Abstract
BackgroundSan-ao decoction (SAD) has been widely used in Chinese medicine against respiratory diseases, such as asthma and rhinallergosis. The bioactive compounds for such pharmacological action remain unknown.MethodsWe developed a methodology to isolate the bioactive compounds of SAD. The assay involved the immobilization of beta 2-adrenoceptor (β2-AR) onto magnetic fine particles, the capture of target compounds by the immobilized receptor, the identification of the receptor bound compounds by reversed-phase high-performance liquid chromatography coupled with tandem mass spectrometry.ResultsVicenin, shaftoside, isoshaftoside, liquiritin apioside and isoliquiritin apioside were identified as β2-AR ligands in SAD extract. The binding of these compounds to β2-AR occurred on serine169, serine170 and phenylalanine256 of the receptor.ConclusionsThe developed methodology has high stability and specificity for recognizing and isolating target compounds. It is an alternative method for rapidly screening bioactive compounds of immobilized receptor from Chinese prescriptions.
Highlights
San-ao decoction (SAD) has been widely used in Chinese medicine against respiratory diseases, such as asthma and rhinallergosis
Morphological characterization of immobilized β2‐AR The sizes of control F e3O4@NH2 FPs and F e3O4@beta 2-adrenoceptor (β2-AR) FPs were analyzed by dynamic light scattering (DLS) with intensity-weighted size distributions (Fig. 3)
The Fourier transform infrared spectroscopy (FT-IR) adsorption band at 575 cm−1 corresponded to Fe–O bond vibrations in iron oxide
Summary
San-ao decoction (SAD) has been widely used in Chinese medicine against respiratory diseases, such as asthma and rhinallergosis. Beta 2-adrenoceptor (β2-AR) chromatography was established for the identification of bioactive compounds of the receptor [16]. This strategy has limitations of time-intensity and labor-intensity in preparation of chromatographic columns. We established a bio-affinity technique by the utilization of magnetic material to remove the isolation and identification bottlenecks in identifying procedures [17,18,19,20,21]. Magnetic fine particle (MFP)-based ligand isolation methods are highly desirable because they utilize the unique properties of MFPs, such as convenient solid–liquid separation, high surface area and good biocompatibility [22,23,24]. With the molecular docking technology, we predicted the potential activity of bioactive compounds obtained from ligand capture [25]
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