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Rapid Screening of Potential Phosphodiesterase Inhibitors from the Roots of Ilex pubescens Hook. et Arn. Using a Combination of Ultrafiltration and LC – MS

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ABSTRACT

The cyclic nucleotide phosphodiesterase (PDE) plays an important role in regulating the levels of second messenger molecules cAMP and cGMP. Various PDE inhibitors have been successfully developed into drugs for targeted diseases. In addition, PDE inhibitors can also be found in different foods and natural medicines. In this study, ultrafiltration liquid chromatography–diode-array detector–electrospray ionization–ion-trap–time-of-flight–mass spectrometry (ultrafiltration LC–DAD–ESI–IT–TOF–MS) was applied to screen PDE inhibitors from the roots of Ilex pubescens Hook. et Arn. As a result, 11 major compounds were identified in I. pubescens roots, with nine compounds as potential PDE inhibitors, among which five were further confirmed to be active against PDEI and PDE5A dose-dependently in vitro, with ilexsaponin A1 and ilexsaponin B2 being the strongest. HPLC quantification of these bioactive compounds suggested that they are major components in the plant. The results demonstrate that ultrafiltration LC–DAD–ESI–IT–TOF–MS is an efficient method for rapid screening of PDE inhibitors from natural medicines.

No MeSH data available.


Structures of the 11 compounds identified in Radix Ilicis Pubescentis.
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fig2: Structures of the 11 compounds identified in Radix Ilicis Pubescentis.

Mentions: LC–PDA–ESI–IT–TOF–MS analysis was used to identify the 11 major compounds in I. pubescens roots. The mass spectral data in negative ion mode was used for characterization. The MS fragmentations of compounds together with their retention times (tR) are summarized in Table 1. Their structures were shown in Figure 2. The ultrafiltrate was analyzed under the same condition. The retention times and MS data (see Table S1 in the Supplementary Material available online at https://doi.org/10.1155/2017/2749643) of the 11 compounds obtained from the ultrafiltrate were identical to those from I. pubescens roots extract. The EIC chromatograms of each compound in negative ion mode of the extract, ultrafiltrate, and blank sample were also shown in supplementary Figures S1–9. Compounds 1, 3, 5, 8, 9, 10, and 11 were further identified unambiguously by comparing their chromatographic and MS behaviors to corresponding standard compounds as shown in Table S1.


Rapid Screening of Potential Phosphodiesterase Inhibitors from the Roots of Ilex pubescens Hook. et Arn. Using a Combination of Ultrafiltration and LC – MS
Structures of the 11 compounds identified in Radix Ilicis Pubescentis.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5382325&req=5

fig2: Structures of the 11 compounds identified in Radix Ilicis Pubescentis.
Mentions: LC–PDA–ESI–IT–TOF–MS analysis was used to identify the 11 major compounds in I. pubescens roots. The mass spectral data in negative ion mode was used for characterization. The MS fragmentations of compounds together with their retention times (tR) are summarized in Table 1. Their structures were shown in Figure 2. The ultrafiltrate was analyzed under the same condition. The retention times and MS data (see Table S1 in the Supplementary Material available online at https://doi.org/10.1155/2017/2749643) of the 11 compounds obtained from the ultrafiltrate were identical to those from I. pubescens roots extract. The EIC chromatograms of each compound in negative ion mode of the extract, ultrafiltrate, and blank sample were also shown in supplementary Figures S1–9. Compounds 1, 3, 5, 8, 9, 10, and 11 were further identified unambiguously by comparing their chromatographic and MS behaviors to corresponding standard compounds as shown in Table S1.

View Article: PubMed Central - PubMed

ABSTRACT

The cyclic nucleotide phosphodiesterase (PDE) plays an important role in regulating the levels of second messenger molecules cAMP and cGMP. Various PDE inhibitors have been successfully developed into drugs for targeted diseases. In addition, PDE inhibitors can also be found in different foods and natural medicines. In this study, ultrafiltration liquid chromatography–diode-array detector–electrospray ionization–ion-trap–time-of-flight–mass spectrometry (ultrafiltration LC–DAD–ESI–IT–TOF–MS) was applied to screen PDE inhibitors from the roots of Ilex pubescens Hook. et Arn. As a result, 11 major compounds were identified in I. pubescens roots, with nine compounds as potential PDE inhibitors, among which five were further confirmed to be active against PDEI and PDE5A dose-dependently in vitro, with ilexsaponin A1 and ilexsaponin B2 being the strongest. HPLC quantification of these bioactive compounds suggested that they are major components in the plant. The results demonstrate that ultrafiltration LC–DAD–ESI–IT–TOF–MS is an efficient method for rapid screening of PDE inhibitors from natural medicines.

No MeSH data available.