Therapeutic antibodies are valued for their high specificity and selectivity in immu-notherapy.However,the potential toxicity they may elicit underscores the necessity of assessing their preclinical efficacy and safet...Therapeutic antibodies are valued for their high specificity and selectivity in immu-notherapy.However,the potential toxicity they may elicit underscores the necessity of assessing their preclinical efficacy and safety using suitable animal models.In this context,we review the various categories and applications of humanized mice,which have been engrafted with human cells or tissues to mimic the human immune system.These models are extensively utilized in the nonclinical assessment and development of various antibody drugs,acting as a conduit to clinical research.However,several challenges remain,including the limited lifespan of humanized mice,inadequate en-graftment of human cells,and the rudimentary nature of the immune environment in these models.The development of humanized immune system models in mice pre-sents both opportunities and challenges,potentially leading to new insights into the evolution and application of antibody therapeutics.展开更多
In our previous study, we have elucidated the chemical profile ofYGS40, a fraction of Yi-Gan San （YGS）, used for the treatment of Alzheimer＇s disease （AD）. Oxidative stress-induced apoptosis is implicated in neur...In our previous study, we have elucidated the chemical profile ofYGS40, a fraction of Yi-Gan San （YGS）, used for the treatment of Alzheimer＇s disease （AD）. Oxidative stress-induced apoptosis is implicated in neurodegenerative disorders such as AD. The aim of the present study was to explore the protective effects of YGS40 against hydrogen peroxide （H202）-induced apoptosis in PC12 cells and the underlying mechanisms. PC12 cells were exposed to 100 μmol·L 1 of H202 for 12 h with or without YGS40 pretreatment. Cytotoxicity was determined by MTT （3, （4, 5-dimethylthiazole-2-yl） 2, 5-diphenyl-tetrazolium bromide） and lactate dehydrogenase （LDH） release assays; apoptosis was detected by Annexin V/propidium iodide coupled staining and by determining caspase-3 activity and Bax and Bcl-2 protein levels. Mitochondrial membrane potential （MMP） was assessed by the retention of rhodamine123; and the activities of superoxide dismutase （SOD） and malondialdehyde （MDA） were measured using commercially available enzymatic kits. Pretreatment with YGS40 significantly prevented H2O2-induced cytotoxicity and protected the cells against H2O2-triggered apoptosis characterized by extemalization of membrane phosphatidylserine and caspase-3 activation and the increased ratio of Bax/Bcl-2 in PC12 cells. Further studies showed that YGS40 suppressed H2O2-induced MMP loss, increased SOD activity, and decreased MDA level. These findings suggest that YGS40 may be beneficial for the prevention and treatment of oxidative stress-mediated disorders.展开更多
Shikimic acid(SA) is the key synthetic material for the chemical synthesis of Oseltamivir, which is prescribed as the front-line treatment for serious cases of influenza. Multi-gene expression vector can be used for e...Shikimic acid(SA) is the key synthetic material for the chemical synthesis of Oseltamivir, which is prescribed as the front-line treatment for serious cases of influenza. Multi-gene expression vector can be used for expressing the plurality of the genes in one plasmid, so it is widely applied to increase the yield of metabolites. In the present study, on the basis of a shikimate kinase genetic defect strain Escherichia coli BL21(?aro L/aro K, DE3), the key enzyme genes aro G, aro B, tkt A and aro E of SA pathway were co-expressed and compared systematically by constructing a series of multi-gene expression vectors. The results showed that different gene co-expression combinations(two, three or four genes) or gene orders had different effects on the production of SA. SA production of the recombinant BL21-GBAE reached to 886.38 mg·L^(-1), which was 17-fold(P < 0.05) of the parent strain BL21(?aro L/aro K, DE3).展开更多
During the last few years, the preparation of novel fluorescent probes for the detection of carbon dioxide has attracted considerable attention since carbon dioxide plays extremely important roles in widespread fields...During the last few years, the preparation of novel fluorescent probes for the detection of carbon dioxide has attracted considerable attention since carbon dioxide plays extremely important roles in widespread fields including chemical, environmental, clinical analysis, and agri-food industry. This review focuses on the recent advances in the design principles, recognition mechanisms, and preparation of small-molecule fluorescent probes for the selective detection and monitoring of CO;. Moreover, their properties and functions will be discussed detailedly as well.展开更多
During the past few years, the construction of fluorescent supramolecular metallocycles has attracted extensive attention due to their diverse applications such as sensing, photoelectric devices, and mimicking complic...During the past few years, the construction of fluorescent supramolecular metallocycles has attracted extensive attention due to their diverse applications such as sensing, photoelectric devices, and mimicking complicated natural photo-processes. In this review, we will discuss how we entered the field of fluorescent supramolecular metallacycles and what we investigated in this field. The preparation of various fluorescent supramolecular metallacycles and their applications in monitoring the dynamics of coordination-driven self-assembly, sensing, catalysts, and supramolecular gels will be summarized.展开更多
A series of novel amide derivatives bearing an indazole moiety were synthesized and evaluated for their in vitro S-adenosylL-homocysteine hydrolase（SAHase） inhibitory activity. Among these compounds, 8b,8m, 8r and 8...A series of novel amide derivatives bearing an indazole moiety were synthesized and evaluated for their in vitro S-adenosylL-homocysteine hydrolase（SAHase） inhibitory activity. Among these compounds, 8b,8m, 8r and 8w showed better or similar inhibitory effects compared to the positive control aristeromycin. These results provide a novel lead for the discovery of more potent non-adenosine analogs as SAHase inhibitors.展开更多
A novel series of pyrazolo[3,4-c]pyrazol and pyrrolo[2,3-c]pyrazol derivatives were designed,synthesized and biologically evaluated as potential Bruton's tyrosine kinase(BTK)inhibitors.Ten compounds exhibited vari...A novel series of pyrazolo[3,4-c]pyrazol and pyrrolo[2,3-c]pyrazol derivatives were designed,synthesized and biologically evaluated as potential Bruton's tyrosine kinase(BTK)inhibitors.Ten compounds exhibited variant inhibitory activities against BTK in vitro,and 8a showed the highest potency(IC50=127 nmol/L against BTK enzyme).The molecular docking of compound 8a was performed to understand the probable binding mode for this novel pyrazolo[3,4-c]pyrazol derivatives with BTK,which provided a comprehensive guide for further structural modification.展开更多
The title compound（2S,3S,E）-4-（3,4-dihydroxybenzylidene）-3-（3,4-dihydroxyphenyl）-5-oxotetrahydrofuran-2-carboxylic acid was synthesized and the 2D structure was characterized by ^1H-NMR ^13C-NMR and LCMS. The ab...The title compound（2S,3S,E）-4-（3,4-dihydroxybenzylidene）-3-（3,4-dihydroxyphenyl）-5-oxotetrahydrofuran-2-carboxylic acid was synthesized and the 2D structure was characterized by ^1H-NMR ^13C-NMR and LCMS. The absolute configuration was determined by single-crystal X-ray diffraction of the key intermediate（2S,3S,E）-ethyl 4-（3,4-dimethoxybenzylidene）-3-（3,4-dimethoxyphenyl）-5-oxotetrahydrofuran-2-carboxylate. The crystal belongs to tetragonal system, space group P41 with a = 13.0665（2）, b = 13.0665（2）, c = 13.4735（2）A, V = 2300.4（6） A^3, Z = 4, Dc = 1.278 g/cm^3, μ（CuKα） = 0.801 mm^-1, F（000） = 936, S = 1.050, R = 0.0364 and wR（I 〉 2σ（I）） = 0.1071. X-ray diffraction results showed that the molecular structure is stabilized totally by Van der Waals force. The antibacterial activity tests showed that the title compound possesses slight synergistic effect against MRSA and Acinetobacter baumanii.展开更多
A novel route of enzalutamide was developed in five steps.Starting from 4-amino-2-(trifluoromethyl)benzonitrile(7)and Boc-2-aminoisobutyric acid(16),condensation,deprotection,Ullmann coupling,cyclization and amination...A novel route of enzalutamide was developed in five steps.Starting from 4-amino-2-(trifluoromethyl)benzonitrile(7)and Boc-2-aminoisobutyric acid(16),condensation,deprotection,Ullmann coupling,cyclization and amination provided enzalutamide in 41.0%total yield.This route avoids the using of toxic chemical,unstable intermediate and high-risk reaction.It is a potential efficient and economical procedure for industrialization.展开更多
Epirubicin,an important anthracycline-type antitumor drug,is industrially produced through a chemical semisynthetic process.The production of epirubicin by genetic engineering microorganism directly is a more sustaina...Epirubicin,an important anthracycline-type antitumor drug,is industrially produced through a chemical semisynthetic process.The production of epirubicin by genetic engineering microorganism directly is a more sustainable alternative.But its yield was below 1mg/L in previous studies.Here the epirubicin was formed by introducing heterologous Streptomyces avermitilis aveBIV gene into high-doxorubicin producing S.peucetius SIPI-11 dnrn V mutant blocked in the biosynthesis of daunosamine,the deoxysugar component of doxorubicin,and a recombinant strain EPI-1 was constructed.The recombinant strain EPI-1 produced 47mg/L epirubicin,which was more than 47-fold higher than the engineered strain ever reported,indicating using overproduction strains as hosts is an efficient way to enhance epirubicin yield.Furthermore,to increase the yield of epirubicin in recombinant S.peucetius,we overexpressed the related genes of daunosamine formation and gtycosylation in the epirubicin biosynthesis pathway.The engineered strain EPI-2 produced 95mg/L epirubicin,which increased by 102%compared with EPI-1,which also demonstrated that daunosamine formation and glycosylation vcere the rate-limiting steps in the biosynthesis of epirubicin.Finally,the EPI-2 was cultured in a 5L fermenter,and the maximum titer of epirubicin reached 110mg/L at 168h.The results showed that S.peucetius EPI-2 has potential industrial application in epirubicin production by one-step fermentation.展开更多
Four new 3,4-secocycloartane triterpenoids,pseudolactones A-D(1-4),were isolated from the ethanol extract of the cones of Pseudol arixamabilis.Their structures were established by extensive 1D-and 2D-NMR experiments.T...Four new 3,4-secocycloartane triterpenoids,pseudolactones A-D(1-4),were isolated from the ethanol extract of the cones of Pseudol arixamabilis.Their structures were established by extensive 1D-and 2D-NMR experiments.The cones of P.arixamabilis are enriched in the ring-expanded or cleaved cycloartane triterpenoids.This work provides new insight into cycloartane triterpenoids from the cones of P.arixamabilis.展开更多
AIM: A quantitative ELISA kit for the detection of human secreted CD306/LAIR-2 was developed using monoclonal and polyclonal antibodies which were raised against a highly purified recombinant human secreted CD306/LAIR...AIM: A quantitative ELISA kit for the detection of human secreted CD306/LAIR-2 was developed using monoclonal and polyclonal antibodies which were raised against a highly purified recombinant human secreted CD306/LAIR-2. METHODS: Anti-human secreted CD306/LAIR-2 monoclonal and polyclonal antibodies were raised by immunizing mouse or rabbit with recombinant human secreted CD306/LAIR-2. The monoclonal antibody was purified by protein G affinity, whereas the polyclonal antibody was purified by protein A affinity. The best match pair of antibodies were found and used to develop a double antibody sandwich ELISA kit for the detection of human secreted CD306/LAIR-2 in human samples. RESULTS: A human secreted CD306/LAIR-2 ELISA kit was formulated with highly purified recombinant human secreted CD306/LAIR-2, highly specific monoclonal and polyclonal antibodies. This kit realized the quantitative measurement of recombinant human CD306/LAIR-2 and natural CD306/LAIR-2 in human serum samples. CONCLUSIONS: The developed human secreted CD306/LAIR-2 ELISA kit is a reliable quantitation immunoassay kit.展开更多
Imaging detection of interlinked dual proteases is imperative for precise tumor imaging,which remains challenging due to limited modification position of specific substrate and possible steric hindrance.Herein,we have...Imaging detection of interlinked dual proteases is imperative for precise tumor imaging,which remains challenging due to limited modification position of specific substrate and possible steric hindrance.Herein,we have developed a unimolecular chemiluminescent probe(LGP-CL)tandemly activated by two proteases interlinked with liver cancer to achieve precise tumor imaging.Probe LGP-CL consists of a phenoxy-dioxetane scaffold caged by a tripeptide substrate(LGP,leucine-glycine-proline)as the sensing layer,which can be cleaved sequentially by aminopeptidase N(APN)and dipeptidyl peptidase IV(DPPIV)to turn on a strong chemiluminescent signal,and silenced by specific inhibitor of each enzyme,which accounts for an integrated logic gate(AND,OR and INHIBIT).The successful cleavage of dual proteases on the metabolic site depends on the proper structure of the tripeptide substrate,as confirmed by two probes design.Probe LGP-CL(LGP as the substrate)enables the excellent“dual-lock-dual-key”fit with a382-fold enhancement of chemiluminescent emission while no obvious signal is observed by using GPLCL(GPL as the substrate).By virtue of its rapid response(several minutes),high sensitivity and good cell viability,probe LGP-CL has been utilized to evaluate upregulated levels of proteases in vitro and in living systems,especially to distinguish liver tumor cells(Hep G2)from others(LO_(2),MCF-7,MCF-10a and RAW264.7).Overall,the newly developed CL probe may facilitate rapid investigation into the role played by proteases in liver diseases,enabling timely selection appropriate treatment.Therefore,our work not only sheds light on the rational design of optical probes for dual protease imaging,but provides a promising tool for clinical diagnosis and even drug discovery.展开更多
Gnaphalium affine D. Don, a medicinal and edible plant, has been used to treat gout in traditional Chinese medicine and popularly consumed in China for a long time. A detailed phytochemical investigation on the aerial...Gnaphalium affine D. Don, a medicinal and edible plant, has been used to treat gout in traditional Chinese medicine and popularly consumed in China for a long time. A detailed phytochemical investigation on the aerial part of G. affine led to the isolation of two new esters of caffeoylquinic acid named(-) ethyl 1, 4-di-O-caffeoylquinate(1) and(-) methyl 1, 4-di-O-caffeoylquinate(2), together with 35 known compounds(3-37). Their structures were elucidated by spectroscopic data and first-order multiplet analysis. All the isolated compounds were tested for their xanthine oxidase inhibitory activity with an in vitro enzyme inhibitory screening assay. Among the tested compounds, 1(IC_(50) 11.94 μmol·L^(-1)) and 2(IC_(50) 15.04 μmol·L^(-1)) showed a good inhibitory activity. The current results supported the medical use of the plant.展开更多
AIM: To study the chemical constituents of the roots and stem bark of Kadsura coccinea. METHOD: Compounds were isolated by column chromatography on silica gel and Sephadex LH-20, and finally purified by prep-HPLC. The...AIM: To study the chemical constituents of the roots and stem bark of Kadsura coccinea. METHOD: Compounds were isolated by column chromatography on silica gel and Sephadex LH-20, and finally purified by prep-HPLC. Their structures were elucidated by extensive spectroscopic methods, including 1D- and 2D-NMR, and HR-ESI-MS. RESULTS: Two compounds were determined as(7′S,8′S,8R)-(8β,8′α)- dimethyl-4,4′-dihydroxy-5,3′-dimethoxy-5′-cyclolignan glucoside(1) and micrandiactone H(2), respectively. CONCLUSION: Compunds 1 and 2 are new and neither showed inhibitory effects on nitric oxide(NO) production in lipopolysaccharide-induced RAW264.7 macrophages.展开更多
Bufalin is one of the main pharmacological and toxicological components of Venenum Bufonis and many traditional Chinese medicine preparations.The cardiotoxicity clearly limits its application to patients living in cou...Bufalin is one of the main pharmacological and toxicological components of Venenum Bufonis and many traditional Chinese medicine preparations.The cardiotoxicity clearly limits its application to patients living in countries.Hence,an investigation of its toxicological mechanism is helpful for new drug development and treatment of the related clinical adverse reactions.We investigate the cardiotoxicity of bufalin using human induced pluripotent stem cells-derived cardiomyocytes(hiPSC-CMs)(0.003–0.1μmol·L–1),human induced pluripotent stem cells-derived cardiomyocytes(hiPSC-CMs)(0.03–0.3μmol·L–1)and eight human cardiac ion channel currents(0.01–100μmol·L–1)combined with an impedance-based bioanalytical and patch clamp method.Biphasic effect of bufalin on the contractility in hiPSC-CMs,which has been shown to strengthen myocardial contractility,accelerate conduction,and increase beating rate at the earlier stage of administration,whereas weakened myocardial contractility,abolished conduction,and ceased beating rate at the later stage of administration.Bufalin decreased the action potential duration(Action potential duration at 30%,50%and 90%repolarization),cardiac action potential amplitude,and maximal depolarization rate and depolarized the resting membrane potential of hiPSC-CMs.Spontaneous beating rates of hiPSC-CMs were markedly increased at 0.03μmol·L–1,while were weakened at 0.3μmol·L–1 after application.Bufalin blocks INav1.5 in a concentration-dependent manner with half maximal inhibitory concentration of 74.5μmol·L–1.Bufalin respectively increased the late sodium current and Na+-Ca2+exchange current with a concentration for 50%of maximal effect of 2.48 and 66.06μmol·L–1 in hiPSC-CMs.Whereas,bufalin showed no significant effects on other cardiac ion channel currents.The enhancement of the late sodium current is one of the main mechanism for cardiotoxicity of bufalin.展开更多
Cerebral ischemia-reperfusion injury(CI/RI)remains the main cause of disability and death in stroke patients due to lack of effective therapeutic strategies.One of the main issues related to CI/RI treatment is the pre...Cerebral ischemia-reperfusion injury(CI/RI)remains the main cause of disability and death in stroke patients due to lack of effective therapeutic strategies.One of the main issues related to CI/RI treatment is the presence of the blood-brain barrier(BBB),which affects the intracerebral delivery of drugs.Ginkgolide B(GB),a major bioactive component in commercially available products of Ginkgo biloba,has been shown significance in CI/RI treatment by regulating inflammatory pathways,oxidative damage,and metabolic disturbance,and seems to be a candidate for stroke recovery.However,limited by its poor hydrophilicity and lipophilicity,the development of GB preparations with good solubility,stability,and the ability to cross the BBB remains a challenge.Herein,we propose a combinatorial strategy by conjugating GB with highly lipophilic docosahexaenoic acid(DHA)to obtain a covalent complex GB-DHA,which can not only enhance the pharmacological effect of GB,but can also be encapsulated in liposomes stably.The amount of finally constructed Lipo@GB-DHA targeting to ischemic hemisphere was validated 2.2 times that of free solution in middle cerebral artery occlusion(MCAO)rats.Compared to the marketed ginkgolide injection,Lipo@GB-DHA significantly reduced infarct volume with better neurobehavioral recovery in MCAO rats after being intravenously administered both at 2 h and 6 h post-reperfusion.Low levels of reactive oxygen species(ROS)and high neuron survival in vitro was maintained via Lipo@GB-DHA treatment,while microglia in the ischemic brain were polarized from the pro-inflammatory M1 phenotype to the tissue-repairing M2 phenotype,which modulate neuroinflammatory and angiogenesis.In addition,Lipo@GB-DHA inhibited neuronal apoptosis via regulating the apoptotic pathway and maintained homeostasis by activating the autophagy pathway.Thus,transforming GB into a lipophilic complex and loading it into liposomes provides a promising nanomedicine strategy with excellent CI/RI therapeutic efficacy and industrialization prospects.展开更多
Romipeptides A and B(1 and 2),two new romidepsin derivatives,and three known compounds,chromopeptide A(3),romidepsin(4)and valine-leucine dipeptide(5)were isolated from the fermentation broth of Chromobacterium violac...Romipeptides A and B(1 and 2),two new romidepsin derivatives,and three known compounds,chromopeptide A(3),romidepsin(4)and valine-leucine dipeptide(5)were isolated from the fermentation broth of Chromobacterium violaceum No.968.Their structures were elucidated by interpretation of their UV,HR-ESI-MS and NMR spectra.The absolute configuration of compound 1 and 2 were established by single crystal X-ray diffraction analysis.Compounds 1–5 were evaluated for their anti-proliferative activities against three human cancer cell lines,SW620,HL60,and A549.The results showed most of these compounds exhibited antitumor activities in vitro,in which compound 2 displayed potent cytotoxicity to SW620,HL60 and A549 cell lines,with IC_(50) of 12.5,6.7 and 5.7 nmol·L^(–1),respectively.展开更多
Two new flavonoid glycosides, named viscumneoside XⅡ(1), and viscumneoside XⅢ(2);a new dihydrogen flavonoid glycoside product named viscumneoside XⅣ(3), were isolated from the aerial part of Viscum album, along wit...Two new flavonoid glycosides, named viscumneoside XⅡ(1), and viscumneoside XⅢ(2);a new dihydrogen flavonoid glycoside product named viscumneoside XⅣ(3), were isolated from the aerial part of Viscum album, along with seven known compounds(4-10). Their structures were identified by analysis of spectroscopic data. In addition, cytotoxicity assay showed that 1, 2 and 3 possessed significant inhibitory activities against C6, A549 and MDA-MB-231(the inhibition rate arrived about 50%, 70% and74% respectively with IC50 ≤ 60.00 μmol·L^-1), while the inhibition of TF-1 and Hela was not significant.展开更多
Our continued works on the chemical constituents of Ginkgo biloba(G.biloba)leaves has led to the isolation of two novel phenylbutenoids(1,2),along with five previously unidentified terpene glycosides(3−7).Among them,c...Our continued works on the chemical constituents of Ginkgo biloba(G.biloba)leaves has led to the isolation of two novel phenylbutenoids(1,2),along with five previously unidentified terpene glycosides(3−7).Among them,compounds 1 and 2 represent unique(Z)-phenylbutenoids,3−6 are megastigmane glycosides,and 7 is identified as a rare bilobanone glycoside(Fig.1).This study marks the first reported isolation of phenylbutenoid and bilobanone glycoside from G.biloba.The chemical structures of these compounds were elucidated through extensive spectroscopic analysis,including HR-ESI-MS and various 1D and 2D NMR experiments.Furthermore,the absolute configurations of these molecules were determined using Mosher’s method,ECD experiments,and Cu-KαX-ray crystallographic analyses.展开更多
基金supported by the Independent Research and Development Projects Foundation of Shanghai InnoStar Bio-Techology Co.,Ltd.(H23ZZYF01 and H24ZZYF01).
文摘Therapeutic antibodies are valued for their high specificity and selectivity in immu-notherapy.However,the potential toxicity they may elicit underscores the necessity of assessing their preclinical efficacy and safety using suitable animal models.In this context,we review the various categories and applications of humanized mice,which have been engrafted with human cells or tissues to mimic the human immune system.These models are extensively utilized in the nonclinical assessment and development of various antibody drugs,acting as a conduit to clinical research.However,several challenges remain,including the limited lifespan of humanized mice,inadequate en-graftment of human cells,and the rudimentary nature of the immune environment in these models.The development of humanized immune system models in mice pre-sents both opportunities and challenges,potentially leading to new insights into the evolution and application of antibody therapeutics.
基金supported by the National Natural Science Foundation of China(Nos.81274046 and 81373956)
文摘In our previous study, we have elucidated the chemical profile ofYGS40, a fraction of Yi-Gan San （YGS）, used for the treatment of Alzheimer＇s disease （AD）. Oxidative stress-induced apoptosis is implicated in neurodegenerative disorders such as AD. The aim of the present study was to explore the protective effects of YGS40 against hydrogen peroxide （H202）-induced apoptosis in PC12 cells and the underlying mechanisms. PC12 cells were exposed to 100 μmol·L 1 of H202 for 12 h with or without YGS40 pretreatment. Cytotoxicity was determined by MTT （3, （4, 5-dimethylthiazole-2-yl） 2, 5-diphenyl-tetrazolium bromide） and lactate dehydrogenase （LDH） release assays; apoptosis was detected by Annexin V/propidium iodide coupled staining and by determining caspase-3 activity and Bax and Bcl-2 protein levels. Mitochondrial membrane potential （MMP） was assessed by the retention of rhodamine123; and the activities of superoxide dismutase （SOD） and malondialdehyde （MDA） were measured using commercially available enzymatic kits. Pretreatment with YGS40 significantly prevented H2O2-induced cytotoxicity and protected the cells against H2O2-triggered apoptosis characterized by extemalization of membrane phosphatidylserine and caspase-3 activation and the increased ratio of Bax/Bcl-2 in PC12 cells. Further studies showed that YGS40 suppressed H2O2-induced MMP loss, increased SOD activity, and decreased MDA level. These findings suggest that YGS40 may be beneficial for the prevention and treatment of oxidative stress-mediated disorders.
基金supported by National Important Special Foundation of the New Drug Development(2012ZX 09301002-003 and 2014ZX09201001-012)Shanghai Innovation Action Plan of Science and Technology(14431905900)
文摘Shikimic acid(SA) is the key synthetic material for the chemical synthesis of Oseltamivir, which is prescribed as the front-line treatment for serious cases of influenza. Multi-gene expression vector can be used for expressing the plurality of the genes in one plasmid, so it is widely applied to increase the yield of metabolites. In the present study, on the basis of a shikimate kinase genetic defect strain Escherichia coli BL21(?aro L/aro K, DE3), the key enzyme genes aro G, aro B, tkt A and aro E of SA pathway were co-expressed and compared systematically by constructing a series of multi-gene expression vectors. The results showed that different gene co-expression combinations(two, three or four genes) or gene orders had different effects on the production of SA. SA production of the recombinant BL21-GBAE reached to 886.38 mg·L^(-1), which was 17-fold(P < 0.05) of the parent strain BL21(?aro L/aro K, DE3).
基金the National Natural Science Foundation of China (Nos. 21871092, 21672070, 31570360)Shanghai Pujiang Program (No. 18PJD015)+1 种基金STCSM (Nos. 16XD1401000, 17XD14230000)Shanghai Rising-Star Program (No. 16QB1403800) for the financial support
文摘During the last few years, the preparation of novel fluorescent probes for the detection of carbon dioxide has attracted considerable attention since carbon dioxide plays extremely important roles in widespread fields including chemical, environmental, clinical analysis, and agri-food industry. This review focuses on the recent advances in the design principles, recognition mechanisms, and preparation of small-molecule fluorescent probes for the selective detection and monitoring of CO;. Moreover, their properties and functions will be discussed detailedly as well.
基金the National Natural Science Foundation of China (Nos.21871092 and 21672070)Shanghai Pujiang Program (No.18PJD015)the State Key Laboratory of Fine Chemicals (No.KF1801) for the financial support
文摘During the past few years, the construction of fluorescent supramolecular metallocycles has attracted extensive attention due to their diverse applications such as sensing, photoelectric devices, and mimicking complicated natural photo-processes. In this review, we will discuss how we entered the field of fluorescent supramolecular metallacycles and what we investigated in this field. The preparation of various fluorescent supramolecular metallacycles and their applications in monitoring the dynamics of coordination-driven self-assembly, sensing, catalysts, and supramolecular gels will be summarized.
基金supported by National Natural Science Foundation of China (No. 81560653)Guangxi Natural Science Foundation of China (No. 2015GXNSFBA139124)
文摘A series of novel amide derivatives bearing an indazole moiety were synthesized and evaluated for their in vitro S-adenosylL-homocysteine hydrolase（SAHase） inhibitory activity. Among these compounds, 8b,8m, 8r and 8w showed better or similar inhibitory effects compared to the positive control aristeromycin. These results provide a novel lead for the discovery of more potent non-adenosine analogs as SAHase inhibitors.
基金This research work was financially supported by Shanghai Rising-Star Program(Grant No.16QB1403900).
文摘A novel series of pyrazolo[3,4-c]pyrazol and pyrrolo[2,3-c]pyrazol derivatives were designed,synthesized and biologically evaluated as potential Bruton's tyrosine kinase(BTK)inhibitors.Ten compounds exhibited variant inhibitory activities against BTK in vitro,and 8a showed the highest potency(IC50=127 nmol/L against BTK enzyme).The molecular docking of compound 8a was performed to understand the probable binding mode for this novel pyrazolo[3,4-c]pyrazol derivatives with BTK,which provided a comprehensive guide for further structural modification.
基金financially sponsored by the Shanghai Rising-Star Program(B)(16QB1403800)
文摘The title compound（2S,3S,E）-4-（3,4-dihydroxybenzylidene）-3-（3,4-dihydroxyphenyl）-5-oxotetrahydrofuran-2-carboxylic acid was synthesized and the 2D structure was characterized by ^1H-NMR ^13C-NMR and LCMS. The absolute configuration was determined by single-crystal X-ray diffraction of the key intermediate（2S,3S,E）-ethyl 4-（3,4-dimethoxybenzylidene）-3-（3,4-dimethoxyphenyl）-5-oxotetrahydrofuran-2-carboxylate. The crystal belongs to tetragonal system, space group P41 with a = 13.0665（2）, b = 13.0665（2）, c = 13.4735（2）A, V = 2300.4（6） A^3, Z = 4, Dc = 1.278 g/cm^3, μ（CuKα） = 0.801 mm^-1, F（000） = 936, S = 1.050, R = 0.0364 and wR（I 〉 2σ（I）） = 0.1071. X-ray diffraction results showed that the molecular structure is stabilized totally by Van der Waals force. The antibacterial activity tests showed that the title compound possesses slight synergistic effect against MRSA and Acinetobacter baumanii.
基金supported by Scientific Research Cadre Project(AMSCP)2021National Key Research and Development Program of China(No.2020YFA0509200 to C.Sheng)。
文摘A novel route of enzalutamide was developed in five steps.Starting from 4-amino-2-(trifluoromethyl)benzonitrile(7)and Boc-2-aminoisobutyric acid(16),condensation,deprotection,Ullmann coupling,cyclization and amination provided enzalutamide in 41.0%total yield.This route avoids the using of toxic chemical,unstable intermediate and high-risk reaction.It is a potential efficient and economical procedure for industrialization.
文摘Epirubicin,an important anthracycline-type antitumor drug,is industrially produced through a chemical semisynthetic process.The production of epirubicin by genetic engineering microorganism directly is a more sustainable alternative.But its yield was below 1mg/L in previous studies.Here the epirubicin was formed by introducing heterologous Streptomyces avermitilis aveBIV gene into high-doxorubicin producing S.peucetius SIPI-11 dnrn V mutant blocked in the biosynthesis of daunosamine,the deoxysugar component of doxorubicin,and a recombinant strain EPI-1 was constructed.The recombinant strain EPI-1 produced 47mg/L epirubicin,which was more than 47-fold higher than the engineered strain ever reported,indicating using overproduction strains as hosts is an efficient way to enhance epirubicin yield.Furthermore,to increase the yield of epirubicin in recombinant S.peucetius,we overexpressed the related genes of daunosamine formation and gtycosylation in the epirubicin biosynthesis pathway.The engineered strain EPI-2 produced 95mg/L epirubicin,which increased by 102%compared with EPI-1,which also demonstrated that daunosamine formation and glycosylation vcere the rate-limiting steps in the biosynthesis of epirubicin.Finally,the EPI-2 was cultured in a 5L fermenter,and the maximum titer of epirubicin reached 110mg/L at 168h.The results showed that S.peucetius EPI-2 has potential industrial application in epirubicin production by one-step fermentation.
基金supported by NSFC(31870327,81230090,81520108030,1302658)National Major Project of China(2018ZX09731016-005)+2 种基金The Key Research and Development Program of China(2017YFC1702002,2017YFC1700200)Professor of Chang Jiang Scholars Program,Scientific Foundation of Shanghai China(17431902800,16401901300)Shanghai Engineering Research Center for the Preparation of Bioactive Natural Products(10DZ2251300).
文摘Four new 3,4-secocycloartane triterpenoids,pseudolactones A-D(1-4),were isolated from the ethanol extract of the cones of Pseudol arixamabilis.Their structures were established by extensive 1D-and 2D-NMR experiments.The cones of P.arixamabilis are enriched in the ring-expanded or cleaved cycloartane triterpenoids.This work provides new insight into cycloartane triterpenoids from the cones of P.arixamabilis.
基金supported by the National Important Special Foundation of the New Drug Development(No.2010ZX 09401-403)the Shanghai Industry-University-Research-Medical Cooperation Project(No.12DZ1931903)
文摘AIM: A quantitative ELISA kit for the detection of human secreted CD306/LAIR-2 was developed using monoclonal and polyclonal antibodies which were raised against a highly purified recombinant human secreted CD306/LAIR-2. METHODS: Anti-human secreted CD306/LAIR-2 monoclonal and polyclonal antibodies were raised by immunizing mouse or rabbit with recombinant human secreted CD306/LAIR-2. The monoclonal antibody was purified by protein G affinity, whereas the polyclonal antibody was purified by protein A affinity. The best match pair of antibodies were found and used to develop a double antibody sandwich ELISA kit for the detection of human secreted CD306/LAIR-2 in human samples. RESULTS: A human secreted CD306/LAIR-2 ELISA kit was formulated with highly purified recombinant human secreted CD306/LAIR-2, highly specific monoclonal and polyclonal antibodies. This kit realized the quantitative measurement of recombinant human CD306/LAIR-2 and natural CD306/LAIR-2 in human serum samples. CONCLUSIONS: The developed human secreted CD306/LAIR-2 ELISA kit is a reliable quantitation immunoassay kit.
基金financial support by National Key R&D Program of China,MOST(No.2023YFC2510000)Shanghai Science and Technology(No.21N31900500)+2 种基金Shanghai Municipal Health Commission Project(No.202140016)Training Program for Outstanding Young Medical and Pharmaceutical Talents of Minhang District Health System(No.mwyjyx08)the Project of Basic Medicine funded by Fudan-Minhang Health Consortium(Nos.2021MHJC10 and 2023FM09)。
文摘Imaging detection of interlinked dual proteases is imperative for precise tumor imaging,which remains challenging due to limited modification position of specific substrate and possible steric hindrance.Herein,we have developed a unimolecular chemiluminescent probe(LGP-CL)tandemly activated by two proteases interlinked with liver cancer to achieve precise tumor imaging.Probe LGP-CL consists of a phenoxy-dioxetane scaffold caged by a tripeptide substrate(LGP,leucine-glycine-proline)as the sensing layer,which can be cleaved sequentially by aminopeptidase N(APN)and dipeptidyl peptidase IV(DPPIV)to turn on a strong chemiluminescent signal,and silenced by specific inhibitor of each enzyme,which accounts for an integrated logic gate(AND,OR and INHIBIT).The successful cleavage of dual proteases on the metabolic site depends on the proper structure of the tripeptide substrate,as confirmed by two probes design.Probe LGP-CL(LGP as the substrate)enables the excellent“dual-lock-dual-key”fit with a382-fold enhancement of chemiluminescent emission while no obvious signal is observed by using GPLCL(GPL as the substrate).By virtue of its rapid response(several minutes),high sensitivity and good cell viability,probe LGP-CL has been utilized to evaluate upregulated levels of proteases in vitro and in living systems,especially to distinguish liver tumor cells(Hep G2)from others(LO_(2),MCF-7,MCF-10a and RAW264.7).Overall,the newly developed CL probe may facilitate rapid investigation into the role played by proteases in liver diseases,enabling timely selection appropriate treatment.Therefore,our work not only sheds light on the rational design of optical probes for dual protease imaging,but provides a promising tool for clinical diagnosis and even drug discovery.
基金supported by the Natural Science Foundation of Shanghai(No.15ZR1440100)the National Natural Science Foundation of China(No.81603279)
文摘Gnaphalium affine D. Don, a medicinal and edible plant, has been used to treat gout in traditional Chinese medicine and popularly consumed in China for a long time. A detailed phytochemical investigation on the aerial part of G. affine led to the isolation of two new esters of caffeoylquinic acid named(-) ethyl 1, 4-di-O-caffeoylquinate(1) and(-) methyl 1, 4-di-O-caffeoylquinate(2), together with 35 known compounds(3-37). Their structures were elucidated by spectroscopic data and first-order multiplet analysis. All the isolated compounds were tested for their xanthine oxidase inhibitory activity with an in vitro enzyme inhibitory screening assay. Among the tested compounds, 1(IC_(50) 11.94 μmol·L^(-1)) and 2(IC_(50) 15.04 μmol·L^(-1)) showed a good inhibitory activity. The current results supported the medical use of the plant.
基金supported by the Program for New Century Excellent Talents in University(NCET-12-0977)the National New Drug Innovation Great Project of China(2011ZX09307-002-02)+1 种基金the Program for Changjiang Scholars and Innovative Research Team in a University(PCSIRT-IRT1193)the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
文摘AIM: To study the chemical constituents of the roots and stem bark of Kadsura coccinea. METHOD: Compounds were isolated by column chromatography on silica gel and Sephadex LH-20, and finally purified by prep-HPLC. Their structures were elucidated by extensive spectroscopic methods, including 1D- and 2D-NMR, and HR-ESI-MS. RESULTS: Two compounds were determined as(7′S,8′S,8R)-(8β,8′α)- dimethyl-4,4′-dihydroxy-5,3′-dimethoxy-5′-cyclolignan glucoside(1) and micrandiactone H(2), respectively. CONCLUSION: Compunds 1 and 2 are new and neither showed inhibitory effects on nitric oxide(NO) production in lipopolysaccharide-induced RAW264.7 macrophages.
基金supported by the National Foundation of Science and Technology(2018ZX09201017-008)。
文摘Bufalin is one of the main pharmacological and toxicological components of Venenum Bufonis and many traditional Chinese medicine preparations.The cardiotoxicity clearly limits its application to patients living in countries.Hence,an investigation of its toxicological mechanism is helpful for new drug development and treatment of the related clinical adverse reactions.We investigate the cardiotoxicity of bufalin using human induced pluripotent stem cells-derived cardiomyocytes(hiPSC-CMs)(0.003–0.1μmol·L–1),human induced pluripotent stem cells-derived cardiomyocytes(hiPSC-CMs)(0.03–0.3μmol·L–1)and eight human cardiac ion channel currents(0.01–100μmol·L–1)combined with an impedance-based bioanalytical and patch clamp method.Biphasic effect of bufalin on the contractility in hiPSC-CMs,which has been shown to strengthen myocardial contractility,accelerate conduction,and increase beating rate at the earlier stage of administration,whereas weakened myocardial contractility,abolished conduction,and ceased beating rate at the later stage of administration.Bufalin decreased the action potential duration(Action potential duration at 30%,50%and 90%repolarization),cardiac action potential amplitude,and maximal depolarization rate and depolarized the resting membrane potential of hiPSC-CMs.Spontaneous beating rates of hiPSC-CMs were markedly increased at 0.03μmol·L–1,while were weakened at 0.3μmol·L–1 after application.Bufalin blocks INav1.5 in a concentration-dependent manner with half maximal inhibitory concentration of 74.5μmol·L–1.Bufalin respectively increased the late sodium current and Na+-Ca2+exchange current with a concentration for 50%of maximal effect of 2.48 and 66.06μmol·L–1 in hiPSC-CMs.Whereas,bufalin showed no significant effects on other cardiac ion channel currents.The enhancement of the late sodium current is one of the main mechanism for cardiotoxicity of bufalin.
基金This research was funded by the National Natural Science Foundation of China(No.81773911,81690263 and 81573616)the Development Project of Shanghai Peak Disciplines-Integrated Medicine(No.20180101).
文摘Cerebral ischemia-reperfusion injury(CI/RI)remains the main cause of disability and death in stroke patients due to lack of effective therapeutic strategies.One of the main issues related to CI/RI treatment is the presence of the blood-brain barrier(BBB),which affects the intracerebral delivery of drugs.Ginkgolide B(GB),a major bioactive component in commercially available products of Ginkgo biloba,has been shown significance in CI/RI treatment by regulating inflammatory pathways,oxidative damage,and metabolic disturbance,and seems to be a candidate for stroke recovery.However,limited by its poor hydrophilicity and lipophilicity,the development of GB preparations with good solubility,stability,and the ability to cross the BBB remains a challenge.Herein,we propose a combinatorial strategy by conjugating GB with highly lipophilic docosahexaenoic acid(DHA)to obtain a covalent complex GB-DHA,which can not only enhance the pharmacological effect of GB,but can also be encapsulated in liposomes stably.The amount of finally constructed Lipo@GB-DHA targeting to ischemic hemisphere was validated 2.2 times that of free solution in middle cerebral artery occlusion(MCAO)rats.Compared to the marketed ginkgolide injection,Lipo@GB-DHA significantly reduced infarct volume with better neurobehavioral recovery in MCAO rats after being intravenously administered both at 2 h and 6 h post-reperfusion.Low levels of reactive oxygen species(ROS)and high neuron survival in vitro was maintained via Lipo@GB-DHA treatment,while microglia in the ischemic brain were polarized from the pro-inflammatory M1 phenotype to the tissue-repairing M2 phenotype,which modulate neuroinflammatory and angiogenesis.In addition,Lipo@GB-DHA inhibited neuronal apoptosis via regulating the apoptotic pathway and maintained homeostasis by activating the autophagy pathway.Thus,transforming GB into a lipophilic complex and loading it into liposomes provides a promising nanomedicine strategy with excellent CI/RI therapeutic efficacy and industrialization prospects.
文摘Romipeptides A and B(1 and 2),two new romidepsin derivatives,and three known compounds,chromopeptide A(3),romidepsin(4)and valine-leucine dipeptide(5)were isolated from the fermentation broth of Chromobacterium violaceum No.968.Their structures were elucidated by interpretation of their UV,HR-ESI-MS and NMR spectra.The absolute configuration of compound 1 and 2 were established by single crystal X-ray diffraction analysis.Compounds 1–5 were evaluated for their anti-proliferative activities against three human cancer cell lines,SW620,HL60,and A549.The results showed most of these compounds exhibited antitumor activities in vitro,in which compound 2 displayed potent cytotoxicity to SW620,HL60 and A549 cell lines,with IC_(50) of 12.5,6.7 and 5.7 nmol·L^(–1),respectively.
基金supported by the National Science and Technology Development Fund(No.2009ZX09301-007)the Scientific Research Projects of Shannxi Academy of Sciences(No.2016K-17)+4 种基金the Projects of Shaanxi Provincial Science and Technology Department(No.2016SF-264)Special Fund for Agro-scientific Research in The Public Interest(Grant 389,No.201203062)Support Plan for Young 392 Talents of Science and Technology Association of Colleges and Universities in Shaanxi Province 393(No.20150105)Key Research and Development Plan Project of Shaanxi Province(No.2018SF-034)Scientific Research from Shaanxi Provincial Department of Education(Nos.14JS100 and 17JK0243)
文摘Two new flavonoid glycosides, named viscumneoside XⅡ(1), and viscumneoside XⅢ(2);a new dihydrogen flavonoid glycoside product named viscumneoside XⅣ(3), were isolated from the aerial part of Viscum album, along with seven known compounds(4-10). Their structures were identified by analysis of spectroscopic data. In addition, cytotoxicity assay showed that 1, 2 and 3 possessed significant inhibitory activities against C6, A549 and MDA-MB-231(the inhibition rate arrived about 50%, 70% and74% respectively with IC50 ≤ 60.00 μmol·L^-1), while the inhibition of TF-1 and Hela was not significant.
基金supported by the National Natural Science Foundation of China(Nos.82141203,82003624,82004003,82004215)the Science and Technology Commission of Shanghai Municipality(Nos.20YF1458700,20YF1459000)+1 种基金the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(No.ZYYCXTDD-202004)Shanghai Frontiers Science Center of TCM Chemical Biology,Shanghai Engineering Research Center for the Preparation of Bioactive Natural Products(No.16DZ2280200).
文摘Our continued works on the chemical constituents of Ginkgo biloba(G.biloba)leaves has led to the isolation of two novel phenylbutenoids(1,2),along with five previously unidentified terpene glycosides(3−7).Among them,compounds 1 and 2 represent unique(Z)-phenylbutenoids,3−6 are megastigmane glycosides,and 7 is identified as a rare bilobanone glycoside(Fig.1).This study marks the first reported isolation of phenylbutenoid and bilobanone glycoside from G.biloba.The chemical structures of these compounds were elucidated through extensive spectroscopic analysis,including HR-ESI-MS and various 1D and 2D NMR experiments.Furthermore,the absolute configurations of these molecules were determined using Mosher’s method,ECD experiments,and Cu-KαX-ray crystallographic analyses.
