Hydroxysteroid Dehydrogenase-Catalyzed Highly Regio-, Chemo-, and Enantioselective Hydrogenation of 3-Keto in Steroids.

A highly selective hydrogenation of 3-keto in steroids to 3-hydroxyl steroids catalyzed by hydroxysteroid dehydrogenases (HSDHs) was demonstrated. The Ct3α-HSDH-catalyzed hydrogenation generated 3α-hydroxyl steroids as the main enantiopure isomers in high yields, while the Ss3ß-HSDH catalytic system afforded 3ß-hydroxyl steroids in excellent yields. In both catalytic systems, the hydrogenation proceeded regioselectively at 3-keto with 7-, 11-, 17-, and 20-keto almost unreacted, and chemoselectively with the C═C bond and ester group unattacked. Our HSDH-promoted hydrogenation showed advantages like high regio-, chemo-, and enantioselectivity, good yields, mild conditions, a wide substrate scope, and being suitable for gram-scale synthesis. Notably, bioactive molecules like dehydroepiandrosterone, brienolone, and alfaxalone were obtained facilely in high yields via our hydrogenation approach.

Tumor cell-intrinsic SETD2 inactivation sensitizes cancer cells to immune checkpoint blockade through the NR2F1-STAT1 pathway.

BACKGROUND: Cancer immunotherapies can induce durable tumor regression, but most patients do not respond. SETD2 mutation has been linked to the efficacy of immune checkpoint inhibitors (ICIs) immunotherapy. The functional importance of the SETD2 inactivation and how to modulate immunotherapy response remains unclear. METHODS: To explore the function of SETD2 in immunotherapy, knockout and subsequent functional experiments were conducted. Bulk RNA-seq, ATAC-seq, Chip-seq and single-cell RNA-seq were performed to dissect the mechanism and explore the immune microenvironment of mouse tumor. Flow cytometry was used to assess cell surface antigen and intratumoral T cell levels. RESULTS: We comprehensively determine the effect of SETD2 inactivation in ICIs therapy and elucidate the mechanistic impact on tumor immunity. Murine syngeneic tumors harboring Setd2 inactivation are sensitive to ICIs. By bulk and single-cell RNA-seq, we further reveal that SETD2 inactivation reprograms intratumoral immune cells and inflames the tumor microenvironment, which is characterized by high infiltration of T cells and enhanced antigen presentation to activate CD8+ T cell-mediated killing. Mechanistically, via an integrated multiomics analysis using ATAC-seq, ChIP-seq and RNA-seq, we demonstrate that SETD2 inactivation reduces NR2F1 transcription by impairing H3K36me3 deposition and chromatin accessibility, which activates the STAT1 signaling pathway to promote chemokines and programmed cell death protein-1 (PD-1) expression and enhance antigen presentation. All these regulatory mechanisms synergistically promote the effects of anti-programmed cell death ligand 1 immunotherapy in Setd2-knockout syngeneic mouse models. The SETD2-NR2F1-STAT1 regulatory axis is conserved in human and murine cancers. Finally, cancer patients harboring SETD2 mutations who received ICIs show increased durable clinical benefits and survival. CONCLUSIONS: These findings provide novel insights into the biology of SETD2 inactivation regulation and reveal a new potential therapeutic biomarker for ICIs immunotherapy in various refractory cancers.

Thioalkynes in Ring Forming Reactions.

Organic cycles play an important role in chemistry, pharmacology and material science for their unique properties. Construction of organic cycles from thioalkynes attracted increasing attention due to the facile access of thioalkynes. 2H-Azirines were synthesized successfully from thioalkynyl oxime ethers. Cyclobutanes were formed through chiral titanium catalyzed cycloaddition of thioalkynes. Cyclopentenes were afforded by annulation of thioalkynes. Thioalkynes could be also applied to synthesize thiophenes, oxazoles, benzo[b]thiophenes, 2H-chromenes, 2-phenylbenzothiazoles, diazacyclobutene, etc. In this review, construction of organic cycles from thioalkynes were highlighted. Firstly, the property and application of organic cyclic compounds were simply introduced. After presenting the general methods to access organic cycles, applications of thioalkynes as synthons to prepare organic cycles were classified and presented in detail. Based on different kinds of organic cycles obtained from thioalkynes, organic reactions for synthesis of three-, four-, five-, six-membered as well as fused cycles would be summarized and the plausible reaction mechanisms could be presented if available.

Optimization and Impurity Control Strategy for Lithocholic Acid Production Using Commercially Plant-Sourced Bisnoralcohol.

In this study, lithocholic acid (LCA) was prepared using commercially available plant-sourced bisnoralcohol (BA), and the overall yield of the product was 70.6% for five steps. To prevent process-related impurities, the isomerizations of catalytic hydrogenation in the C4-C5 double bond and reduction of the 3-keto group were optimized. The double bond reduction isomerization was improved (5ß-H:5α-H = 97:3) using palladium-copper nanowires (Pd-Cu NWs) instead of Pd/C. The reduction of the 3-keto group was 100% converted to a 3α-OH product by 3α-hydroxysteroid dehydrogenase/carbonyl reductase catalysis. Moreover, the impurities during the optimization process were comprehensively studied. Compared with the reported synthesis methods, our developed method significantly improved the isomer ratio and overall yield, affording ICH-grade quality of LCA, and it is more cost-effective and suitable for large-scale production of LCA.

Activity of Anlotinib in the Second-Line Therapy of Metastatic Gastrointestinal Stromal Tumors: A Prospective, Multicenter, In Vitro Study.

BACKGROUND: Anlotinib is a multi-target tyrosine kinase inhibitor that can effectively inhibit tumor cell proliferation after receptor kinase activation caused by KIT gene mutation. METHODS: We tested the inhibitory effect of anlotinib in GIST cell lines with different gene mutations and evaluated the efficacy of anlotinib for patients with metastatic GIST after imatinib failure in a multicenter, single-arm, phase II study. RESULTS: In vitro, V654A mutation encoded by KIT exon 13 was intermediately sensitive to anlotinib. Moreover, anlotinib was able to partly suppress the activation loop mutation D820A from exon 17 while another activation loop mutation N822K, also from exon 17, was resistant to anlotinib. From September 2018 to October 2020, 64 patients from 9 Chinese medical centers were enrolled in this study. Seven patients had partial response and 39 patients had stable disease. The median PFS was 8.0 months. There was no statistical significance comparing with PFS of sunitinib second-line therapy at the same period. The most common adverse events related to anlotinib treatment were hypertension, neutropenia, and fatigue. CONCLUSION: Anlotinib showed moderate antitumor activity in drug-resistant GIST cell lines in vitro, and good PFS and better tolerance in second-line therapy study.

Research on the ozone formation sensitivity indicator of four urban agglomerations of China using Ozone Monitoring Instrument (OMI) satellite data and ground-based measurements.

Near surface ozone is a typical secondary pollutant, and is mostly generated by a series of complex photochemical reactions of volatile organic compounds (VOCs) and nitrogen oxides (NOx) in the air under sunlight. At present, a large number of studies have applied FNR (a ratio of formaldehyde (HCHO) to nitrogen dioxide (NO2) retrieved by satellite) indicator to study the ozone formation sensitivity (OFS). OFS analysis is critical for taking targeted ozone pollution prevention and control measures. Regional OFS can be more accurately diagnosed by utilizing localized FNR threshold. In this study, localized FNR thresholds were established for four severe ozone polluted urban agglomerations in China (Beijing-Tianjin-Hebei (BTH) region, Yangtze River Delta (YRD) region, Pearl River Delta (PRD) region, and Chengdu-Chongqing (CY) region), based on the statistical analysis between FNR (obtained from OMI observation, with daily transit time of approximately 13:45 local standard time) and ΔO3/ΔNO2 (the ratio of ozone change to nitrogen dioxide change between two consecutive months, obtained from ground measurements) from 2014 to 2016. And these thresholds were verified by the statistical analysis between FNR and ΔO3/O3 (ozone change rate between two consecutive months), and between FNR and O3 concentration during the OFS significant shift months. Furthermore, the results were also compared and verified with the method proposed by previous studies. The results indicate that there are significant regional dependences in the FNR threshold, and the lower-upper limits for the four urban agglomerations are as follows: 0.65-1.21 for BTH, 0.64-1.48 for the YRD, 1.25-2.39 for the PRD, and 1.44-3.69 for CY (FNR < lower limit indicates VOCs-limited regime; lower limit < FNR < upper limit indicates transitional regime; FNR > upper limit indicates NOx-limited regime). This method eliminates the problems associated with the undifferentiated use of FNR thresholds in different regions and significantly reduces the deviations for OFS.

[Coating-derived VOCs Emission Characteristics and Environmental Impacts from the Furniture Industry in Guangdong Province].

To determine the differences in emissions among different types of coatings, such as solvent-based, water-based, solvent-based ultra-violet(UV), water-based UV, and powder coatings, representative furniture manufacturing companies were selected for analysis. The emission concentrations and compositional characteristics of volatile organic compounds(VOCs) in different types of coatings were compared and studied. The ozone formation potential(OFP) and secondary organic aerosol formation potential(SOAFP) of the different types of coatings were also analyzed. Solvent-based coatings has higher TVOC concentrations, OFPs, and SOAFPs than water-based, solvent-based UV, water-based UV, and powder coatings. The concentrations and composition of VOCs emitted from the different types of coatings were also different. The main VOC groups of the solvent-based and solvent-based UV coatings were aromatic hydrocarbons and oxygenated volatile organic compounds(OVOCs). Specifically, the proportions of aromatic hydrocarbons are 41.91%-60.67% and 42.51%-43.00%, respectively, and the proportions of OVOCs were 24.75%-41.29% and 41.34%-43.21%, respectively. OVOCs accounted for the highest proportion of VOCs in the water-based, water-based UV, and powder coatings, at 54.02%-62.10%, 55.23%-64.81%, and 42.98%-46.45%, respectively. The major VOC compound of the solvent-based coatings was styrene(14.68%), and the main component of the water-based coatings was methylal(14.61%). The main species of VOCs from the solvent-based UV and water-based UV coatings were butyl acetate(15.36% and 20.56%, respectively). The most abundant species from the powder coatings was ethyl 3-ethoxy propionate(20.19%). Aromatic hydrocarbons were the most important contributor to the OFP of the solvent-based and solvent-based UV coatings, accounting for 79.84% and 80.32%, respectively. Aromatic hydrocarbons(51.48% and 36.71%) and OVOCs(42.30% and 41.03%) were the major contributors to the OFP of the water-based and water-based UV coatings, respectively. Aromatic hydrocarbons(43.46%), OVOCs(28.06%), and olefins(25.24%) were the main factors affecting the OFP of the powder coatings. Aromatic hydrocarbons dominate the SOAFP of solvent-based, water-based, solvent-based UV, water-based UV, and powder coatings, accounting for more than 99%.

E3 ligase MKRN3 is a tumor suppressor regulating PABPC1 ubiquitination in non-small cell lung cancer.

Central precocious puberty (CPP), largely caused by germline mutations in the MKRN3 gene, has been epidemiologically linked to cancers. MKRN3 is frequently mutated in non-small cell lung cancers (NSCLCs) with five cohorts. Genomic MKRN3 aberrations are significantly enriched in NSCLC samples harboring oncogenic KRAS mutations. Low MKRN3 expression levels correlate with poor patient survival. Reconstitution of MKRN3 in MKRN3-inactivated NSCLC cells directly abrogates in vitro and in vivo tumor growth and proliferation. MKRN3 knockout mice are susceptible to urethane-induced lung cancer, and lung cell-specific knockout of endogenous MKRN3 accelerates NSCLC tumorigenesis in vivo. A mass spectrometry-based proteomics screen identified PABPC1 as a major substrate for MKRN3. The tumor suppressor function of MKRN3 is dependent on its E3 ligase activity, and MKRN3 missense mutations identified in patients substantially compromise MKRN3-mediated PABPC1 ubiquitination. Furthermore, MKRN3 modulates cell proliferation through PABPC1 nonproteolytic ubiquitination and subsequently, PABPC1-mediated global protein synthesis. Our integrated approaches demonstrate that the CPP-associated gene MKRN3 is a tumor suppressor.

Relationships of ozone formation sensitivity with precursors emissions, meteorology and land use types, in Guangdong-Hong Kong-Macao Greater Bay Area, China.

Due to the influences of precursors emissions, meteorology, geography and other factors, ozone formation sensitivity (OFS) is generally spatially and temporally heterogeneous. This study characterized detailed spatial and temporal variations of OFS in Guangdong-Hong Kong-Macao Greater Bay Area (GBA) from 2012 to 2016 based on OMI satellite data, and analyzed the relationships of OFS with precursors emissions, meteorology and land use types (LUTs). From 2012 to 2016, the OFS tended to be NOx-limited in GBA, with the value of FNR (HCHO/NO2) increasing from 2.04 to 2.22. According to the total annual emission statistics of precursors, NOx emissions decreased by 33.1% and VOCs emissions increased by 35.2% from 2012 to 2016, directly resulting in OFS tending to be NOx-limited. The Grey Relation Analysis results show that total column water (TCW), surface net solar radiation (SSR), air temperature at 2 m (T2) and surface pressure (SP) are the top four meteorological factors with the greatest influences on OFS. There are significant positive correlations between FNR and T2, SSR, TCW, and significant negative correlations between FNR and SP. In GBA, the OFS tends to be NOx-limited regime in wet season (higher T2, SSR, TCW and lower SP) and VOCs-limited regime in dry season (lower T2, SSR, TCW and higher SP). The FNR displays obvious gradient variations on different LUTs, with the highest in "Rural areas", second in "Suburban areas" and lowest in "Urban areas".

Spectrum of activity of dasatinib against mutant KIT kinases associated with drug-sensitive and drug-resistant gastrointestinal stromal tumors.

BACKGROUND: The majority of GISTs express mutationally activated KIT. Imatinib and sunitinib are approved KIT-inhibiting therapies. Their efficacy is usually hampered by the acquired multiple secondary drug-resistance KIT mutations. The most problematic resistance subset is GISTs with acquisition of secondary mutations in the KIT activation loop. Here, we establish the spectrum of activity of dasatinib against a comprehensive collection of clinically relevant KIT mutants associated with drug-sensitive and drug-resistant GIST. METHODS: The cellular and in vitro activities of tyrosine kinase inhibitors (TKIs) against mutant KIT were assessed using a panel of engineered and GIST-derived cell lines. The in vivo activities of dasatinib were determined using TKI-resistant xenograft models. RESULTS: In engineered and GIST-derived cell lines, dasatinib potently inhibited KIT with primary mutations in exon 11 or 9 and a range of secondary imatinib-resistant mutations in exons 13 and 14, encoding the ATP-binding pocket, and in exons 17 and 18, encoding the activation loop, with the exception of a substitution at codon T670. Our data show that dasatinib is more potent than imatinib or sunitinib at inhibiting the activity of drug-resistant KIT mutants. Dasatinib also induces regression in GIST-derived xenograft models containing these secondary mutations. A major determinant of the efficacy of dasatinib for the treatment of advanced GIST is the activity of this inhibitor against KIT mutants. CONCLUSION: Dasatinib shows efficacy in cancer models, inhibiting a wide range of oncogenic primary and drug-resistant KIT mutants. These results have implications for the further development of dasatinib precision therapy in GIST patients.

[Volatile Organic Compound Emission Characteristics of Furniture Manufacturing Enterprises and the Influence on the Atmospheric Environment].

The furniture manufacturing industry is a typical industry with high pollution, low added value, relatively outdated technology and low levels of pollution control. The process of furniture manufacturing uses a large number of paints and adhesives, which emit a great quantity of volatile organic compounds (VOCs). The furniture manufacturing industry is a key industry for the control of VOCs in China. The VOCs emission characteristics and environmental impact of the furniture manufacturing industry has been studied in this work, which could be helpful for the Chinese government when formulating VOCs pollution control policy for this industry. In this study, a typical furniture manufacturing enterprise was chosen as the object. The emission concentration level and source profile of VOCs in a typical enterprise was obtained, and an assessment of the environmental impact of furniture manufacturing was developed. The results showed that the concentration of VOCs in the workshop ranged from 9.18 to 181.58 mg·m-3, the concentration of VOCs in the stack was 30.64-155.94 mg·m-3, and the treatment efficiency was 7.43%-67.14%. The main species of VOCs in the workshop were aromatic hydrocarbons, esters, and aldehydes and ketones; the main species of VOCs in the stack are esters and aromatic hydrocarbons, followed by alkanes, and the main VOCs in the industry are sec-butyl acetate, toluene, m-xylene, methylal and ethylbenzene. The average ozone generation potential (OFP) of workshop and stack VOCs was 258.01 and 289.14 mg·m-3, respectively, and the average secondary organic aerosol generation potential (SOAFP) of workshop and stack VOCs was 148.66 and 165.31 mg·m-3, respectively. The most important contribution to the OFP and SOAFP in each emission sector is aromatic hydrocarbons. The OFP and SOAFP in the edge-sealing workshop are large and the VOCs should be controlled. The main malodorous substances at the shop boundary are sec-butyl acetate, m-xylene, butyl acetate, p-xylene, ethylbenzene, 1-ethyl-3-methylbenzene, o-xylene, and toluene; the VOCs at the factory boundary produce almost no odor pollution. Targeted enhanced control of aromatic hydrocarbons and esters should be adopted to achieve effective emission reduction of VOCs in furniture manufacturing.

[Structure-activity relationship of diosgenin derivatives as Bcl-2 antagonists].

The purpose of this paper is to clarify the structure-activity relationship of anti-tumor activity of diosgenin derivatives in vitro. Study has found that diosgenin can inhibit the reproduction of tumor cells by inducing apoptosis and the main target spot of this effect is Bcl-2. Based on the characteristics of pharmacophoric points' of the three-dimensional pharmacophore for Bcl-2 inhibitors, we have docked lots of diosgenin derivatives with Bcl-2, then synthesized 31 compounds of them, finally assessed the anti-tumor activity of the diosgenin derivatives in vitro against A375, A549, HepG-2 and K562. Preliminary studies of SAR have indicated that the aliphatic esters, and aromatic esters of diosgenin without F ring have no anti-tumor activity in vitro. The triazole bromides of diosgenin all achieve fairly good anti-tumor activity in vitro, and those with larger hydrophobic group have the better activity. The stronger is the hydrogen bonding interaction and dipole-dipole interaction of the heterocyclic of diosgenin and diosgenin without F ring and the acid ester of diosgenin without F ring, the better is the activity of derivatives.