s Chemis ct try u d & o r R P e s l e a r a Muñoz et al., Nat Prod Chem Res 2015, 3:4 r u t c h a N Natural Products Chemistry & Research DOI: 10.4172/2329-6836.1000182 ISSN: 2329-6836 Research Article Open Access Antifungal and Insecticidal properties of the Phytoconstituents of Drimys winteri (Winteraceae) growing in Chiloe Island (Chile) Muñoz O1*, Gutierrez M2, Gonzalez R3, Hammann S4 and Vetter W4 1Departamento de Química, Facultad de Ciencias, Universidad de Chile, Casilla 653, Santiago, Chile 2Laboratorio de Síntesis Orgánica, Instituto de Quimica de Recursos Naturales, Universidad de Talca, Talca, Chile 3Facultad de Ciencias Agronomicas, Universidad de Chile 4University of Hohenheim, Institute of Food Chemistry (170b), Garbenstr, 28, D-70593 Stuttgart, Germany Abstract Seven drimane-type sesquiterpenes: polygodial, drimenol, drimendiol, drimenin, isodrimininol, isodrimenin and proximadiol (cryptomeridiol); three lignans: sesamin, cubebin and its epimer eudesmin, and one sterol: (β-sitosterol) were isolated from Drimys winteri (Winteraceae) from Chiloe Island, Chile. Eudesmin and its phytosterols were isolated from the plant for the first time, and sesamin was found to be the major product in the plant’s bark by a significant margin. Furthermore, qualitative and quantitative differences were detected between some products. Structures were established using 1D and 2D NMR (COSY, HSQC, and HMBC) spectra, HRESIMS and comparison with published data. Some isolated compounds were tested against the phytopathogenic fungi Botritys cinerea, Dothiorella sp., Penicillium sp. and Cladosporium cladosporoides; additionally, a study was carried out to determine the contact toxic action of leaf and stem bark extracts upon phloematic insect pests from the Pseudococcus viburni (Signoret) (Hemiptera: Pseudococcidae) mealy bug complex. Keywords: Drimys winteri; Lignans; Sterols; Sesquiterpenoids; sesquiterpenes reported from the bark and leaves collected in Brazil Antifungal activity; Insecticidal properties [13] were not considered because D. winteri is not present in Brazil [3]. The main characteristics and properties of this tree have been Introduction attributed to the presence of certain sesquiterpenoids. The reactivity Drimys winteri J. R and G. Forster var. chilensis (DC.) A. Gray is a of the unsaturated dialdehyde functionality towards biological tree native to Central and Southern Chile, which grows in continental nucleophiles is considered responsible for the antifeedant activity of Chile from Coquimbo to Aysén, including Chiloé Island. It is abundant those compounds (polygodial and epipolygodial) [14,15]. The chemical in wet swampy localities from sea level to an altitude of 1700 m [1]. In composition of the essential oils obtained by hydrodistillation of the continental Chile, Winteraceous plants are represented by two species stem bark and leaves of continental Chile (Santiago) D. winteri was also of Drimys: D. winteri J. R. Forster and G. Forster and D. andina Reiche studied using GC and GC/MS for which results have been published [2]. D. winteri is a tree whereas D. andina is a shrub [3]. Marticorena [16]. The essential oils from the stem bark and leaves of D. winteri from and Quezada also distinguish the chilensis (DC.) A. Gray variety of D. Chiloé Island and continental Chile (Santiago) were studied revealing winteri [4]. Drimys winteri is a sacred plant to the Mapuche indigenous certain differences in their chemical composition. Sesquiterpene people, who uses its aerial parts for the treatment of dermatitis, hydrocarbons constituted the main chemical groups in the stem bark stomach pain, toothache, tumors, and other illnesses [5]. Extracts oils (in which α-santalene, trans -β- bergamotene curcumenes are the of the stem bark have long been used to treat human and bovine major components), whereas monoterpenes (in which (α) - pinene, diseases [6]. In the past, the stem bark of D. winteri was exported to β-pinene, and linalool are the main components) constituted the Europe as an antiscorbutic medicine [1,5]. The tree is also used for main chemical groups in the leaves of Island plants. Sesquiterpenes commercial purposes, in wood production, crafts, manufacturing and (germacrene) and phenylpropanoids are most abundant in the leaves the pulp industry due to the high quality of its fibers. Its architectural of continental plants [16]. The aim of the present study was to compare characteristics and natural resistance to insects and microorganisms the chemical profile of insular D. winteri from Chiloé Island (growing are advantageous for playgrounds in parks and gardens in a wide range and flourishing in climate conditions of high humidity, abundant of weather conditions. The wood of this species is highly valued and water and very scarce volcanic soil) that differ greatly from those where used to manufacture furniture and musical instruments as well as to the continental population is found. We also separately investigated protect crops [7]. Furthermore, extracts of aerial parts of D. winteri have the components present in the bark and leaves and evaluated their industrial applications in products such as cosmetics, phytonutrients and pest repellent agents [7-9]. Previous reports on the chemical composition of continental D. winteri revealed the presence of tannins, *Corresponding author: Orlando Muñoz, Professor, Universidad de Chile, Chemistry Las Palmeras 3425 Ñuñoa Santiago, Santiago 7800003, Chile, Tel: 56-2-92343886; flavonoids and essential oils containing sesquiterpene lactones [5]. The Fax: 56-2-22713888; E-mail: [email protected] drimane-type sesquiterpenoids possess a wide variety of biological May 28, 2015; July 07, 2015; July 15, 2015 activities, including antimicrobial, pungency, antibacterial, antifungal, Received Accepted Published antifeedant, cytotoxic, molluscicidal, piscicidal, growth regulation and Citation: Muñoz O, Gutierrez M, Gonzalez R, Hammann S, Vetter W (2015) Antifungal phytotoxic properties [9]. The pungency of several drimanes and their and Insecticidal properties of the Phytoconstituents of Drimys winteri (Winteraceae) growing in Chiloe Island (Chile). Nat Prod Chem Res 3: 182. doi:10.4172/2329- irritant properties on the skin (allergies) has also attracted attention [6- 6836.1000182 10]. To the best of our knowledge, as of the present, fourteen drimane- type sesquiterpenoids have been isolated from continental D. winteri Copyright: © 2015 Muñoz O, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted and probably also from D. andina [5,6], as well as one aromadendrane use, distribution, and reproduction in any medium, provided the original author and derivative [11] and one drimane-type sesquiterpene trimer [12]. The source are credited. Nat Prod Chem Res Volume 3 • Issue 4 • 1000182 ISSN: 2329-6836 NPCR, an open access journal Citation: Muñoz O, Gutierrez M, Gonzalez R, Hammann S, Vetter W (2015) Antifungal and Insecticidal properties of the Phytoconstituents of Drimys winteri (Winteraceae) growing in Chiloe Island (Chile). Nat Prod Chem Res 3: 182. doi:10.4172/2329-6836.1000182 Page 2 of 6 insecticidal and antifungal properties for potential industrial use. The (b) and another compound identified as sesamin c (0.95 g) (b). investigation also showed an unusual concentration of the sesamin 2. Powdered leaves (350 g) were Soxhlet-extracted using the same lignan in bark from Chiloe canelo populations in comparison with procedure as described for stem bark and produced 25.9 g of yellow continental populations, which would partially explain its extensive residue. Part of this (17.5 g) was mixed with basic aluminum oxide use by the local population as a treatment for certain conditions [17]; 90 (Merck) and subjected to chromatographic separation using an and the absence of bacteria and insect attacks on the species [18]. Until n-hexane-ethyl acetate solvent mixture. Elution with n-hexane-ethyl now, neither the existence of the lignan eudesmin nor its phytosterol acetate (9:1) yielded 0.65 g of a yellow residue consisting of a mixture content had been reported. This study also evaluates its possible use as of four compounds. Further separation using the Chromatotron (film a dietary supplement. thickness 2 mm) and elution with n-hexane-ethyl acetate (8:2) produced Materials and Methods three main compounds which were further purified by CC (silica gel 230-400 mesh, 600 g) and fractionated by n-hexane-ethyl acetate (9:1 to General 8:2) gradient elution yielding cubebin (0.12 g), its epimer (epi-cubebin (0.061 g) and eudesmin (0.041 g); Sesamin was also isolated. Column chromatography (CC) was carried out using silica gel 60G (Merck 7734, Germany). Thin layer chromatography (TLC) took Polygodial (Figure 1) (Scheme a): [α] D and spectroscopic place on silica gel GF254 (Merck 5554) with i) hexane-ethyl acetate data (NMR, MS) were identical to data previously reported on this 8:2 and ii) hexane-acetone 8:2. Spots were detected by UV light or compound [6]. with a Liebermann-Burchard reagent and heated to 110ºC for 1 min. Drimenol (Figure 2) (Scheme a): The MS, 1HNMR and 13CNMR Prep. TLC was performed on 2-mm thick silica gel F plates (Merck 254 data on this compound correlated well with that reported elsewhere 7731) and Chromatotron (Harrison-research 7924 T) 1 or 2 mm thick [12,19,20]. discs, using silica gel 60 PF 254 (Merck 7749). Flash chromatography took place on silica gel 60 H (Merck 7739) with a hexane-ethyl acetate Drimendiol (Figure 3) (Scheme a): The1 HNMR and 13CNMR gradient of 0-100% EtOAc. Mps are uncorrected. Optical rotations spectral data on this compound correlated well with that reported were measured with a Perkin Elmer 241 MC polarimeter. 1HNMR elsewhere [6]. spectra at 500MHz were recorded in CDCl3 on a Bruker-DRX 500 MHz 1 13 1H Larmor frequency equipped with a 5-mm probe. 1H, 13C and 2D Drimenin: The MS, HNMR and CNMR spectral data on this (COSY, HMQC, HMBC) experiments were performed using standard compound correlated well with reported data [20,21]. Bruker pulse sequences. Capillary NMR spectra were recorded on a Isodrimeninol: Already identified in D.