ABSTRACT
Nonalcoholic steatohepatitis (NASH), a chronic liver disease without an approved therapy, is associated with lipotoxicity and insulin resistance and is a major cause of cirrhosis and hepatocellular carcinoma. Aramchol, a partial inhibitor of hepatic stearoyl-CoA desaturase (SCD1) improved steatohepatitis and fibrosis in rodents and reduced steatosis in an early clinical trial. ARREST, a 52-week, double-blind, placebo-controlled, phase 2b trial randomized 247 patients with NASH (n = 101, n = 98 and n = 48 in the Aramchol 400 mg, 600 mg and placebo arms, respectively; NCT02279524 ). The primary end point was a decrease in hepatic triglycerides by magnetic resonance spectroscopy at 52 weeks with a dose of 600 mg of Aramchol. Key secondary end points included liver histology and alanine aminotransferase (ALT). Aramchol 600 mg produced a placebo-corrected decrease in liver triglycerides without meeting the prespecified significance (-3.1, 95% confidence interval (CI) -6.4 to 0.2, P = 0.066), precluding further formal statistical analysis. NASH resolution without worsening fibrosis was achieved in 16.7% (13 out of 78) of Aramchol 600 mg versus 5% (2 out of 40) of the placebo arm (odds ratio (OR) = 4.74, 95% CI = 0.99 to 22.7) and fibrosis improvement by ≥1 stage without worsening NASH in 29.5% versus 17.5% (OR = 1.88, 95% CI = 0.7 to 5.0), respectively. The placebo-corrected decrease in ALT for 600 mg was -29.1 IU l-1 (95% CI = -41.6 to -16.5). Early termination due to adverse events (AEs) was <5%, and Aramchol 600 and 400 mg were safe, well tolerated and without imbalance in serious or severe AEs between arms. Although the primary end point of a reduction in liver fat did not meet the prespecified significance level with Aramchol 600 mg, the observed safety and changes in liver histology and enzymes provide a rationale for SCD1 modulation as a promising therapy for NASH and fibrosis and are being evaluated in an ongoing phase 3 program.
Subject(s)
Cholic Acids/administration & dosage , Liver/drug effects , Non-alcoholic Fatty Liver Disease/drug therapy , Stearoyl-CoA Desaturase/genetics , Alanine Transaminase , Biopsy , Cholic Acids/adverse effects , Double-Blind Method , Female , Humans , Liver/metabolism , Liver/pathology , Magnetic Resonance Spectroscopy , Male , Middle Aged , Non-alcoholic Fatty Liver Disease/genetics , Non-alcoholic Fatty Liver Disease/pathology , Triglycerides/metabolismABSTRACT
BACKGROUND: Nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3 (NLRP3) plays an important role in mediating inflammatory responses during ischemic stroke. Bile acid receptor Takeda-G-protein-receptor-5 (TGR5) has been identified as an important component in regulating brain inflammatory responses. In this study, we investigated the mechanism of TGR5 in alleviating neuroinflammation after middle cerebral artery occlusion (MCAO). METHODS: Sprague-Dawley rats were subjected to MCAO and TGR5 agonist INT777 was administered intranasally 1 h after MCAO. Small interfering RNAs (siRNA) targeting TGR5 and Pellino3 were administered through intracerebroventricular injection 48 h before MCAO. Infarct volumes and neurologic scores were evaluated, and ELISA, flow cytometry, immunofluorescence staining, immunoblotting, and co-immunoprecipitation were used for the evaluations. RESULTS: Endogenous TGR5 and Pellino3 levels increased after MCAO. TGR5 activation by INT777 significantly decreased pro-inflammatory cytokine, cleaved caspase-8, and NLRP3 levels, thereby reducing brain infarctions; both short- and long-term neurobehavioral assessments showed improvements. Ischemic damage induced the interaction of TGR5 with Pellino3. Knockdown of either TGR5 or Pellino3 increased the accumulation of cleaved caspase-8 and NLRP3, aggravated cerebral impairments, and abolished the anti-inflammatory effects of INT777 after MCAO. CONCLUSIONS: TGR5 activation attenuated brain injury by inhibiting neuroinflammation after MCAO, which could be mediated by Pellino3 inhibition of caspase-8/NLRP3.
Subject(s)
Caspase 8/metabolism , Infarction, Middle Cerebral Artery/metabolism , Inflammation Mediators/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Receptors, G-Protein-Coupled/metabolism , Ubiquitin-Protein Ligases/metabolism , Administration, Intranasal , Animals , Brain/drug effects , Brain/metabolism , Cholic Acids/administration & dosage , Infarction, Middle Cerebral Artery/prevention & control , Inflammation Mediators/antagonists & inhibitors , Injections, Intraventricular , Male , NLR Family, Pyrin Domain-Containing 3 Protein/antagonists & inhibitors , RNA, Small Interfering/administration & dosage , Rats , Rats, Sprague-Dawley , Receptors, G-Protein-Coupled/agonists , Ubiquitin-Protein Ligases/antagonists & inhibitorsABSTRACT
BACKGROUND & AIMS: Fontan surgery for single ventricle congenital heart disease leads to Fontan-associated liver disease (FALD). Typical laboratory tests, imaging, and histopathology cannot predict clinical severity in FALD. HepQuant SHUNT is a proprietary serum test of hepatic function and physiology that has not yet been evaluated in FALD. METHODS: Fourteen adult FALD patients at a single urban tertiary care center who underwent a Fontan procedure in childhood received HepQuant SHUNT testing between September 2015 and April 2018. The HepQuant SHUNT disease severity index (DSI) assesses global liver function and physiology from systemic and portal hepatic filtration rates (HFRs, clearances adjusted for body mass) of orally and intravenously administered cholates labeled with deuterium or 13C. The SHUNT parameter of the test measures portal systemic shunting from the ratio of Systemic HFR to Portal HFR. Chart review included laboratory tests, imaging, and clinical findings. Data from FALD patients were compared with data from healthy controls. RESULTS: The average DSI and SHUNT values for the FALD patients were 17.5% and 36.1%, respectively, compared to 9.2% and 24.1%, respectively, for controls. Twelve (85.7%) FALD patients had a DSI >15 (upper limit of normal). Seven (50.0%) FALD patients had SHUNT values >30% (upper limit of normal), while three FALD patients (21.4%) had SHUNT values >49%. One FALD patient with preoperative SHUNT of 69%, who underwent a combined heart-liver transplant, had confirmed cirrhotic morphology within the liver explant. CONCLUSIONS: This pilot study demonstrated that most FALD patients had hepatic impairment detected by abnormal DSI, with a smaller number having markedly elevated SHUNT values >49% suggesting intrinsic liver disease. The HepQuant SHUNT test may be useful in detecting and quantifying liver disease severity in FALD patients.
Subject(s)
Cholic Acids/administration & dosage , Fontan Procedure/adverse effects , Heart Defects, Congenital/surgery , Liver Circulation , Liver Diseases/diagnosis , Liver Function Tests , Administration, Intravenous , Adult , Case-Control Studies , Cholic Acids/blood , Cross-Sectional Studies , Female , Heart Defects, Congenital/diagnostic imaging , Heart Defects, Congenital/physiopathology , Hepatobiliary Elimination , Humans , Liver Diseases/etiology , Liver Diseases/physiopathology , Male , Middle Aged , Pilot Projects , Predictive Value of Tests , Proof of Concept Study , Severity of Illness Index , Young AdultABSTRACT
Positron emission tomography (PET) with 11C-cholylsarcosine (11C-CSar), a radiolabelled synthetic N-methylglycine (sarcosine) conjugate of cholic acid, is a novel molecular imaging technique that enables quantitative assessment of the individual transport steps involved in hepatic secretion of conjugated bile acids. Here, we present the method and discuss its potential clinical and scientific applications based on findings in the first human study of healthy subjects and patients with cholestasis. We also present a clinical example of a patient studied during and six months after an episode of drug-induced cholestatic liver injury.
Subject(s)
Bile Acids and Salts/metabolism , Bile Ducts/diagnostic imaging , Chemical and Drug Induced Liver Injury/diagnostic imaging , Cholestasis/diagnostic imaging , Positron-Emission Tomography/methods , Anti-Bacterial Agents/adverse effects , Bile Ducts/metabolism , Carbon Radioisotopes/administration & dosage , Carbon Radioisotopes/chemistry , Chemical and Drug Induced Liver Injury/etiology , Cholestasis/etiology , Cholic Acids/administration & dosage , Cholic Acids/chemistry , Feasibility Studies , Female , Humans , Liver/diagnostic imaging , Liver/metabolism , Liver/pathology , Middle Aged , Molecular Imaging/methods , Pneumonia/drug therapy , Radioactive Tracers , Sarcosine/administration & dosage , Sarcosine/analogs & derivatives , Sarcosine/chemistryABSTRACT
OBJECTIVES: Early diagnosis of bile acid synthesis disorders (BASDs) is important because, untreated, these conditions can be fatal. Our objectives were to screen children with cholestasis or unexplained liver disease for BASD and in those with confirmed BASD to evaluate the effectiveness of cholic acid therapy. METHODS: A routine serum total bile acid measurement was performed on children with cholestasis, liver cirrhosis, and liver failure. Patients were screened for BASD by fast atom bombardment ionization-mass spectrometry (FAB-MS) analysis of urine, and molecular analysis confirmed diagnosis. Treatment response to oral cholic acid (10-15 mg/kg bw/day) was assessed from liver function tests and fat-soluble vitamin levels. FAB-MS analysis of urine was used to monitor compliance and biochemical response. RESULTS: Between 2007 and 2016, 626 patients were evaluated; 450 with infantile cholestasis. Fifteen cases of BASD were diagnosed: 12 presented with infantile cholestasis (2.7%, 7 boys), an 8-year-old boy presented with cirrhosis, and two 18-month-old boys presented with hepatomegaly and rickets. Eleven were caused by 3ß-hydroxy-Δ-C27-steroid oxidoreductase dehydrogenase deficiency, 3 from Δ-3-oxosteroid 5ß-reductase deficiency, and 1 had Zellweger spectrum disorder. In all but 1, serum total bile acids were normal or low. With cholic acid therapy, 10 are alive and healthy with their native liver. Liver failure developed in 3 infants despite therapy; 2 died and 1 underwent liver transplantation. CONCLUSIONS: BASDs are rare but treatable causes of metabolic liver disease in Saudi Arabia. BASD should be considered in infants with cholestasis and low or normal serum total bile acid concentrations.
Subject(s)
Adrenal Hyperplasia, Congenital/drug therapy , Arabs , Bile Acids and Salts/blood , Cholic Acids/administration & dosage , Gastrointestinal Agents/administration & dosage , Liver Diseases/diagnosis , Administration, Oral , Adrenal Hyperplasia, Congenital/blood , Adrenal Hyperplasia, Congenital/urine , Bile Acids and Salts/urine , Child , Child, Preschool , Gas Chromatography-Mass Spectrometry , Humans , Infant , Liver/physiopathology , Liver Diseases/drug therapy , Liver Function Tests , Longitudinal Studies , Saudi Arabia , Spectrometry, Mass, Secondary IonABSTRACT
Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. Different methods are employed to prepare the LMWHs that are clinically approved for the market currently. In particular, enoxaparin, which has a reducing sugar moiety at the end-site of polysaccharide, is prepared by alkaline depolymerization. Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. The end-site-specific conjugation is important for polysaccharide drug development because of the heterogeneity of polysaccharides. This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. Considering the importance of end-specific conjugation, these results suggest that EnoxaTD could be a drug candidate for oral heparin development.
Subject(s)
Cholic Acids/pharmacology , Drug Delivery Systems , Enoxaparin/pharmacology , Venous Thrombosis/drug therapy , Administration, Oral , Animals , Cell Survival/drug effects , Cholic Acids/administration & dosage , Cholic Acids/chemistry , Dogs , Dose-Response Relationship, Drug , Enoxaparin/administration & dosage , Enoxaparin/chemistry , Glycosylation , Madin Darby Canine Kidney Cells/drug effects , Male , Mice , Mice, Inbred C57BL , Models, Molecular , Molecular Structure , Rats , Rats, Sprague-Dawley , Structure-Activity RelationshipABSTRACT
A novel formulation for amphotericin B (AmB) delivery has been developed using micelle-forming 5 kDa monomethoxy-polyethylene glycol functionalized with cholanic acid (PEG 5kDa-cholane). This polymer was found to increase 10(3) times the AmB solubility with a 12:1 AmB/PEG5 kDa-cholane molar ratio (2:1 w/w ratio). Dynamic light scattering and transmission electron microscopy analyses showed that PEG5 kDa-cholane associated with AmB to form 30 nm micelles. Isothermal titration calorimetry analyses performed at different pH showed that PEG 5kDa-cholane interacts with AmB according to multiple-site association profiles. Affinity constants and enthalpy and entropy changes were found to depend on pH, suggesting that the polymer interaction depends on the AmB ionization and aggregation. The freeze-dried product could be promptly re-dispersed forming a colloidal dispersion with the biopharmaceutical features of the freshly prepared micelles, namely AmB solubility and micelle size. The dispersion was stable over one month incubation at room temperature. FT-infrared spectrometry, differential scanning calorimetry and X-ray diffractometry showed that in the freeze-dried product, AmB intimately interacts with PEG 5kDa-cholane. In presence of serum albumin, AmB formulated with PEG 5kDa-cholane was found to undergo less extensive and slower disaggregation than in Fungizone(®). Antifungal activity studies performed using Candida albicans showed that AmB/PEG 5kDa-cholane was 15% more active than AmB in buffer.
Subject(s)
Amphotericin B/administration & dosage , Cholic Acids/administration & dosage , Cholic Acids/chemistry , Drug Delivery Systems , Polyethylene Glycols/administration & dosage , Polyethylene Glycols/chemistry , Amphotericin B/chemistry , Amphotericin B/pharmacology , Antifungal Agents/administration & dosage , Antifungal Agents/chemistry , Antifungal Agents/pharmacology , Chemical Phenomena , Chemistry, Pharmaceutical , Circular Dichroism , Drug Liberation , Drug Stability , Humans , Micelles , Microbial Sensitivity Tests , Nanotechnology , Serum Albumin/chemistryABSTRACT
The objective of this study was to investigate the potential of liposomes containing bile salts as an ophthalmic delivery system for tacrolimus to improve corneal permeability. Liposomes containing bile salts, including sodium taurocholate, sodium deoxycholate, and sodium glycocholate, were produced by the thin-film dispersion method with a particle size of approximately 100 nm and an entrapment efficiency of more than 90%. Less than 5% tacrolimus was released from conventional liposomes and from liposomes containing sodium taurocholate, sodium deoxycholate, or sodium glycocholate over 12 hours. The cellular uptake of conventional liposomes was significantly higher than that of liposomes containing bile salts. However, liposomes containing bile salts exerted a 3-4-fold increase of tacrolimus in ex vivo corneal transport of tacrolimus compared with conventional liposomes. When rabbit eyes were treated with a DiI perchlorate-loaded liposome suspension, liposomes containing bile salts showed fast and sustained penetration across the cornea. Unfortunately, liposomes containing sodium deoxycholate caused toxicity or irritation to both spontaneously derived human corneal epithelial cells and the rabbit cornea. Therefore, liposomes containing sodium taurocholate and sodium glycocholate are potential carriers in ocular drug delivery systems, given their low toxicity and vastly improved permeability.
Subject(s)
Cholic Acids/pharmacokinetics , Drug Carriers/pharmacokinetics , Liposomes/pharmacokinetics , Tacrolimus/pharmacokinetics , Administration, Ophthalmic , Analysis of Variance , Animals , Cell Survival/drug effects , Cholic Acids/administration & dosage , Cholic Acids/chemistry , Cholic Acids/toxicity , Cornea/drug effects , Cornea/pathology , Drug Carriers/administration & dosage , Drug Carriers/chemistry , Drug Carriers/toxicity , Liposomes/administration & dosage , Liposomes/chemistry , Liposomes/toxicity , Particle Size , Rabbits , Tacrolimus/administration & dosage , Tacrolimus/chemistryABSTRACT
PURPOSE: A phase I trial of intravesical recombinant adenovirus mediated interferon-α2b gene therapy (rAd-IFNα) formulated with the excipient SCH Syn3 was conducted in patients with nonmuscle invasive bladder cancer who had disease recurrence after treatment with bacillus Calmette-Guérin. The primary objective was to determine the safety of rAd-IFNα/Syn3. Secondary end points were demonstrated effective rAd-IFNα gene expression and preliminary evidence of clinical activity at 3 months. MATERIALS AND METHODS: A total of 17 patients with recurrent nonmuscle invasive bladder cancer after bacillus Calmette-Guérin treatment were enrolled in the study. A single treatment of rAd-IFNα (3 × 10(9) to 3 × 10(11) particles per ml) formulated with the excipient Syn3 was administered. Patient safety was evaluated for 12 or more weeks. Efficacy of gene transfer was determined by urine IFNα protein concentrations. Preliminary drug efficacy was determined at 3 months. RESULTS: Intravesical rAd-IFNα/Syn3 was well tolerated as no dose limiting toxicity was encountered. Urgency was the most common adverse event and all cases were grade 1 or 2. rAd-IFNα DNA was not detected in the blood. However, transient low serum IFNα and Syn3 levels were measured. High and prolonged dose related urine IFNα levels were achieved with the initial treatment. Of the 14 patients treated at doses of 10(10) or more particles per ml with detectable urine IFNα, 6 (43%) experienced a complete response at 3 months and 2 remained disease-free at 29.0 and 39.2 months, respectively. CONCLUSIONS: Intravesical rAd-IFNα/Syn3 was well tolerated with no dose limiting toxicity encountered. Dose dependent urinary IFNα concentrations confirmed efficient gene transfer and expression. Intravesical rAd-IFNα/Syn3 demonstrated clinical activity in nonmuscle invasive bladder cancer recurring after bacillus Calmette-Guérin.
Subject(s)
Carcinoma, Transitional Cell/therapy , Genetic Therapy/methods , Interferon-alpha/administration & dosage , Neoplasm Recurrence, Local/mortality , Neoplasm Recurrence, Local/therapy , Urinary Bladder Neoplasms/therapy , Adenoviridae/genetics , Administration, Intravesical , Adult , Aged , Aged, 80 and over , BCG Vaccine/administration & dosage , Carcinoma, Transitional Cell/mortality , Carcinoma, Transitional Cell/pathology , Cholic Acids/administration & dosage , Disaccharides/administration & dosage , Dose-Response Relationship, Drug , Drug Administration Schedule , Female , Genetic Vectors , Humans , Interferon alpha-2 , Interferon-alpha/genetics , Male , Maximum Tolerated Dose , Middle Aged , Neoplasm Invasiveness/pathology , Neoplasm Recurrence, Local/pathology , Prognosis , Recombinant Proteins/administration & dosage , Recombinant Proteins/genetics , Risk Assessment , Survival Analysis , Treatment Failure , Treatment Outcome , Urinary Bladder Neoplasms/mortality , Urinary Bladder Neoplasms/pathologyABSTRACT
In recent studies we showed that gliclazide has no hypoglycemic effect on type 1 diabetic (T1D) rats while MKC does, and their combination exerted a better hypoglycemic effect than MKC alone. We also showed that the most hypoglycemic effect was noticed when T1D rats were treated with probiotics then gavaged with MKC + gliclazide (blood glucose decreased from 24 ± 3 to 10 ± 2 mmol/l). The aim of this study is to investigate the influence of probiotics on MKC pharmacokinetics when coadministered with gliclazide, in T1D rats. 80 male Wistar rats (weight 350 ± 50 g) were randomly allocated into 8 groups (10 rats/group), 4 of which were injected with alloxan (30 mg/kg) to induce T1D. Group 1 was healthy and group 2 was diabetic. Groups 3 (healthy) and 4 (diabetic) were gavaged with probiotics (75 mg/kg) every 12 h for 3 days and 12 h later all groups received a single oral dose of MKC + gliclazide (4 and 20 mg/kg respectively). The remaining 4 groups were treated in the same way but administered MKC + gliclazide via the i.v. route. Blood samples collected from T1D rats prior to MKC + gliclazide revealed that probiotic treatment alone reduced blood glucose levels twofold. When coadministered with gliclazide, the bioavailability of MKC was reduced in healthy rats treated with probiotics but remained the same in diabetic pretreated rats. The decrease in MKC bioavailability, when administered with gliclazide, caused by probiotic treatment in healthy but not diabetic rats suggests that probiotic treatment induced MKC metabolism or impaired its absorption, only in healthy animals. The different MKC bioavailability in healthy and diabetic rats could be explained by different induction of presystemic elimination of MKC in the gut by probiotic treatment.
Subject(s)
Cholic Acids/pharmacokinetics , Diabetes Mellitus, Experimental/drug therapy , Gliclazide/pharmacokinetics , Hypoglycemic Agents/pharmacokinetics , Probiotics/pharmacology , Administration, Oral , Animals , Biological Availability , Blood Glucose/drug effects , Chenodeoxycholic Acid/analogs & derivatives , Cholic Acids/administration & dosage , Chromatography, High Pressure Liquid/methods , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 1/drug therapy , Diabetes Mellitus, Type 1/metabolism , Drug Therapy, Combination , Food-Drug Interactions , Gliclazide/administration & dosage , Hypoglycemic Agents/administration & dosage , Injections, Intravenous , Male , Random Allocation , Rats , Rats, WistarABSTRACT
The 11ß-hydroxysteroid dehydrogenases (11ß-HSDs) play a pivotal role in glucocorticoid (GC) action. 11ß-HSD1 is a predominant reductase, activating GCs from inert metabolites, whereas 11ß-HSD2 is a potent dehydrogenase inactivating GCs. Knowing the metabolic effects of GCs, a selective inhibition of 11ß-HSD1 represents a potential target for therapy of impaired glucose tolerance, insulin insensitivity and central obesity. In vitro, 11ß-HSD1 is selectively inhibited by chenodesoxycholic acid (CDCA) and upregulated under GC exposure. Therefore, we aimed to investigate the effects of CDCA and prednisolone on hepatic 11ß-HSD1 activity in vivo by measuring 11-reduction of orally given cortisone (E) acetate to cortisol (F). CDCA or placebo was given to 5 male healthy volunteers within a randomised cross-over trial before and after oral administration of 12.5 mg E acetate at 8:00 h. For measurement of in vivo effects of GCs on 11ß-HSD1 activity, hepatic reduction of 25 mg E acetate before and after treatment with prednisolone (30 mg for 6 days) was determined in 7 healthy males. Serum GC levels were determined using a fully automated liquid chromatographic system. CDCA had no effect on the activity of 11ß-HSD1 in vivo. Prednisolone therapy leads to a marked rise in serum F concentrations and an elevated F/E serum ratio. This proves GC-induced activation of hepatic 11ß-HSD1, which could not be extinguished by a parallel increase of IGF-1 under prednisolone. CDCA does not affect in vivo activity of 11ß-HSD1 when given in therapeutic dosages. During GC treatment, increased hepatic activation of E to F may aggravate metabolic side effects of GCs such as seen in the metabolic syndrome.
Subject(s)
11-beta-Hydroxysteroid Dehydrogenase Type 1/metabolism , Cholic Acids/administration & dosage , Enzyme Inhibitors/administration & dosage , Prednisolone/administration & dosage , 11-beta-Hydroxysteroid Dehydrogenase Type 1/antagonists & inhibitors , 11-beta-Hydroxysteroid Dehydrogenase Type 1/genetics , Adult , Cortisone/metabolism , Glucocorticoids/metabolism , Humans , Hydrocortisone/metabolism , Liver/enzymology , Liver/metabolism , Male , Young AdultABSTRACT
Oral drug delivery offers an attractive method of needle-free drug administration. Unfortunately, oral delivery is often hampered by the poor permeability of drugs across the intestinal epithelium. Although several single chemical permeation enhancers have been shown to alleviate permeability difficulties, this often occurs at the expense of safety. This in vitro study demonstrates the use of binary and ternary combinations of permeation enhancers to create synergistic enhancer formulations (SEFs) that offer a high level of potency while inducing very little toxicity in Caco-2 cells. Although relatively rare in the explored formulation space, SEFs were abundant enough to significantly increase the repertoire of permeation enhancers that are safe and effective in vitro. The most promising enhancers from the binary study led to easily identifiable ternary SEFs, thus increasing the efficiency of the discovery process. Some of the best performers of the study included binary combinations of hexylamine and chembetaine and ternary combinations of sodium laureth sulfate, decyltrimethyl ammonium bromide, and chembetaine, all at a total concentration of 0.1% (w/v). Furthermore, several SEFs were shown to be capable of increasing mannitol and 70 kDa dextran permeability across Caco-2 monolayers 15- and 8-fold, respectively. These results encourage further exploration of several leading formulations for in vivo applications in oral drug delivery.
Subject(s)
Adjuvants, Pharmaceutic/pharmacology , Cell Membrane Permeability/drug effects , Adjuvants, Pharmaceutic/administration & dosage , Administration, Oral , Amines/administration & dosage , Amines/pharmacology , Bicyclic Monoterpenes , Caco-2 Cells , Cell Survival/drug effects , Cholic Acids/administration & dosage , Cholic Acids/pharmacology , Decanoic Acids/administration & dosage , Decanoic Acids/pharmacology , Dextrans/metabolism , Drug Combinations , Drug Synergism , Electric Impedance , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Humans , Intestinal Absorption/drug effects , Intestinal Mucosa/drug effects , Intestinal Mucosa/metabolism , Mannitol/metabolism , Pyrrolidinones/administration & dosage , Pyrrolidinones/pharmacology , Quaternary Ammonium Compounds/administration & dosage , Quaternary Ammonium Compounds/pharmacology , Sarcosine/administration & dosage , Sarcosine/analogs & derivatives , Sarcosine/pharmacology , Sodium Dodecyl Sulfate/administration & dosage , Sodium Dodecyl Sulfate/analogs & derivatives , Sodium Dodecyl Sulfate/pharmacology , Surface-Active Agents/administration & dosage , Surface-Active Agents/pharmacology , Terpenes/administration & dosage , Terpenes/pharmacologyABSTRACT
Fatty acid bile acid conjugates (FABACs) prevent and dissolve cholesterol gallstones and prevent diet induced fatty liver, in mice. The present studies aimed to test their hypocholesterolemic effects in mice. Gallstone susceptible (C57L/J) mice, on high fat (HFD) or regular diet (RD), were treated with the conjugate of cholic acid with arachidic acid (FABAC; Aramchol). FABAC reduced the elevated plasma cholesterol levels induced by the HFD. In C57L/J mice, FABAC reduced plasma cholesterol by 50% (p<0.001). In mice fed HFD, hepatic cholesterol synthesis was reduced, whereas CYP7A1 activity and expression were increased by FABAC. The ratio of fecal bile acids/neutral sterols was increased, as was the total fecal sterol excretion. In conclusion, FABACs markedly reduce elevated plasma cholesterol in mice by reducing the hepatic synthesis of cholesterol, in conjunction with an increase of its catabolism and excretion from the body.
Subject(s)
Anticholesteremic Agents/administration & dosage , Anticholesteremic Agents/chemistry , Bile Acids and Salts/administration & dosage , Cholic Acids/administration & dosage , Animals , Anticholesteremic Agents/therapeutic use , Bile Acids and Salts/therapeutic use , Body Weight/drug effects , Body Weight/genetics , Cholesterol/analysis , Cholesterol/blood , Cholic Acids/therapeutic use , Dietary Fats/administration & dosage , Eicosanoic Acids/administration & dosage , Feces/chemistry , Gallstones/enzymology , Gallstones/genetics , Gallstones/physiopathology , Gallstones/prevention & control , Genetic Predisposition to Disease , Lipoproteins/blood , Mice , Mice, Inbred C57BL , Microsomes, Liver/chemistry , Microsomes, Liver/drug effects , Microsomes, Liver/enzymology , Organ Size/drug effects , Organ Size/genetics , Triglycerides/bloodABSTRACT
Ganoderma lucidum is a well-known mushroom with various pharmacological effects that has been used for health and longevity purposes. The objective of this study was to investigate the anti-invasive effect of lucidenic acids isolated from a new G. lucidum strain (YK-02) against human hepatoma carcinoma (HepG(2)) cells. Triterpenoid components in the ethanol extract of G. lucidum (YK-02) were separated by means of a semi-preparative RP HPLC. Four major peaks were separated and crystallized from triterpenoids fraction, and were identified as lucidenic acids A, B, C, and N according to their spectroscopic values of (1)H NMR and MS. Treatment of the lucidenic acids (50 microM) in the presence of 200 nM phorbol 12-myristate 13-acetate (PMA) after 24 h of incubation all resulted in significant inhibitory effects on PMA-induced MMP-9 activity and invasion of HepG(2 )cells. The results indicate that the lucidenic acids isolated from G. lucidum (YK-02) are anti-invasive bioactive components on hepatoma cells.
Subject(s)
Cholic Acids/administration & dosage , Cholic Acids/isolation & purification , Ganoderma/chemistry , Neoplasm Invasiveness/prevention & control , Terpenes/administration & dosage , Terpenes/isolation & purification , Carcinoma, Hepatocellular , Cell Line, Tumor , Chromatography, High Pressure Liquid , Humans , Liver Neoplasms , Matrix Metalloproteinase 9/metabolism , Matrix Metalloproteinase Inhibitors , Terpenes/chemistry , Tetradecanoylphorbol Acetate/pharmacology , Triterpenes/administration & dosage , Triterpenes/isolation & purificationABSTRACT
Intravesical administration of interferon alpha-2b protein (IFN) has been successfully used in the treatment of patients with superficial bladder tumors. Local dosing of IFN minimizes well-known systemic side effects of the drug, but exposure to bladder tumors is limited by the duration of instillation and transient concentrations achieved in the urothelium. Intravesical delivery of the gene encoding interferon results in an alternative strategy for IFN-based therapy of the disease, enabling sustained exposure of IFN protein that results from production by tumor and non-tumor cells in the urothelium. Efficient gene delivery and expression of IFN has been achieved using a recombinant adenovirus gene delivery system (rAd-IFN) in conjunction with the novel small molecule excipient Syn3. Studies with rAd-IFN/Syn3 in animal models result in urine concentrations of IFN that persisted for weeks and correlated with potent anti-tumor effects. The objective of this review is to communicate the rationale and preclinical findings that support ongoing clinical investigation of intravesical rAd-IFN/Syn3 in superficial bladder cancer.
Subject(s)
Cholic Acids/administration & dosage , Disaccharides/administration & dosage , Excipients/administration & dosage , Interferon-alpha/administration & dosage , Interferon-alpha/genetics , Urinary Bladder Neoplasms/therapy , Adenoviridae/genetics , Administration, Intravesical , Animals , Cholic Acids/therapeutic use , Disaccharides/therapeutic use , Excipients/therapeutic use , Genetic Therapy , Humans , Interferon alpha-2 , Interferon-alpha/therapeutic use , Recombinant Proteins , Urinary Bladder Neoplasms/metabolism , Urothelium/metabolismABSTRACT
Primary biliary cirrhosis (PBC) is a chronic cholestatic autoimmune liver disease that predominantly affects middle-aged women. It is characterized by slowly progressive destruction of the small intrahepatic bile ducts together with portal inflammation, and this initially leads to fibrosis and later to cirrhosis. It is currently accepted that the pathogenesis of PBC is multifactorial with genetic and environmental factors interplaying to determine the disease onset and progression. In addition to antimitochondrial antibody (AMA), which is the hallmark of PBC and is detected in at least 90% of the patients, other autoantibodies (antinuclear antibody, anti-smooth muscle antibody and rheumatoid factor, etc.) may also be found in the patients. There is no correlation between the titer of AMAs and the disease severity. Most patients are diagnosed either during the asymptomatic phase of PBC or after presenting with non-specific symptoms. Pruritus and fatigue are the most common symptoms of PBC. The prognosis of PBC has improved significantly during the last few decades. Patients are now diagnosed earlier in its clinical course, they are more likely to be asymptomatic at diagnosis and they are more likely to receive medical treatment. A wide variety of drugs have been assessed for the treatment of this condition: such immunosuppressive agents as corticosteroids, cyclosporine and azathioprine have a weak effect on the disease's natural history. Ursodeoxycholic acid (UDCA) is the only currently approved medical treatment. For PBC patients with end-stage liver disease or an unacceptable quality of life, liver transplantation is the only accepted therapeutic option. Early diagnosis and treatment of PBC are important because effective treatment with UDCA has been shown to delay disease progression and improve rate survival in the early stage.
Subject(s)
Autoimmune Diseases/diagnosis , Autoimmune Diseases/drug therapy , Cholagogues and Choleretics/therapeutic use , Liver Cirrhosis, Biliary/diagnosis , Liver Cirrhosis, Biliary/drug therapy , Autoimmune Diseases/epidemiology , Cholestadienes/administration & dosage , Cholestadienes/therapeutic use , Cholic Acids/administration & dosage , Cholic Acids/therapeutic use , Female , Humans , Liver Cirrhosis, Biliary/epidemiology , Male , Middle Aged , Prevalence , Rifampin/administration & dosage , Rifampin/therapeutic useABSTRACT
A liver-selective glucocorticoid (GC) receptor antagonist (A-348441) was used to determine the effect of reduced hepatic GC signaling on hepatic glucose production. Fasted conscious dogs were studied in the presence (GRA, n = 6) or absence (CON, n = 6) of the intraduodenally administered GC receptor antagonist (100 mg/kg). All dogs were maintained on a pancreatic clamp and in a euglycemic state for 7 hours to ensure that any changes in glucose metabolism were the direct result of the effects of A-348441, which was given at the start of a 5-hour experimental period. In the GRA group, the arterial plasma insulin level was 4.6 +/- 0.7 and 4.8 +/- 0.6 microU/mL during the basal and the last 30 minutes of the experimental periods, respectively. In the CON group, it was 4.0 +/- 0.3 and 4.5 +/- 0.5 microU/mL in the 2 periods, respectively. The arterial plasma glucagon level was 49 +/- 4 and 46 +/- 3 pg/mL in the 2 periods in the GRA group, and 45 +/- 3 and 42 +/- 3 pg/mL in the CON group. Net hepatic glucose balance progressively decreased in the GRA group from 1.31 +/- 0.18 to 0.49 +/- 0.30 mg/kg per minute, whereas in the CON group, net hepatic glucose balance was 1.17 +/- 0.09 and 1.43 +/- 0.18 mg/kg per minute during the basal and last 30 minutes of the experimental periods, respectively. No significant change in net renal or gut glucose balance or nonhepatic glucose uptake was observed in either group. This study demonstrates that the GC receptor plays an important role in the regulation of basal hepatic glucose production and represents a significant potential therapeutic target.
Subject(s)
Glucose/biosynthesis , Liver/metabolism , Receptors, Glucocorticoid/antagonists & inhibitors , Animals , Blood Glucose/analysis , Cholic Acids/administration & dosage , Cholic Acids/pharmacology , Dogs , Estrone/administration & dosage , Estrone/analogs & derivatives , Estrone/pharmacology , Glucagon/blood , Glucose Clamp Technique , KineticsABSTRACT
There is a need to improve the treatment of superficial bladder cancer. One area which holds promise is intravesical gene therapy. Recently, studies undertaken by us have shown that marked tumor regression of bladder cancers occurred after two daily intravesical administrations of an adenovirus encoding human interferon alpha (Ad-IFNalpha) using a mouse superficial bladder cancer model in which human bladder tumors are growing. A dose of 1 x 10(11) particles/ml (P/ml) was used along with 1 mg/ml of Syn3, a gene transfer-enhancing agent. Since clinical studies are being planned using this approach, it became critical to determine if one exposure and lower particle number could be equally effective. We report that indeed a single dose of Ad-IFNalpha in Syn3 at doses of 1 x 10(10)-1 x 10(11) P/ml is highly effective in reducing the size of the tumors, whereas 1 x 10(9) P/ml was not. Efficacy was also correlated with the level of IFN produced in the urine after treatment. Based on the results of the present studies, a Phase I trial is being planned for superficial bladder cancer, which will involve a single initial treatment with Ad-IFNalpha/Syn3 and measurement of IFN in the urine over time as an indicator of adequate gene transfer and expression.
Subject(s)
Cholic Acids/administration & dosage , Disaccharides/administration & dosage , Genetic Therapy/methods , Genetic Vectors/genetics , Interferon-alpha/therapeutic use , Interferon-alpha/urine , Urinary Bladder Neoplasms/therapy , Administration, Intravesical , Animals , Cell Line, Tumor , Dose-Response Relationship, Drug , Genetic Vectors/administration & dosage , Green Fluorescent Proteins , Humans , Interferon-alpha/genetics , MiceABSTRACT
Primary biliary cirrhosis (PBC) is a chronic cholestatic autoimmune liver disease that predominantly affects middle-aged women. It is characterized by slowly progressive destruction of the small intrahepatic bile ducts together with portal inflammation, and this initially leads to fibrosis and later to cirrhosis. It is currently accepted that the pathogenesis of PBC is multifactorial with genetic and environmental factors interplaying to determine the disease onset and progression. In addition to antimitochondrial antibody (AMA), which is the hallmark of PBC and is detected in at least 90% of the patients, other autoantibodies (antinuclear antibody, anti-smooth muscle antibody and rheumatoid factor, etc.) may also be found in the patients. There is no correlation between the titer of AMAs and the disease severity. Most patients are diagnosed either during the asymptomatic phase of PBC or after presenting with non-specific symptoms. Pruritus and fatigue are the most common symptoms of PBC. The prognosis of PBC has improved significantly during the last few decades. Patients are now diagnosed earlier in its clinical course, they are more likely to be asymptomatic at diagnosis and they are more likely to receive medical treatment. A wide variety of drugs have been assessed for the treatment of this condition: such immunosuppressive agents as corticosteroids, cyclosporine and azathioprine have a weak effect on the disease's natural history. Ursodeoxycholic acid (UDCA) is the only currently approved medical treatment. For PBC patients with end-stage liver disease or an unacceptable quality of life, liver transplantation is the only accepted therapeutic option. Early diagnosis and treatment of PBC are important because effective treatment with UDCA has been shown to delay disease progression and improve rate survival in the early stage.
Subject(s)
Female , Humans , Male , Middle Aged , Autoimmune Diseases/diagnosis , Cholagogues and Choleretics/therapeutic use , Cholestadienes/administration & dosage , Cholic Acids/administration & dosage , Liver Cirrhosis, Biliary/diagnosis , Prevalence , Rifampin/administration & dosageABSTRACT
Watchful waiting is the most important alternative to cholecystectomy in patients with uncomplicated gallstone disease. Symptom relief occurs in a large percentage of the patients, and the risk of developing complications is relatively small. However, in patients with a long life expectancy the cumulative risk may be significant, which must be taken into consideration. Extracorporal shock wave lithotripsy and medical treatment play a very small role in the treatment, and only on very special indications. The results of these methods are relatively poor and associated with a high recurrence rate.