Safety, Tolerability, and Pharmacokinetics of the Novel Adenosine A2A Antagonist/Inverse Agonist KW-6356 Following Single and Multiple Oral Administration in Healthy Volunteers.

KW-6356 is an adenosine A2A receptor-selective antagonist and inverse agonist. We conducted 2 studies: study 6356-001 (no NCT number), a randomized, double-blind, placebo-controlled phase 1 trial of single ascending (1, 3, 10 mg) and multiple (6 mg once daily) oral doses of KW-6356 in healthy Japanese subjects; and study 6356-004 (NCT03830528), including a randomized, double-blind, placebo-controlled phase 1 trial of single ascending (21, 42, 60 mg) and multiple (24 mg once daily) oral doses of KW-6356, and a phase 1 open-label trial of multiple oral doses (6 mg once daily) of KW-6356 in healthy Japanese and White subjects, to evaluate the safety, tolerability, and pharmacokinetics of KW-6356. KW-6356 was well tolerated after administration of single doses of up to 60 mg and multiple doses of up to 24 mg once daily for 14 days. The pharmacokinetics of KW-6356 were linear after a single dose of up to 60 mg KW-6356. The mean terminal elimination half-life of KW-6356 ranged from 18.4 to 43.1 hours following administration of single doses of 1-60 mg. There was no clear difference in the safety or pharmacokinetics of KW-6356 between healthy Japanese and White subjects.

Assessment of CYP-Mediated Drug Interactions for Evocalcet, a New Calcimimetic Agent, Based on In Vitro Investigations and a Cocktail Study in Humans.

Evocalcet is a novel calcimimetic agent for the treatment of secondary hyperparathyroidism (SHPT). This study evaluated the effects of evocalcet on inhibition and induction of cytochrome P450 (CYP) isozymes. Although drug interactions arising from reversible inhibition of CYP isozymes by evocalcet were considered unlikely based on the results of in vitro studies and static model analyses, the potential for evocalcet to cause time-dependent inhibition of CYP3A or induction of several CYP isozymes could not be ruled out. Therefore, a clinical drug-drug interaction (DDI) study to evaluate the effects of evocalcet on the pharmacokinetics (PKs) of probe substrates for CYP isozymes (CYP1A2, CYP2B6, CYP2C8, CYP2C9, and CYP3A) was conducted in healthy male volunteers using a novel cocktail combination. Evocalcet did not significantly affect the PKs of the probe substrates, confirming that CYP-mediated interactions were unlikely.

Significant effect of the posterior tibial slope and medial/lateral ligament balance on knee flexion in total knee arthroplasty.

PURPOSE: The intra-operative femorotibial joint gap and ligament balance, the predictors affecting these gaps and their balances, as well as the postoperative knee flexion, were examined. These factors were assessed radiographically after a posterior cruciate-retaining total knee arthroplasty (TKA). The posterior condylar offset and posterior tibial slope have been reported as the most important intra-operative factors affecting cruciate-retaining-type TKAs. The joint gap and balance have not been investigated in assessments of the posterior condylar offset and the posterior tibial slope. METHODS: The femorotibial gap and medial/lateral ligament balance were measured with an offset-type tensor. The femorotibial gaps were measured at 0°, 45°, 90° and 135° of knee flexion, and various gap changes were calculated at 0°-90° and 0°-135°. Cruciate-retaining-type arthroplasties were performed in 98 knees with varus osteoarthritis. RESULTS: The 0°-90° femorotibial gap change was strongly affected by the posterior condylar offset value (postoperative posterior condylar offset subtracted by the preoperative posterior condylar offset). The 0°-135° femorotibial gap change was significantly correlated with the posterior tibial slope and the 135° medial/lateral ligament balance. The postoperative flexion angle was positively correlated with the preoperative flexion angle, γ angle and the posterior tibial slope. Multiple-regression analysis demonstrated that the preoperative flexion angle, γ angle, posterior tibial slope and 90° medial/lateral ligament balance were significant independent factors for the postoperative knee flexion angle. The flexion angle change (postoperative flexion angle subtracted by the preoperative flexion angle) was also strongly correlated with the preoperative flexion angle, posterior tibial slope and 90° medial/lateral ligament balance. CONCLUSION: The postoperative flexion angle is affected by multiple factors, especially in cruciate-retaining-type TKAs. However, it is important to pay attention not only to the posterior tibial slope, but also to the flexion medial/lateral ligament balance during surgery. A cruciate-retaining-type TKA has the potential to achieve both stability and a wide range of motion and to improve the patients' activities of daily living.

Effect of a centrally active angiotensin converting enzyme inhibitor, perindopril, on cognitive performance in chronic cerebral hypo-perfusion rats.

We have previously demonstrated that perindopril, an angiotensin-converting enzyme (ACE) inhibitor, ameliorated the cognitive deficits in Alzheimer's disease model animals, independently of its anti-hypertensive effect. In this study, we again investigated the effects of perindopril on cognitive function in a vascular dementia model animal, comparing it with other ACE inhibitors. We also determined ACE activity in the brain and extracellular acetylcholine (ACh) concentration in the perirhinal cortex in order to elucidate the mechanism(s) responsible for the effects of these ACE inhibitors on cognitive function. Perindopril was suggested to be more centrally active than imidapril and enalapril, in consideration of the relative distribution of their active metabolites in the brain. This property was at least partially attributed to the lipophilicity of the compound. While the 3 day treatment with perindopril, imidapril or enalapril lowered blood pressure to the same level in spontaneous hypertensive rats, only perindopril reversed the decline in the recognition index in chronic cerebral hypo-perfusion rats, regarded as an animal model of vascular dementia, during an object recognition task. Using the same dosing regimen, perindopril inhibited the brain ACE activities of rats more than imidapril or enalapril. Moreover, a single treatment with perindopril enhanced the extracellular level of ACh in the perirhinal cortex of normal rats. Therefore, we confirmed that only centrally active ACE inhibitors, such as perindopril, can inhibit the ACE in the brain, augmenting cholinergic neurotransmission and thereby ameliorating cognitive impairment in the animal model of vascular dementia.

Effect of a centrally active angiotensin-converting enzyme inhibitor, perindopril, on cognitive performance in a mouse model of Alzheimer's disease.

Angiotensin-converting enzyme (ACE) inhibitors have clinically been widely used as anti-hypertensive agents. In the present study, we compared the effects of a centrally active ACE inhibitor, perindopril, with those of non-centrally active ACE inhibitors, imidapril and enalapril, on cognitive performance in amyloid beta(Abeta) (25-35)-injected mice, a rodent model of Alzheimer's disease. We also determined the brain ACE activity in order to elucidate the relationship between the cognitive function and ACE inhibition in the brain. Abeta(25-35)-injected mice showed a cognitive impairment in spontaneous alteration and object recognition tests, the indices of immediate working memory and relatively long-term recognition memory, respectively. As indicated by these tests, the oral administration of perindopril (0.1, 0.3 or 1mg/kg/day) significantly reversed the cognitive impairment in these mice, whereas neither imidapril (0.3, 1 or 3mg/kg/day) nor enalapril (1, 3 or 10mg/kg/day) had any effect on cognitive performance. Perindopril (1mg/kg/day), imidapril (3mg/kg/day), or enalapril (10mg/kg/day) all inhibited the plasma ACE activities by more than 90%. Using the same dosing regimen, only perindopril inhibited the brain ACE activities by more than 50%, whereas imidapril and enalapril showed much less potent effects. These results suggest that perindopril ameliorated the cognitive impairment in the Alzheimer's disease model mice through the inhibition of brain ACE activity, but not peripheral ACE activity. Based on our observations, we concluded that a centrally active ACE inhibitor, perindopril, may therefore have a beneficial effect on Alzheimer's disease as well as hypertension.

P-glycoprotein limits the brain penetration of olopatadine hydrochloride, H1-receptor antagonist.

Olopatadine, a new second-generation antihistamine, is widely used in the treatment of allergic disorders. The low levels of histamine H1 receptor occupancy in human brain by olopatadine, which is related to its minimal sedation, suggest its low penetration into the brain. The present study evaluates the impact of P-glycoprotein (P-gp) on brain penetration and plasma concentration of olopatadine. The uptake amount of olopatadine in human P-gp transfected LLC-PK1 cells (LLC-GA5-COL150) was lower than that in LLC-PK1. The uptake of olopatadine in LLC-GA5-COL150 was increased in the same level as that in LLC-PK1 in the presence of cyclosporine A, a P-gp inhibitor. After intravenous or oral administration of olopatadine to wild type (WT) and mdr1a/1b knockout (KO) mice at a dose of 1 mg/kg, the brain concentration in KO mice was higher than that in WT mice. On the other hand, the plasma concentration of olopatadine after either route of administration was not different between WT and KO mice. These results suggest that olopatadine is a substrate of P-gp, and that P-gp limits the brain penetration but dose not affect the plasma concentration of olopatadine.

Dislocation of the shoulder joint with ipsilateral humeral shaft fracture: two case reports.

INTRODUCTION: A combination of the shaft fracture of the humerus with shoulder dislocation is a rarely documented injury. Moreover, few reports describe a long-term outcome after a follow-up period over 10 years. The present article is the first report of long-term clinical results obtained with this combined injury. MATERIALS AND METHODS: We present two cases of the dislocation of the shoulder joint with ipsilateral humeral shaft fracture. One is an anterior dislocation and the other is posterior. The anterior dislocation was closely reduced on the date of injury, but the posterior dislocation required open reduction 6 weeks after the injury because it was missed at the initial diagnosis and identified 4 weeks after the injury. In the latter case, shoulder pain and limitation of the motion persisting after the first surgery had to be treated by anterior acromioplasty and arthrolysis of the shoulder. In both cases, the humeral fracture was fixed by retrograde intramedullary nailing using a Küntscher nail and successful bone union was achieved. RESULTS: At over 10-year follow-up examination, both patients had no limitation in activities of daily living and no restrictions to their normal occupation. The radiographs of the shoulder joint demonstrated good clinical results, with no degenerative change or osteonecrosis. CONCLUSION: We have experienced two cases of dislocation of the shoulder joint with ipsilateral humeral shaft fracture and reported long-term results over 10 years. Good clinical results were demonstrated in both cases.

Involvement of rat organic anion transporter 3 in the uptake of an organic herbicide, 2,4-dichlorophenoxyacetate, by the isolated rat choroid plexus.

2,4-Dichlorophenoxyacetic acid (2,4-D) is an anionic herbicide. The purpose of the present study is to examine whether organic anion transporter 3 (Oat3; Slc22a8) is solely responsible for the uptake of 2,4-D by the isolated rat choroid plexus (CP). When expressed in LLC-PK1 cells, rOat3 was mainly localized to the basolateral membrane. Although there was no vectorial transport of 2,4-D in the control LLC-PK1 cells, expression of rOat3 increased the basal-to-apical transport of 2,4-D fourfold without affecting the transcellular transport in the opposite direction. The basal-to-apical transport of 2,4-D in rOat3-LLC was saturable with a K(m) value of 20 microM. The uptake of 2,4-D by the isolated rat CP was determined using the centrifugal filtration method. Saturable uptake of 2,4-D was observed in the isolated rat CP with a K(m) value of 22 microM. Probenecid and substrates of rOat3, such as p-aminohippurate, benzylpenicillin, and cimetidine, inhibited the uptake of 2,4-D by the isolated rat CP. Their K(i) values were comparable with those for the uptake of benzylpenicillin by the isolated rat CP, which is mainly mediated by rOat3. Furthermore, benzylpenicillin was a competitive inhibitor for the uptake of 2,4-D by the isolated rat CP. These results suggest that 2,4-D and benzylpenicillin share the same transporter for their uptake by the isolated rat CP, and rOat3 is the most likely candidate transporter.

Carrier-mediated uptake of H2-receptor antagonists by the rat choroid plexus: involvement of rat organic anion transporter 3.

The choroid plexus (CP) acts as a site for the elimination of xenobiotic organic compounds from the cerebrospinal fluid (CSF). The purpose of the present study is to investigate the role of rat organic anion transporter 3 (rOat3; Slc22a8) in the uptake of H(2)-receptor antagonists (cimetidine, ranitidine, and famotidine) by the isolated rat CP. Saturable uptake of cimetidine and ranitidine was observed in rOat3-LLC with K(m) values of 80 and 120 microM, respectively, whereas famotidine was found to be a poor substrate. The steady-state concentration of the H(2)-receptor antagonists in the CSF was significantly increased by simultaneously administered probenecid, although it did not affect their brain and plasma concentrations. Saturable uptake of cimetidine and ranitidine was observed in the isolated rat CP with K(m) values of 93 and 170 microM, respectively, whereas 50% of the uptake of famotidine remained at the highest concentration examined (1 mM). The K(i) value of ranitidine for the uptake of cimetidine by the isolated CP (50 microM) was similar to its own K(m) value, suggesting that they share the same transporter for their uptake. The inhibition potency of organic anions such as benzylpenicillin, estradiol 17beta-glucuronide, p-aminohippurate, and estrone sulfate for the uptake of cimetidine by the isolated rat CP was similar to that for benzylpenicillin, the uptake of which has been hypothesized to be mediated by rOat3, whereas a minimal effect by tetraethylammonium excludes involvement of organic cation transporter(s). These results suggest that rOat3 is the most likely candidate transporter involved in regulating the CSF concentration of H(2)-receptor antagonists at the CP.

Expression and functional involvement of organic anion transporting polypeptide subtype 3 (Slc21a7) in rat choroid plexus.

PURPOSE: It has been shown that transport(s) are involved in the uptake of estradiol 17beta glucuronide (E217betaG) by the choroid plexus (CP). The purpose of this study is to compare the substrate specificity of the transporter in the CP with those of rat organic anion transporting polypeptide 1 (rOatp1) and rOatp3. METHODS: The expression of rOatp1 and rOatp3 in rat CP was confirmed by RT-PCR and Western blot analyses. The substrate specificity of rOatp1 and rOatp3 was compared using cDNA-transfected LLC-PK1 cells. The uptake of E217betaG by rat isolated CP was determined by centrifugal filtration technique. RESULTS: PCR analyses demonstrated that the mRNA expression of rOatp3 was abundant in the CP, whereas that of rOatp1 was low. Immunohistochemical staining revealed that rOatp3 is expressed on the apical membrane of the CP. Kinetic parameters (Km and Ki values) of rOatp3 were similar to those for rOatp1. The results of mutual inhibition study suggest that E217betaG and taurocholate share the same mechanism in the CP. Corticosterone, estrone-3-sulfate and indomethacin are moderate inhibitors, but no effects by digoxin, p-aminohippurate, benzylpenicillin and cimetidine were observed. CONCLUSIONS: rOatp3 is most possible candidate transporter involved in the uptake of organic anions on the brush border membrane of the choroid epithelial cells.

The intestinal first-pass metabolism of substrates of CYP3A4 and P-glycoprotein-quantitative analysis based on information from the literature.

It is suggested that the bioavailability of CYP3A4 substrates might be low due to first-pass metabolism in the small intestine, and it is possible that P-glycoprotein (P-gp) may influence first-pass metabolism in a co-operative manner. We have collected information of the pharmacokinetics of CYP3A4 substrates to evaluate the fraction absorbed (Fa), intestinal availability (Fg) and hepatic availability (Fh) and have investigated the intestinal first-pass metabolism and the effect of P-gp on this. The pharmacokinetic data involved ten compounds metabolized by CYP3A4 in humans, with and without an inhibitor or inducer. FaFg, which is the product of Fa and Fg, and Fh were calculated using three liver blood flow rates (17.1, 21.4, 25.5 mL/min/kg) in consideration of variations in the liver flow rate. Co-administration with an inhibitor of CYP3A4 and treatment of an inducer of CYP3A4 caused an increase and decrease in the FaFg of CYP3A4 substrates, regardless of the liver blood flow, indicating that CYP3A4 substrates exhibit a first-pass effect in their metabolism. This holds true regardless of whether the compounds are P-gp substrates or not. No relationship was observed between FaFg and Fh, regardless of the hepatic blood flow rate and the P-gp substrates. The FaFg of both P-gp and non P-gp substrates decreased as the hepatic intrinsic clearance increased. FaFg was markedly reduced when the hepatic intrinsic clearance was more than 100 mL/min/kg. This in vivo intrinsic clearance corresponds to an in vitro intrinsic clearance of 78 muL/min/mg human hepatic microsomal protein, equivalent to a half-life of 8.9 min for the substrate in a commonly used metabolic stability test with human microsomes (1 mgMs protein/mL). This phenomenon was not observed in substrates of CYP isoforms other than CYP3A4. In conclusion, it is suggested that CYP3A4 substrates which have a hepatic intrinsic clearance of 100 mL/min/kg exhibit a low bioavailability due to intestinal first-pass metabolism, regardless of whether they are substrates of P-gp or not.

Expression and functional characterization of rat organic anion transporter 3 (rOat3) in the choroid plexus.

We reported previously that an efficient efflux system for benzylpenicillin (PCG) is located on the choroid plexus (CP). In this study, we investigated the involvement of rat organic anion transporter 1 (rOat1; Slc22a6) and rOat3 (Slc22a8) in the uptake of PCG and p-aminohippurate (PAH) by the CP. Western blot analysis indicates the expression of rOat3, but not rOat1, on the CP, and immunohistochemical staining shows that rOat3 is localized on the brush border membrane of the choroid epithelial cells. PCG and PAH were found to be taken up by isolated rat CP, with K(m) values of 111 and 354 microM, respectively. A mutual inhibition study suggests that the same transporter is responsible for the uptake of PCG and PAH by isolated rat CP. This was confirmed by examining the effect of organic anions and cimetidine on their uptake. Estradiol-17beta-glucuronide and cimetidine were found to be selective inhibitors of rOat3. The inhibition constants of the inhibitors including estradiol-17beta-glucuronide and cimetidine were comparable for the uptake of PAH and PCG by isolated rat CP. In addition, these values were also comparable with those for rOat3, but not with those for rOat1. These results suggest that rOat3 is mainly responsible for the uptake of PCG and PAH by isolated rat CP, and it functions as one of the detoxification systems on the CP by removing its substrates from the cerebrospinal fluid.