Print version ISSN 0103-5053
On-line version ISSN 1678-4790
J. Braz. Chem. Soc. vol.16 no.4 São Paulo July/Aug. 2005
Aamer Saeed*; Somia Ehsan
Department of Chemistry, Quaid-I-Azam University 45320 Islamabad, Pakistan
A simple synthesis of racemic 8-hydroxy-6,7-dimethoxy-3-methyl-3,4-dihydroisocoumarin or kigelin, a metabolite of Aspergillus terreus and Kigelia pinnata, exhibiting several bioactivities, is described. 3,4,5-Trimethoxyhomophthalic acid was refluxed with acetic anhydride in dry pyridine to afford 2,3,4-trimethoxy-6-(2-oxopropyl)benzoic acid. Reduction of the latter followed by cyclodehydration yielded 6,7,8-trimethoxy-3-methyl-3,4-dihydroisocoumarin which on regioselective demethylation furnished the (±)-kigelin.
Keywords: dihydroisocoumarin, Kigelia pinnata, Aspergillus terreus
Descreve-se a síntese da 8-hidróxi-6,7-dimetóxi-3-metil-3,4-diidroisocumarina (±) kigelina, um metabólito bioativo encontrado em Aspergillus terreus e Kigelia pinnata. O ácido 3,4,5-trimetóxi-homoftálico após tratamento com nidrido acético na presença de piridina, sob refluxo, forneceu o ácido 2,3,4-trimetóxi-6-(2-oxopropril)benzóico. Redução deste último, seguido de ciclodesidratação resultou na formação da 6,7,8-trimetóxi-3-metil-3,4-diidroisocumarina, que sofreu desmetilação regiosseletiva fornecendo a (±) kigelina.
Kigelia pinnata (Syn. Kigelia africana) is a highly variable mono-specific genus of the family Bignoniaceae. K. pinnata (also known as the 'sausage tree' or Worsboom due to its large fruits) is cultivated in many parts of India as an ornamental and roadside tree1 and in Africa where it grows as an endemic species in many areas and finds a variety of medicinal uses.2 The plant has deep chocolate red flowers and gourd like fruit.
Govindachari et al. isolated kigelin3 as the major constituent of plant from the root heartwood of Kigelia pinnata DC. The structure of kigelin was established by chemical methods and spectroscopic techniques as 8-hydroxy-6,7-dimethoxy-3-methyl-3,4-dihydroisocoumarin (1) and it was concluded that the absolute configuration at C-3 was R on the basis of CD spectral analysis (Figure 1).
Chaudary et al. have recently reported 4 the isolation of (3R)-kigelin from the extract of fungus Aspergillus terreus culture medium. Kigelin was shown to display significant activity against human pathogenic dermatophytes, Microsporum canis and Trichophyton longifusus and also exhibited potent xanthine oxidase (XO) inhibitor activity even better than those of 3-t-butyl-4-hydroxyanisole (BHA) and propylgallate (PG).
Kigelin is an active ingredient of cosmetics and skin lotions and is likely to be responsible for skin healing properties. It is used in the removal of solar keratosis lesions on the skin, particularly the face, neck, hands and arms.2,5,6. Kigelia cream containing kigelin as major constituent is used successfully against melanoma. Kigelin is known to cure certain types of eczemas and is also an active constituent of Bust Force Multi Toner Serum.7
The great biological and commercial importance of kigelin together with its simple structure has made kigelin an attractive target for synthesis.8-14 The aim of present study was to devise an efficient synthesis of commercially important and bioactive dihydroisocoumarin 1.
Results and Discussion
We envisaged the synthesis of 1 using 3,4,5-trimethoxyhomophthalic acid (4) as a key starting material. The requisite acid 4 was prepared in two steps in good yield from the commercially available 3-(3,4,5-trimethoxyphenyl)propionic acid (2) using the method of Bauta et al.15 Thus, cyclodehydration of 2 using polyphosphoric acid gave 5,6,7-trimethoxy-1-indanone (3). A toluene solution of indanone 3 and diethyl oxalate was treated with a suspension of sodium methoxide in toluene followed by the addition of potassium hydroxide and 30% H2O2 to furnish 4 (Scheme1).
Reaction of 4 with acetic anhydride in dry pyridine afforded 2,3,4-trimethoxy-6-(2-oxopropyl)benzoic acid (5).16 Spectral data for 5 include the signal for benzylic protons at d 3.99 and that for benzylic carbon at d 43.7, characteristic M+-H2O ion at m/z 250, and carboxylic and ketonic carbonyl absorptions at 1685 and 1720 cm-1, respectively.
Reduction of 5 with sodium borohydride gave the racemic hydroxy acid (6) which was cyclodehydrated using refluxing acetic anhydride to furnish 6,7,8-trimethoxy-3-methyl-3,4-dihydroisocoumarin (7). In the IR spectrum of 7, the lactonic carbonyl absorption was observed at 1725 cm-1. The protons of the C-4 methylene group adjacent to the stereogenic centre (C3) exhibited the diastereotopic effect characteristic of an ABX system.17 The hydrogen cis to side chain methyl resonated slightly upfield at d 2.81 and the trans hydrogen slightly downfield, at d 2.95 respectively. The H-3 (X) showed a multiplet at d 4.56.The carbon signals for C-3 and C-4 appeared at d 80.2 and 68.1.
Regioselective demethylation of trimethoxy-dihydroisocoumarin 7 was attained using boron tribromide under mild conditions (-78 °C, 20 min) to furnish kigelin 1. The presence of phenolic hydroxyl was confirmed by purple ferric chloride test and solubility in dilute aqueous sodium hydroxide. The lactonic carbonyl absorption was lowered to 1665 cm-1 due to chelation with 8-hydroxyl which appeared at 3440 cm-1.
In summary, an efficient synthesis of racemic kigelin, a natural bioactive dihydroisocoumarin has been accomplished.
Melting points were recorded using a MEL TEMP MP-D apparatus and are uncorrected. 1H NMR and the 13C NMR spectra were determined as CDCl3 or CD3OCD3 solutions at 400 MHz (Bruker AM-400) and 100 MHz (Bruker AM-100), respectively. FT-IR spectra were recorded on an FTS 3000 MX spectrophotometer; mass spectra (EI, 70 eV) on a MAT 312 instrument and elemental analyses were conducted using a CHN-Rapid Heräus apparatus. All compounds were purified by thin layer chromatography using silica gel from Merck having a fluorescent indicator F254 (Merck) and the spots were detected under ultraviolet light.
5, 6,7-Trimethoxyindan-1-one (3)
3-(3,4,5-Trimethoxyphenyl)propionic acid (2) (1 g, 4.5 mmol) was dissolved in polyphosphoric acid (12.5 g) and the resulting yellow solution was heated along with stirring at 90 °C for 2 h. The cooled solution was added to 200 mL ice-water and extracted with ethyl acetate (4x 100 mL). The combined extracts were washed with 5% sodium bicarbonate solution and then with water until the washings were neutral. The organic layer was dried (MgSO4), filtered and rotary evaporated to dryness. Recrystallization using ethyl acetate gave (3) as light brown crystals. (0.758 g, 76%); mp 107-108 °C; Rf = 0.46 (pet. ether-ethyl acetate 6:4); IR (KBr) nmax/cm-1: 1685, 1590; 1H NMR (400 MHz, CDCl3): d 2.65 (m, 2H, H-3), 3.0 (m, 2H, H-2), 3.86 (s, 6H, OMe) 3.89 (s, 3H, OMe), 6.45 (s, 1H, H-4); MS (70 eV): m/z (rel. int.): 222 (M+, 26), 194 (53), 180 (70), 179 (82). Anal. Calc. for C12H14O4: C 64.85, H 6.35; Found: C 64.78, H 6.25%.
3,4,5-Trimethoxyhomophthalic acid (4)
A solution of indanone (3) (0.49 g, 2.20 mmol) and diethyl oxalate (0.52 mL) in toluene (5.3 mL) was added slowly to a suspension of NaOMe (0.27 g) in toluene (0.3 mL) at 0 °C. After addition was complete, the mixture was stirred for 1 h at ambient temperature. The solvent was removed in vacuo and the residue was suspended in MeOH (13 mL). Solid KOH (85%, 1.3 g) was slowly added portion wise keeping the temperature below 50 °C; then H2O2 (30%, 2.5 mL) was added slowly, keeping the temperature below 64 °C. The mixture was stirred further at ambient temperature for 16 h. The mixture was filtered and the filtrate was partially evaporated in vacuo to remove MeOH. The remaining aqueous filtrate was washed with ether and the organic layer was discarded. The aqueous layer was acidified with 12 mol L-1 HCI to pH < 2. The acidic aqueous layer was extracted with ethyl acetate and the combined organic portions were dried (MgSO4). The solvent was removed in vacuo and the residue was crystallized to give (4) (0.143 g, 72%); mp 151-153 °C; Rf =0.08 (pet. ether-ethyl acetate 6:4); IR (KBr) nmax/cm-1: 3350, 2600, 1720, 1685, 1590; 1H NMR (400 MHz, CDCl3): d 3.77 (s, 2H, ArCH2), 3.86 (s, 6H, OMe), 3.89 (s, 3H, OMe), 6.47 (s, 1H, H-2); 13C NMR (100 MHz, CDCl3): d 171.7 (COOH), 168.1 (COOH), 162.6 (C3), 159.9 (C5), 137.6 (C1), 116.2 (C2), 109.3 (C6), 56.2 (OMe) 55.6 (OMe), 39.7 (ArCH2); MS (70 eV): 286 (M+, 30), 268 (38), 242 (53), 198 (90), 184 (35). Anal. Calc. for C13H18O7: C 55.54, H 6.34; Found: C, 55.51, H 6.39%.
2,3,4-Trimethoxy-6-(2-oxopropyl)benzoic acid (5)
To a stirred mixture of 3,4,5-trimethoxyhomophthalic acid 4 (0.2 g, 0.70 mmol) in acetic anhydride (0.80 mL), pyridine (0.44 mL) was added under argon. The solid was dissolved instantly and after 2 min. precipitate was observed. Dry ether (6.5 mL) was added to facilitate the stirring. The solid was filtered and washed with ether after being stirred overnight. It was then suspended in water (8 mL) and heated at 60 °C. 10% aqueous sodium hydroxide solution was added dropwise until it was completely dissolved and pH 11 was established. Next, the reaction mixture was treated with diluted hydrochloric acid until pH 2 was attained; then extracted with ethyl acetate (4x100 mL), followed by drying of the extracts (MgSO4) and concentration. The residue was purified by preparative thin layer chromatography using pet. ether and ethyl acetate (6:4) as eluent and finally by recrystallization from ethyl acetate to afford 5 (0.164 g, 82%) as white scales. mp 176-180 °C; Rf =0.13 (pet. ether-ethyl acetate 6:4); IR (KBr) nmax/cm-1: 3194, 1730, 1594, 1695, 1244, 1047; 1H NMR (400 MHz, CDCl3): d 2.29 (s, 3H, CH3), 3.86 (s, 6H, MeO), 3.89 (s, 3H, MeO), 3.99 (s, 2H, CH2), 6.45 (d, J 2.2 Hz, 1H, H5), 11.2 (1H, br s, COOH); 13C NMR (100 MHz, CDCl3): d 195.5 (C2'), 168.1 (COOH), 132.7 (C5), 131.8 (C6), 127.4 (C7), 77.7 (C1'), 55.9 (MeO), 55.6 (MeO x2), 42.9 (C3'); EIMS: m/z (rel.int.): 268 (M+, 30), 250 (41), 196 (53), 178 (70), 150 (32). Anal. Calc. for C13H16O6: C, 58.20; H, 6.01; Found: C, 58.14; H 6.13%.
(±)-2,3,4-Trimethoxy-6-(2-hydroxypropyl) benzoic acid (6)
Keto acid (5) (0.634 mmol, 0.17 g) was heated under reflux with sodium borohydride (0.15 g) in absolute ethanol (15.2 mL) for 4 h. Ethanol was then rotary evaporated, the residue diluted with cold water and acidified with diluted sulfuric acid to give a precipitate, which was extracted with ethyl acetate. Finally the solvent was rotary evaporated to afford 6 as a semi solid (0.13 g, 0.475 mmol, 75%).
(±)-6,7,8-Trimethoxy-3-methyl-3,4-dihydroisocou marin (7)
The hydroxy acid (6) (0.12 g, 0.44 mmol) was dissolved in acetic anhydride (1 mL) and heated under reflux for 2 h. The reaction mixture was then cooled and water (16 mL) was added. The oil, which separated on stirring, was extracted with dichloromethane. The extracts were combined, treated with 5% aqueous sodium bicarbonate solution washed with water, dried over sodium sulphate anhydrous and filtered. The solvent was stripped off on rotary evaporator to leave 7 (0.088 g, 80%) as a colorless crystalline solid mp 103 104 °C (lit.10 105 °C). IR (KBr) nmax/cm-1: 2924, 2852, 1725, 1595, 1426,1 235, 1121; 1H NMR (CD3OD): d 6.54 (1H, s, H-5), 4.56 (IH, m, H-3), 3.87 (3H, s, 6-OMe), 3.93 (3H, s, 8-OMe), 3.90 (3H, s, 7-OMe), 2.94 (1H, dd, J 16.1 Hz, J 3.4 Hz, H-4), 2.81 (1H, dd, J 16.1 Hz, J 11.1 Hz, H-4), 1.46 (3H, d, J 6.3 Hz, 3-Me); 13C NMR (CD3OD): 170.9 (C1), 80.2 (C3), 68.1 (C4), 139.3 (4a), 104.3 (C5), 160.2 (C6), 134.2 (C7), 156.7 (C8), 102.9 (C8a), 60.9 (7 OMe), 56.8 (6 OMe), 16.3 (3 Me); EIMS: m/z (rel. int.): 252 (76) [M]+, 223 (25), 209 (16), 195 (34), 191 (21), 179 (46), 177 (40), 151 (37). Anal. Calc. for C13H16O5: C 61.90, H 6.39; Found: C 61.86, H 6.46%.
(±)-8-Hydroxy-6,7-dimethoxy-3-methyl-3,4 -dihydroisocoumarin (1)
A 1 mol L-1 solution of boron tribromide in dry dichloromethane (0.55 mL) was injected into a stirred solution of (7) (0.1 g, 0.4 mmol) in dry dichloromethane (4 mL) at -78 ºC under argon. After stirring for 20 min., the mixture was poured into ice water (20 mL) and stirred for 30 min. The layers were separated and the aqueous layer was extracted with dichloromethane and then with ethyl acetate. The combined organic phases were dried using anhydrous sodium sulphate and concentrated. It was further purified by preparative TLC using petroleum ether and ethyl acetate (8:2) as eluent and finally by recrystallization from ethyl acetate to afford (1) (0.071 g, 75%) as a colorless crystalline solid. mp 141-143 ºC (lit. 5 144 ºC). IR (CHCl3) n max/cm-1: 2924, 2852, 1663, 1426, 1269, 1121; 1H NMR (CD3OD): d 6.51 (1H, s, H 5), 4.71 (IH, m, H 3), 3.77 (3H, s, 6 OMe), 3.90 (3H, s, 7 OMe), 2.96 (1H, dd, J 16.2 Hz, J 3.4 Hz, H-4), 2.87 (1H, dd, J 16.2 Hz, J 11.1 Hz, H-4), 1.47 (3H, d, J 6.3, 3-Me); 13C NMR (CD3OD): d 170.9 (C1), 79.8 (C3), 67.7 (C4), 139.3 (4a), 104.3 (C5), 160.2 (C6), 134.2 (C7), 156.7 (C8), 102.9 (8a), 60.9 (7 OMe), 56.8 (6 OMe), 16.3 (3 Me); EIMS: m/z (rel. int.): 238 (M+, 100), 223 (25), 179 (46), 209 (16), 195 (34), 177 (40), 151 (37), 136 (15), 106 (19), 77 (27), 51 (62). Anal. Calc. for C12H14O5: C 60.50, H 5.92; Found: C 60.47, H 6.05%.
The authors gratefully acknowledge the research grant under URF project, from the department of chemistry, Quaid-I-Azam University,Islamabad, Pakistan.
1. Belsare, D.P.; Pal S.C.; Mandal, S.C., Abstracts of the International Society of Ethnobiology, 9th International Congress, University of Kent, Canterburry, UK, 2004. [ Links ]
3. Govindachari, T.R.; Patankar, S.J.; Viswanathan, N.; Phytochemistry 1971, 10, 1603. [ Links ]
4. Choudhary, M.I.; Musharraf, S.G.; Mukhmoor, T.; Shaheen, F.; Ali, S.; Atta-ur-Rahman; Z. Naturforsch., C: J. Biosci. 2004, 59, 324. [ Links ]
8. Govindachari, T.R.; Patankar, S.J.; Viswanathan, N.; Indian J. Chem. 1971, 9, 507. [ Links ]
9. Chatterjea, J.N.; Bhakta, C.; Sinha, N.D.; J. Indian Chem. Soc. 1975, 52, 158. [ Links ]
10. Bhide, B.H.; Gupta, V.P.; Narasimhan, N.S.; Mali, R.S.; Chem. Ind. (London) 1975, 519. [ Links ]
11. Sibi, M. P.; Miah, M. A. J. ; Snieckus, V.; J. Org. Chem. 1984, 49, 737. [ Links ]
12. Napolitano, E.; Morsani, M.; Fiaschi, R.; Gazz. Chim. Ital. 1991, 121, 249. [ Links ]
13. Narasimhan, N.S.; Bapat, C.P.; J. Chem. Soc., Perkin Trans. I 1982, 2099. [ Links ]
14. Viswanathan, N.; Patankar, S. J.; Indian J. Chem. Sect. B 1987, 26B, 564. [ Links ]
15. Bauta, W.E.; Lovett, D.P.; Cantrell, W.R.; Burke, B.D.; J. Org. Chem. 2003, 68, 5967. [ Links ]
16. Lewis, C.N.; Staunton, J.; Sunter, D.C.; J. Chem. Soc., Perkin Trans. I 1988, 747. [ Links ]
17. Saeed, A.; Helv. Chim. Acta 2003, 86, 377. [ Links ]
Received: October 15, 2004
Published on the web: May 18, 2005