Natural products from scleroderma sp and leptoscyphus expansus (lehm ) grolle

Natural Products from Scleroderma sp and Leptoscyphus expansus (Lehm.) Grolle Kong Chiak Wu A Thesis Submitted for the Degree of Master of Science Department of Chemistry National University of Singapore 2008 I ACKNOWLEDGEMENT I would like to acknowledge the enormous help given to me during the course of my research I wish to thank Prof Leslie Harrison for his guidance and advice for the isolation and especially the elucidation of the isolated compounds Special thanks to Xiaowei, Mingxing and Li Wei for their suggestions, encouragement and helps I also want to thank Ms Han Yanhui and Ler Peggy for recording NMR spectra; Wong Lai Kwai and Lai Hui Ngee for Mass Spectra II Abstract The natural products from an un-identified Scleroderma sp and Leptoscyphus expansus (Lehm.) Grolle were studied A total of five known compounds were isolated from Scleroderma sp Of the five compounds isolated, four were pulvinic acid derivatives which were responsible for the distinctive colour of the fungus The four pulvinic acid derivatives are methyl-4,4’-di-O-methylatromentate (56), methyl per-O-methyl-4Eatromentate (57), methyl per-O-methyl-4Z-atromentate (58) and methyl 3’,5’-dichloro4,4’-di-O-methylatromentate (66) The fifth compound from this fungus was a triterpenoid, (22R,23E)-22-Acetoxylanosta-8,23-diene-3α,25-diol (70) All of these compounds have been reported from an unknown species of Lycoperdon sp These results showed that the two fungi share similar biosynthesis pathway despite the fact that they are from two different families A total of seven compounds were isolated from the liverwort Leptoscyphus expansus (Lehm.) Grolle Five of these were known flavones, all of which were reported elsewhere and they show bioactivities against various diseases The flavones are 5-hydroxyl-7-methoxyflavone (92), 5-hydroxy-7,4’-dimethoxyflavone (95), 5,7-dimethoxyflavone (97), 5-hydroxy-6, 7-dimethoxyflavone (98) and 5, 6, 7trimethoxyflavone (Baicalein trimethyl ether) (102) A Chromene, methyl 5, 7dihydroxy-2, 2-dimethyl-2H-chromene-6-carboxylate (103) and a bibenzyls, 2, 2- dimethyl-5-methoxy-8-carboxy-7-(2-phenylethyl) chromene (108) were also isolated from the liverwort Keywords: Scleroderma sp., Leptoscyphus expansus (Lehm.) Grolle, pulvinic acid derivatives, flavones III Table of Contents Chapter Chapter Chapter Reference General Introduction 1.1 Primary Metabolites and Secondary Metabolites 1.2 The Building Blocks 1.3 Natural Products as Drug Leads 1.4 Current State of Industrial Natural Product Research 1.5 Natural Products versus Synthetic Compounds 1.6 NPs and NP-Derived Drugs in Various Disease Area 10 12 Chemistry of an unidentified Scleroderma sp 2.1 Introduction 2.2 Biosynthesis of pulvinic acid derivatives 2.3 Results and Discussion 2.4 Experimental 21 24 26 45 Chemistry of Leptoscyphus expansus (Lehm.) Grolle 3.1 Introduction 2.2 Biosynthesis and bioactivity of flavones 3.3 Results and Discussion 3.4 Experimental 52 54 60 102 111 IV List of Table Table Table Table Table Table Table Table Table Table Table 10 Table 11 Table 12 Table 13 Table 14 Table 15 Table 16 Natural product derived immunosuppressant drugs Oncology drugs derived from plant, microorganism and marine organism H (300 MHz) and 13C (125 MHz) NMR data for methyl 4,4’-di-Omethylatromentate (56) in CHCl3 H (300 MHz) and 13C (125 MHz) NMR data for methyl per-O-methyl4E-atromentate (57) in CHCl3 H (300 MHz) and 13C (125 MHz) NMR data for methyl per-O-methyl4Z-atromentate (58) in CHCl3 H (300 MHz) and 13C (125 MHz) NMR data for methyl 3’,5’-dichloro4,4’-O-methylatromentate (66) in CHCl3 H (300 MHz) and 13C (125 MHz) NMR data for (22R,23E)-22Acetoxylanosta-8,23-diene-3α,25-diol (70) in CHCl3 13 C NMR spectrum of 93 H (300 MHz), 13C (125 MHz) NMR and HMBC (125 MHz) data for 5-hydroxy-7-methoxyflavone (92) in CHCl3 H (500 MHz) and 13C (125 MHz) NMR data for 5-hydroxy-7,4’dimethoxyflavone (95) in CHCl3 H (300 MHz) and 13C (125 MHz) NMR data for 5,7,-dimethoxyflavone (97) in CHCl3 H (500 MHz) and 13C (125 MHz) NMR data for 5-hydroxy-6,7dimethoxyflavone (98) in CHCl3 H (300 MHz) and 13C (125 MHz) NMR data for 5,6,7trimethoxyflavone (102) in CHCl3 H (500 MHz) and 13C (125 MHz) NMR data for Methyl 5, 7dihydroxy-2, 2-dimethyl-2H-chromene-6-carboxylate (103) in CHCl3 H (500 MHz) and 13C (125 MHz) NMR data for Methyl 5, 7dimethoxy-2, 2-dimethyl-2H-chromene-6-carboxylate (104) in CHCl3 H (500 MHz) and 13C (125 MHz) NMR data for 2,2-dimethyl-5methoxy-8-carboxy-7-(2-phenylethyl) chromene (108) in CHCl3 17 18 29 32 35 40 44 64 65 71 75 80 83 88 90 97 V List of Figures Figure Figure Figure Figure Figure Figure Figure Figure Figure Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Figure 16 Figure 17 Figure 18 Figure 19 Figure 20 Figure 21 Figure 22 Figure 23 Figure 24 Figure 25 Biosynthesis of atromentic acid New numbering system for pulvinic acid derivatives Biosynthesis of 56 from its precursor 55 The possible biosynthetic pathway of 58 Biosynthesis of the basic skeleton of flavonoids, naringenin (81) and liquiritigenin (83) Biosynthesis of corresponding flavonol, flavone and 2hydroxyisoflavone from 81 and 83 Intramolecular hydrogen bond of 92 The structure and numbering of flavone (93) HMBC correlation of 92 Complexation of 5-hydroxy-7,4’-dimethoxyflavone (95) with various metallic ions 5-hydroxy-6,7-dimethoxy and 5-hydroxy-7,8-dimethoxy substitution pattern of A ring Alnustin and 5-hydroxy-3, 7, 8-trimethoxyflavone with their 1H resonance at H-8 and H-6 5,6,7-trimethoxy and 6, 7,8-trimethoxy substitution pattern of A ring Chromene moiety Phloroglucinol moiety HMBC for Methyl 5, 7-dihydroxy-2, 2-dimethyl-2H-chromene-6carboxylate (103) Selected NOE for Methyl 5, 7-dimethoxy-2, 2-dimethyl-2Hchromene-6-carboxylate (104) HMBC correlation of Methyl 5, 7-dimethoxy-2, 2-dimethyl-2Hchromene-6-carboxylate (103) Possible synthetic pathway of Methyl 5, 7-dihydroxy-2, 2-dimethyl2H-chromene-6-carboxylate (103) Photochromism of Chromene HMBC correlation of 2,2-dimethyl-5-methoxy-8-carboxy-7-(2phenylethyl) chromene (108) Selected NOE correlation of 2,2-dimethyl-5-methoxy-8-carboxy-7(2-phenylethyl) chromene (108) ORTEP of 2,2-dimethyl-5-methoxy-8-carboxy-7-(2-phenylethyl) chromene (108) Possible biosynthetic pathway of 2,2-dimethyl-5-methoxy-8carboxy-7-(2-phenylethyl) chromene (108) Cyclisation of prenyl group 25 29 31 36 56 57 63 64 65 70 79 80 82 86 86 87 90 91 93 94 97 98 98 100 101 VI List of Symbols and Abbreviations CHI COESY EIMS F3H FNS HMBC HMQC HREIMS IFS IR MS NMR NOE NOESY NP UV Chalcone Isomerase Correlation Spectroscopy Electron Impact Mass Spectrometry Flavanone 3β-hydroxylase Flavone Synthases Heteronuclear Multiple Bond Correlation Heteronuclear Multiple Quantum Correlation High Resolution Electron Impact Mass Spectrometry Isoflavone Synthase Infrared Spectroscopy Mass Spectrometry Nuclear Magnetic Resonance Nuclear Overhauser Enhancement Nuclear Overhauser Enhancement Spectroscopy Natural Product Ultraviolet Spectroscopy VII Chapter General Introduction The origin, properties and purpose of natural product has fascinated researchers for many years For centuries, drugs were entirely of natural origin and composed of herbs, animal products, and inorganic materials Early remedies were mostly derived from plant (herbs) Natural drugs derived from microorganisms have a much shorter history, and their major impact on medicine goes back only about 60 years to the introduction of the antibiotic penicillin Since then microorganisms have played an important part in natural product chemistry Despite of this fact, plants still form the most important sources of natural product-derived drugs Fungi and marine organism have also received attention from researchers and represent new sources of natural products Natural products are generally divided into two broad classes, primary metabolites and secondary metabolites 1.2 Primary Metabolites and Secondary Metabolites Primary metabolites comprise molecules that are essential for life; principally proteins, carbohydrates, fats and nucleic acids, and these molecules are produced by metabolic pathways common to most organisms Thus these common metabolic pathways demonstrate the fundamental unity of all living matter For example, the Krebs cycle is utilized by most organisms to produce energy in the form of adenosine triphosphate (ATP) [1], whereas photosynthesis is used by all green plants to produce food (glucose) [2] The study of primary metabolites (catabolism and anabolism) is known as biochemistry Secondary metabolites are found in only specific organisms, or groups of organisms, and are an expression of the individuality of species [3] Thus, secondary metabolites are of limited distribution and some of them are unique to specific species Secondary metabolites are not necessarily produced under all conditions, and in the vast majority of cases the function of these compounds and their benefit to the organism is not yet known Secondary metabolites are usually not essential for the survival of the producing organism; however, some of them are nevertheless produced with specific purpose For example, cinnamolide (1) is produced by the mollusk Dendrodoris denisoni as a defensive, predator deterrent metabolite [4] Some of the secondary metabolites are produced at specific development periods of the life cycle These include metabolites made during starvation (e.g carbapenem antibiotics produced by Pseudomonas bacteria), in development (e.g antibiotics made when Streptomycetes enter cellular differentiation pathways), and signaling (such as quorum-sensing molecules biosynthesized at particular culture densities of microbes) [24] O O (1) The study of secondary metabolites is known as natural product (NP) chemistry, which provides us with the pharmacologically active NPs 1.2 The Building Blocks The extremely diverse structures of natural products seem confusing to the beginners Fortunately, there are only a few building blocks which are employed in the biosynthesis of NPs This demonstrates the fundamental unity of all living organism The most important building blocks are acetyl coenzyme A (acetyl-CoA) (2), shikimic acid (3), mevalonic acid (4), and 1-deoxyxylusose 5-phosphate (5) These building blocks are actually produced from primary metabolic pathway and shunted into the secondary pathways when a particular metabolic channel is opened They are employed respectively in the acetate, shikimate, mevalonate and deoxyxylusose phosphate pathways [3] Important natural products formed from acetate pathway include phenolic compound [e.g orsellinic acid (6)], prostaglandins [e.g prostaglandin PGF2α (7)] and various fatty acids On the other hand, the shikimate pathway leads to a variety of phenols [e.g ferulic acid (8)], cinnamic acid (9), lignins and alkaloids The mevalonate and deoxyxylulose phosphate pathways are together responsible for the biosynthesis of a vast array of terpenoids Examples of terpenoids are trans citral (10), farnesene (11) and vitamin A (12) CO2H OH OH O CoA S (2) HO OH OH OH (3) OP CO2H (4) O OH (5) Fraction The fourth fraction (1.6 g) was chromatographed on Sephadex LH-20 to afford a fraction (ca 685 mg), which was further chromatographed on silica gel column (60 % EtOAcisooctane) to afford one compound, 5, 6, 7-Trimethoxyflavone (Baicalein trimethyl ether) (102) (137 mg) Fraction The fifth fraction (1.5 g) was chromatographed on Sephadex LH-20 to afford a fraction (450 mg), which was further chromatographed on silica gel column (60 % EtOAcisooctane) and DIOL (30 % EtOAc-isooctane) to give a compound, 5,7Dimethoxyflavone (97) (211.2 mg) Fraction The sixth fraction (1 g) was chromatographed on Sephadex LH-20 to afford a fraction (400 mg), which was further chromatographed on silica gel column (60 % EtOACisooctane) and reversed phase column (80 % MeOH-H2O) to afford a compound, 5Hydroxy-6, 7-dimethoxyflavone (98) (20 mg) 5-Hydroxy-7-methoxyflavone (92) (12.5 mg) Yellow solid; UV (methanol) λmax nm: 269, 306 and; FI-IR νmax (CHCl3) cm-1: 3408, 1668, 1600, 1587 and 1456, 1200 and 1160, 762 and 680; EI-MS m/z (rel int.): 269 [M + H]+ (18), 268 [M]+ (100), 267 [M – H]+ (11), 239 [M-C-OH]+ (33), 225 [M-C-OCH3]+ (11); HREI-MS: m/z 268.0730 105 (C16H12O4 requires m/z 268.0732); 1H NMR (300 MHz, CDCl3): δ 12.71 (1H, br s, 5OH), 7.88 (2H, m, H-2’ and H-6’), 7.53 (3H, m, H-3’, H-4’ and H-5’), 6.66 (1H, s, H-3), 6.49 (1H, d, J = 2.07 Hz, H-8), 6.36 (1H, d, J = 2.07 Hz) and 3.88 (3H, s, 7-OCH3); 13C NMR (75 MHz, CDCl3) and HMBC (125 MHz): δC 182.48 (C-4), 165.60 (C-7), 163.98 (C-2), 162.17 (C-5), 157.78 (C-9), 131.81 (C-4’), 131.30 (C-1’), 129.06 (C-3’ and C-5’), 126.27 (C2’ and C-6’), 105.85 (C-3), 105.69 (C-10), 98.19 (C-6), 92.66 (C-8), and 55.79 (7-OCH3) 5-Hydroxy-7,4’-dimethoxyflavone (95) (2 mg) Yellow solid; UV (methanol) λmax nm: 272, 324 and; FI-IR νmax (CHCl3) cm-1: 3405, 3075, 2925, 1660, 1600, 1587 and 1445, 1182 and 1035; EI-MS m/z (rel int.): 299 [M + H]+ (13), 298 [M]+ (100), 297 [M – H]+ (11), 269 [M-C-OH]+ (30), 255 [M-C-OCH3]+ (12); HREI-MS: m/z 298.0841 (C17H14O5 requires m/z 298.0837); 1H NMR (500 MHz, CDCl3): δ 7.85 (2H, m, H-2’ and H-6’), 7.02 (3H, m, H-3’, H-4’ and H-5’), 6.58 (1H, s, H-3), 6.49 (1H, d, J = 2.07 Hz, H-8), 6.37 (1H, d, J = 2.07 Hz), 3.90 (3H, s, 4’-OCH3 or 7-OCH3) and 3.88 (3H, s, 4’-OCH3 or 7OCH3); 13 C NMR (125 MHz, CDCl3): δC 182.47 (C-4)], 165.40 (C-7), 163.92 (C-2), 162.50 (C-4’), 162.13 (C-5), 157.61 (C-9), 128.05 (C-2’ and C-6’), 123.38 (C-1’), 114.50 (C-3’ and C-5’), 105.50 (C-10), 104.21 (C-3), 98.04 (C-6), 92.63 (C-8), 55.79 (-OCH3), 55.53 (-OCH3) 5,7-Dimethoxyflavone (97) (211.2 mg) Yellow solid; UV (methanol) λmax nm: 264, 307 and; FI-IR νmax (CHCl3) cm-1: 3012, 1643, 1610, 1600, and 1460, 1213 and 1117, 3012; EI-MS m/z (rel int.): 283 [M + H]+ (18), 282 [M]+ (100), 281 [M – H]+ (62), 264 [M- 106 H2O]+ (7), 253 (48), 236 (47), 224 (13), 209 (28); HREI-MS: m/z 282.0882 (C17H14O4 requires m/z 282.0888); 1H NMR (300 MHz, CDCl3): δ 7.81 (2H, m, H-2’ and H-6’), 7.45 (3H, m, H-3’, H-4’ and H-5’), 6.63 (1H, s, H-3), 6.52 (1H, d, J = 2.0 Hz, H-8), 6.31 (1H, d, J = 2.0 Hz, H-6), 3.89 (3H, s, 5-OCH3 or 7-OCH3) and 3.86 (3H, s, 5-OCH3 or 7OCH3); 13C NMR (75 MHz, CDCl3): δC 177.66 (C-4), 164.06 (C-7), 161.0 (C-2), 160.71 (C-5), 159.75 (C-9), 131.24 (C-1’), 131.12 (para, C-4’), 128.79 (meta, C-3’ and C-5’), 125.82 (ortho, C-2’ and C-6’), 108.69 (C-3), 108.67 (C-10), 96.09 (C-6), 92.73 (C-8), 56.19 (-OCH3), and 55.64 (-OCH3) 5-Hydroxy-6, 7-dimethoxyflavone (98) (20 mg) Yellow solid; UV (methanol) λmax nm: 280, 310 and; FI-IR νmax (CHCl3) cm-1: 3515, 1685, 1630, 1600, and 1456, 1200 and 1158, 780 and 685; EI-MS m/z (rel int.): 299 [M + H]+ (8), 298 [M]+ (100), 297 [M – H]+ (10), 283 [M-CH3]+ (70), 269 (11), 255(10), 181 (15), 153 (36); HREI-MS: m/z 298.0837 (C17H14O5 requires m/z 298.0841); 1H NMR (500 MHz, CDCl3): δ 12.67 (1H, br s, 5-OH), 7.89 (2H, m, H-2’ and H-6’), 7.53 (3H, m, H-3’, H-4’ and H-5’), 6.68 (1H, s, H-3), 6.57 (1H, s, H-8), 3.97 (3H, s, 6-OCH3 or 7-OCH3) and 3.93 (3H, s, 6-OCH3 or 7OCH3); 13 C NMR (125 MHz, CDCl3): δC 182.72 (C-4), 163.96 (C-2), 158.90 (C-7), 153.32 (C-9), 153.02 (C-5), 132.68 (C-6), 131.83 (C-4’), 131.29 (C-1’), 129.08 (C-3’ and C-5’), 126.24 (C2’ and C-6’), 106.29 (C-10), 105.61 (C-3), 90.63 (C-8), 60.85 (-OCH3) and 56.32 (-OCH3) 5, 6, 7-Trimethoxyflavone (Baicalein trimethyl ether) (102) (137 mg) Yellow solid; UV (methanol) λmax nm: 263, 301 and; FI-IR νmax (CHCl3) cm-1: 3000, 2945, 1625, 1595, 107 and 1348, 1232 and 1118, 770 and 690; EI-MS m/z (rel int.): 313 [M + H]+ (5), 312 [M]+ (13), 297 [M – CH3]+ (100), 284 [M-CO]+ (10), 269 (14), 254 (16), 167 (12); HREI-MS: m/z 312.0998 (C18H16O5 requires m/z 312.0993); 1H NMR (300 MHz, CDCl3): δ 7.83 (2H, m, H-2’ and H-6’), 7.46 (3H, m, H-3’, H-4’ and H-5’), 6.78 (1H, s, H-3), 6.65 (1H, d, s, H-8), 3.93 (3H, s, 5-OCH3 , 6-OCH3 or 7-OCH3), 3.92 (3H, s, 5-OCH3 , 6-OCH3 or 7-OCH3) and 3.86 3.93 (3H, s, 5-OCH3 , 6-OCH3 or 7-OCH3); 13 C NMR (75 MHz, CDCl3): δC 177.42 (C-4), 161.23 (C-2), 157.80 (C-7), 154.43 (C-9), 152.25 (C-5), 140.22 (C-6), 131.50 (C-1’), 131.22 (C-4’), 128.80 (C-3’ and C-5’), 125.84 (C2’ and C-6’), 112.55 (C-10), 107.94 (C-3), 96.16 (C-8), 62.01 (5- or 6-OCH3), 61.34 (5- or 6-OCH3), 56.16 (7-OCH3) Methyl 5, 7-dihydroxy-2, 2-dimethyl-2H-chromene-6-carboxylate (103) (14.5 mg) Yellow oil; UV (methanol) λmax nm: 262, 333 and; FI-IR νmax (CHCl3) cm-1: 3437, 3022, 2976 1649, 1588, 1461, and 1368, 1213, 1156; EI-MS m/z (rel int.): 251 [M + H]+ (5), 250 [M]+ (20), 235 [M – CH3]+ (37), 217 (4), 203[M-2xCH3-OH]+ (100), 175 (2), 147 (1) 135 (2), 91 (3), 69 (5), 55 (2); HREI-MS: m/z 250.0840 (C13H14O5 requires m/z 250.0837); 1H NMR (500 MHz, CDCl3): δ 6.61 (1H, d, J = 10.11 Hz, H-4), 5.96 (1H, s, H-8), 5.46 (1H, d, J = 10.11 Hz, H-3), 4.03 (3H, s, 11-OCH3), 1.42 (6H, s, H3-12 and H313); 13 C NMR (125 MHz, CDCl3) and HMBC (125 MHz, CDCl3): δC 169.79 (C-11), 160.60 (C-5, C-7 and C-9), 125.90 (C-3), 115.91 (C-4), 102.31 (C-10), 96.75 (C-8), 93.51 (C-6), 77.74 (C-2), 52.48 (-OCH3) and 28.29 (C-12 and C-13) 108 Methylation of Methyl 5, 7-dihydroxy-2, 2-dimethyl-2H-chromene-6-carboxylate (104) Seven milligram of 103 was reacted with iodomethane (CH3I) in boiling acetone in the presence of excess potassium carbonate anhydrous (K2CO3) The reaction was carried out under reflux at 60 oC and was monitored with TLC analysis CH3I was added to the reaction mixture in 30 minutes interval until the reaction was completed The product, methyl 5,7-dimethoxy-2,2-dimethyl-2H-chromene-6-carboxylate (104) was separated from the reaction mixture by column chromatography in a Pasteur pipette using acetone as the mobile phase Only the product 104 was eluted and it was pure (based on TLC and H NMR analysis) 6.8 mg of product was obtained Methyl 5,7-dimethoxy-2,2-dimethyl-2H-chromene-6-carboxylate (104) (6.8 mg) Yellow oil; UV (methanol) λmax nm: 232, 286 and; FI-IR νmax (CHCl3) cm-1: 3022, 2952, 1645, 1607, 1466, and 1370, 1213 and 1156; EI-MS m/z (rel int.): 279 [M + H]+ (2), 278 [M]+ (16), 277 [M-H]+, 263 [M – CH3]+ (100), 247 [M-OCH3]+ (6), 217 (12), 189 (5), 161 (8) 116 (5), 44 (17); HREI-MS: m/z 278.1144 (C15H18O5 requires m/z 278.1149); 1H NMR (500 MHz, CDCl3): δ 6.48 (1H, d, J = 10.1 Hz, H-4), 6.20 (1H, s, H-8), 5.51 (1H, d, J = 10.1 Hz, H-3), 3.89 (3H, s, 11-OCH3), 3.80 (3H, s, 5-OCH3), 3.78 (3H, s, 7-OCH3) and 1.42 (6H, s, H3-12 and H3-13); 13C NMR (125 MHz, CDCl3) and HMBC (125 MHz, CDCl3): δC 166.73 (C-11), 157.92 (C-7), 156.18 (C-9), 154.73 (C-5), 127.72 (C-3), 116.48 (C-4), 76.81 (C-2), 110.19 (C-6), 107.83 (C-10), 96.16 (C-8), 62.88 (-OCH3), 56.01 (-OCH3), 52.30 (-OCH3), 27.82 (C-12 and C-13) 109 2, 2-dimethyl-5-methoxy-8-carboxy-7-(2-phenylethyl) chromene (108) (46.1 mg) White solid; ; UV (methanol) λmax nm: 231, 282 and; FI-IR νmax (CHCl3) cm-1: 3450 (br), 2977, 2934, 1727, 1596, 1459, and 1373, 1115 and 1040; EI-MS m/z (rel int.): 339 [M + H]+ (2), 338 [M]+ (6), 323 [M – CH3]+ (48), 305 [M-CH3-H2O]+ (7), 294 [M-CO2]+ (12), 279 [M-CH3-CO2]+ (100), 188 (44) 145 (8), 115 (5), 91 (6); HREI-MS: m/z 338.1517 (C21H22O4 requires m/z 338.1512); 1H NMR (500 MHz, CDCl3): δ 7.27 (2H, m, H-3” and H-5”), 7.25 (2H, m, H-2” and H-6”), 7.17 (1H, m, H-4”), 6.65 (1H, d, J = 10.1 Hz, H-4), 6.24 (1H, s, H-6), 5.63 (1H, d, J = 10.1 Hz, H-3), 3.77 (3H, s, 5-OCH3), 3.31 (2H, m, H2α), 2.92 (2H, m, H2-β) and 1.54 (6H, s, H3-1’ and H3-2’); 13 C NMR (125 MHz, CDCl3) and HMBC (125 MHz, CDCl3): δC 165.58 (Carboxyl Carbon), 156.89 (C-5), 152.83 (C9), 148.52 (C-7), 142.02 (C-1”), 128.76 (C-2” and C-6”), 128.22 (C-3” and C-5”), 128.01 (C-3), 125.77 (C-4”), 116.53 (C-4), 110.14 (C-8), 108.23 (C-10), 107.99 (C-6), 79.54 (C2), 55.64 (-OCH3), 38.65 (C-α), 37.65 (C-β), and 27.51 (C-1’ and C-2’) Crystallographic data for 2, 2-dimethyl-5-methoxy-8-carboxy-7-(2-phenylethyl) chromene C21H22O4, Mr 338.39, orthorhombic, space group Pna2(1), a = 20.6556(8) Å, b = 9.0133(4) Å, c = 19.5612(8) Å, α = β = γ = 90o, V = 1335.2(2) Å, Z = 8, Density (calculated) = 1.234 Mg/m3, F(000) = 1440, λ = 0.71073 Å, µ = 0.085 mm-1 Data were collected using a crystal of size ca 0.60×0.36×0.20 mm3 (CCDC 242768) 110 Reference Berg, JM; JL Tymoczko, L Stryer (2002) Biochemistry - 5th Edition WH Freeman and Company, 465-484, 498-501 Raven, Peter H.; Ray F Evert, Susan E Eichhorn (2005) Biology of Plants, 7th Edition New York: W.H Freeman and Company Publishers, 124-127 Dewick, P M (2005) Medicinal Natural Products: A Biosynthetic Approach 2nd ed John Wiley and Sons Grkovic, T., Appleton, D R., and Copp, B R (2005) Chemistry in New Zealand Dec 2005, 12-15 Stanforth, S P (2006) Natural Product Chemistry at a Glance Blackwell Publishing Pp 2-6 Newman, D.J., Cragg, G.M., Snader, K.M (2000) Nat Prod Rep 17, 215-234 Mann, J (2000) Murder, Magic and Medicin., 2nd ed Oxford University Press: Oxford, UK Newman, D.J., Cragg, G.M and Snader, K.M (2003) J Nat Prod 66, 10021037 Butler, M S (2004) J Nat Prod 67, 2141-2153 10 Shu, Y.Z (1998) J Nat Prod 61, 1053-1071 11 Koehn, F E., and Carter, G T (2005) Nature Rev Drug Discovery 4, 206-220 12 Rawlins, M D (2004) Nature Rev Drug Discovery 3, 360-364 13 Rouchi, A M (2003) Chem Eng New October 13, 77-91 14 Cordell, G A (2002) Phytochem Rev 1, 261-273 15 Strohl, W R (2000) Drug Discovery Today 5, 39-41 111 16 Lee, A., Breitenbucher, J G (2003) Curr Opin Drug Discovery Dev 6, 494-508 17 Horton, D A., Bourne, G T Smythe, M L (2003) Chem Rev 103, 893-930 18 Newman, D., Cragg, G., Kingston, D (Academic, London, 2003) The Practice of Medicinal Chemistry (ed Wermuth, C G.) 91-109 19 Feher, M and Schmidt, J M (2003) J Chem Inf Comput Sci 43, 218-227 20 Henkel, T., Brunne, R., Muller, H and Reichel, F (1999) Angew Chem Int Ed Engl 38, 643-647 21 Stahura, F., Godden, J W., Ling, X and Bajorath, J (2000) J Chem Inf Comput Sci 40, 1245-1252 22 Martin, Y C and Critchlow, R E (1999) J Comb Chem 1, 32-45 23 Butler, M S (2005) Nat Prod Rep 22, 162-195 24 Clardy, J and Walsh, C (2004) Nature 432, 829-837 25 Bonfiglio, G., Russo, G and Nicoletti, G (2002) Expert Opin Investig Drugs 11, 529-544 26 Chopra, I (2002) Drug Resist Update 5, 119-125 27 Oki, T., Hirano, M., Tomatsu, K., Numata, K and Kamei, H (1989) J Antibit 42, 1756-1762 28 Molina, J M., Goguel, J., Sarfati, C., Michiels, J F., Desportes-Livage, I., Balkan, S., Chastang, C., Cotte, L., Maslo, C., Struxiano, A., Drouin, F., and Decazes, J M (2000) AIDS (London) 14, 1341-1348 29 Graul, A I (2001) Drug News Perspect 14, 12-31 30 Yang, S S., Cragg, G M., Newman, D J and Bader, J P (2001) J Nat Prod 64, 265-277 112 31 Buss, A D., Cox, B and Waigh, R D (2003) in Burger’s Medicinal Chemistry and Drug Discovery, 6th Ed, Vol 1: Drug Discovery, ed Abraham, D J., Wiley, Hoboken, New Jersey Pp 847-900 32 ML Laboratories: Further Information Available at http://www.mllabs.com 33 Liu, J S., Zhu, Y L., Yu, C M., Zhou, Y Z., Han, Y Y., Wu, G W and Qi, B F (1986) Can J Chem 64, 837-839 34 Kaminska, B., Gaweda-Walerych, K and Zawadzka, M., (2004) J Cell Mol Med 8, 45-58 35 Donnelly, R., Idris, I and Forrester, J V (2004) Br J Ophthalmol 88, 145-151 36 Kiuchi, M., Adachi, K., Kohara, T., Minoguchi, M., Hanano, T., Aoki, Y., Mishina, T., Arita, M., Nakao, N., Ohtsuki, M., Hoshino, Y., Teshima, K., Chiba, K., Sasaki, S and Fujita, T (2000) J Med Chem 43, 2946-2961 37 Hall, I R., Stephenson, S L., Buchanan, P K., Yun, W., Cole, A L (2003) Journal of Edible and Poisonous Mushrooms of the World, New Zealand Institute for Crop and Food Research Limited: Christchurch, New Zealand 38 Sims, K P., Watling, R., Jeffries, P (1995) Mycotaxon LVI, 403-420 39 Ramsey, R (2003) Trial Field Key to the Species of Sclerodermataceae in the Pacific Northwest prepared for the Pacific Northwest key Council 40 Bresinsky, A., Besl, H (1990) A Colour Atlas of Poisonous Fungi: A Handbook for Pharmacists, Doctors and Biologists Wolfe Publishing Ltd 41 Wang, C Y., Gao, J M., Yang, X., Dong, Z J and Liu, J K (2002) Chinese Traditional and Herbal Drugs 33 (9), 778-780 113 42 Winner, M., Giménez, A., Schmidt, H., Sontag, B., Steffan, B and Steglich, W (2004) Angew Chewm Int Ed 43, 1883-1886 43 Kanokmedhakul, S., Kanokmendhakul, K., Prajuabsuk, T., Soytong, K., Kongsaeree, P and Suksamrarn, A (2003) Planta Medica 69, 568-571 44 Steffan, B., Steglich, W (1984) Angew Chem Int Ed Engl 23, 445-447 45 Gill, M., Lally, D A (1985) Phytochemistry 24, 1351-1354 46 Letcher, R M and Eggers, S H (1967) Tetrahedron Letter 3541-3546 47 Arnold, N., Steglich, W., and Besl, H Z (1996) Mykologie 62, 69-73 48 Marumoto, R., Kilpert, C., Steglich, W (1986) Z Naturforsch., C: J Biosci 41, 363-365 49 Foden, F R., McCormick, J., O’Mant, D.M (1975) Journal of Medicinal Chemistry 18, 199-203 50 Sar, S A., Blunt, J W., Cole, A L J., Din, L B and Munro, M H G (2005) Journal of Natural Products 68, 1799-1801 51 Foden F R., McCormick, J., and O’Mant, D M (1975) Journal of Medicinal Chemistry 18, 199-203 52 Marumoto, R., Pilzlaboratorium, R., Hyogo, A., Kilpert, C., and Steglich, W (1986) Z Naturforsch 41C, 363-365 53 Heurtaux, B., Lion, C., Gall, T L and Mioskowski, C (2004) Journal of Organic Chemistry 70, 1474-1477 54 Singh, P and Anchel, M (1971) Phytochemistry 10, 3259-3262 55 Culberson, W L and Culberson, C F (1970) The Bryologist 73 (1), 1-31 114 56 Shin, Y., Tamai, Y and Terazawa, M (2000) International Journal of Medicinal Mushroom 2, 201-207 57 Shin, Y., Tamai, Y and Terazawa, M (2000) Eurasian Journal of Forest Research 1, 43-50 58 Gedge, D R., Pattenden, G and Smith, A G (1986) J Chem Soc Perkin Transaction I 2127-2131 59 Gill M (2001) Australian Journal of Chemistry 54, 721-734 60 Dhami, K S and Stothers, J B (1966) Can J Chem 44, 2855 61 Agrawal, P K., Thakur, R S and Bansal, M C (ed.) (1989) Carbon-13 NMR of Flavonoids, edited by Agrawal, P K Chapter pp 95-182 Elsevier 62 Agrawal, P K and Markham, K R (ed.) (1989) Carbon-13 NMR of Flavonoids, edited by Agrawal, P K Chapter pp 1-40 Elsevier 63 Hofer, O (1975) Tetrahedron Lett 3415 64 Asakawa, Y (1971) Bull Chem Soc Japan 44, 2761-2766 65 Wagner, H., Maurer, G., Farkas, L., Hansel, R and Ohlendorf, D (1971) Chem Ber 104 2381 66 Asaka, Y., Kondo, K., Tori, M., Hashimoto, T., and Ogawa, H (1991) Phytochemistry 30, 219-234 67 Geissman, T A (ed.) (1962) The Chemistry of Flavonoid Compounds p 416 Macmillan, New york 68 Jaipetch, T., Reutrakul, V., Tuntiwachwuttikul, P and Santisuk, T (1982) Phytochemistry 22 (2) 625-626 115 69 Ritter, J K., Kessler, F K., Thompson, M T., Grove, A D., Auyeung, D J., and Fisher, R A (1999) Hepatology 30, 476-484 70 Malfatti, M A and Felton, J S (2001) Carcinogenesis 22, 1087-1093 71 Walle, U K and Walle, T (2002) Drug Metabolism and Disposition 30, 564569 72 Sutthanut, K., Sripanidkulchai, B., Yenjai, C and Jay, M (2007) Journal of Chromatograhphy A 1143, 227-233 73 Ahmad, F., Cheng, L L and Mat Ali, R (2003) ACGC Chemical Research Communications 16, 48-52 74 Mayer, R (1992) Planta Medica 59, 269-271 75 Wen, X., Walle, U K and Walle, T (2005) Carcinogenesis 26, 803-809 76 Zahir, A., Jossang, A., and Bodo, B (1996) Journal of Natural Products 59, 701703 77 Silva, M and Mundaca, J M (1971) Phytochemical Reports 10, 1942-1943 78 Khamis, S., Bibby, M C., Brown, J E., Cooper, P A., Scowen, I and Wright, C W (2004) Phytotherapy Research 18, 507-510 79 Panichpol, K and Waterman, P G (1978) Phytochemistry 17, 1363-1367 80 Ahmad, S (1983) Planta Medica 68, 62-63 81 Wang, S X., Zhang, F J., Feng, Q P and Li, Y L (1992) Journal of Inorganic Biochemistry 46, 251-257 82 Hayashi, K., Hayashi, T., Otsuka, H and Takeda, Y (1997) Journal of Antimicrobial Chemotherapy 39, 821-824 83 Mues, R (1985) Abstracta Botanica 9, Suppl 2, 171 116 84 Markham, K R (1988) in The Flavonoids Advances in Research Since 1980, edited by Harborne, J B p 427 London: Chapman and Hall 85 Asakawa, Y (1995) Progress in the Chemistry of Organic Natural Products 65, p 1-618 Springer-Verlag Wien New York 86 Grolle, R (1962) Nova Acta Leopoldina 25 (161), 1-143 87 Engel, J J (1991) Studies on Geocalycaceae (Hepaticae) IV Lamellocolea, a new genus of Leptoscyphoideae from New Zealand J Hatt Bot Lab 70, 63-78 88 Gradstein, S R., Churchill, S P., Salazar Allen, N (2001) Mem NY Bot Gard 86, 1-577 89 Ralston, L., Subramanian, S., Matsuno, M., and Yu, O (2005) Plant Physiology 137, 1375-1388 90 Shimada, N., Aoki, T., Sato, S., Nakamura, Y., Tbata, S and Ayabe, S (2003) Plant Physiology 131, 941-951 91 Sawada, Y., Kinoshita, K., Akashi, T., Aoki, T., and Ayabe, S (2002) Plant Journal 31, 555-564 92 Takahama, U., and Yoshitama, K (1998) Journal of Plant Research 111, 97 93 Hiraga, S., Sasaki, K., Ito, H., Ohashi, Y., Matsui, H (2001) Plant Cell Physiology 42, 462 94 Klebanoff, S J (2005) Journal of Leukocyte Biology 77, 598 95 Garcia-Argáez, A N., Apan, T O R., Delgado, H P., Velázquez, G., MartinezVázquez, M (2000) Planta Medica 66, 279 96 Bors, W., Heller, W., Michel, C., and Saran, M (1990) Methods Enzymology 186,343 117 97 Kabeya, L M., de Marchi, A A., Kanashiro, A., Lopes, N P., da Silva, C H T P., Pupo, M T and Lucisano-Valim, Y M (2006) Bioorganic and Medicinal Chemistry 15, 1516-1524 98 Bisignano, G., Sanogo, R., Marino, R., Aquino, R., D’Angelo, V., Germano, M P., De Pasquale, R., and Pizza, C (2000) Letters in Applied Microbiology 30, 105-108 99 Cai, L., Wu, C D (1996) Journal of Natural Products 59, 987-990 100 Williamson, A., Day, A J., Plumb, G W., and Couteau, D (2000) Biochemical Society Transactions 28, 16-22 101 Steinbeck, C., Spitzer, V., Starosta, M., von Poser, G (1997) Journal of Natural Products 60, 627 102 Gomez, F., Quijano, L., Calderon, A., Perales, A., and Rios, T (1982) Phytochemistry 21, 2095 103 Ashan, M., Armstrong, J A., Gibbons, S., Gray, A I and Waterman, P G (1994) Phytochemistry 37, 259 104 Feld, H., Rycroft, D S and Zapp, J (2003) Z Naturforsch B: Journal of Chemical Sciences 59B, 825-828 105 Oliveira, M M Carvalho L M., Moustrou, C., Samat, A., Guglielmetti, R and Oliveira-Campos, A M E (2001) Helvetica Chimica Acta 84, 1163-1171 106 Bouas-Laurent, H and Dürr, H (2001) Pure Appl Chem 73 (4), 639-665 107 Morita, M., Takahashi, I., Kanai, M., Okafuji, F., Iwashima, M., Hayashi, T., Watanabe, S., Hamazaki, T., Shimozawa, N., Suzuki, Y., Furuya, H., Yamada, T., and Imanaka, T (2005) FEBS Letters 579, 409-414 118 108 Paladini, A C., Marder, M., Viola, H., Wofman, C., Wasowski, C and Medina, J H (1999) Journal of Pharmacy and Pharmacology 51, 519-526 109 Lee, H –H (1981) Journal of Chemical Society: Perkin Transaction I 32053213 110 Bowie, J H and Cameron, D W (1966) Australian Journal of Chemistry 19, 1627 111 Rukachaisirikul, V., Tadpetch, K., Watthanaphanit, A., Saengsanae, N., and Phongpaichit, S (2005) Journal of Natural Product 68, 1218-1221 112 Knaggs, A R (2001) Natural Products Report 18, 334-355 113 Leong, Y W (1997) Isolation and Structure Elucidation of Natural Products A thesis submitted for the degree of Doctor of Philosophy, National University of Singapore 114 Sugeng, K L B., Triadi, B., Rizna, T and Kazuko, K (2002) Eurasian Journal of Forest Research 5, 33-37 115 Lobo, A M., Macedo de Abreu, P., Prabhakar, S., Gondinho, L S., Jones, R., Rzepa, H S and Williams, D J (1988) Phytochemistry 27, 3569 116 Cullmann, F and Becker, H (1999) Zeitschrift fuer Naturforschung C-A Journal of Biosciences 54 (3/4), 147-150 117 Panichpol, K and Waterman, P G (1978) Phytochemistry 17, 1363-1367 118 Kutney, J P and Hanssen, H W (1971) Phytochemistry 10, 3298 119 [...]... 6-OCH 3); δC 171.79 (C- 6) and 54.36 (6-OCH 3)] , a γ-lactone carbonyl [δC 166.13 (C- 1)] , an enolic hydroxyl proton [δH 13.56 (1H, s, 3-OH)], two para-disubstituted benzene rings [δH 8.13 (2H, d, J = 9.06 Hz, H-2’ and H-6 ), 7.20 (2H, d, J = 8.7 Hz, H-8 and H-1 2), 6.96 (2H, d, J = 9.06 Hz, H-3’ and H-5 ) and 6.94 (2H, d, J = 8.7 Hz, H-9 and H-1 1); δC 129.23 (2×, C-2’ and C-6 ), 131.37 (2×, C-8 and C-1 2), ... origin and examples include lysergic acid (1 5) (shikimic acid and terpenoid origin) and the insecticide rotenone (1 6) (acetyl CoA, shikimic acid and terpenoid origin) This is advantageous because it adds diversity to the range of NPs produced by the organism CO2H OCH3 H N CH3 CH3O O H H O O O H N H H (1 5) (1 6) 5 1.3 Natural Products as Drug Leads For thousands of years medicine and natural products (NPs)... chloride test Its UV spectrum exhibits absorption bands at 230, 260, 305 and 385 nm and its IR spectrum showed strong bands for hydroxyl group [3547 cm-1 (OH)], benzene rings (1600, 1513, 1456 cm- 1) as well as γ-lactone (1770 cm- 1) and unsaturated ester [1680 cm-1 (C=O), 1250 and 1068 cm-1 (C-O)] groups H3CO 6 7 5 O 5' OCH3 OH 3 11 3' 1' O H3CO 1 O 9 56 The 1H and 13 C NMR spectra (see Table 3) contain signals... OH OH ( 6) ( 7) CO2H CH3 O CO2 H HO CHO ( 8) ( 9) (1 0) (1 1) OH (1 2) In addition to acetyl-CoA, shikimic acid, mevalonic acid, and deoxyxylulose phosphate, other building blocks based on amino acids are frequently employed in NP biosynthesis Examples include the alkaloid laburnine (1 3) and the tripeptide (1 4) (precursor of many types of penicillin) 4 H H2N H H N SH CO2H O O NH OH HO2C N (1 3) (1 4) It is... biochemical specificity and other molecular properties that make them favourable as lead structures for drug discovery, and which serve to differentiate them from libraries of synthetic and combinatorial compounds [18] NPs-derived drugs [e.g Taxol (2 1) and Rapamycin (2 2)] show structural difference from that of the synthetic drugs [e.g Viagra (2 3) and Prozac (2 4)] [24] Studies reveal that natural products. .. traditional medicines and natural poison [6] Some of the plant derived NPs used as early medicine are aspirin (1 7), morphine (1 8), quinine (1 9), and pilocarpine (2 0) [6-7] Aspirin and morphine are used as pain killers Unfortunately, morphine also induces a state of euphoria and mental detachment, together with nausea, vomiting, constipation, tolerance, and addiction On the other hand, quinine has been... (75 MHz) NMR data for methyl 4,4’-di-Omethylatromentate (5 6) in CHCl3 (J in Hz in parentheses) Position δH (CDCl 3) δC (CDCl 3) 1 166.13 2 104.97 3 158.60 4 154.58 5 115.00 6 171.79 7 124.96 8 7.20 d (8. 7) 131.37 9 6.94 d (8. 7) 113.47 10 -159.40 11 6.94 d (8. 7) 113.47 12 7.20 d (8. 7) 131.37 1’ -121.67 2’ 8.13 d (9.0 6) 129.23 3’ 6.96 d (9.0 6) 113.82 4’ -159.60 5’ 6.96 d (9.0 6) 113.82 6’ 8.13 d (9.0 6) 129.23... N O O O O HO O (2 5) N HO H H OH OH HO OH O CONH2 (2 6) (2 7) (2 8) O O H O O CO2H CO2H H2N H2N O H O OCH3 O O O O O OH O CO2H (2 9) O (3 0) (3 1) 14 Some of the earliest NP derived drugs used for the treatment of pain and central nervous system (neurological disease) diseases include the opiate alkaloids from the opium poppy, Papaver somniferum, ergot from the fungus, Claviceps purpurea, and the tropane alkaloids... oncology are mainly from three sources: plant, 17 microorganism and marine organism Some of the NPs and NP-derived drugs with anticancer properties are presented in Table 2 Table 2 Oncology Drugs derived from plant, microorganism and marine organism [23] Name (Synonym) Plant derived compounds Exatecan (DX-891f) (3 7) XRP-9881 (RPR-109881A) (3 8) Vinflunine ditartrate (Javlor ) (3 9) Mechanism of Action... C-1 2), 113.82 (2×, C-3’ and C-5 ) and 113.47 (2×, C-9 and C-1 1)] , two non ortho-disubstituted methoxyl groups [δH 3.82 (6H, s, 4’-OCH3 and 10-OCH 3); δC 55.19 (2x, 4’-OCH3 and 10-OCH 3)] and eight substituted sp2 carbons, four of which are oxygenated Therefore the compound has thirteen units of unsaturation which can be accounted for by two benzene rings, an ester, 27 a lactone and two C—C double bonds ... Yanhui and Ler Peggy for recording NMR spectra; Wong Lai Kwai and Lai Hui Ngee for Mass Spectra II Abstract The natural products from an un-identified Scleroderma sp and Leptoscyphus expansus (Lehm.). .. important sources of natural product-derived drugs Fungi and marine organism have also received attention from researchers and represent new sources of natural products Natural products are generally... H (15) (16) 1.3 Natural Products as Drug Leads For thousands of years medicine and natural products (NPs) have been closely linked through the use of traditional medicines and natural poison [6]