The β2-adrenergic receptor (β2AR) has been a model system for understanding regulatory mechanisms of G-protein–coupled receptor (GPCR) actions and plays a significant role in cardiovascular and pulmonary diseases. Because all known β-adrenergic receptor drugs target the orthosteric binding site of the receptor, we set out to isolate allosteric ligands for this receptor by panning DNA-encoded small-molecule libraries comprising 190 million distinct compounds against purified human β2AR. Here, we report the discovery of a small-molecule negative allosteric modulator (antagonist), compound 15 [([4-((2S)-3-(((S)-3-(3-bromophenyl)-1-(methylamino)-1-oxopropan-2-yl)amino)-2-(2-cyclohexyl-2-phenylacetamido)-3-oxopropyl)benzamide], exhibiting a unique chemotype and low micromolar affinity for the β2AR. Binding of 15 to the receptor cooperatively enhances orthosteric inverse agonist binding while negatively modulating binding of orthosteric agonists. Studies with a specific antibody that binds to an intracellular region of the β2AR suggest that 15 binds in proximity to the G-protein binding site on the cytosolic surface of the β2AR. In cell-signaling studies, 15 inhibits cAMP production through the β2AR, but not that mediated by other Gs-coupled receptors. Compound 15 also similarly inhibits β-arrestin recruitment to the activated β2AR. This study presents an allosteric small-molecule ligand for the β2AR and introduces a broadly applicable method for screening DNA-encoded small-molecule libraries against purified GPCR targets. Importantly, such an approach could facilitate the discovery of GPCR drugs with tailored allosteric effects.

There are many karst collapse columns in coal seam roof in the southern coal field in China, which are different from those in coal seam floor in the northern coal field, due to the stratum characteristics. The karst collapse column in coal seam roof tends to reactivate and conduct water and induce the serious water inrush disaster, when the karst collapse column communicates with the overlying aquifer. In order to reveal the evolution mechanism of water-conducting channel of collapse column in karst mining area of southwest China, the aquifers and water inflow rule in 1908 working face in Qianjin coal mine are analyzed. Besides, the particle size distribution and mineral component of collapse column are researched by the X-ray diffraction test and the screening method, which are the basis for researching the water inrush mechanism in karst collapse column. On this basis, the water inrush of roof collapse column under the influence of mining is researched by establishing the numerical calculation model with the UDEC numerical software. The results show that the water flowing into the 1908 working face comes from the Changxing formation aquifer and Yulongshan formation aquifer above the coal seam, and the proportion of coarse particles and fine particles in collapse column is 89.86% and 10.14%, respectively. With the advance of working face, the water-conducting channel connected the working face with the aquifer, or the surface is formed by collapse pits, karst caves, and collapse column. The research results can be treated as an important basis for the water-preserved mining in southern coal field in China.

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Layering of filling body is common in large-scale filling areas. In this paper, the cement–sand ratio of 1:8 is based on the configuration of 68%, 70%, 72% and 74%; four concentrations; and filling layers of one-, two-, three- and four-layered cemented filling samples. Combined with a uniaxial compression test and two-dimensional particle flow software (PFC2D), the mechanical properties and failure modes were explored. The results show that the concentration can strengthen the uniaxial compressive strength of the filling body while increasing the filling times weakens its power; therefore, the constitutive damage model was constructed. It was found that the initial layered damage existed in the layered filling, and the total damage showed an upward trend of first fast and then slow; the main failure modes of one-layer and two-layer backfills showed prominent shear failure characteristics, and the three-layer and four-layer fillings were closer to tensile failure. From the crack generation to the specimen failure, there is a mutual conversion between different energies.