Discovery of Brain-Penetrant Glucosylceramide Synthase Inhibitors with a Novel Pharmacophore.

Inhibition of glucosylceramide synthase (GCS) is a major therapeutic strategy for Gaucher's disease and has been suggested as a potential target for treating Parkinson's disease. Herein, we report the discovery of novel brain-penetrant GCS inhibitors. Assessment of the structure-activity relationship revealed a unique pharmacophore in this series. The lipophilic ortho-substituent of aromatic ring A and the appropriate directionality of aromatic ring B were key for potency. Optimization of the absorption, distribution, metabolism, elimination, toxicity (ADMETox) profile resulted in the discovery of T-036, a potent GCS inhibitor in vivo. Pharmacophore-based scaffold hopping was performed to mitigate safety concerns associated with T-036. The ring opening of T-036 resulted in another potent GCS inhibitor with a lower toxicological risk, T-690, which reduced glucosylceramide in a dose-dependent manner in the plasma and cortex of mice. Finally, we discuss the structural aspects of the compounds that impart a unique inhibition mode and lower the cardiovascular risk.

CE: Understanding the Nurse's Role in Managing Gaucher Disease.

: Lysosomal storage disorders (LSDs) are a group of inherited metabolic conditions, the overall incidence of which is estimated to range from one in 5,000 to one in 7,000 live births. Gaucher disease, the most common LSD, is of autosomal recessive inheritance. It results from a deficiency of acid ß-glucocerebrosidase and can affect the spleen, liver, bone, bone marrow, and central nervous system. Gaucher disease is clinically classified into one of three phenotypes, depending on the absence or presence of neurodegenerative disease and the rate of disease progression. Although there is no cure for Gaucher disease, it may be treated with enzyme replacement and substrate reduction therapy. With the development of enzyme testing through dried blood spots, Gaucher disease may now be detected at birth through newborn screening. The purpose of this article is to review the epidemiology and pathophysiology of Gaucher disease, update nurses on advances in newborn screening, diagnosis, and management of this genetic disorder, and highlight the role of nurses in the diagnosis and care of patients with Gaucher disease.

[Effects of artificial microRNA targeting glucosylceramide synthase on drug sensitivity of breast cancer cells].

OBJECTIVE: To explore the inhibitory effects on glucosylceramide synthase (GCS) expression and drug sensitivity in breast cancer cells by transfecting artificial microRNA targeting GCS. METHODS: Two microRNA expression vectors targeting GCS were constructed and transfected into MCF-7/ADR cells via Lipofectamine 2000. The levels of GCS mRNA and protein were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively. Methyl thiazolyl tetrazolium (MTT) assay was used to assess the chemosensitivity of MCF-7/ADR cells to adriamycin (ADM) and vincristine. RESULTS: After transfection of two microRNA expression vectors, the expression of GCSmRNA in MCF-7/ADR cells was 0.098 ± 0.005 and 0.143 ± 0.007 respectively. Compared with the control cells (0.875 ± 0.008), the difference was significant (P < 0.01). The expression of GCS protein (0.127 ± 0.004, 0.165 ± 0.008) in MCF-7/ADR cells was lower than that in the control cells (0.765 ± 0.007; P < 0.01). Furthermore, in comparison with the control cells, the resistance factor to adriamycin significantly dropped to 4.06 and 6.06 while the drug resistance to vincristine decreased to 8.30 and 12.67 respectively (P < 0.01). CONCLUSION: Artificial microRNA targeting GCS inhibits the GCS expression and restores significantly the sensitivity of breast cancer cells to anticancer drugs. These findings may provide a novel strategy of enhancing the chemotherapy sensitivity of breast cancer.

Effects of transglucosidase on diabetes, cardiovascular risk factors and hepatic biomarkers in patients with type 2 diabetes: a 12-week, randomized, double-blind, placebo-controlled trial.

In this 12-week, randomized, double-blind, placebo-controlled trial, the efficacy and safety of transglucosidase (TGD) were compared with placebo in patients with type 2 diabetes mellitus (T2DM). At 12 weeks, TGD 300 mg/day and TGD 900 mg/day significantly reduced HbA1c (0.18 and 0.21%) and insulin concentration (19.4 and 25.0 pmol/l), respectively, vs. placebo. TGD 300 mg/day and TGD 900 mg/day also significantly reduced low-density lipoprotein cholesterol (0.22 and 0.17 mmol/l, respectively). TGD 900 mg/day significantly reduced triglyceride by 0.24 mmol/l and diastolic blood pressure by 8 mmHg. Placebo was associated with a significant increase from baseline in body mass index, alanine aminotransferase and aspartate aminotransferase (0.17 kg/m(2) , 3 and 2 U/l, respectively), whereas TGD was not. TGD 300 mg/day significantly increased high-molecular-weight adiponectin by 0.6 µg/ml. Adverse events did not differ significantly between the groups. TGD resulted in lowering of HbA1c and blood insulin level and improvements in metabolic and cardiovascular risk factors in T2DM.

Anidulafungin.

Anidulafungin is a novel antifungal agent which, like other echinocandins, inhibits beta-(1,3)-D-glucan synthase and disrupts fungal cell-wall synthesis. It has marked antifungal activity against a broad spectrum of Candida spp. and Aspergillus spp., including amphotericin B- and triazole-resistant strains. In clinical trials, anidulafungin has primarily been evaluated in patients with oesophageal and invasive candidiasis. Preliminary data are emerging for other indications such as invasive aspergillosis. In a large, multicentre, double-blind, double-dummy, randomised trial in patients with oesophageal candidiasis, intravenous anidulafungin 50 mg/day was as effective as oral fluconazole 100 mg/day regarding end-of-treatment rates of endoscopic cure and clinical and microbiological success. Duration of treatment was approximately 2-3 weeks, and patients in both groups received a loading dose of study drug (twice the daily maintenance dose) on day 1. Anidulafungin is generally well tolerated. Across the dosage range 50-100 mg/day, adverse events appear not to be dose- or infusion-related. In the largest clinical trial to date, the most common treatment-related adverse events were phlebitis/thrombophlebitis, headache, nausea, vomiting and pyrexia.

Intranasal immunization of humans with Streptococcus mutans antigens.

To evaluate the effectiveness of a low dose of soluble or liposomal (L) glucosyltransferase-enriched preparation (E-GTF) in inducing mucosal immune responses after intranasal immunization, 12 adults were immunized on days 0 and 7 by the IN route with 62.5 microg of soluble E-GTF or L-E-GTF. An increase in the mean salivary IgA anti-E-GTF response (P < 0.03) was seen in the L-E-GTF but not the soluble E-GTF group. A significant increase (P < 0.05) in the mean specific IgA antibody activity was also seen in nasal wash from both groups. Although the nasal wash responses were higher in the L-E-GTF than in the soluble E-GTF group, they were not significantly different. The soluble E-GTF immunized group showed a higher serum IgG response than the L-E-GTF immunized group on day 90 (P < 0.05). These results indicate that as little as 62.5 microg of E-GTF, when given by the intranasal route, induced an IgA response in secretions.

[Glucosyltransferase (GTF) and immunization against dental caries in humans]. / Glykosyltranspherase (GTF) kai anosopoiese kata tes teredonas ston anthropo.

Glucosyltransferase (GTF) is an extracellular or cell-associated enzyme synthesized by the "mutans" group of streptococci as well as the S. sanguis and is responsible for the biosynthesis of extracellular polysaccharides. Glucan formation in dental plaque mediates binding of S. mutans and S. sanguis to one another as well as to other bacteria. Although the production of organic acids is an indispensable property of the cariogenic strains of bacteria, their ability to stick and accumulate on tooth surfaces is an almost equally significant parameter of virulence. Furthermore, GTF has been proven to be an effective antigen in eliciting caries-protective secretory IgA antibodies in rodent models. That led to the identification of GTF as a potential antigen for use in a human caries vaccine. The results of recent studies suggest that oral immunization with GTF, has the potential to elicit a secretory IgA antibody response and to interfere with accumulation and permanent colonization of S. mutans on smooth tooth surfaces. However, it cannot be inferred from the available data whether such a vaccine is effective in caries prevention in human beings. Although many modifications seem to be necessary with regard to the composition, dose, frequency and route of administration of the antigen, the preparation of a safe and effective vaccine against human caries is considered to be feasible in the near future.

Immunization with dextransucrases, levansucrases, and glycosidic hydrolases from oral streptococci. II. Immunization with glucosyltransferases, fructosyltransferases, and glycosidic hydrolases from oral streptococci in monkeys.

The feasibility of immunizing monkeys with enzymes from oral streptococci in an attempt to reduce dental caries was investigated. Forty rhesus monkeys, Macaca mulatta, were used. Cariogenic streptococci, S mutans, were implanted into all the monkeys' mouths. There was no pathological effect resulting from immunization. Of the 40 animals, 30 retained the implanted flora throughout the experiment; the remaining 10 were reimplanted until the streptococci remained. In six months, gross carious lesions were evident with plaque. Inhibitiors present in the monkey sera after immunization inhibited glucosyltransferase, fructosyltransferase, and neuraminidase activities. It was presumed the inhibitors were antibodies. There was a reduction of 68.6% in the total carious lesions in the animals immunized intraorally with glucosyltransferase, 62.4% reduction in those injected with fructosyltransferase, and 57.4% reduction in total lesions in those immunized with glycosidic hydrolases after 19 months, as compared to the control group. There were no gross lesions apparent in the group immunized with glycosidic hydrolases. It appears that immunization with enzymes significantly reduces carries and is feasible in a primate model.