LC-MS guided isolation of three pairs of enantiomeric alkaloids from Macleaya cordata and their enantioseparations, antiproliferative activity, apoptosis-inducing property

Abstract

(±)-Macleayins F-H (1-3), three pairs of new enantiomeric alkaloid dimers, along with four known alkaloids (4-7) as their plausible biogenetic precursors, were isolated from the aerial parts of Macleaya cordata. Compounds 1-3 were obtained under the guidance of LC-MS investigation, and their structures were elucidated by analysis of the 1D and 2D NMR spectroscopic data. The racemic mixtures were successfully separated by chiral HPLC, and the absolute configurations of enantiomers were determined by electronic circular dichroism (ECD) spectroscopy. Compounds 1-7 showed antiproliferative activity against HL-60 with IC 50 values of 1.34-41.30 μM, especially compounds 1-2 exhibited the best inhibitory activity against HL-60 cell lines. In addition, the preliminary mechanism investigation for compound 2 using Annexin V/7-AAD double-staining assay, DAPI staining assay and JC-1 staining method, indicated that 2 inhibited cancer cell proliferation potentially through inducing apoptosis via the mitochondria-related pathway and arrested cell cycle of HL-60 cells at S phase.Benzophenanthridine and protopine alkaloids occur in Macleaya cordata (Willd) R. Br. and have been investigated for their intriguing bioactivity, such as anticancer, anti-bacterial, insecticidal, and anti-inflammatory effects 1,2 . Macleaya cordata is a perennial plant in the family of Papaveraceae, which has been used in folk medicine for the treatment of cervical cancer, thyroid cancer, inflamed wounds, ringworm infection, arthritis, and trichomonas vaginalis 3,4 . In our previous investigation, two pairs of enantiomeric alkaloid dimers (macleayins A and B) with cytotoxicity were isolated from M. cordata 5 . In our present study, LC-MS-guided fractionation (Fig. 1) led to the isolation of three pairs of analogous dimers, macleayins F-H (1-3) biogenetically derived from a benzophenanthridine and protopine alkaloid through C6-C 13′ linkage. Four possible biogenetic precursors, sanguinarine, chelerythrine, protopine, and allocryptopine (4-7) were obtained (Fig. 2). Herein, we reported the isolation, and structural elucidation of new compounds. In addition, the antiproliferative properties and action mechanism were also investigated. Results and DiscussionStructural Elucidation of Compounds 1a/1b-3a/3b. Macleayin F (1) was isolated as white amorphous powder, with the molecular formula of C 41 H 36 N 2 O 9 , deduced from HRESIMS [M + H] + (m/z 701.2491, calcd for 701.2494), appropriate for 25 degrees of unsaturation. The IR spectrum indicated the presence of ketone carbonyl (1656 cm −1 ), methylenedioxyl group (2792, 941 cm −1 ), and aromatic ring (1616, 1486, 1462 cm −1 ). In the UV spectrum, the absorption maxima at 227 and 287 nm were detected. The HRESIMS/MS spectrum exhibited the

Systematic characterization of alkaloids in Eomecon chionantha Hance using ultrahigh‐performance liquid chromatography–tandem quadrupole Exactive Orbitrap mass spectrometry with a four‐step screening strategy

“…Based on the previous strategy and comparison with the literature, compound 93 was identified as (±)‐bocconarborine A. Similarly, compounds 42 , 43 , 91 , 92 , 94 and 95 were identified as (±)‐macleayins A, (+)‐macleayin G, (±)‐macleayins B, chelidimerine, sanguinarine‐methylene‐chelerythrine and chelerythrine‐methylene‐chelerythrine, respectively, which are in agreement with the literature 45,46 …”

Oregano Oil Combined with Macleaya Cordata Oral Solution Improves the Growth Performance and Immune Response of Broilers

“…A growing number of studies have shown that diets supplemented with oregano oil can promote performance and improve the antioxidant status and immunity of broilers [ 9 , 10 ]. Macleaya cordata is a perennial herb of the poppy family, widely distributed in southern China [ 11 ]. Previous studies have also shown that sanguinarine from macleaya cordata is the main bioactive substance [ 12 ].…”

Efficient extraction and purification of benzo[c]phenanthridine alkaloids from Macleaya cordata (Willd) R. Br. by combination of ultrahigh pressure extraction and pH‐zone‐refining counter‐current chromatography with anti‐breast cancer activity in vitro

“…13 C‐NMR (100 MHz, MeOH‐ d 4 ) δ C : 52.9 ( N ‐CH 3 ), 57.5 (8‐OCH 3 ), 62.8 (7‐OCH 3 ), 104.3 (‐OCH 2 O‐2,3), 105.0 (C‐4), 107.0 (C‐1), 119.4 (C‐11), 119.8 (C‐6a), 120.8 (C‐10), 121.7 (C‐4a), 127.0 (C‐10b), 127.3 (C‐9), 129.9 (C‐10a), 132.5 (C‐4b), 133.3 (C‐12), 134.2 (C‐12a), 147.4 (C‐7), 150.7 (C‐3), 150.9 (C‐2), 151.7 (C‐8), 152.0 (C‐6). Compared with the literature, 18,29 compounds 1 (C 20 H 14 NO 4 ) and 2 (C 21 H 18 NO 4 ) were identified as sanguinarine and chelerythrine, respectively.…”