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MTBSTFA derivatization-LC-MS/MS approach for the quantitative analysis of endogenous nucleotides in human colorectal carcinoma cells

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Journal of Pharmaceutical Analysis 2022年第1期12卷 77-86页

作者： Huixia Zhang Yan Li Zheng Li Christopher Wai-Kei Lam Peng Zhu Caiyun Wang Hua Zhou Wei Zhang state key laboratory of quality research in chinese medicines Macao Institute for Applied Research in Medicine and HealthMacao University of Science and TechnologyTaipaMacaoChina

Endogenous ribonucleotides(RNs)and deoxyribonucleotides(dRNs)are important metabolites related to the pathogenesis of many *** light of their physiological and pathological significances,a novel and sensitive pre-column derivatization method with N-(t-butyldimethylsilyl)-N-methyltrifluoroacetamide(MTBSTFA)was developed to determine RNs and dRNs in human cells using high-performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS).A one-step extraction of cells with 85%methanol followed by a simple derivatization reaction within 5 min at room temperature contributed to shortened analysis *** derivatives of 22 nucleoside mono-,di-and tri-phosphates were retained on the typical C;column and eluted by ammonium acetate and acetonitrile in 9 *** these optimal conditions,good linearity was achieved in the tested calibration *** lower limit of quantitation(LLOQ)was determined to be 0.1-0.4μM for the tested RNs and 0.001-0.1μM for *** addition,the precision(CV)was<15%and the RSD of stability was lower than 10.4%.Furthermore,this method was applied to quantify the endogenous nucleotides in human colorectal carcinoma cell lines HCT 116 exposed to *** conclusion,our method has proven to be simple,rapid,sensitive,and *** may be used for specific expanded studies on intracellular pharmacology in vitro.

关键词： Ribonucleotides Deoxyribonucleotides Derivatization N-(t-butyldimethylsilyl)-N-methyltrifluoroacetamide High-performance liquid chromatography tandem mass spectrometry

Discovery of a novel EGFR ligand DPBA that degrades EGFR and suppresses EGFR-positive NSCLC growth

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Signal Transduction and Targeted Therapy 2020年第1期5卷 495-507页

作者： Nan Yao Chen-Ran Wang Ming-Qun Liu Ying-Jie Li Wei-Min Chen Zheng-Qiu Li Qi Qi Jin-Jian Lu Chun-Lin Fan Min-Feng Chen Ming Qi Xiao-Bo Li Jian Hong Dong-Mei Zhang Wen-Cai Ye College of Pharmacy Jinan UniversityGuangzhouChina Guangdong Province key laboratory of Pharmacodynamic Constituents of Traditional chinese medicine and New Drugs research Jinan UniversityGuangzhouChina School of medicine Jinan UniversityGuangzhouChina state key laboratory of quality research in chinese medicine Institute of Chinese Medical SciencesUniversity of MacaoMacaoChina

Epidermal growth factor receptor(EGFR)activation plays a pivotal role in EGFR-driven non-small cell lung cancer(NSCLC)and is considered as a key target of molecular targeted *** tyrosine kinase inhibitors(TKIs)have been canonically used in NSCLC ***,prevalent innate and acquired resistances and EGFR kinase-independent pro-survival properties limit the clinical efficacy of EGFR ***,the discovery of novel EGFR degraders is a promising approach towards improving therapeutic efficacy and overcoming drug ***,we identified a 23-hydroxybetulinic acid derivative,namely DPBA,as a novel EGFR small-molecule *** exerted potent in vitro and in vivo anticancer activity in both EGFR wild type and mutant NSCLC by degrading *** studies disclosed that DPBA binds to the EGFR extracellular domain at sites differing from those of EGF and *** did not induce EGFR dimerization,phosphorylation,and ubiquitination,but it significantly promoted EGFR degradation and repressed downstream survival *** analyses showed that DPBA induced clathrin-independent EGFR endocytosis mediated by flotillin-dependent lipid rafts and unaffected by EGFR *** of the early and late endosome markers rab5 and rab7 but not the recycling endosome marker rab11 was involved in DPBA-induced EGFR lysosomal *** present study offers a new EGFR ligand for EGFR pharmacological degradation and proposes it as a potential treatment for EGFR-positive NSCLC,particularly NSCLC with innate or acquired EGFR TKI *** can also serve as a chemical probe in the studies on EGFR trafficking and degradation.

关键词： NSCLC EGFR positive

Modern research thoughts and methods on bio-active components of TCM formulae

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chinese Journal of Natural medicines 2022年第7期20卷 481-493页

作者： TANG Yu-Ping XU Ding-Qiao YUE Shi-Jun CHEN Yan-Yan FU Rui-Jia BAI Xue key laboratory of Shaanxi Administration of Traditional chinese medicine for TCM Compatibility State Key Laboratory of Research&Development of Characteristic Qin Medicine Resources(Cultivation)Shaanxi Key Laboratory of Chinese Medicine Fundamentals and New Drug ResearchShaanxi Collaborative Innovation Center of Chinese Medicinal Resource IndustrializationShaanxi University of Chinese MedicineXi’an 712046China

TCM formulae are the important guidances for clinical application of traditional chinese medicines,which follow the principles of diagnosis and treatment in *** the bio-active components of TCM formulae is the key to the modernization and internationalization of traditional chinese *** the rapid development of modern instruments and technology,many new theories,methods and strategies are emerging,which upgrade the research of TCM formulae into a higher *** when the medicinal efficacy,bio-active components,function mechanism of TCM formulae are understood,we can guarantee TCM safety and quality *** this paper,we summarized the latest modern research thoughts and methods on bio-active components of TCM formulae including formula decomposition study,serum pharmacology and serum pharmacochemistry,association analysis,biochro-matography,network pharmacology,metabolomics and proteomics,so as to provide reference for the research and development of TCM in the future.

关键词： chinese medicine TCM formulae Bio-active component Morden research methods

A robust luminescent assay for screening alkyladenine DNA glycosylase inhibitors to overcome DNA repair and temozolomide drug resistance

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Journal of Pharmaceutical Analysis 2023年第5期13卷 514-522页

作者： Ying-Qi Song Guo-Dong Li Dou Niu Feng Chen Shaozhen Jing Vincent Kam Wai Wong Wanhe Wang Chung-Hang Leung state key laboratory of quality research in chinese medicine Institute of Chinese Medical SciencesUniversity of MacaoMacao999078China Department of Biomedical Sciences Faculty of Health SciencesUniversity of MacaoMacao999078China Institute of Medical research Northwestern Polytechnical UniversityXi'an710072China Dr.Neher's Biophysics laboratory for Innovative Drug Discovery State Key Laboratory of Quality Research in Chinese MedicineMacao University of Science and TechnologyMacao999078China

Temozolomide(TMZ)is an anticancer agent used to treat glioblastoma,typically following radiation therapy and/or surgical ***,despite its effectiveness,at least 50%of patients do not respond to TMZ,which is associated with repair and/or tolerance of TMZ-induced DNA *** have demonstrated that alkyladenine DNA glycosylase(AAG),an enzyme that triggers the base excision repair(BER)pathway by excising TMZ-induced N3-methyladenine(3meA)and N7-methylguanine lesions,is overexpressed in glioblastoma tissues compared to normal ***,it is essential to develop a rapid and efficient screening method for AAG inhibitors to overcome TMZ resistance in ***,we report a robust time-resolved photoluminescence platform for identifying AAG inhibitors with improved sensitivity compared to conventional steady-state spectroscopic *** a proof-of-concept,this assay was used to screen 1440 food and drug administration-approved drugs against AAG,resulting in the repurposing of sunitinib as a potential AAG *** restored glioblastoma(GBM)cancer cell sensitivity to TMZ,inhibited GBM cell proliferation and stem cell characteristics,and induced GBM cell cycle ***,this strategy offers a new method for the rapid identification of small-molecule inhibitors of BER enzyme activities that can prevent false negatives due to a fluorescent background.

关键词： Drug screening Alkyladenine DNA glycosylase N3-methyladenine Glioblastoma Temozolomide Sunitinib

Supramolecular Manipulation of Intracellular Acidic Vesicles to Improve Cellular Self-Repair and Self-Defense

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CCS Chemistry 2024年第6期6卷 1487-1498页

作者： Ziyi Wang Chen Sun Yu Dong Yuan-Fu Ding Chen Zhao Ian Wa Ho Jiahong Lu Ruibing Wang state key laboratory of quality research in chinese medicine Institute of Chinese Medical SciencesUniversity of MacaoTaipaMacao 999078China

Lysosomes are the“cell stomach”that digests various species including macromolecules and pathogens for self-repair and self-defense upon fusion with endosomes,autophagosomes,and ***,artificially manipulating intracellular vesicles could potentially promote lysosome-mediated intracellular digestion for disease prevention and ***,a supramolecular strategy to efficiently tether and fuse acidic vesicles(i.e.,lysosomes,late endosomes,autolysosomes,and phagosomes)has been developed for the first time to enhance the digestion of cellular wastes and foreign matter,including pathogens.A linear polyethylene glycol(PEG)derivative dually tagged with morpholine(MOR)and adamantane(ADA)on the opposite side,namely,MOR-PEG-ADA,was designed to target intracellular acidic vesicles via MOR and to decorate their surfaces with ***,the addition of cucurbit[7]uril(CB[7])grafted hyaluronic acid(HA)induced supramolecular tethering and fusion of acidic vesicles via strong host-guest interactions between CB[7]of CB[7]-HA and ADA residing on the surface of acidic *** a proof-ofconcept,the overall cellular metabolism,including endogenous autophagy and the exogenous endocytosis and phagocytosis,was effectively upregulated,demonstrating that supramolecular regulation of the dynamics of intracellular acidic vesicles may promote the lysosome’s digestion to significantly improve cellular self-repair and self-defense.

关键词： supramolecular chemistry organelle regulation intracellular acidic vesicles lysosomes selfdefense self-repair

Taohong Siwu Decoction:a classical chinese prescription for treatment of orthopedic diseases

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chinese Journal of Natural medicines 2024年第8期22卷 711-723页

作者： SHI Yunzhen WANG Shengpeng DENG Disi WANG Yitao Institute of chinese Medical Sciences State Key Laboratory of Quality Research in Chinese MedicineUniversity of MacaoMacao SAR 999078China Gynaecology Hospital of Chengdu University of Traditional Chinese MedicineChengdu 611137China Clinical Medical College Chengdu University of Traditional Chinese MedicineChengdu 610075China

The pathogenesis of orthopedic diseases is intimately linked to blood stasis,frequently arising from damage to primary and secondary blood *** disruption can lead to“blood leaving the meridians”or Qi stagnation,resulting in blood stasis *** Siwu Decoction(THSWD)is a renowned classical chinese medicinal formula extensively used to promote blood circulation and mitigate blood *** studies have demonstrated its significant therapeutic effects on various orthopedic conditions,particularly its anti-inflammatory and analgesic properties,as well as its efficacy in preventing deep vein thrombosis *** these findings,research on THSWD remains fragmented,and its interdisciplinary impact is *** review aims to provide a comprehensive evaluation of the efficacy and pharmacological mechanisms of THSWD in treating common orthopedic ***,we employ bibliometric analysis to explore research trends and hotspots related to *** hope this review will enhance the recognition and application of THSWD in orthopedic treatments and guide future research into its pharmacological mechanisms.

关键词： Taohong Siwu Decoction Classical prescriptions Orthopedic diseases Fracture Femoral head necrosis Joint replacement

Development of genipin crosslinked gelatin matrices on surface interaction: Enhancing the biocompatibility by attenuating sterile inflammation

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chinese Journal of Chemical Engineering 2021年第10期34卷 205-215页

作者： Qiang-Song Wang Gui-Fang Wang Hai-Yun Zhang Fa-Quan Zhao Yuan-Lu Cui Tianjin key laboratory of Biomedical Materials Institute of Biomedical EngineeringChinese Academy of Medical Science&Peking Union Medical CollegeTianjin 300192China state key laboratory of Component-based chinese medicine Research Center of Traditional Chinese MedicineTianjin University of Traditional Chinese MedicineTianjin 301617China

The inflammation can be stimulated by the surgical implantation and biomaterial presence through the foreign body via *** play a key role in the interaction of host tissue to implant *** present study,the immuno-inflammatory responses of genipin crosslinked gelatin matrices(GCGM)to macrophages in vitro and the host tissue surfaces in rats were *** results showed that the mechanical properties,swelling and degradation of gelatin matrices were improved by the crosslinking of genipin at physiological *** macrophage on the surface of GCGM could avoid to be *** interaction of macrophage and GCGM suggested that GCGM could reduce the inflammatory response with downregulating the production and m RNA expression of pro-inflammatory *** anti-inflammatory effect of GCGM was demonstrated to be related to inhibit nuclear factor kappa-B(NF-κB)signaling ***,gelatin matrices crosslinked with genipin could decrease the acute and chronic inflammatory response interacting with host tissue surfaces to enhance the biocompatibility in *** results showed that GCGM could avoid to active macrophages and were endowed with anti-inflammatory properties,suggesting the significant potential for clinical success with the development of immunomodulatory biomaterials.

关键词： Genipin Gelatin matrices Anti-inflammation Biocompatibility NF-κB

Liver metabolomics study reveals protective function of Phyllanthus urinaria against CCl_4-induced liver injury

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chinese Journal of Natural medicines 2017年第7期15卷 525-533页

作者： GUO Qing ZHANG Qian-Qian CHEN Jia-Qing ZHANG Wei QIU Hong-Cong ZHANG Zun-Jian LIU Bu-Ming XU Feng-Guo MOE key laboratory of Drug quality Control and Pharmacovigilance China Pharmaceutical University Nanjing 210009 China state key laboratory of natural medicine China Pharmaceutical University Nanjing 210009 China state key laboratory for quality research in chinese medicines Macao University of Science & Technology Avenida Wai Long Taipa Macao China Guangxi key laboratory of Traditional chinese medicine quality Standards Nanning 530022 China

Phyllanthus Urinaria L.(PUL) is a traditional chinese medicine used to treat hepatic and renal disorders. However, the mechanism of its hepatoprotective action is not fully understood. In the present study, blood biochemical indexes and liver histopathological changes were used to estimate the extent of hepatic injury. GC/MS and LC/MS-based untargeted metabolomics were used in combination to characterize the potential biomarkers associated with the protective activity of PUL against CCl_4-induced liver injury in rats. PUL treatment could reverse the increase in ALT, AST and ALP induced by CCl_4 and attenuate the pathological changes in rat liver. Significant changes in liver metabolic profiling were observed in PUL-treated group compared with liver injury model group. Seventeen biomarkers related to the hepatoprotective effects of PUL against CCl_4-induced liver injury were screened out using nonparametric test and Pearson's correlation analysis(OPLS-DA). The results suggested that the potential hepatoprotective effects of PUL in attenuating CCl_4-induced hepatotoxicity could be partially attributed to regulating L-carnitine, taurocholic acid, and amino acids metabolism, which may become promising targets for treatment of liver toxicity. In conclusion, this study provides new insights into the mechanism of the hepatoprotection of Phyllanthus Urinaria.

关键词： Metabolomics Hepatic protection Phyllanthus Urinaria Carbon tetrachloride

Paeoniflorin Promotes Angiogenesis in A Vascular Insufficiency Model of Zebrafish in vivo and in Human Umbilical Vein Endothelial Cells in vitro

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chinese Journal of Integrative medicine 2018年第7期24卷 494-501页

作者： XIN Qi-qi YANG Bin-rui ZHOU He-feng WANG Yan YI Bo-wen CONG Wei-hong LEE Simon Ming-Yuen CHEN Ke-ji Clinical Medical College Beijing University of Chinese Medicine state key laboratory of quality research in chinese medicine Institute of Chinese Medical Sciences University of Macao laboratory of Cardiovascular Diseases Xiyuan Hospital China Academy of Chinese Medical Sciences Department of Pharmaceutical Preparation Xiyuan Hospital China Academy of Chinese Medical Sciences

Objective： To investigate the pro-angiogenic effects of paeoniflorin（PF） in a vascular insufficiency model of zebrafish and in human umbilical vein endothelial cells（HUVECs）. Methods： In vivo, the pro-angiogenic effects of PF were tested in a vascular insufficiency model in the Tg（fli-1：EGFP）y1 transgenic zebrafish. The 24 h post fertilization（hpf） embryos were pretreated with vascular endothelial growth factor（VEGF） receptor tyrosine kinase inhibitor Ⅱ（VRI） for 3 h to establish the vascular insufficiency model and then post-treated with PF for 24 h. The formation of intersegmental vessels（ISVs） was observed with a fluorescence microscope. The m RNA expression of fms-like tyrosine kinase-1（flt-1）, kinase insert domain receptor（kdr）, kinase insert domain receptor like（kdrl） and von Willebrand factor（v WF） were analyzed by real-time polymerase chain reaction（PCR）. In vitro, the pro-angiogenic effects of PF were observed in HUVECs in which cell proliferation, migration and tube formation were assessed. Results： PF（6.25–100 μmol/L） could rescue VRI-induced blood vessel loss in zebrafish and PF（25–100 μmol/L）, thereby restoring the m RNA expressions of flt-1, kdr, kdrl and v WF, which were down-regulated by VRI treatment. In addition, PF（0.001–0.03 μmol/L） could promote the proliferation of HUVECs while PF stimulated HUVECs migration at 1.0–10 μmol/L and tube formation at 0.3 μmol/L. Conclusion： PF could promote angiogenesis in a vascular insufficiency model of zebrafish in vivo and in HUVECs in vitro.

关键词： angiogenesis paeoniflorin zebrafish human umbilical vein endothelial cell