Preparative isolation and purification of steroidal glycoalkaloid from the ripe berries of Solanum nigrum L. by preparative HPLC–MS and UHPLC–TOF‐MS/MS and its anti‐non‐small cell lung tumors effects in vitro and in vivo

Abstract

Overcoming epidermal growth factor receptor resistance is a critical problem that needs to be solved in clinical practice. Drugs that downregulate the fatty acid synthaseepidermal growth factor receptor will become novel treatments for non-small cell lung cancer. Solanum nigrum, extracted with water at 4 • C, shows strong cytotoxic activity and inhibits tumor growth in Lewis tumor bearing-mice in a dose-dependent manner. A novel active compound in S. nigrum, solaoiacid, was successfully separated and purified from S. nigrum by preparative high-performance liquid chromatography with mass spectrometry and ultra high performance liquid chromatography with timeof-flight tandem mass spectrometry. The IC 50 of solaoiacid on lung cancer cells was 2.3 μmol/L, which was significantly lower than that of the known steroidal glycoalkaloid. Label-free proteomics and STRING Network analysis were used to identify significantly deregulated proteins in lung cancer cells that were treated with the fresh ripe fruit extracts of S. nigrum. S. nigrum regulates multiple signal pathways, including the epidermal growth factor receptor pathway. S. nigrum downregulated 24 main proteins with direct roles in fatty acid biosynthesis. Both S. nigrum and solaoiacid showed strong downregulation of the fatty acid synthase-epidermal growth factor receptor and anti-non-small cell lung cancer effects, and thus will become a novel drug for the treatment of non-small cell lung cancer. K E Y W O R D S epidermal growth factor receptor, fatty acid synthase, non-small cell lung cancer, proteomics, Solanum nigrum berries Article Related Abbreviation: EGFR, epidermal growth factor receptor; FASN, fatty acid synthase; NSCLC, non-small cell lung cancer; Pre-HPLC-MS, preparative liquid chromatography high-performance liquid chromatography-mass spectrometry; SNDA, aqueous extracts of the dried ripe berries of Solanum nigrum; SNFA, aqueous extracts of the fresh ripe Targeted therapy has become the model paradigm of precision medicine in non-small cell lung cancer (NSCLC) berries of Solanum nigrum; SNLH, aqueous extracts of the whole plants of Solanum nigrum L.

Homoisoflavonoids profiling of Ophiopogon japonicus by off‐line coupling high‐speed countercurrent chromatography with high‐performance liquid chromatography–diode array detector‒quadrupole time‐of‐flight tandem mass spectrometry

“…It was reported that about 50 homoisoflavoniods were purified and identified by repeated column fractionations [1]. HPLC with diode array detector (DAD) and MS/MS has been developed for rapid isolation and characterization of bioactive components from complex natural products [8–11]. However, information about comprehensive analysis of homoisoflavonoids, especially for minors, in O. japonicus by HPLC–DAD–MS/MS is restricted because of the existence of multicomponents, restriction of detection sensitivity, and coelution of minor and abundant compounds [12,13].…”

Anticancer activity of glycoalkaloids from Solanum plants: A review

“…Both S. nigrum L. extract and the newly identified GA of this plant, solaoiacid, inhibit the FASN-EGFR signaling pathway and affect the migration, autophagy, apoptosis, and immune response of non-small cell lung cancer (NSCLC) cells. Furthermore, solaoiacid acts with an IC 50 = 2.3 μmol/L, which is significantly lower than that of SM (Shi et al, 2019). The extract of S. nigrum L. had antitumor effects on rat C6 MG in vitro and in vivo.…”

Analysis of steroidal glycoalkaloids and their metabolites in Solanum nigrum fruits based on liquid chromatography‐tandem mass spectrometry and molecular networking

“…According to the literature [15][16][17], the steroidal glycoalkaloids from S. nigrum are composed of seven basic types of aglycones (including isomers), namely, solasodine (S), hydroxysolasodine (HS), hydroxymethylsolasodine (HMS), dihydroxymethylsolasodine (DHMS), carboxylsolasodine (CS), and hydroxycarboxylsolasodine (HCS) linked with glycosyl groups, which are made up of hexoses, such as glucose (Glc) and galactose (Gal), 6-deoxyhexoses, such as rhamnose (Rha), pentoses, such as xylose, and uronic acids, such as glucuronic acid (GlcA) and galacturonic acid (GalA), through ether glycosidic bonds at carbon-3 of the aglycones. Substitution of groups on the steroid skeleton often occurs at carbon-7α (B-ring), carbon-12β (C-ring), and carbon-27 (F-ring) sites.…”

“…At present, most of the steroidal glycoalkaloids identified in Solanum nigrum fruit are monoglycosides, diglycosides, and triglycosides with molecular weights less than 1000 Da with solasodine or its substitute as the aglycone [7 -10]. The alkaloids found include solasodine, khasianine, solasonine, solamargine, solaoiacid, solanine A, γ-solasonine, 7α-OH solamargine, 7α-OH khasianine, 7α-OH solasonine, β 2 -solasonine, 12β, 27dihydroxysolasodine (27-DHS) [11][12][13][14], which have been proven to be cytotoxic to a variety of cancer cells [11][12][13][14][15]. They are mainly present in immature fruits and decrease gradually to disappearance with the ripening of the fruits [16,17].…”