Determine equilibrium dissociation constant of drug-membrane receptor affinity using the cell membrane chromatography relative standard method

Ma, Weina; Yang, Lei; Lv, Yanni; Jia, Fu; Zhang, Yanmin; He, Langchong

doi:10.1016/j.chroma.2017.04.053

Cited by 49 publications

(28 citation statements)

References 43 publications

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“…The frontal analysis method was performed to determine the equilibrium dissociation constant ( K D ) between ligand and the affinity binding site on the receptor, which had been described in our previous work . The breakthrough curves of 5, 10, 20, 40, 60, 80 and 100 n m analyte binding to MRGPRX2 were determined by conditions described as below: mobile phases were consisted by solvent A and solvent B. Solvent A was prepared by 1 m m PBS (pH 7.4) dissolving with 100 n m analyte, and solvent B was prepared by 1 m m PBS (pH 7.4).…”

Section: Methodsmentioning

confidence: 98%

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Discovery and analysis the anti-pseudo-allergic components from Perilla frutescens leaves by overexpressed MRGPRX2 cell membrane chromatography coupled with HPLC-ESI-IT-TOF system

Yang

Zeng

Wang

et al. 2020

Journal of Pharmacy and Pharmacology

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Objectives Screen and identify the anti-pseudo-allergic activity components of Perilla frutescens leaves that interacted with MRGPRX2 (a new reported pseudoallergic reaction-related receptor). Methods An overexpressed MRGPRX2 cell membrane chromatography (CMC) coupled with HPLC-ESI-IT-TOF system has been established to screen and identify the effective components from P. frutescens leaves. A frontal analysis method was performed to investigate the binding affinity between ligands and MRGPRX2. Their activity of relieving pseudo-allergic reaction was evaluated in vitro by histamine release assay, β-hexosaminidase release assay and intracellular Ca 2+ mobilization assay. Key findings Extract of P. frutescens leaves was proved to be effective in antipseudo-allergic reaction by inhibiting MRGPRX2. Apigenin (API) and rosmarinic acid (ROS) were confirmed to be the potential anti-allergy compounds that could bind with MRGPRX2. The binding affinity (K D ) of ROS and API with MRGPRX2 was (8.79 ± 0.13) 9 10 −8 M and (6.54 ± 1.69) 9 10 −8 M, respectively. The IC 50 of API inhibiting laboratory of allergic disease 2 cells degranulation was also determined to be (51.96 ± 0.18) μM. Conclusions A MRGPRX2/CMC coupled with HPLC-ESI-IT-TOF system was successfully established and applied to discover the effective components from P. frutescens leaves.

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Section: Methodsmentioning

confidence: 98%

“…The cell membrane stationary phase (CMSP) was prepared by agitating the mixture for 30 min and standing overnight at 4°C. Membranes were made into stationary phases by the physical adsorption with silica gel . Then, the CMC column (L × I.D., 10 mm × 2.0 mm) was then packed up through a wet packing method (1 ml/min, 10 MPa, 10 min).…”

Section: Methodsmentioning

confidence: 99%

Discovery and analysis the anti-pseudo-allergic components from Perilla frutescens leaves by overexpressed MRGPRX2 cell membrane chromatography coupled with HPLC-ESI-IT-TOF system

Yang

Zeng

Wang

et al. 2020

Journal of Pharmacy and Pharmacology

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“…Sorafenib was ordered from Dalian Meilun Biotech Co., Ltd (Dalian, China) and a novel agent, B10 (purity >98%), was designed and synthesized by Jie Zhang's group in the Research and Engineering Center for Natural Medicine Xi'an Jiaotong University . All compounds were dissolved in DMSO for use in biochemical assays.…”

Section: Methodsmentioning

confidence: 99%

HMQ‐T‐B10 induces human liver cell apoptosis by competitively targeting EphrinB2 and regulating its pathway

Dai

Shi

et al. 2018

J Cellular Molecular Medi

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Hepatocellular carcinoma (HCC) is a highly prevalent cancer worldwide and it is necessary to discover and develop novel preventive strategies and therapeutic approaches for HCC. Herein, we report that EphrinB2 expression is correlated with liver cancer progression. Moreover, by using phosphorylated proteomics array, we reveal a pro‐apoptosis protein whose phosphorylation and activation levels are up‐regulated upon EphrinB2 knockdown. These results suggest that EphrinB2 may act as an anti‐apoptotic protein in liver cancer cells. We also explored the therapeutic potential of HMQ‐T‐B10 (B10), which was designed and synthesized in our laboratory, for HCC and its underlying mechanisms in vitro and in vivo. Our data demonstrate that B10 could bind EphrinB2 and show inhibitory activity on human liver cancer cells. Moreover, induction of human liver cancer cell apoptosis by B10 could be augmented upon EphrinB2 knockdown. B10 inhibited HCC cell growth and induced HCC cell apoptosis by repressing the EphrinB2 and VEGFR2 signalling pathway. Growth of xenograft tumours derived from Hep3B in nude mice was also significantly inhibited by B10. Collectively, these findings highlight the potential molecular mechanisms of B10 and its potential as an effective antitumour agent for HCC.

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“…Cell membrane chromatography (CMC) is a bio‐affinity chromatographic technique that has developed rapidly in recent years [1–4]. The interaction process between drug molecules and membrane receptors can be simulated in vitro by using membrane and chromatographic carrier as stationary phase and chromatographic analysis method.…”

Section: Introductionmentioning

confidence: 99%

Preparation and application of polyvinyl alcohol‐decorated cell membrane chromatography for screening anti‐osteoporosis components from Liuwei Dihuang decoction‐containing serum

Wang²,

Xu³

et al. 2020

J of Separation Science

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Cell membrane chromatography is a powerful tool for screening active components from complex matrices. New cell membrane carriers need to be developed to increase the coverage of cell membranes on the surface of stationary phases, thereby improving cell membrane chromatographic retention. In this work, we prepared polyvinyl alcohol-poly dimethyl diallyl ammonium chloride modified silica gel as a cell membrane carrier. Osteoblast cell was used as cell membrane source, which was widely used to evaluate the osteogenic activity of drugs. The new cell membrane chromatographic stationary phase was used to screen anti-osteoporosis components in Liuwei Dihuang decoction-containing serum. The chemical structures of the new modified materials were characterized by scanning electronic microscopy, Fourier transform infrared spectroscopy, and elemental analysis characterization.Compared with the common cell membrane column, the cell membrane coverage of this modified material was increased by 30%, and thus provided a stronger retention effect in positive drugs. Nineteen metabolites in rat serum samples were retained on the cell membrane chromatography and identified by ultra-high-performance liquid chromatography/time-of-flight mass spectrometry. Among those, four components (morroniside, catalpol, loganin, and acteoside) were selected for in vitro pharmacodynamics validation. They significantly increased the osteoblast proliferation. The new modified material was successfully applied to screen anti-osteoporosis components from Liuwei Dihuang Decoction-containing serum. K E Y W O R D Sosteoblast cell membrane chromatography, polyvinyl alcohol-decorated, metabolites, rat serum, traditional Chinese medicineArticle Related Abbreviations: CMC, cell membrane chromatography; OM, osteoblast cell membrane; PDAC, poly dimethyl diallyl ammonium chloride; PVA, polyvinyl alcohol; TCM, traditional Chinese medicine.

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Determine equilibrium dissociation constant of drug-membrane receptor affinity using the cell membrane chromatography relative standard method

Cited by 49 publications

References 43 publications

Discovery and analysis the anti-pseudo-allergic components from Perilla frutescens leaves by overexpressed MRGPRX2 cell membrane chromatography coupled with HPLC-ESI-IT-TOF system

Discovery and analysis the anti-pseudo-allergic components from Perilla frutescens leaves by overexpressed MRGPRX2 cell membrane chromatography coupled with HPLC-ESI-IT-TOF system

HMQ‐T‐B10 induces human liver cell apoptosis by competitively targeting EphrinB2 and regulating its pathway

Preparation and application of polyvinyl alcohol‐decorated cell membrane chromatography for screening anti‐osteoporosis components from Liuwei Dihuang decoction‐containing serum

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