Background: Electronic platform surveys are ideal for self-administration in adolescent populations, as nearly all adolescents in the US report using or having access to smartphones. This technology-savvy population seems prepared to graduate away from paper surveys. Despite predicted advantages of using smartphones for data collection, there is a surprising lack of data regarding the use of this mode for surveillance of substance use trends among adolescent populations. Objectives: The objective of this study was to evaluate completion rates, times, and responses of high school students taking the Secondary Student Life Survey: Nevada (SSLS:NV) on their own personal smartphones compared to provided computers or tablets. Methods: The SSLS:NV is a web-based survey designed to assess adolescent beliefs, attitudes, and use-trends surrounding substance use. The SSLS:NV was self-administered via self-selected device (personal vs. provided) within one class period to approximately one thousand 9th-12th grade students in December 2016. Data was collected and analyzed to compare outcomes by computer, tablet, or smartphone. Statistical analysis was performed in SPSS 23.0 using χ2 or Fisher's exact test for categorical data and one-way ANOVA for continuous data. Results: SSLS:NV completion times averaged 21 min overall (p = .193). Differences were seen with completion rates of 86% smartphone 94% tablet, and 95% computer (p < .001), while responses to lifetime substance use were similar across all groups. Conclusions/Importance: The current study provides proof of concept that personal smartphones are effective in achieving more comprehensive adolescent substance use surveillance within a relatively short amount of time, while retaining robust response rates.

The glutamatergic antagonist, RO‐25‐6981, has previously been shown to exert both rapid and sustained antidepressant‐like activity. The purpose of the current study is to delineate its putative antidepressant mechanisms of NR2B‐selective NMDA receptor antagonism and monoamine reuptake transporter inhibition. RO‐25‐6981 and structural analogs with varying degrees of NR2B binding affinity were tested for antidepressant‐like behavior in wild‐type mice and animals deficient in NR2B subunit expression. In the tail suspension test, four RO‐25‐6981 analogs (TR‐2, TR‐4, TR‐5, and TR‐6) were found to exhibit antidepressant‐like activity in wild‐type mice following acute administration (30 mg/kg, i.p., 30 min) with maximal reductions in immobility by approximately half compared to vehicle‐treated controls. By contrast, RO‐25‐6981 (10 mg/kg, i.p., 30 min) reduced immobility by approximately 90%, an effect comparable to that exhibited by the traditional monoaminergic antidepressants fluoxetine and desipramine. However, unlike RO‐25‐6981, TR‐2, TR‐4, TR‐5 and TR‐6 profoundly limited generalized locomotor activity suggesting increased activity in the tail suspension test was related to psychotropic vs. generalized drug effects. In contrast, other TR analogs tested showed no antidepressant‐like activity in the tail suspension test, despite possessing robust NMDA receptor antagonist activity via mid‐ to low‐nanomolar binding affinity at the NR2B subunit. Interestingly, RO‐25‐6981 and TR‐5 exhibited similar antidepressant‐like activity in wild‐type and NR2B‐deficient mice, despite possessing low‐nanomolar NR2B binding. In addition, cellular serotonin transport assays showed functional inhibitory activity of both agents. Taken together, these data suggest that the antidepressant‐like activity of RO‐25‐6981 and its analogs does not correlate with the degree of NMDA receptor antagonism. Furthermore, these data point to monoamine reuptake inhibition contributing to the overall antidepressant‐like activity of RO‐25‐6981 in animal models of mood.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Ro 25-6981 is a highly potent and selective blocker of N-methyl-d-aspartate receptors that has been shown to possess both rapid and sustained antidepressant activity. In the present study, we report the biopharmaceutical characterization of Ro 25-6981 by evaluating gastrointestinal stability, transepithelial permeability, stability in human liver microsomes, and in silico metabolic prediction. Moreover, in vivo efficacy of Ro 25-6981 after oral administration was evaluated in animal models of depression. When mixed with 5 different simulated gastrointestinal fluids, no loss of parent compound was observed after 6 h, indicating compound stability in the gastrointestinal environment. At the tested concentrations, Ro 25-6981 was shown to have transepithelial permeability with apparent permeability (Papp) values comparable to highly permeable drugs. Ro 25-6981 was metabolized within 30 min in human liver microsomes, and the metabolic prediction data showed glucuronidation and sulfation as potential metabolic pathways. The in vivo efficacy data suggested that Ro 25-6981, when administered orally at 30 mg/kg, exhibits antidepressant-like activity following oral administration with efficacy comparable to traditional antidepressants that is both dose- and time-dependent. Overall, due to optimal gastrointestinal stability, oral permeability, and oral efficacy, Ro 25-6981 can be a potential therapeutic option for the treatment of depression.

The increasing number of people afflicted with diabetes throughout the world is a major health issue. Inhibitors of the sodium-dependent glucose cotransporters (SGLT) have appeared as viable therapeutics to control blood glucose levels in diabetic patents. Herein we report the discovery of LX2761, a locally acting SGLT1 inhibitor that is highly potent in vitro and delays intestinal glucose absorption in vivo to improve glycemic control.

LX2761 is a potent sodium/glucose cotransporter 1 inhibitor restricted to the intestinal lumen after oral administration. Studies presented here evaluated the effect of orally administered LX2761 on glycemic control in preclinical models. In healthy mice and rats treated with LX2761, blood glucose excursions were lower and plasma total glucagon-like peptide-1 (GLP-1) levels higher after an oral glucose challenge; these decreased glucose excursions persisted even when the glucose challenge occurred 15 hours after LX2761 dosing in ad lib-fed mice. Further, treating mice with LX2761 and the dipeptidyl-peptidase 4 inhibitor sitagliptin synergistically increased active GLP-1 levels, suggesting increased LX2761-mediated release of GLP-1 into the portal circulation. LX2761 also lowered postprandial glucose, fasting glucose, and hemoglobin A1C, and increased plasma total GLP-1, during long-term treatment of mice with either early- or late-onset streptozotocin-diabetes; in the late-onset cohort, LX2761 treatment improved survival. Mice and rats treated with LX2761 occasionally had diarrhea; this dose-dependent side effect decreased in severity and frequency over time, and LX2761 doses were identified that decreased postprandial glucose excursions without causing diarrhea. Further, the frequency of LX2761-associated diarrhea was greatly decreased in mice either by gradual dose escalation or by pretreatment with resistant starch 4, which is slowly digested to glucose in the colon, a process that primes the colon for glucose metabolism by selecting for glucose-fermenting bacterial species. These data suggest that clinical trials are warranted to determine if LX2761 doses and dosing strategies exist that provide improved glycemic control combined with adequate gastrointestinal tolerability in people living with diabetes.

Rational design of lead compounds targeting monoamine transporters is critical to developing novel therapeutics for treating psychiatric and substance abuse disorders. A 3‐D dopamine transporter (DAT) computer model was used to virtually screen a small molecule library for high DAT affinity drug‐like compounds. One hit, coded “MI‐4,” inhibited human dopamine, norepinephrine, and serotonin transporters in vitro . In vivo administration in mice (i.p.) induced robust, dose‐dependent antidepressant‐like effects in behavioral models of mood responsive to acute administration (tail suspension and forced swim tests) and chronic administration (novelty‐induced hypophagia test, chronic social‐defeat stress). Importantly, MI‐4 exhibited minimal abuse liability in behavioral and neurological models (conditioned place preference and dopamine in vivo microdialysis). To facilitate pharmacokinetic analysis, a rapid and highly selective reversed‐phase HPLC method for of MI‐4 was developed and validated according to ICH guidelines. Analysis in human, rat and mouse plasma resulted in a distinct single peak of MI‐4 with USP tailing factor < 2.0 using the MI‐4 analog ifenprodil as an internal standard. The stability of MI‐4 incubated in simulated fasted and fed‐state gastric fluid was 98–100% after 3 hrs and 91–99% after 6 hrs. The Papp value of MI‐4 (0.5 mM) across caco‐2 monolayers after one hour was 33.2×10 −6 cm/s, indicating high membrane permeability. Moreover, oral administration (p.o.) of MI‐4 in mice resulted in dose‐ and time‐dependent reductions in tail suspension test immobility scores comparable to the established antidepressants fluoxetine and desipramine, indicating oral efficacy for antidepressant‐like effects. MI‐4 was found to be Ro‐25‐6981, an ifenprodil analog and reputed NMDA antagonist. These data suggest that Ro‐25‐6981, previously known for rapid‐acting antidepressant activity, may also functionally inhibit monoamine reuptake and produce sustained antidepressant effects in vivo . This demonstrates, as proof of principle, the viability of combining glutamatergic and monoaminergic mechanisms to produce rapid and sustained antidepressant‐like effects.

Sodium-glucose cotransporter 2 (SGLT2) inhibitors are a new class of oral anti-diabetic agents that improve glycemic control by inhibiting SGLT2-mediated renal glucose reabsorption. Currently available agents increase urinary glucose excretion (UGE) to <50% of maximal values because they do not inhibit SGLT1, which reabsorbs >50% of filtered glucose when SGLT2 is completely inhibited. This led us to test whether LP-925219, a small molecule dual SGLT1/SGLT2 inhibitor, increases UGE to maximal values in wild-type (WT) mice. We first tested LP-925219 inhibition of glucose transport by HEK293 cells expressing SGLT1 or SGLT2, and then characterized LP-925219 pharmacokinetics. We found that LP-925219 was a potent inhibitor of mouse SGLT1 (IC50 = 22.6 nmol/L) and SGLT2 (IC50 = 0.5 nmol/L), and that a 10 mg/kg oral dose was bioavailable (87%) with a long half-life (7 h). We next delivered LP-925219 by oral gavage to WT, SGLT1 knockout (KO), SGLT2 KO, and SGLT1/SGLT2 double KO (DKO) mice and measured their 24-h UGE. We found that, in vehicle-treated mice, DKO UGE was maximal and SGLT2 KO, SGLT1 KO, and WT UGEs were 30%, 2%, and 0.2% of maximal, respectively; we also found that LP-925219 dosed at 60 mg/kg twice daily increased UGE of SGLT1 KO, SGLT2 KO, and WT mice to DKO UGE levels. These findings show that orally available dual SGLT1/SGLT2 inhibitors can maximize 24-h UGE in mammals, and suggest that such agents merit further evaluation for their potential, in diabetic patients, to achieve better glycemic control than is achieved using selective SGLT2 inhibitors.

The first allosteric, type III inhibitor of LIM-kinase 2 (LIMK2) is reported. A series of molecules that feature both an N-phenylsulfonamide and tertiary amide were not only very potent at LIMK2 but also were extremely selective against a panel of other kinases. Enzymatic kinetic studies showed these molecules to be noncompetitive with ATP, suggesting allosteric inhibition. X-ray crystallography confirmed that these sulfonamides are a rare example of a type III kinase inhibitor that binds away from the highly conserved hinge region and instead resides in the hydrophobic pocket formed in the DFG-out conformation of the kinase, thus accounting for the high level of selectivity observed.

The structure of LX7101, a dual LIM-kinase and ROCK inhibitor for the treatment of ocular hypertension and associated glaucoma, is disclosed. Previously reported LIM kinase inhibitors suffered from poor aqueous stability due to solvolysis of the central urea. Replacement of the urea with a hindered amide resulted in aqueous stable compounds, and addition of solubilizing groups resulted in a set of compounds with good properties for topical dosing in the eye and good efficacy in a mouse model of ocular hypertension. LX7101 was selected as a clinical candidate from this group based on superior efficacy in lowering intraocular pressure and a good safety profile. LX7101 completed IND enabling studies and was tested in a Phase 1 clinical trial in glaucoma patients, where it showed efficacy in lowering intraocular pressure.

An ultra high-pressure liquid chromatography/mass spectrometry (UHPLC/MS) separation and analysis method has been devised for open access analysis of synthetic reactions used in the production of DNA-encoded chemical libraries. The aqueous mobile phase is 100mM hexafluoroisopropanol and 8.6mM triethylamine; the organic mobile phase is methanol. The UHPLC separation uses a C18 OST column (50mm×2.1mm×1.7μm) at 60°C, with a flow rate of 0.6mL/min. Gradient concentration is from 10 to 40% B in 1.0min, increasing to 95% B at 1.2min. Cycle time was about 5min. This method provides a detection limit of a 20-mer oligonucleotide by mass spectrometry of better than 1pmol on-column. Linear UV response for 20-mer extends from 2 to 200pmol/μL in concentration, same-day relative average deviations are less than 5% and bias (observed minus expected) is less than 10%. Deconvoluted mass spectra are generated for components in the predicted mass range using a maximum entropy algorithm. Mass accuracy of deconvoluted spectra is typically 20ppm or better for isotopomers of oligonucleotides up to 7000Da.

Thirty-six patients with type 2 diabetes mellitus (T2DM) were randomized 1:1:1 to receive a once-daily oral dose of placebo or 150 or 300 mg of the dual SGLT1/SGLT2 inhibitor LX4211 for 28 days. Relative to placebo, LX4211 enhanced urinary glucose excretion by inhibiting SGLT2-mediated renal glucose reabsorption; markedly and significantly improved multiple measures of glycemic control, including fasting plasma glucose, oral glucose tolerance, and HbA(1c); and significantly lowered serum triglycerides. LX4211 also mediated trends for lower weight, lower blood pressure, and higher glucagon-like peptide-1 levels. In a follow-up single-dose study in 12 patients with T2DM, LX4211 (300 mg) significantly increased glucagon-like peptide-1 and peptide YY levels relative to pretreatment values, probably by delaying SGLT1-mediated intestinal glucose absorption. In both studies, LX4211 was well tolerated without evidence of increased gastrointestinal side effects. These data support further study of LX4211-mediated dual SGLT1/SGLT2 inhibition as a novel mechanism of action in the treatment of T2DM.

The prevalence of diabetes throughout the world continues to increase and has become a major health issue. Recently there have been several reports of inhibitors directed toward the sodium-dependent glucose cotransporter 2 (SGLT2) as a method of maintaining glucose homeostasis in diabetic patients. Herein we report the discovery of the novel O-xyloside 7c that inhibits SGLT2 in vitro and urinary glucose reabsorption in vivo.