A novel cyclobutane class of nonpeptidic glucagon-like peptide-1 (GLP-1) receptor agonists, exemplified by 3, was identified using receptor binding and multiple response element/cAMP response element (MRE/CRE)-driven reporter gene assays. The structures of 3 and its three isomers were elucidated by NMR, HRESIMS, and X-ray crystallography. A series of structural modifications were also made based on the core structure of 3 with different substitution groups at the west and east ends. Among these analogues, compound 16 was found to be 4- to 5-fold more potent than 3 both in vitro and in vivo.

Background Glucagon-like peptide-1 (GLP-1) is recognized as an important regulator of glucose homeostasis. Efforts to utilize GLP-1 mimetics in the treatment of diabetes have yielded clinical benefits. A major hurdle for an effective oral therapy has been the difficulty of finding a non-peptidic GLP-1 receptor (GLP-1R) agonist. While its oral bioavailability still poses significant challenges, Boc5, one of the first such compounds, has demonstrated the attainment of GLP-1R agonism in diabetic mice. The present work was to investigate whether subchronic Boc5 treatment can restore glycemic control and induce sustainable weight loss in diet-induced obese (DIO) mice, an animal model of human obesity and insulin resistance. Methodology/Principal Findings DIO mice were treated three times a week with Boc5 (0.3, 1 and 3 mg) for 12 weeks. Body weight, body mass index (BMI), food intake, fasting glucose, intraperitoneal glucose tolerance and insulin induced glucose clearance were monitored regularly throughout the treatment. Glucose-stimulated insulin secretion, β-cell mass, islet size, body composition, serum metabolic profiles, lipogenesis, lipolysis, adipose hypertrophy and lipid deposition in the liver and muscle were also measured after 12 weeks of dosing. Boc5 dose-dependently reduced body weight, BMI and food intake in DIO mice. These changes were associated with significant decreases in fat mass, adipocyte hypertrophy and peripheral tissue lipid accumulation. Boc5 treatment also restored glycemic control through marked improvement of insulin sensitivity and normalization of β-cell mass. Administration of Boc5 (3 mg) reduced basal but enhanced insulin-mediated glucose incorporation and noradrenaline-stimulated lipolysis in isolated adipocytes from obese mice. Furthermore, circulating leptin, adiponectin, triglyceride, total cholesterol, nonesterified fatty acid and high-density lipoprotein/low-density lipoprotein ratio were normalized to various extents by Boc5 treatment. Conclusions/Significance Boc5 may produce metabolic benefits via multiple synergistic mechanisms and may represent an attractive tool for therapeutic intervention of obesity and diabetes, by means of non-peptidic GLP-1R agonism.

A novel "signal-on" assay for sensitive and selective detection of Hg(2+) in aqueous solution based on mismatched DNA and a molecular light switch complex, [Ru(phen)(2)(dppz)](2+), was reported in the present work. This Hg(2+) specific sensor system composed of two label-free DNA probes with eight T-T mismatches, which could form stable DNA duplexes by thymine-Hg(2+)-thymine (T-Hg(2+)-T) in the presence of Hg(2+). The luminescence of [Ru(phen)(2)(dppz)](2+) is very weak in aqueous solution, and significant luminescence can be observed when intercalating into DNA duplexes. By monitoring Hg(2+)-dependent luminescence intensity enhancement, highly sensitive and selective determination of Hg(2+) has been achieved. Under the optimum conditions, the luminescence intensity was proportional to the concentration of Hg(2+) in the range of 1.0x10(-9)-1.5x10(-7) M. A detection limit of 3.5x10(-10) M Hg(2+) was achieved using 3sigma. The presence of other metal ions did not interfere with the detection of Hg(2+) even at high concentrations. This approach is simple and rapid, which can be used to monitor the Hg(2+) concentration in drinking water and natural media within 30 min.

无机汞在天然水体中可被微生物转化为毒性更强的甲基汞,在生物体内大量富集后经过食物链进入人体,危害人体健康.由于Hg~(2+)不能被牛物降解,一旦进入环境,只能被稀释或转移,因此实现高灵敏度、高选择性地检测Hg~(2+)具有重要意义.

BACKGROUND:Our recent discovery of the substituted cyclobutane Boc5, one of the first non-peptidic agonists at glucagon-like peptide-1 receptors, offers the potential of combining oral availability with full agonism capable of eliciting antidiabetic and antiobesity effects. The present study was aimed at determining the in vivo pharmacologic properties of Boc5 in both normal and diabetic mice following chronic administration, with emphasis on glycemic control and weight loss.METHODOLOGY/PRINCIPAL FINDINGS:C57BL/6J and db/db mice were treated daily with Boc5 for 4 weeks and a range of pharmacologic parameters, including hemoglobin A1c, intraperitoneal glucose tolerance, insulin tolerance, fasting insulin and leptin levels, food intake, body weight and fat mass, were assessed before and after the treatment. Effects on food intake, gastric emptying, and insulinogenic index were also investigated in animals acutely administered with Boc5. Boc5 (3 mg) was able to induce a durable restoration of glycemic control (normalization of both hemoglobin A1c and intraperitoneal glucose tolerance) in db/db mice, following 4 weeks of daily administration. As with peptidic glucagon-like peptide-1 receptor agonists, its glycemic benefit and weight (fat) loss were associated with dose-dependent effects that included reduction in food intake, slowing of gastric emptying (both of which reduce nutrient-drive at beta-cells), stimulation of insulin secretion (which was glucose-dependent), and elevation in insulin sensitivity. There was little effect on normal mice treated in the same manner.CONCLUSIONS/SIGNIFICANCE:Our findings suggest that Boc5 is the only non-peptidic molecule reported thus far to simultaneously activate this spectrum of antidiabetic effects.

AIM:To develop a homogeneous binding assay for high-throughput screening (HTS) of hit compounds at human neuromedin U receptor (hNMU-R) 1 and to identify non-peptidic small molecule hNMU-R modulators through functional assessments and structure-activity relationship (SAR) analyses.METHODS:Membrane preparations of Chinese hamster ovary cells (CHO-K1) stably expressing hNMU-R1, [125I]hNMU-25, and wheat germ agglutinin-coupled microbeads were used to develop an HTS assay based on scintillation proximity assay (SPA) technology. This method was applied to a large-scale screening campaign against a diverse library of 36,000 synthetic compounds or natural products and subsequent confirmation studies. CHO-K1 cells stably expressing full-length hNMU-R1 or hNMU-R2 and a calcium-sensitive dye were employed to functionally measure intracellular calcium mobilization upon ligand stimulation. Preliminary SAR was determined based on limited structural modifications.RESULTS:The Ki value (0.7 nmol/L) of hNMU-25 (the natural ligand) at hNMU-R1 measured by the SPA method was consistent with that reported in the literature, and the Z'factor for this HTS assay was 0.81. A total of 100 hits, showing more than 30% competitive inhibition on [125I]hNMU-25 binding to hNMU-R1, were identified initially, 3 of which were confirmed thereafter to have reasonable hNMU-R1-binding affinities and similar chemical structures. Based on their common molecular skeleton, 203 analogs were synthesized and tested. Among the 16 analogs that retained variable hNMU-R1- binding abilities, 2 elicited calcium influx in both hNMU-R1 and hNMU-R2-expressing cells, but none displayed antagonist activity.CONCLUSION:The homogeneous hNMU-R1 binding assay is an efficient and robust tool for screening potential hNMU-R modulators. Two non-selective hNMU-R agonists discovered are of low molecular weight nature with novel chemical structures. The preliminary SAR investigation suggests that both the triphenyl and guanidinol groups are crucial to the bioactivities observed.

British Journal of Pharmacology (2008) 154, 1161; doi:10.1038/bjp.2008.251

A sensitive label-free bio-barcode assay provided a PCR-free method for quantitative detection of two nucleic acid targets (HTLV-I and HTLV-II) simultaneously. This DNA biosensor was fabricated with two-component oligonucleotide-modified gold nanoparticles (AuNPs) and two-component oligonucleotide-modified magnetic beads (MBs), which can sandwich a specific target. After liberating the adsorbed thiolated barcode DNA strands (poly A and poly G) from the AuNPs surface with dithiothreitol (DTT) and acidic dipurinization, the electrochemical measurements were directly performed based on the redox activity of guanine (G) and adenine (A) nucleobases. Under the optimal assembling and detection conditions, a good linearity for simultaneous detection was obtained in the range from 4.4 × 10−11 to 2.0 × 10−9 M, and the detection limit (3σ) was estimated to be 1.71 × 10−12 M for T1-DNA and 1.55 × 10−12 M for T2-DNA.

Aim: Melatonin (MT) is a neurohormone produced and secreted primarily by the pineal gland in a circadian manner, and mainly acts through 2 receptor subtypes: MT 1 and MT 2 in humans. The diversity in their tissue distribution is in favor of different functions for each receptor subtype. Selective modulators are therefore required to determine the physiological roles of these melatonin receptor subtypes and their implications in pathological processes. Methods: A homogenous MT 1 /MT 2 receptor binding assay was established for high-throughput screening of new ligands at the hMT 1 and/or hMT 2 receptors. The functional properties (agonists or antagonists) were assessed by a conventional guanosine-5′[γ- 35 S] triphosphate (GTP-γS) assay. Results: Three hMT 1 receptor-selective small molecule antagonists and 1 hMT 2 receptor-selective small molecule antagonist with novel structural features were identified following a high-throughput screening campaign of 48 240 synthetic and natural compounds. Conclusion: The findings may assist in the expansion of chemical probes to these 2 receptor subtypes.