In this work,a new pyrylium derivatization-assisted liquid chromatography-mass spectrometry(LC-MS)method was developed for metabolite profiling of the glutathione anabolic pathway(GAP)in cancer tissues and cells.The p...In this work,a new pyrylium derivatization-assisted liquid chromatography-mass spectrometry(LC-MS)method was developed for metabolite profiling of the glutathione anabolic pathway(GAP)in cancer tissues and cells.The pyrylium salt of 6,7-dimethoxy-3-methyl isochromenylium tetrafluoroborate(DMMIC)was used to label the amino group of metabolites,and a reductant of dithiothreitol(DTT)was employed to stabilize the thiol group.By combining DMMIC derivatization with LC-MS,it was feasible to quantify the 13 main metabolites on the GAP in complex biological samples,which had good linearity(R^(2)=0.99810.9999),precision(interday precision of 1.6%e19.0%and intraday precision of 1.4%e19.8%)and accuracy(83.4%-115.7%).Moreover,the recovery assessments in tissues(82.5%e107.3%)and in cells(98.1%e118.9%)with GSH-^(13)C2,^(15)N,and Cys-^(15)N demonstrated the reliability of the method in detecting tissues and cells.Following a methodological evaluation,the method was applied successfully to investigate difference in the GAP between the carcinoma and para-carcinoma tissues of esophageal squamous cell carcinoma(ESCC)and the effect of p-hydroxycinnamaldehyde(CMSP)on the GAP in KYSE150 esophageal cancer cells.The results demonstrate that the developed method provides a promising new tool to elucidate the roles of GAP in physiological and pathological processes,which can contribute to research on drugs and diseases.展开更多
A rhodium(Ⅲ)-catalyzed hydrosilylation/cyclization reaction of cyclohexadienone-tetheredα,β-unsaturated aldehydes(1,6-dienes)with triethylsilane is described,providing a series of cishydrobenzofurans,cis-hydroindol...A rhodium(Ⅲ)-catalyzed hydrosilylation/cyclization reaction of cyclohexadienone-tetheredα,β-unsaturated aldehydes(1,6-dienes)with triethylsilane is described,providing a series of cishydrobenzofurans,cis-hydroindoles,and cishydroindenes bearing silyl enol ether in good to excellent yields and excellent stereoselectivities.Additionally,the versatility of this method was demonstrated through a gram-scale experiment and various downstream transformations,highlighting its utility.展开更多
Taking advantage of the Warburg effect in cancer cells, glucose conjugation has emerged as a useful strategy for targeted delivery of anticancer agents. Pristimerin is a naturally occurring triterpenoid that displays ...Taking advantage of the Warburg effect in cancer cells, glucose conjugation has emerged as a useful strategy for targeted delivery of anticancer agents. Pristimerin is a naturally occurring triterpenoid that displays potent but non-selective cytotoxicity. We developed a convergent and modular approach to construction of glucose-payload conjugates featuring copper-mediated azide-alkyne cycloaddition and prepared a glucose conjugate of pristimerin through this approach. The anticancer activity of this conjugate was evaluated in cancer cells and normal cells;however, the selectivity toward cancer cells was not significantly improved. We then examined the extracellular stability of the conjugate and found that its ester linkage was cleaved rapidly in Dulbecco’s Modified Eagle’s Medium at 37 °C, which resulted in the release of pristimerin. In fact, the inorganic components in this medium were sufficient to induce the cleavage.Given that the subtle difference between intrinsic stability and extracellular stability of the conjugate linker is often underappreciated, this work highlights the importance of the latter in the development of target-selective conjugates.展开更多
HuR(human antigen R), an mRNA-binding protein responsible for poor prognosis in nearly all kinds of malignancies, is a potential anti-tumor target for drug development. While screening HuR inhibitors with a fluorescen...HuR(human antigen R), an mRNA-binding protein responsible for poor prognosis in nearly all kinds of malignancies, is a potential anti-tumor target for drug development. While screening HuR inhibitors with a fluorescence polarization(FP) based high-throughput screening(HTS) system, the clinically used drug eltrombopag was identified. Activity of eltrombopag on molecular level was verified with FP, electrophoretic mobility shift assay(EMSA), simulation docking and surface plasmon resonance(SPR). Further, we showed that eltrombopag inhibited in vitro cell proliferation of multiple cancer cell lines and macrophages, and the in vivo anti-tumor activity was also demonstrated in a 4T1 tumor-bearing mouse model. The in vivo data showed that eltrombopag was efficient in reducing microvessels in tumor tissues. We then confirmed the HuR-dependent anti-angiogenesis effect of eltrombopag in 4T1 cells and RAW264.7 macrophages with qRT-PCR, HuR-overexpression and HuR-silencing assays, RNA stability assays, RNA immunoprecipitation and luciferase assays. Finally, we analyzed the in vitro anti-angiogenesis effect of eltrombopag on human umbilical vein endothelial cells(HUVECs) mediated by macrophages with cell scratch assay and in vitro Matrigel angiogenesis assay. With these data, we revealed the HuR-dependent anti-angiogenesis effect of eltrombopag in breast tumor, suggesting that the existing drug eltrombopag may be used as an anti-cancer drug.展开更多
Vinylphosphates play an important role in the fields of synthetic chemistry,pharmaceuticals and material science.It is a long-standing challenge to control the geometry of multi-substituted vinylphosphates.Herein,an e...Vinylphosphates play an important role in the fields of synthetic chemistry,pharmaceuticals and material science.It is a long-standing challenge to control the geometry of multi-substituted vinylphosphates.Herein,an efficient method for the synthesis of 2,2-disubstituted vinylphosphonates has been reported.The reaction proceeded via aryl to vinyl 1,4-palladium migration progress,in which two difficulties that the deactivation of palladium catalyst by the coordination of excess secondary phosphine oxide and the competitive cross-coupling to form arylphosphonates are successfully overcome.The judicious selection of phosphine ligand is the key to success.This protocol features excellent regio-and stereoselectivitiy,generally excellent reaction yields and broad substrate socpe.展开更多
Kosinostatin(KST)contains an uncommon aminopyrrole moiety,whose biosynthesis has remained elusive.Herein,aminopyrrolinic acid,which was generated by an L-ectoine synthase-like enzyme KstB3 via cyclization of L-glutami...Kosinostatin(KST)contains an uncommon aminopyrrole moiety,whose biosynthesis has remained elusive.Herein,aminopyrrolinic acid,which was generated by an L-ectoine synthase-like enzyme KstB3 via cyclization of L-glutamine,was identified to be the real substrate of adenylation enzyme KstB1.Subsequently,a FAD-dependent dehydrogenase KstB4 along with a transglutaminase-like enzyme KstB6 were also involved in formation of aminopyrrole.These results provided an unusual pathway for 2-aminopyrrole formation in KST biosynthesis.展开更多
Different from the established trans-endo-selective cyclization of alkyne-tethered electrophiles that involve an E/Z isomerization process,herein,the authors present a novel strategy to allow trans-exo-selective aryla...Different from the established trans-endo-selective cyclization of alkyne-tethered electrophiles that involve an E/Z isomerization process,herein,the authors present a novel strategy to allow trans-exo-selective arylative cyclization of 1,6-enynes.Through initiation of rhodium(III)-catalyzed C-H activation,a diverse range of N-heterocyclic directing groups,including pyridine,pyrazole,imidazo[1,2-a]pyridine,benzoxazole,benzothiazole,and purine,was feasible for the cascade transformation,exhibiting high efficiency(up to 92%yield),broad substrate scope,and excellent functional group compatibility.Moreover,the modification of natural products and pharmaceutical compounds was also demonstrated to showcase its synthetic utility.Based on density functional theory(DFT)calculations,a key three-membered ring intermediate through the insertion relay,rather than the direct E/Z isomerization of alkenyl rhodium species,controlled the stereochemical outcome for this trans-exo-selective cyclization.The subsequent ring-opening protonation of the more favored rotamer led to exclusive trans-exo-selectivity.展开更多
Molecular aggregation affects the electronic interactions between molecules and has emerged as a powerful tool in material science.Aggregate effect finds wide applications in the research of new physical phenomena;how...Molecular aggregation affects the electronic interactions between molecules and has emerged as a powerful tool in material science.Aggregate effect finds wide applications in the research of new physical phenomena;however,its value for chemical reaction development has been far less explored.Herein,we report the development of aggregation-enabled alkene insertion into carbon–halogen bonds.The spontaneous cleavage of C–X(X=Cl,Br,or I)bonds generates an intimate ion pair,which can be quickly captured by alkenes in an aggregated state.Additional catalysts or promoters are not necessary under such circumstances,and solvent quenching experiments indicate that the aggregated state is critical for achieving such sequences.The ionic insertion mode is supported by mechanistic studies,density functional theory calculations,and symmetry-adapted perturbation theory analysis.Results also show that the non-aggregated state may quench the transition state and terminate the insertion process.展开更多
基金We thank the Shanghai Municipal Committee of Science and Technology(Grant Nos.:20XD1423400,23ZR1460900 and 20DZ2201100)Shanghai Municipal Health Commission/Shanghai Municipal Administration of Traditional Chinese Medicine(Grant No.:ZY(2021e2023)-0501)+2 种基金Shanghai Science and Technology Development Fund from Central Leading Local Government(Grant No.:YDZX20223100001004)National Natural Science Foundation of China(Grant No.:21672249)Expenditure Budget Program of Shanghai University of Traditional Chinese Medicine(Grant Nos.:2020LK051,and 2021LK001).
文摘In this work,a new pyrylium derivatization-assisted liquid chromatography-mass spectrometry(LC-MS)method was developed for metabolite profiling of the glutathione anabolic pathway(GAP)in cancer tissues and cells.The pyrylium salt of 6,7-dimethoxy-3-methyl isochromenylium tetrafluoroborate(DMMIC)was used to label the amino group of metabolites,and a reductant of dithiothreitol(DTT)was employed to stabilize the thiol group.By combining DMMIC derivatization with LC-MS,it was feasible to quantify the 13 main metabolites on the GAP in complex biological samples,which had good linearity(R^(2)=0.99810.9999),precision(interday precision of 1.6%e19.0%and intraday precision of 1.4%e19.8%)and accuracy(83.4%-115.7%).Moreover,the recovery assessments in tissues(82.5%e107.3%)and in cells(98.1%e118.9%)with GSH-^(13)C2,^(15)N,and Cys-^(15)N demonstrated the reliability of the method in detecting tissues and cells.Following a methodological evaluation,the method was applied successfully to investigate difference in the GAP between the carcinoma and para-carcinoma tissues of esophageal squamous cell carcinoma(ESCC)and the effect of p-hydroxycinnamaldehyde(CMSP)on the GAP in KYSE150 esophageal cancer cells.The results demonstrate that the developed method provides a promising new tool to elucidate the roles of GAP in physiological and pathological processes,which can contribute to research on drugs and diseases.
基金financial support from the National Key R&D Program of China(No.2022YFF1202600)the National Natural Science Foundation of China(Nos.22001172,22071155,and22371188)+4 种基金the Science and Technology Commission of Shanghai Municipality(Nos.20XD1403600 and 20400750300)the Shanghai Municipal Education Commission(No.2019-01-07-00-10-E00072)the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(No.ZYYCXTD-202004)the Shanghai Municipal Health Commission/Shanghai Municipal Administration of Traditional Chinese Medicine[No.ZY(2021-2023)-0501]Organizational Key R&D Program of SHUTCM(No.2023YZZ01)。
文摘A rhodium(Ⅲ)-catalyzed hydrosilylation/cyclization reaction of cyclohexadienone-tetheredα,β-unsaturated aldehydes(1,6-dienes)with triethylsilane is described,providing a series of cishydrobenzofurans,cis-hydroindoles,and cishydroindenes bearing silyl enol ether in good to excellent yields and excellent stereoselectivities.Additionally,the versatility of this method was demonstrated through a gram-scale experiment and various downstream transformations,highlighting its utility.
基金supported by Ministry of Science and Technology (National Key Research and Development Program of China, No. 2018YFA0901900)National Natural Science Foundation of China (Nos. 21931014, U2002221, 21772225, 21572064, 81502956 and 21621002)+3 种基金Chinese Academy of Sciences (Strategic Priority Research Program, No. XDB20000000International Partner Program, No. 121731KYSB20190039Key Research Program of Frontier Sciences, No. QYZDB-SSW-SLH040)Science and Technology Commission of Shanghai Municipality (Nos. 20430713400, 17XD1404600 and JCYJ-SHFY-2022–005)。
文摘Taking advantage of the Warburg effect in cancer cells, glucose conjugation has emerged as a useful strategy for targeted delivery of anticancer agents. Pristimerin is a naturally occurring triterpenoid that displays potent but non-selective cytotoxicity. We developed a convergent and modular approach to construction of glucose-payload conjugates featuring copper-mediated azide-alkyne cycloaddition and prepared a glucose conjugate of pristimerin through this approach. The anticancer activity of this conjugate was evaluated in cancer cells and normal cells;however, the selectivity toward cancer cells was not significantly improved. We then examined the extracellular stability of the conjugate and found that its ester linkage was cleaved rapidly in Dulbecco’s Modified Eagle’s Medium at 37 °C, which resulted in the release of pristimerin. In fact, the inorganic components in this medium were sufficient to induce the cleavage.Given that the subtle difference between intrinsic stability and extracellular stability of the conjugate linker is often underappreciated, this work highlights the importance of the latter in the development of target-selective conjugates.
基金supported by National Natural Science Foundation of China(81573579)the New Interdisciplinary Subject Funding Program for Shanghai Traditional Chinese Medicine(E2-F18003,China)+3 种基金Shanghai Municipal Education Commission(SMEC,2019-01-07-00-10-E00072,China)Science and Technology Commission of Shanghai Municipality(STCSM,18401933500,China)Postdoctoral Science Foundation of China(A2-X1802408)Shanghai Municipal Commission of Health and Family Planning(2018YQ003,China)
文摘HuR(human antigen R), an mRNA-binding protein responsible for poor prognosis in nearly all kinds of malignancies, is a potential anti-tumor target for drug development. While screening HuR inhibitors with a fluorescence polarization(FP) based high-throughput screening(HTS) system, the clinically used drug eltrombopag was identified. Activity of eltrombopag on molecular level was verified with FP, electrophoretic mobility shift assay(EMSA), simulation docking and surface plasmon resonance(SPR). Further, we showed that eltrombopag inhibited in vitro cell proliferation of multiple cancer cell lines and macrophages, and the in vivo anti-tumor activity was also demonstrated in a 4T1 tumor-bearing mouse model. The in vivo data showed that eltrombopag was efficient in reducing microvessels in tumor tissues. We then confirmed the HuR-dependent anti-angiogenesis effect of eltrombopag in 4T1 cells and RAW264.7 macrophages with qRT-PCR, HuR-overexpression and HuR-silencing assays, RNA stability assays, RNA immunoprecipitation and luciferase assays. Finally, we analyzed the in vitro anti-angiogenesis effect of eltrombopag on human umbilical vein endothelial cells(HUVECs) mediated by macrophages with cell scratch assay and in vitro Matrigel angiogenesis assay. With these data, we revealed the HuR-dependent anti-angiogenesis effect of eltrombopag in breast tumor, suggesting that the existing drug eltrombopag may be used as an anti-cancer drug.
基金supported by the National Natural Science Foundation of China(21871284 and 91956113)the fellowship of China Postdoctoral Science Foundation(2021M692152)+2 种基金the Science and Technology Commission of Shanghai Municipality(20XD1423400,21ZR1482100 and 22ZR1458900)the Shanghai Municipal Education Commission(2019-01-07-0010-E00072)the Shanghai Frontiers Science Center for Traditional Chinese Medicine Chemical Biology.
文摘Vinylphosphates play an important role in the fields of synthetic chemistry,pharmaceuticals and material science.It is a long-standing challenge to control the geometry of multi-substituted vinylphosphates.Herein,an efficient method for the synthesis of 2,2-disubstituted vinylphosphonates has been reported.The reaction proceeded via aryl to vinyl 1,4-palladium migration progress,in which two difficulties that the deactivation of palladium catalyst by the coordination of excess secondary phosphine oxide and the competitive cross-coupling to form arylphosphonates are successfully overcome.The judicious selection of phosphine ligand is the key to success.This protocol features excellent regio-and stereoselectivitiy,generally excellent reaction yields and broad substrate socpe.
基金We thank Prof.Zixin Deng's Lab.at Shanghai Jiao Tong University and Dr.Chao Peng of the Mass Spectrometry System at the National Facility for Protein Science in Shanghai(NFPS),Zhangjiang Lab,China for obtaining MS data of proteins.Supporting grants including the National Natural Science Foundation of China(21632007 and 21621002)the Chinese Academy of Sciences(QYZDJ-SSW-SLH037)+1 种基金the Academy of Finland(285971)the Sigrid Juselius Foundation are highly acknowledged.
文摘Kosinostatin(KST)contains an uncommon aminopyrrole moiety,whose biosynthesis has remained elusive.Herein,aminopyrrolinic acid,which was generated by an L-ectoine synthase-like enzyme KstB3 via cyclization of L-glutamine,was identified to be the real substrate of adenylation enzyme KstB1.Subsequently,a FAD-dependent dehydrogenase KstB4 along with a transglutaminase-like enzyme KstB6 were also involved in formation of aminopyrrole.These results provided an unusual pathway for 2-aminopyrrole formation in KST biosynthesis.
基金Financial support was generously provided by the National Natural Science Foundation of China(nos.21871184,21871284,21702182,and 21873081),the Shanghai Municipal Education Commission(no.2019-01-07-00-10-E00072),the Science and Technology Commission of Shanghai Municipality(no.18401933500),the Strategic Priority Research Program of the Chinese Academy of Sciences(no.XDB 20020100),the Key Research Program of Frontier Science(no.QYZDYSSW-SLH026),the Fundamental Research Funds for the Central Universities(no.2019QNA3009),and the China Postdoctoral Science Foundation(no.2018M640546).Calculations were performed on the high-performance computing system at the Department of Chemistry,Zhejiang University.The authors thank Jie Sun(Shanghai Institute of Organic Chemistry)for X-ray crystallographic analysis.
文摘Different from the established trans-endo-selective cyclization of alkyne-tethered electrophiles that involve an E/Z isomerization process,herein,the authors present a novel strategy to allow trans-exo-selective arylative cyclization of 1,6-enynes.Through initiation of rhodium(III)-catalyzed C-H activation,a diverse range of N-heterocyclic directing groups,including pyridine,pyrazole,imidazo[1,2-a]pyridine,benzoxazole,benzothiazole,and purine,was feasible for the cascade transformation,exhibiting high efficiency(up to 92%yield),broad substrate scope,and excellent functional group compatibility.Moreover,the modification of natural products and pharmaceutical compounds was also demonstrated to showcase its synthetic utility.Based on density functional theory(DFT)calculations,a key three-membered ring intermediate through the insertion relay,rather than the direct E/Z isomerization of alkenyl rhodium species,controlled the stereochemical outcome for this trans-exo-selective cyclization.The subsequent ring-opening protonation of the more favored rotamer led to exclusive trans-exo-selectivity.
基金National Natural Science Foundation of China,Grant/Award Numbers:91940305,21933009,81874181,22271195,21871284Natural Science Foundation of Fujian Province,Grant/Award Number:2021J01525+3 种基金Major Scientific and Technological Special Project for“Significant New Drugs Creation”,Grant/Award Number:2019ZX09301158The Emerging Frontier Program of Hospital Development Centre,Grant/Award Number:SHDC12018107Shanghai Science and Technology Development Fund from Central Leading Local Government,Grant/Award Number:YDZX20223100001004Shanghai Municipal Health Commission/Shanghai Municipal Administration of Traditional Chinese Medicine,Grant/Award Number:ZY(2021-2023)−0501。
文摘Molecular aggregation affects the electronic interactions between molecules and has emerged as a powerful tool in material science.Aggregate effect finds wide applications in the research of new physical phenomena;however,its value for chemical reaction development has been far less explored.Herein,we report the development of aggregation-enabled alkene insertion into carbon–halogen bonds.The spontaneous cleavage of C–X(X=Cl,Br,or I)bonds generates an intimate ion pair,which can be quickly captured by alkenes in an aggregated state.Additional catalysts or promoters are not necessary under such circumstances,and solvent quenching experiments indicate that the aggregated state is critical for achieving such sequences.The ionic insertion mode is supported by mechanistic studies,density functional theory calculations,and symmetry-adapted perturbation theory analysis.Results also show that the non-aggregated state may quench the transition state and terminate the insertion process.
