The subcutaneous injection of CCK-8 (2 micrograms/kg) stimulated a physiological level of pancreatic enzyme secretion in normal control rats, but did not lead to any biochemical or histological evidence of acute pancreatitis. A higher dose of CCK-8 (8 micrograms/kg), however, did produce both biochemical and histological evidence of acute pancreatitis in the normal control rats. When 2 micrograms/kg of CCK-8 was injected subcutaneously in rats 6 and 12 h after the creation of the CDL, neither the biochemical nor the histological findings of acute pancreatitis showed any progression compared with the changes in controls given no CCK-8. Serum CCK levels, measured by radio-immunoassay, increased significantly from mean levels of 5.39 pg/ml (+/- 0.95 SD) before creation of the CDL to 42.06 pg/ml (+/- 2.27 SD) 6 h after, and 41.95 pg/ml (+/- 1.88 SD) 12 h after its creation (P < 0.01). The difference between serum CCK levels at 6 and 12 h was not statistically significant. Following the release of the loop, serum CCK levels decreased gradually, especially in rats in which the loop was released 6 h after being created. Although no marked biochemical and histological changes of acute pancreatitis were observed following the administration of 2 micrograms/kg of CCK-8 to rats upon release of the loop 6 h and 12 h after its creation, a higher dose of CCK-8 (8 micrograms/kg) in these rats adversely affected both the biochemical and histological findings of acute pancreatitis. Based on these findings, it was concluded that neither endogenous CCK release, as a result of the CDL, nor physiological stimulation of the pancreas by exogenous CCK-8 administration, caused progression from edematous to hemorrhagic acute pancreatitis, and neither CCK treatment had any adverse effect on the early recovery phase of CDL-induced acute pancreatitis. A pharmacological dose of CCK, however, exacerbated the acute pancreatitis, even in the early recovery stage.

Quantitative electron-microscopic analyses of pulpal nerve fibres in the mouse lower incisor after neonatal capsaicin treatment.

A single dose of capsaicin (50 mg/kg) was injected subcutaneously into four mice on day 2 of life; four untreated mice were used as controls. Six months later, a drop of 30 microM capsaicin was instilled on to the cornea of all the mice and the number of times the eyes were wiped was counted to assess the effect of capsaicin on trigeminal sensory neurones. Ultrathin cross-sections were made of the apical pulp of the incisors on both sides of control (n = 8) and capsaicin-treated animals (n = 8). Electron micrographs of pulp nerves were taken and enlarged to a final magnification of x34,000. The numbers of unmyelinated axons in the pulps of all 16 incisors and of unmyelinated axons per Schwann cell in the pulps of four incisors each from the control and capsaicin-treated groups were counted. The short diameters of unmyelinated axons were measured with a computer-operated image analyser. The number of eye wipings was eight-fold less in the capsaicin-treated than in the normal group. This finding clearly indicated that capsaicin irreversibly affected the chemogenic nociceptive trigeminal neurones. The mean number of unmyelinated axons was 345 in controls and 217 (37.1% reduction) in capsaicin-treated animals. The number of unmyelinated axons of less than 0.6 microns dia was 41.5% less in capsaicin-treated mice than in controls. Thus, fine unmyelinated axons in the mouse incisor pulp are capsaicin sensitive, and they are assumed to be nociceptive fibres conveying pain stimuli from the tooth. Capsaicin affected Schwann cells, even those with few unmyelinated axons.(ABSTRACT TRUNCATED AT 250 WORDS)