Locomotor activity.

<p>Distance travelled during 2 weeks of treatment with TP or SARM-2f was calculated by the formula described in the Materials and methods section (A). (B) Activity during the light or dark phase in day 13–14. (C) Levator ani muscle and body weight measured after 2-week dosing. Food intake was calculated during 2-week dosing. The drug was administered to castrated mice with DHEA for 2 weeks. The measurement technique has been described in the Methods section. Data are shown as the means ± SD (n = 8). ^##P < 0.005 vs. ORX (vehicle treated castrated mice) group using a Williams test. *P < 0.05, **P < 0.01 vs. ORX group using a student t-test.</p

Induction of sexual behavior by SARM-2f and TP in castrated male rats.

<p>(A) Sexual behavior scores. (B) Lean body mass change and levator ani muscle weight (g), lean body mass. The drug was administered to castrated male rats for three weeks. Sexual behavior was determined by observation of the estrous cycle of female rats that had been mated with male rats treated with SARM-2f or TP at the end of the treatment period. (n = 10).*P < 0.05 vs. the control group with a Fishers' exact-test. **P < 0.005 vs. control group with the Williams test. Ctrl, Control vehicle group, TP, testosterone propionate.</p

Cofactor recruitment analysis in 293T cells with DHT, testosterone, TSAA-291, and SARM-2f.

<p>Luciferase activity was measured after 24 h treatment with varying concentrations of DHT, testosterone, TSAA-291, and SARM-2f (30, 100, and 300 nM).</p

Effects of SARM-2f on mouse voluntary running.

<p>The outline of the voluntary running test is shown in Fig 6A. Running distance during 2 weeks of treatment with TP or SARM-2f was calculated by the formula described in the Materials and methods section (B). (B) Running count average in day13–14. (C) Levator ani muscle weight measured after 2-week dosing. The drug was administered to castrated mice with DHEA for 2 weeks. The measurement technique is described in the Materials and methods section. Data are shown as the means ± SD (n = 8). *P < 0.025 vs. the ORX (vehicle treated castrated mice) group using the Williams test. #P < 0.05 vs the ORX by student t-test.</p

Concentrations of SARM-2f in tissues after oral administration at a dose of 0.75 mg/kg to rats.

<p>Data are shown as the means ± SD. (n = 3).</p

Cell context-dependent AR transcriptional activity of SARM-2f and DHT.

<p>The ability of compounds to influence AR-mediated transcriptional activation was examined in skeletal muscle cells (SkMC) (A) and prostate epithelial cells (PrEC) (B) by reporter assays as described in Material and methods. Data are shown as the means ± SD (n = 3).</p

Tissue specificity of SARM-2f and testosterone propionate (TP) in castrated rats.

<p>Effect of SARM-2f (A) and TP (B) on the levator ani muscle weight and prostate weight. The y axis represents percent (%) of the sham group. The drug was administered to castrated male rats for three weeks. Data are shown as the means ±SD (n = 4–5).</p

Discovery of 5‑Chloro-1-(5-chloro-2-(methylsulfonyl)benzyl)-2-imino-1,2-dihydropyridine-3-carboxamide (TAK-259) as a Novel, Selective, and Orally Active α_1D Adrenoceptor Antagonist with Antiurinary Frequency Effects: Reducing Human Ether-a-go-go-Related Gene (hERG) Liabilities

A novel structural class of iminopyridine derivative <b>1</b> was identified as a potent and selective human α_1D adrenoceptor (α_1D adrenergic receptor; α_1D-AR) antagonist against α_1A- and α_1B-AR through screening of an in-house compound library. From initial structure–activity relationship studies, we found lead compound <b>9m</b> with hERG K⁺ channel liability. To develop analogues with reduced hERG K⁺ channel inhibition, a combination of site-directed mutagenesis and docking studies was employed. Further optimization led to the discovery of (<i>R</i>)-<b>9s</b> and <b>9u</b>, which showed antagonistic activity by a bladder strip test in rats with bladder outlet obstruction, as well as ameliorated cystitis-induced urinary frequency in rats. Ultimately, <b>9u</b> was selected as a clinical candidate. This is the first study to show the utility of iminopyridine derivatives as selective α_1D-AR antagonists and evaluate their effects in vivo

Interlaboratory evaluation of LC–MS-basedbiomarker assays: supplementary materials

Validation of biomarker assays is crucial for effective drug development and clinical applications.Interlaboratory reproducibility is vital for reliable comparison and combination of data from differentcenters. This review summarizes interlaboratory studies of quantitative LC–MS-based biomarker assaysusing reference standards for calibration curves. The following points are discussed: trends in reports,reference and internal standards, evaluation of analytical validation parameters, study sample analysisand normalization of biomarker assay data. Full evaluation of these parameters in interlaboratory studiesis limited, necessitating further research. Some reports suggest methods to address variations in biomarkerassay data among laboratories, facilitating organized studies and data combination. Method validationacross laboratories is crucial for reducing interlaboratory differences and reflecting target biomarkerresponses.</p

Potent Body Weight-Lowering Effect of a Neuromedin U Receptor 2‑selective PEGylated Peptide

Neuromedin U (NMU) is a neuropeptide that mediates a variety of physiological functions via its receptors, NMUR1 and NMUR2. Recently, there has been an increased focus on NMU as a promising treatment option for diabetes and obesity. A short form of NMU (NMU-8) has potent agonist activity for both receptors but is metabolically unstable. Therefore, we designed and synthesized NMU-8 analogues modified by polyethylene glycol (PEG; molecular weight, 20 kDa; PEG20k) via a linker. 3-(2-Naphthyl)­alanine substitution at position 19 increased NMUR2 selectivity of NMU-8 analogues with retention of high agonist activity. Compound <b>37</b>, an NMUR2-selective PEG20k analogue containing piperazin-1-ylacetyl linker, exhibited a potent body weight-lowering effect with concomitant inhibition of food intake in a dose-dependent manner (body weight loss of 12.4% at 30 nmol/kg) by once-daily repeated dosing for 2 weeks in mice with diet-induced obesity