Unusual Structures and Cytotoxicities of Chitonoidosides A, A1, B, C, D, and E, Six Triterpene Glycosides from the Far Eastern Sea Cucumber Psolus Chitonoides
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marine drugs Article Unusual Structures and Cytotoxicities of Chitonoidosides A, A1, B, C, D, and E, Six Triterpene Glycosides from the Far Eastern Sea Cucumber Psolus chitonoides Alexandra S. Silchenko, Anatoly I. Kalinovsky, Sergey A. Avilov, Pelageya V. Andrijaschenko, Roman S. Popov, Pavel S. Dmitrenok , Ekaterina A. Chingizova and Vladimir I. Kalinin * G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch of the Russian Academy of Sciences, Pr. 100-letya Vladivostoka 159, 690022 Vladivostok, Russia; [email protected] (A.S.S.); [email protected] (A.I.K.); [email protected] (S.A.A.); [email protected] (P.V.A.); [email protected] (R.S.P.); [email protected] (P.S.D.); [email protected] (E.A.C.) * Correspondence: [email protected]; Tel./Fax: +7-(423)2-31-40-50 Abstract: Six new triterpene tetra-, penta- and hexaosides, chitonoidosides A (1), A1 (2), B (3), C(4), D (5), and E (6), containing one or two sulfate groups, have been isolated from the Far- Eastern sea cucumber Psolus chitonoides, collected near Bering Island (Commander Islands) from the depth of 100–150 m. Three of the isolated compounds (1, 3 and 6) are characterized by the unusual aglycone of new type having 18(20)-ether bond and lacking a lactone in contrast with wide spread holostane derivatives. Another unexpected finding is 3-O-methylxylose residue as a terminal unit in the carbohydrate chains of chitonoidosides B (3), C (4), and E (6), which has never been Citation: Silchenko, A.S.; Kalinovsky, found before in the glycosides from holothurians belonging to the Psolidae family. Moreover, this A.I.; Avilov, S.A.; Andrijaschenko, monosaccharide is sulfated in the compound 4 into unprecedented 3-O-methylxylose 4-O-sulfate P.V.; Popov, R.S.; Dmitrenok, P.S.; residue. Chitonoidoside C (4) is characterized by tetrasaccharide moiety lacking a part of the bottom Chingizova, E.A.; Kalinin, V.I. semi-chain, but having disaccharide fragment attached to C-4 of Xyl1. Such architecture is not Unusual Structures and Cytotoxicities common in sea cucumber glycosides. Cytotoxic activities of the compounds 1–5 against mouse and of Chitonoidosides A, A , B, C, D, 1 human erythrocytes and human cancer cell lines: adenocarcinoma HeLa, colorectal adenocarcinoma and E, Six Triterpene Glycosides from DLD-1, and leukemia promyeloblast HL-60 cells were studied. The cytotoxic effect of chitonoidoside the Far Eastern Sea Cucumber Psolus D (5) was the most significant in this series due to the presence of pentasaccharide disulfated sugar chitonoides. Mar. Drugs 2021, 19, 449. https://doi.org/10.3390/md19080449 chain in combination with holostane aglycone. Surprisingly, the glycosides 1 and 3, comprising the new aglycone without γ-lactone, demonstrated similar activity to the known compounds with Academic Editor: Hitoshi Sashiwa holostane aglycones. Chitonoidoside C (4) was less cytotoxic due to the different architecture of the carbohydrate chain compared to the other glycosides and probably due to the presence of a sulfate Received: 14 July 2021 group at C-4 in 3-O-MeXyl4. Accepted: 3 August 2021 Published: 5 August 2021 Keywords: Psolus chitonoides; triterpene glycosides; chitonoidosides; sea cucumber; cytotoxic activity Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- 1. Introduction iations. Triterpene glycosides, biosynthesized by the sea cucumbers, are natural compounds characterized by the diverse bioactivity [1–5]. Some of them are candidates to be used as new immunomodulators and anti-tumor drugs [6–8], which are urgently needed because cancer is one of the most important global health problems. The isolation of glycosides from Copyright: © 2021 by the authors. different sea cucumber species provides a unique opportunity to expand knowledge on Licensee MDPI, Basel, Switzerland. their structural diversity and discover new unusual substances. The comparative structural This article is an open access article analysis of a significant number of the isolated glycosides leads to the reconstruction of distributed under the terms and the sequences of biosynthetic transformations of their aglycones and carbohydrate chains conditions of the Creative Commons during biosynthesis that can be reflected in the metabolic networks constructed for different Attribution (CC BY) license (https:// species of the sea cucumbers [9,10]. The studies on biological activity of the glycosides creativecommons.org/licenses/by/ reveal the peculiarities of «structure–activity relationships» demonstrating the significance 4.0/). Mar. Drugs 2021, 19, 449. https://doi.org/10.3390/md19080449 https://www.mdpi.com/journal/marinedrugs Mar. Drugs 2021, 19, x 2 of 19 Mar. Drugs 2021, 19, 449 constructed for different species of the sea cucumbers [9,10]. The studies on biological2 of 18 activity of the glycosides reveal the peculiarities of «structure–activity relationships» demonstrating the significance of different structural parts of glycosides for the realization of their physiological action. These data indicate the relevance of searching for new of different structural parts of glycosides for the realization of their physiological action. glycosides and studying their biological activities. These data indicate the relevance of searching for new glycosides and studying their The genus Psolus comprises 58 species, and only three of them were chemically biological activities. investigated up to the present time: P. fabricii [10–14], P. eximius [15], and P. patagonicus The genus Psolus comprises 58 species, and only three of them were chemically inves- tigated[16,17]. upAs result to the of present the stud time:ies onP. fabriciithe fourth[10– representative14], P. eximius [of15 the], and genusP. patagonicus, P. chitonoides[16,, six17]. Asnew result triterpene of the glycosides studies on, chitonoid the fourthoside representatives A (1), A1 (2 of), theB (3 genus,), C (4),P. D chitonoides(5), and E ,(6 six) were new isolated and their chemical structures elucidated by the analyses of the 1H, 13C NMR, 1D triterpene glycosides, chitonoidosides A (1), A1 (2), B (3), C (4), D (5), and E (6) were 1 1 isolatedTOCSY and their2D NMR chemical ( H, H structures-COSY, HMBC, elucidated HSQC, by the ROESY) analyses spectra of the as1 H,well13C as NMR, HR-ESI 1D TOCSYmass spectra. and 2D All NMR of the ( 1originalH,1H-COSY, spectra HMBC, are pres HSQC,ented in ROESY) Figures spectraS1–S48 asin Supplementary well as HR-ESI massdata. The spectra. hemolytic All of theactivities original against spectra mouse are presented and human in Figureserythrocytes, S1–S48 cytotoxic in Supplementary activities data.against The human hemolytic adenocarcinoma activities against HeLa, mouse colorectal and humanadenocarcinoma erythrocytes, DLD cytotoxic-1, and leukemia activities againstpromyeloblast human HL adenocarcinoma-60 cells are reported. HeLa, colorectal adenocarcinoma DLD-1, and leukemia promyeloblast HL-60 cells are reported. 2. Results and Discussion 2.2.1. Results Structural and E Discussionlucidation of the Glycosides 2.1. StructuralThe concentrated Elucidation ethanolic of the Glycosides extract of the sea cucumber Psolus chitonoides was chromatographedThe concentrated on a ethanolic Polychrom extract-1 column of the sea(powdered cucumber Teflon,Psolus Biolar chitonoides, Latvia).was chro-The matographedglycosides were on eluted a Polychrom-1 after using column water (powderedas a mobile Teflon,phase with Biolar, 50% Latvia). EtOH Theand glycosidesseparated wereby repeated eluted afterchromatography using water ason aSi mobile gel columns phase withwith 50% the EtOHeluents and CHCl separated3/EtOH/H by2 re-O peated(100:75:1 chromatography0), (100:100:17),on and Si gel(100:125:25) columns withto give the three eluents fractions CHCl3/EtOH/H (I–III). The2O individual (100:75:10), (100:100:17),compounds 1 and–6 (Figure (100:125:25) 1) were to isolated give three by fractions HPLC of (I–III). the obtained The individual fractions compounds on silica-based1–6 (Figurecolumn1 )Supelcosil were isolated LC-Si by (4.6 HPLC × 150 of themm) obtained and reversed fractions-phase on silica-based semipreparative column column Supel-s cosilPhenomenex LC-Si (4.6 Synergi× 150 Fusion mm) and RP reversed-phase(10 × 250 mm) or semipreparative Supelco Ascentis columns RP-Amide Phenomenex (10 × 250 Synergimm). Fusion RP (10 × 250 mm) or Supelco Ascentis RP-Amide (10 × 250 mm). FigureFigure 1.1. ChemicalChemical structuresstructures ofof glycosidesglycosides isolatedisolated fromfrom PsolusPsolus chitonoideschitonoides:: 1—chitonoidoside—chitonoidoside A; 2—chitonoidoside—chitonoidoside A A11; ; 33—chitonoidoside—chitonoidoside B;B; 44—chitonoidoside—chitonoidoside C;C; 55—chitonoidoside—chitonoidoside D;D; 6—chitonoidoside—chitonoidoside E.E. All the configuration configurationss of monosaccharides in all all glycosides glycosides were were assigned assigned to to D D-series-series along with biogenetical analogies with all known sea sea cucumber cucumber triterpene triterpene glycosides. glycosides. The molecular formulaformula ofof chitonoidosidechitonoidoside AA ((11)) waswas determineddetermined toto be be C C5353HH8383O2323SNaSNa − − fromfrom the [M Na−−NaNa]] ionion peak peak at atm/zm/z 11191119.5049.5049 (calc. (calc. 1119 1119.5051).5051) in the in the(−)HR (−-)HR-ESI-MSESI-MS and + + 1 13 and[MNa [M+NaNa] +Na] ion peakion at peak m/z 1165.4816 at m/z 1165.4816 (calc. 1165.4836) (calc. 1165.4836) in the (+)HR