Application of chromatography technology in the separation of active alkaloids from Hypecoum leptocarpum and their inhibitory effect on fatty acid synthase

Abstract

A method that involved the combination of pH-zone-refining counter-current chromatography and semipreparative reversed-phase liquid chromatography has been established for the preparative separation of alkaloids from Hypecoum leptocarpum. From 1.2 g of crude sample, 31 mg N-feruloyltyramine, 27 mg oxohydrastinine, 47 mg hydroprotopine, 25 mg leptopidine, and 18 mg hypecocarpine have been obtained. The structure of the new compound, hypecocarpine, is confirmed based on the analysis of spectroscopic data, including NMR, UV, and IR spectroscopy and positive electrospray ionization mass spectrometry. The known chemical structures were characterized on the basis of (1) H and (13) C NMR spectroscopy. The purities of the five alkaloids are all over 92.7% as determined by high-performance liquid chromatography. The alkaloids' cytotoxicity in breast cancer cells is assessed by using a Cell Counting Kit assay and their inhibitory effect on fatty acid synthase expression is assessed by a Western blot assay. These results suggest that leptopidine could suppress growth and induce cytotoxicity in breast cancer cells and that the cytotoxicity of leptopidine may be related to its inhibitory effect on fatty acid synthase expression.

Hypecoum spp.—Chemistry and Biological Activity of Alkaloids

“…This section summarizes the alkaloid compounds that have been identified in the Hypecoum spp. since 1972 (Table 2) [2,[7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24]27,[40][41][42][43][44][45][46][47][48][49][50]. So far, a total of 86 naturally occurring alkaloids have been reported for ten plant species-H. procumbens, H. procumbens var.…”

“…glaucescens, H. imberbe, H. lactiflorum, H. pendulum, H. trilobum, H. leptocarpum, H. erectum, H. ponticum and H. parviflorum. Based on their structures, the Hypecoum alkaloids can be divided into eight major groups: protopines (1-11), protoberberines (12)(13)(14)(15)(16)(17), benzophenantridines (18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28), aporphines (29)(30)(31)(32)(33), simple isoquinolines (34)(35)(36)(37)(38), secoberbines , spirobenzilisoquinolines (67)(68)(69)(70)(71)(72)(73) and others (74-86). Two chemical structures have been detected in the literature with the name leptocarpine-protopine alkaloid 8 and alkaloid 82.…”

“…H. leptocarpum [22,23,43] Hypecocarpinine (42) H. leptocarpum [24] Hypecocarpine (43) H. leptocarpum [27] Leptopinine (44) H. leptocarpum [12] H. lactiflorum [7] (S)-Dihydroleptopine (41) (Coryximine)…”

“…In recent years, more than 40 alkaloids have been identified in Hypecoum species, and new alkaloid structures, along with their pharmacological activities, are reported every year, enriching the diversity of natural compounds. Some of them have promising properties for the treatment of important diseases such as inflammation, cancer and microbial infections [18,19,22,23,[26][27][28][29].…”

Mining Therapeutic Efficacy from Treasure Chest of Biodiversity and Chemodiversity: Pharmacophylogeny of Ranunculales Medicinal Plants

“…(165) Leptopidine of H. leptocarpum could suppress growth and induce cytotoxicity in breast cancer cells via inhibiting fatty acid synthase expression. (166) In the subfamily Papaveroideae, Glaucium squamigerum is used by Chinese ethnic minorities against gastrointestinal and respiratory diseases. (3) The most reported bioactivity of Glaucium alkaloids is anticancer and anti-cholinesterase effects.…”