Submit Manuscript | http://medcraveonline.com Abbreviations: 1 H NMR, proton nuclear magnetic resonance; 13 C NMR, carbon nuclear magnetic resonance; 2D, two dimension; δ, chemical shifts; s, singlet; d, doublet; dd, doublet of doublet; t, triplet; m, multiplet; J, coupling constant; Hz, hertz; HRESIMS, high resolution electrospray ionization mass spectrometry; GC/ MS, gas chromatographic mass spectrometry; m/z, mass to charge ratio; MeOH, methanol; EtOAc, ethyl acetate; CH 2 Cl 2 , methylene chloride; CHCl 3 , chloroform; HCl, hydrochloric acid; DFMO, α-difuoromethylornithine; Ac, acetyl; min, minute(s); h, hour(s) THP-1, human monocytic cell line derived from an acute monocytic leukemia patient; IC 50 , concentration that afford 50% inhibition; IC 90 , concentration that afford 90% inhibition Introduction Tunicates are marine invertebrate animals (animals lacking a backbone) that are commonly found attached to rocks, they are three main groups, sessile ascidians, pelagic appendicularians and thaliaceans. 1 They are characterized by the possession of a tunic composed of cellulose. 1 Adult tunicates are flter feeders: the seawater enters a pharynx through an inhalating or oral siphon, in most cases set in motion by ciliary beating, food particles are trapped on a mucous net secreted by the endostyle, and the water and waste exit the body through an exhalating or atrial siphon. 1 Appendicularia, ascidians and some thaliaceans possess a tadpole-like larva with a notochord and metamorphose into sessile adults in the case of ascidians. Tunicates have reversible blood fow. 1 Natural products found in tunicate has began to attract many chemist’s interests; alkaloids, carotenoids, macrolides and tunichromes. 2 The cytotoxic and antibacterial activities of Polyclinum indicum and Polyclinum madrasensis extracts have been tested at various concentrations and showed the results of highest cytotoxicity assay conducted, indicating the presence of cytotoxic compounds in these ascidians. 3,4 The crude extract of ascidian Polyclinum madrasensis showed hemolytic properties against human, cow, goat and chicken bloods that be considered a valuable source for secondary metabolites which could be of pharmaceutical interest. 5 Polyclinal a sulfated polyhydroxy benzaldehyde has been isolated from extracts of the temperate colonial ascidian Polyclinum planum. 6 The highest concentration of this metabolite was found in the zooid-rich outer layers of this ascidian suggesting that it may represent a potential chemical defense against predators. 6 As part of our research dealing with the isolation and biological evaluation of active compounds from marine origin, the chemistry and the biological properties of the tunicate Polyclinum constellatum collected from the Red Sea was investigated, which led to the isolation of nine compounds (1-9), for the frst time from the genus polyclinum. Materials and methods General experimental procedures 1 H-NMR, 13 C-NMR and 2D-NMR spectra were recorded using the residual solvent signal as an internal standard on Bruker BioSpin Gm bH 400 and 500 spectrometers (Bruker, Rheinstetten, Germany). High resolution mass spectrometry were measured using a Bruker BioApex FT mass spectrometer (Bruker, Rheinstetten, Germany). GC/MS analysis was carried out using an HP 6890 series GC (Agilent Pharm Pharmacol Int J. 2017;5(3):108‒113. 108 © 2017 Raslan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and build upon your work non-commercially. Evaluation of secondary metabolites from the red sea tunicate Polyclinum Constellatum Volume 5 Issue 3 - 2017 Ali E Raslan, 1,2 Mohamed M Radwan, 1,3 Safwat A Ahmed, 4 Alaa M Nafady, 2 Amira S Wanas, 1,5 Babu Tekwani, 1,6 Hashim A Hassanean, 4 Mahmoud A ElSohly 1,7 1 National Center for Natural Products Research, School of Pharmacy, University of Mississippi, USA 2 Department of Pharmacognosy, Faculty of Pharmacy, Al-Azhar University, Egypt 3 Department of Pharmacognosy, Faculty of Pharmacy, Alexandria University, Egypt 4 Department of Pharmacognosy, Faculty of Pharmacy, Suez Canal University, Egypt 5 Department of Pharmacognosy, Faculty of Pharmacy, Minia University, Egypt 6 Department of Bimolecular Sciences, School of Pharmacy, University of Mississippi, USA 7 Department of Pharmaceutics and Drug Delivery, School of Pharmacy, University of Mississippi, USA Correspondence: Mohamed M Radwan, National Center for Natural Products Research, School of Pharmacy, University of Mississippi, USA, Tel +16629151708, Fax +16629155287, Email mradwan@olemiss.edu Received: May 02, 2017 | Published: May 26, 2017 Abstract Chemical investigation of the Red Sea tunicate Polyclinum constellatum afforded nine compounds, identified as thymidine (1), uridine (2), adenosine (3), inosine (4), 24-methylene cholesterol (5), dihydrocholesterol (6), cholesterol (7), oleic acid (8) and 1,3-palmityl-2-palmitoleoylglycerol (9). All compounds isolated for the first time from the genus polyclinum. Acetylation of compounds 1, 2, 3, 4 and 5 yielded thymidine 3`,5`-diacetate (1-Ac), uridine 2`,3`,5`-triacetate (2-Ac), adenosine 2`,3`,5`-triacetate (3-Ac), inosine 2`,3`,5`-triacetate (2-Ac) and 24-methylene cholesterol-3-acetate (5- Ac), respectively. Compound 5 showed potent antitrypanocidal activity with IC 50 and IC 90 values of 3.39 and 6.69µg/mL, comparable to α-difluoromethylornithine (3.58 and 8.53μg/mL). The overall findings of the present study have shown that the Red Sea tunicate Polyclinum constellatum revealed different varieties of secondary metabolites with antiprotozoal and cytotoxic activities. Keywords: red sea, tunicate, polyclinum constellatum, antimicrobial, antimalarial, antiprotozoal, cytotoxicity, acetylation of compounds, metabolites, cellulose, appendicularia, natural products Pharmacy & Pharmacology International Journal Research Article Open Access