Abstract
ART‐001 is an orally available selective PI3Kα inhibitor currently being developed for the treatment of slow‐flow vascular malformations (SFVMs). ART‐001 used to be developed for advanced ...solid tumors, but was suspended largely due to significant pharmacokinetic (PK) variability in its phase I studies. This phase I, randomized, double‐blinded, placebo‐controlled study evaluated safety, tolerability and PK of ART‐001 with a newly developed dry syrup formulation, which was designed to optimize PK properties of ART‐001 and to be compliant with the pediatric population. Single and multiple doses of ART‐001 were administered to healthy male adults. ART‐001 was rapidly absorbed after the single and repeated doses, and the exposure of ART‐001 increased with increased dose. The dry syrup formulation substantially improved the intersubject PK variability. Food decreased area under the concentration‐time curve (AUC) and maximum plasma concentration by 12% and 36%, respectively. The plasma concentration had reached a steady‐state on day 5 of the repeated doses of 100 mg and AUC accumulation ratio was 1.9. There were no deaths or serious adverse events. The most frequent adverse event was hyperglycemia. All cases of hyperglycemia were mild to moderate and transient, and required no medical interventions. Serum creatinine increase was observed in 300 mg once daily dosing group leading to dose discontinuation on day 5. In conclusion, it was demonstrated that the single doses and repeated doses of the ART‐001 dry syrup formulation, at up to 400 and 100 mg, respectively, were safe and tolerated with favorable PK profile, supporting further clinical development for the treatment of SFVMs.
Abstract
Non-hormonal therapeutic strategies for endometriosis are needed. The aim of this study was to characterize the effects of prostaglandin (PG)E2 receptor inhibitors to explore their potential ...as novel therapeutic strategies for endometriosis. The expression of PGE2 receptors (EP2 and EP4) in donated tissues from human ovarian endometriosis, adenomyosis and peritoneal endometriosis was examined using immunohistochemistry. Human endometriotic stromal cells (ESC) isolated from ovarian endometriotic tissue and peritoneal macrophages were treated with EP2 and EP4 antagonists. cAMP accumulation and the effect of EP antagonists were measured using cAMP assays. DNA synthesis in ESC was detected using bromodeoxyuridine incorporation analysis. Interleukin (IL)-6 and IL-8 protein levels in ESC supernatants were measured using ELISAs. mRNA expression level for aromatase by ESC, and selected cytokines by peritoneal macrophages was measured using RT–PCR. EP2 and EP4 receptors were expressed in cells derived from control and diseased tissue, ovarian endometriotic, adenomyotic and peritoneal lesions. A selective EP2 antagonist reduced DNA synthesis, cAMP accumulation and IL-1β-induced proinflammatory cytokine secretion and aromatase expression. A selective EP4 antagonist negated IL-1β-induced IL-6 secretion and aromatase expression. In peritoneal macrophages, EP expression was elevated in endometriosis samples but the EP4 antagonist reduced cAMP levels and expression of vascular endothelial growth factor, chemokine ligand 2 and chemokine ligand 3 mRNA. EP2 and EP4 are functioning in endometriosis lesions and peritoneal macrophages, and their selective antagonists can reduce EP-mediated actions, therefore, the EP antagonists are potential therapeutic agents for controlling endometriosis.
The discovery of vibegron, a potent and selective human β3-AR agonist for the treatment of overactive bladder (OAB), is described. An early-generation clinical β3-AR agonist MK-0634 (3) exhibited ...efficacy in humans for the treatment of OAB, but development was discontinued due to unacceptable structure-based toxicity in preclinical species. Optimization of a series of second-generation pyrrolidine-derived β3-AR agonists included reducing the risk for phospholipidosis, the risk of formation of disproportionate human metabolites, and the risk of formation of high levels of circulating metabolites in preclinical species. These efforts resulted in the discovery of vibegron, which possesses improved druglike properties and an overall superior preclinical profile compared to MK-0634. Structure–activity relationships leading to the discovery of vibegron and a summary of its preclinical profile are described.
Aim:
Experimental models of non‐alcoholic steatohepatitis (NASH) are still required for understanding the pathophysiology of this disease. This study aimed to examine whether disease progression is ...accelerated by combining dyslipidemic genetic modification and dietary challenges and develop NASH‐associated hepatic fibrosis, cirrhosis, and carcinoma in a short period.
Methods:
Low‐density lipoprotein receptor knockout mice were fed a modified choline‐deficient amino acid‐defined diet, including 1 w/w% cholesterol and 41 kcal% fat, and was comprehensively profiled over 1 year.
Results:
Microvesicular and macrovesicular steatosis in the liver was observed from the first week after starting the modified choline‐deficient amino acid‐defined diet. Macrovesicular steatosis was exacerbated with time and was observed in almost all hepatocytes at week 8, but slightly decreased at week 16. Infiltration of macrophages and neutrophils, and upregulation of hepatic inflammatory cytokines such as tumor necrosis factor‐α and interleukin‐1β were also observed from week 1. Plasma hepatic transaminase activities were increased at week 1, reached a peak at week 4, and gradually decreased thereafter. In parallel with increases in hepatic gene expression of collagen‐I, the hepatic fibrosis area expanded after week 4 and massively spread all over the liver by week 8. Hepatocellular hyperplasia was observed from week 24. Hepatocellular adenoma and carcinoma were observed from week 31 and 39, respectively.
Conclusion:
These results suggest that, in a rodent NASH model with the combination of genetic modification and dietary challenges, hepatic steatosis, inflammatory cell infiltration and hepatic injury, hepatic fibrosis, hepatocellular hyperplasia, adenoma, and carcinoma can be developed in a relatively short period.
Aim
Liver biopsy is still required for the diagnosis of hepatocellular ballooning and inflammation, which are important histological features of non‐alcoholic steatohepatitis. We undertook this ...multicenter, cross‐sectional study to identify novel blood markers for the diagnosis of hepatocellular ballooning.
Methods
We enrolled 176 patients, of whom 132 were proven by liver biopsy as having non‐alcoholic fatty liver disease (NAFLD) and classified as non‐ballooning (ballooning grade 0) (n = 83) or ballooning (ballooning grade 1 and 2) (n = 49) by a central pathology review. We carried out gas chromatography–mass spectrometry, hydrophilic interaction liquid chromatography tandem mass spectrometry, and lipidomics with plasma.
Results
As correlates of hepatocellular ballooning, among the clinical parameters, serum type IV collagen 7S correlated most significantly with the ballooning grade (correlation coefficient CC = 0.463; P < 0.001). Among the metabolic/lipidomic markers, phosphatidylcholine (PC) (aa‐44:8) correlated most significantly with the ballooning grade (CC = 0.394; P < 0.001). The area under the receiver operating characteristic curve of type IV collagen 7S, choline, and lysophosphatidylethanolamine (LPE) (e‐18:2), was 0.846 (95% confidence interval, 0.772–0.919).
Conclusions
Plasma levels of PC were positively correlated, and those of lysophosphatidylcholine and LPE were negatively correlated with hepatocellular ballooning in NAFLD patients. These non‐invasive metabolic/lipidomic‐based plasma tests might be useful to distinguish between cases of NAFLD with and without hepatocellular ballooning.
Background & Aims The incidence of nonalcoholic steatohepatitis (NASH) is increasing. The pathophysiological mechanisms of NASH and the sequence of events leading to hepatic fibrosis are incompletely ...understood. The aim of this study was to gain insight into the dynamics of key molecular processes involved in NASH and to rank early markers for hepatic fibrosis. Methods A time-course study in low-density lipoprotein–receptor knockout. Leiden mice on a high-fat diet was performed to identify the temporal dynamics of key processes contributing to NASH and fibrosis. An integrative systems biology approach was used to elucidate candidate markers linked to the active fibrosis process by combining transcriptomics, dynamic proteomics, and histopathology. The translational value of these findings were confirmed using human NASH data sets. Results High-fat-diet feeding resulted in obesity, hyperlipidemia, insulin resistance, and NASH with fibrosis in a time-dependent manner. Temporal dynamics of key molecular processes involved in the development of NASH were identified, including lipid metabolism, inflammation, oxidative stress, and fibrosis. A data-integrative approach enabled identification of the active fibrotic process preceding histopathologic detection using a novel molecular fibrosis signature. Human studies were used to identify overlap of genes and processes and to perform a network biology-based prioritization to rank top candidate markers representing the early manifestation of fibrosis. Conclusions An early predictive molecular signature was identified that marked the active profibrotic process before histopathologic fibrosis becomes manifest. Early detection of the onset of NASH and fibrosis enables identification of novel blood-based biomarkers to stratify patients at risk, development of new therapeutics, and help shorten (pre)clinical experimental time frames.
Aim
Low‐density lipoprotein receptor knockout (LDLR‐KO) mice fed a modified choline‐deficient and amino acid‐defined (mCDAA) diet show non‐alcoholic steatohepatitis (NASH)‐like pathophysiology. In ...order to pharmacologically benchmark this model, effects of pioglitazone (a thiazolidinedione) and candesartan cilexetil (an angiotensin II type 1 receptor blocker) on steatosis and liver fibrosis were examined.
Methods
Pioglitazone (10 mg/kg) and candesartan cilexetil (3 mg/kg) were given orally once daily to LDLR‐KO mice under mCDAA diet for 7 weeks. Blood biochemistry and hepatic histology were assessed, and hepatic gene expression levels and triglyceride content were measured.
Results
Pioglitazone suppressed hepatic COL1A1 gene expression by 43% and attenuated hepatic fibrosis areas by 49%. Pioglitazone also decreased plasma alanine aminotransferase levels, liver weight, hepatic triglyceride content, and hepatic expression of other fibrosis‐related genes such as TGFB1, SPP1, TIMP1, and IL6. Candesartan cilexetil suppressed hepatic COL1A1 gene expression by 33%, whereas the other end‐points including hepatic fibrosis areas were not affected.
Conclusions
Pioglitazone showed anti‐fibrotic effects accompanied by improving hepatic transaminase activity and hepatic lipid accumulation, but the effect of candesartan cilexetil was only limited, unlike previous reports for angiotensin II type 1 receptor blockers. As the pharmacological effects of pioglitazone in the current animal model are similar to those reported in patients with NASH, this model may represent some aspects of the pathophysiology of NASH. Further profiling using other agents or mechanisms that have been tested in the clinic will better clarify the utility of the animal model.
A series of conformationally restricted acetanilides were synthesized and evaluated as β3-adrenergic receptor agonists (β3-AR) for the treatment of overactive bladder (OAB). Optimization studies ...identified a five-membered ring as the preferred conformational lock of the acetanilide. Further optimization of both the aromatic and thiazole regions led to compounds such as 19 and 29, which have a good balance of potency and selectivity. These compounds have significantly reduced intrinsic clearance compared to our initial series of pyridylethanolamine β3-AR agonists and thus have improved unbound drug exposures. Both analogues demonstrated dose dependent β3-AR mediated responses in a rat bladder hyperactivity model.
Objectives
A possibility that aging affects (a) expression of the α1D-adrenergic receptor (AR1D), (b) AR1D-mediated contractions and (c) sympathetic innervation in the urinary bladder in rats was ...studied.
Materials and methods
Contraction produced by phenylephrine and inhibition of these contractions by a non-selective α1-adrenergic antagonist prazosin and a selective AR1D antagonist BMY7378 were compared between 6- and 24-month-old Fisher rats. Expressions of VMAT and AR1D in the bladder were assessed by immunofluorescence and Western blot.
Results
Phenylephrine-induced contractions were larger and inhibition of these contractions by BMY7378 was significantly greater in 24-month-old rats. Aging increased expression of AR1D in the bladder. Density of VMAT-immunoreactive neurites was decreased in smooth muscle but elevated in the suburothelial region of 24-month-old rats.
Conclusions
The results suggest that influence of adrenergic activity on bladder contractility increases with aging is due to overexpression of the AR1D. Influence of adrenergic activity on the urothelial function may also be enhanced with aging.
Abstract The inhibition of bladder sensory transmission is critical for the pharmacotherapy of urine storage symptoms. The purpose of this study is to examine the correlation between ...pharmacologically-induced changes in cystometric parameters and spinal c-Fos expression in anesthetized rats with bladder hyperactivity induced by the intravesical infusion of acetic acid. Animals were intravenously infused with either oxybutynin (OXY), a muscarinic receptor antagonist, tamsulosin (TAM), an α1-adrenoceptor antagonist, CL316243 (CL), a β3-adrenoceptor agonist, or saline. Morphine (MOR) treatment served as a positive control to inhibit bladder afferent activity. Intermicturition intervals, micturition pressure and pressure threshold were measured after intravesical acetic acid infusion. Animals were then perfused and spinal cords were removed. Sections from the L6 spinal cord were immunostained with an anti-c-Fos antibody, and c-Fos positive cells in the dorsal region were counted. CL and MOR significantly increased intermicturition intervals, whereas OXY and TAM had no significant effect on intermicturition intervals. While TAM and MOR did not affect the micturition pressure, OXY and CL caused a significant decrease. Pressure threshold was significantly decreased by CL and increased by MOR. All drugs significantly decreased the number of c-Fos positive cells with the following order of efficacy: MOR > CL > OXT > TAM. The number of c-Fos positive cells in each animal from all treatment groups was negatively correlated with its average intermicturition interval and pressure threshold, but not with its micturition pressure. Bladder afferent activity is suppressed by several clinically proven mechanisms as measured by c-Fos expression, despite the varied effects on cystometric parameters of each pharmacological treatment.
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