Screening of mammalian target of rapamycin inhibitors in natural product extracts by capillary electrophoresis in combination with high performance liquid chromatography–tandem mass spectrometry

Zhang, Yanmei; Li, Feng; Li, Mingxia; Kang, Jingwu

doi:10.1016/j.chroma.2015.02.033

Cited by 20 publications

(15 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[29][30][31][32] XO inhibition analysis of screened compounds XO inhibitory activities assays of four screened compounds were carried out to evaluate the inhibition of each compound and verify the effectiveness of CU-LC-MS method. The binding degrees of four screened compounds (3) and biochanin A (4) at the optimum condition were 5.70, 8.28, 6.31 and 37.83%, respectively. These results demonstrated these four compounds possessed inhibition activities on XO.…”

Section: Structural Identificationmentioning

confidence: 98%

“…2 Hence, simple and effective methods aiming at directly screening natural product extracts would be greatly helpful for drug discovery. 3 Centrifugal ultrafiltration (CU) utilized centrifugal force and a semi-permeable membrane to retain suspended solids and high molecular weight solutes, while liquid and low molecular weight solutes were allowed to pass through depending on the nominal molecular weight cut-off of the membrane. 4 Based on these features, active compounds could be retained by membrane together with enzyme due to the binding with enzyme.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Analysis of Xanthine Oxidase Inhibitors fromPuerariae flosUsing Centrifugal Ultrafiltration Coupled with HPLC-MS

Liu¹,

Xiao²,

Ma³

et al. 2016

Journal of the Brazilian Chemical Society

View full text Add to dashboard Cite

In this study, centrifugal ultrafiltration coupled with high performance liquid chromatographymass spectrometry was utilized to screen and identify xanthine oxidase inhibitors from Puerariae flos extract. The experimental conditions of centrifugal ultrafiltration including xanthine oxidase concentration, incubation time, pH and temperature were optimized. At the optimum condition (xanthine oxidase concentration: 30.0 μg mL -1 , incubation time: 20 min, pH 7.0 and temperature: 25 °C), four compounds were successfully screened from P. flos extract and identified as tectoridin, daidzin, ononin and biochanin A. The yields of tectoridin, daidzin, ononin and biochanin A were 0.231, 0.117, 0.303 and 0.089 g from 50.0 g crude P. flos samples. The inhibitory activities of these compounds were verified by xanthine oxidase inhibition assays. The experimental half maximal inhibitory concentration (IC 50 ) values of tectoridin, daidzin, ononin and biochanin A were 88.5, 85.1, 88.8 and 87.0 μmol L -1 , and the binding degree of them were 5. 70, 8.28, 6.31 and 37.83% at the optimum condition, respectively. The proposed method provided a rapid and effective way to screen and analyze active compounds from natural products. Keywords: HPLC-MS, inhibitor, Puerariae flos, ultrafiltration, xanthine oxidase IntroductionNatural products have been used as the most consistently successful resources of new drug discovery for a long time because of their great diversity of the chemical structures and better drug-like properties compared to the synthetic compounds.1 However, natural products resources like plant extracts were very complex and usually contained various kinds of components. Separation and purification of natural compounds were time-consuming and laborious processes.2 Hence, simple and effective methods aiming at directly screening natural product extracts would be greatly helpful for drug discovery. Centrifugal ultrafiltration (CU) utilized centrifugal force and a semi-permeable membrane to retain suspended solids and high molecular weight solutes, while liquid and low molecular weight solutes were allowed to pass through depending on the nominal molecular weight cut-off of the membrane. 4 Based on these features, active compounds could be retained by membrane together with enzyme due to the binding with enzyme. Thus, CU became a useful technique for screening active compounds bound to biomacromolecules such as bovine serum albumin, 5 α-glucosidase, 6 quinone reductase-2, 7 deoxyribonucleic acid (DNA) and liposomes. 8,9 High performance liquid chromatography-mass spectrometry (HPLC-MS or LC-MS) has been widely applied for the simultaneous separation and identification of active compounds in complex mixtures. 10The combination of CU and LC-MS (CU-LC-MS) became a powerful tool in analyzing active compounds due to its simple operation, high speed and low sample consumption. Compared with immobilized enzyme screening assay, complex synthesis procedures could be avoided in CU-LC-MS as well.11 Moreover, the efficient screeni...

show abstract

Section: Structural Identificationmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Analysis of Xanthine Oxidase Inhibitors fromPuerariae flosUsing Centrifugal Ultrafiltration Coupled with HPLC-MS

Liu¹,

Xiao²,

Ma³

et al. 2016

Journal of the Brazilian Chemical Society

View full text Add to dashboard Cite

show abstract

“…This tandem technique integrates advantages of all kinds of apparatuses. Zhang with her teammates created a CE method in conjunction with LC‐MS/MS for screening inhibitors of protein kinase, mammalian target of rapamycin and epidermal growth factor receptor from natural products. This novel approach was proved stable enough to determine other kinds of inhibitors from natural products especially Chinese herbs.…”

Section: Screening Active Compounds Using Cementioning

confidence: 99%

Screening bioactive compounds from natural product and its preparations using capillary electrophoresis

Bai

et al. 2017

Electrophoresis

View full text Add to dashboard Cite

Screening bioactive compounds from natural product and its preparations using capillary electrophoresisBioactive ingredients of natural products can protect human body from harm, as well as prevent and treat disease. Screening bioactive compounds from natural products is attracting particular attention. It is a great challenge to separate and detect the active compounds from complex matrix natural products. CE plays a vital role in screening active compounds because of its unique features such as high-efficiency separation, short-analysis time, minimal sample consumption, and ease to realize automatization etc. Additionally, CE has been developed various modes owing to its abundant advantages in analysis and separation of compounds. The purpose of this work is to review previous developments and applications of CE in screening bioactive compounds derived from natural products from 2007 to 2017. This review does not only summarize the traditional methods of detecting active ingredients but also briefly introduces some novel assays. The trends in the application of CE in active compounds screening are addressed in the article.

show abstract

“…Very recently, Zhang et al. proposed a CE‐LIF precapillary‐based assay combined with HPLC–MS/MS to screen inhibitors of mTorC1 in crude natural product extracts. The 5‐carboxyfluorescein‐labeled peptide F‐4EBP1 (5‐FAM‐Ser‐Thr‐Thr‐Pro‐Gly‐Gly‐Thr‐Leu‐Phe‐Ser‐Thr‐Thr‐Pro‐Gly) was used as substrate.…”

Section: Precapillary Assaysmentioning

confidence: 99%

Advances in capillary electrophoresis for miniaturizing assays on kinase enzymes for drug discovery

Nehmé

Morin

2015

Electrophoresis

View full text Add to dashboard Cite

CE has become a frequently used tool for miniaturizing enzyme assays due particularly to its well-recognized low sample consumption. CE-based enzyme assays cover all aspects of kinetic analysis including the evaluation of enzyme activity, substrate and modulator characterization, and identification. These assays are performed to conduct high-quality primary (hit finding) and secondary (confirmation by determining the half maximal inhibitory concentration, IC ) screening. Nowadays, the kinase family is among the most actively studied pharmaceutical targets in oncology, and in neurodegenerative and inflammation diseases. In this article, we review the fundamentals of the different approaches that may be employed for assaying kinase kinetics. Their advantages and limitations will also be discussed by covering the literature of CE-based assays for purified kinases as well as for kinases in living cells. The last section will be devoted to perspectives by showing some applications of multiplexed and of microchip devices for high-throughput screening kinase assays.

show abstract

Screening of mammalian target of rapamycin inhibitors in natural product extracts by capillary electrophoresis in combination with high performance liquid chromatography–tandem mass spectrometry

Cited by 20 publications

References 44 publications

Analysis of Xanthine Oxidase Inhibitors fromPuerariae flosUsing Centrifugal Ultrafiltration Coupled with HPLC-MS

Analysis of Xanthine Oxidase Inhibitors fromPuerariae flosUsing Centrifugal Ultrafiltration Coupled with HPLC-MS

Screening bioactive compounds from natural product and its preparations using capillary electrophoresis

Advances in capillary electrophoresis for miniaturizing assays on kinase enzymes for drug discovery

Contact Info

Product

Resources

About