Screening of anti-functional dyspepsia compounds in Cynanchum auriculatum: A spectrum-effect relationship analysis, and ATP-binding cassette transporters inhibitor evaluation

Ethnopharmacological relevance: Functional dyspepsia (FD) is a disorder caused by abnormal gut-brain axis regulation and is highly prevalent in China. Cynanchum auriculatum (CA) is often used to treat FD in the ethnic minority areas of Guizhou. Although several CA-based products are currently available in the market, it is un-clear which components of CA are efficacious and what their oral absorption mechanism is.Aim of the study: This study aimed to screen anti-FD components of CA based on the spectrum-effect relationship. In addition, the study evaluated the intestinal absorption mechanism of these components using transporter inhibitors.Materials and methods: The fingerprinting of compounds from CA extract and plasma after oral administration was conducted using ultra-high-performance liquid chromatography quadrupole-time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS). The intestinal contractile parameters were then measured in vitro using the BL-420F Biofunctional Experiment System. Multivariate statistical analysis of the result of spectrum-effect relationship assessment was used to elucidate the correlation between prominent peaks of CA-containing plasma and intestinal contractile activity. The effect of ATP-binding cassette (ABC) transporter inhibitors, such as the P-gp inhibitor verapamil, the MRR inhibitor indomethacin, and the BCRP inhibitor Ko143, on the directional transport of the predicted active ingredients was assessed in vivo.Results: Twenty chromatographic peaks were identified in the CA extract. Of these, three were C21 steroids, four were organic acids, and one was a coumarin, and acetophenone by comparing with reference compounds. Additionally, it is discovered that there are totally 39 migratory components in CA-containing plasma, which was found to significantly promote the contractility of the isolated duodenum. Moreover, multivariate analysis of the spectrum-effect relationship demonstrated that 16 characteristic peaks (3, 6, 8, 10, 11, 13, 14, 18, 21, m1-m4, m7, m15, and m24) in CA-containing plasma were significantly associated with the anti-FD effect. These com-pounds included seven prototype compounds, i.e., cynanoneside A, syringic acid, deacylmetaplexigenin, ferulic acid, scopoletin, baishouwubenzophenone, and qingyangshengenin. The inhibition of ABC transporters demonstrated that the inhibitors verapamil and Ko143 significantly increased (P < 0.05) the uptake of scopoletin and qingyangshengenin. Thus, these compounds may be substrates for P-gp and BCRP. Conclusions: The potential anti-FD components of CA and the effect of ABC transporter inhibitors on these active components were preliminarily clarified. These findings lay a foundation for subsequent in vivo studies.