Supporting Information to Insecticidal and enzyme inhibitory activities ...
Supporting Information to
Insecticidal and enzyme inhibitory activities of sparassol and its analogs against Drosophila suzukii
Junheon Kim1, †, Miyeon Jang 3, †, Kyoung-Tae Lee4, Kyungjae Andrew Yoon5, Chung Gyoo Park1,2,3,*
1
Institute of Agriculture and Life Science, 2Institute of Life Science, and 3Division of Applied Biology (BK 21 plus), Gyeongsang National University, Jinju 52828, Republic of Korea 4
Southern Forest Resources Research Center, National Institute of Forest Science, Jinju 52817, Republic of Korea 5
Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea
†
These authors contributed equally to this study.
*Corresponding author: Prof. Park, Chung Gyoo Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 52828, Republic of Korea. E-mail: [email protected], Tel: +82-55-772-1925, Fax: +82-55-772-1929
Experimental General All the reagents were reagent grade and used without purification. THF was freshly distilled from sodium benzophenone ketyl. Column chromatography was performed using silica gel (Wakogel® C200, Wako Chem. Co., Osaka, Japan). The 1H and 13C NMR spectra were recorded in CDCl3 on 500 MHz and 126 MHz at Center for Research Facilities of Gyeongsang National University.
3,5-dimethoxytoluene. Hydroxy groups were converted to methoxy group following the previously reported method.1 A stirring mixture of orcinol monohydrate (14.2 g, 100 mmol), anhydrous K2CO3 41.4 g (300 mmol) and dimethyl sulfate 20.8 mL (220 mmol) in acetone (200 mL) was refluxed for 4 h. After adding 25 mL of NH4OH, the mixture was heated further 10 min. After cooling, the solvent was removed by evaporator. The residue was diluted with diethyl ether, washed with H2O, 2N NaOH, and brine, and dried over MgSO4. Distillation at 114 °C/ 20-25 mmHg gave desired compound (15.0 g, 99 mmol). Yield; 98.7%; Purity: 99%. 1H NMR (500 MHz, CDCl3, δ): 2.295 (3H, s), 3.759 (6H, s), 6.280 (1H, t, J=2.5 Hz), 6.331 (2H, dd, J1=2.5 Hz, J2=0.5 Hz);
13
C NMR (126 MHz, CDCl3, δ): 21.83,
55.23(×2), 97.54, 107.22 (×2), 140.23, 160.75 (×2).
2-Bromo-1,5-dimeth-3-methylbenzne. It was prepared by following the method of Mondal et al (2007).2 To a solution of 3,5-dimethoxytoluene (15.0 g, 99 mmol) in CHCl3 (150 mL) was added NBS (19.3 g, 109 mmol), and the reaction mixture was refluxed for 10 h. After cooling, water was added in the mixture and the organic layer was extracted with ether. After ethereal solution was washed with brine, it was dried over MgSO4. Silica gel chromatographic S1
purification using a mixture of 2% ether in hexane afforded the target compound (14.5 g, 63 mmol) as a solid. Yield: 63.9%; Purity: 98%; 1H NMR (500 MHz, CDCl3, δ): 6.429 (1H, d, J=2.5), 6.346 (1H, d, J=2.5), 3.861 (3H, s), 3.790 (3H, s), 2.392 (3H, s); 13C NMR (126 MHz, CDCl3, δ): 159.32, 156.65, 139.84, 107.23, 105.13, 97.25, 56.30, 55.49, 23.59.
Methyl 2,4-dimethoxy-6-methylbenzoate (DMB; 3). DMB 3 was synthesized by following the method of Mondal et al. (2007).2 To a stirring solution of 2-bromo-1,5-dimeth-3methylbenzne (14.5 g, 63 mmol) in THF 150 mL at -78 °C was added n-BuLi (1.6 M, 47.3 mL, 76 mmol). After stirring was continued for 1 h at that temperature, methyl chloroformate (15 mL, 190 mmol) was added slowly. After the mixture was stirred at -78 °C for further 1 h, the mixture was allowed to reach room temperature and kept stirring for 1 h. The reaction was quenched by addition of NH4Cl. Ether was added to the reaction mixture, and the separated ethereal solution washed with water and brined and dried over MgSO4. Silica gel chromatographic purification using a mixture of 2% ether in hexane gave 3 (10 g, 48 mmol) as a colorless oil. Yield: 76.5%; Purity: 95.0%; 1H NMR (500 MHz, CDCl3, δ): 6.305 (2H, br. S), 3.867 (3H, s), 3.773 (3H, s), 2.273 (3H, s);
13
C NMR (126 MHz, CDCl3, δ): 168.68,
161.37, 158.20, 138.21, 116.36, 106.66, 96.11, 55.83, 55.26, 51.95, 19.88.
Methyl orsellinate (methyl 2,4-dihydroxy-6-methylbenzoate; 2). To a stirring solution of AlCl3
(38.3 g, 288 mmol) in CH2Cl2 200 mL at 0 °C was added 3 (14.5 g, 63 mmol) in CH2Cl2 150 mL. After stirring for 12 h at room temperature, the reaction mixture was cooled to 0 °C and water was added carefully. After ether extraction, the organic layer was washed with water and brine and dried over MgSO4. Silica gel chromatographic purification using a mixture of 10% ether in hexane gave 1 (3.5 g, 18 mmol; Yield 37.3%, purity: 97.8%), and a mixture of S2
30% ether in hexane gave 2 (4.2 g, 23 mmol; Yield: 47.5%; Purity: 94.3%). Methyl orsellinate (2): 1H NMR (500 MHz, CDCl3, δ): 2.488 (3H, s), 3.925 (3H, s), 5.381 (1H, s), 6.228 (1H, m), 6.279 (1H, d, J=2.5), 11.753 (1H, s); 13C NMR (126 MHz, CDCl3, δ): 24.27, 51.91, 101.28, 105.70, 111.34, 144.03, 160.26, 165.32, 172.14.
Sparassol (methyl 2-hydroxy-4-methoxy-6-methylbenzoate; 1). A mixture of 2 (3.1 g, 17 mmol), K2CO3 (2.6 g, 19 mmol), and methyl iodide (4.8 g, 34 mmol) in acetone 150 mL was refluxed for 4 h. After cooling, the reaction mixture was filtered and the acetone was evaporated. After ether was added to the residue, the organic solution was washed with 2N NaOH, 2N HCl and brine, and dried over MgSO4. Silica gel chromatographic purification using a mixture of 10% ether in hexane gave 1 (2.7 g, 14 mmol). Yield; 82.0%; purity: 98.8%; 1
H NMR (500 MHz, CDCl3, δ): 2.494 (3H, s), 3.796 (3H, s), 3.922 (3H, s), 6.282 (1H, m),
6.330 (1H, d, J=2.5), 11.765 (1H, s);
13
C NMR (126 MHz, CDCl3, δ): 24.34, 51.83, 55.29,
98.71, 105.26, 111.17, 143.12, 163.95, 165.56, 172.22.
S3
Scheme 1. Synthetic scheme for sparassol (1), methyl orsellinate (2), and methyl 2,4dimethoxy-6-methylbenzoate (DMB; 3).
S4
Fig S1. NMR spectrum of sparassol (1).
S5
Fig S2. NMR spectrum of methyl orsellinate (2).
S6
Fig S3. NMR spectrum of methyl 2,4-dimethoxy-6-methylbenzoate (DMB, 3).
S7
References (1)
Mirrington, R. N.; Feutrill, G. I., Orcinol monomethyl ether. Org. Synth. 1988, Coll.
Vol. 6, 859-861. (2)
Mondal, M.; Puranik, V. G.; Argade, N. P., A facile phenol-driven intramolecular
diastereoselective thermal/base-catalyzed dipolar [2 + 2] annulation reactions: An easy access to complex bioactive natural and unnatural benzopyran congeners. J. Org. Chem. 2007, 72, 2068-2076.
S8
Insecticidal and enzyme inhibitory activities of sparassol and its analogs against Drosophila suzukii
Junheon Kim1, †, Miyeon Jang 3, †, Kyoung-Tae Lee4, Kyungjae Andrew Yoon5, Chung Gyoo Park1,2,3,*
1
Institute of Agriculture and Life Science, 2Institute of Life Science, and 3Division of Applied Biology (BK 21 plus), Gyeongsang National University, Jinju 52828, Republic of Korea 4
Southern Forest Resources Research Center, National Institute of Forest Science, Jinju 52817, Republic of Korea 5
Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea
†
These authors contributed equally to this study.
*Corresponding author: Prof. Park, Chung Gyoo Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 52828, Republic of Korea. E-mail: [email protected], Tel: +82-55-772-1925, Fax: +82-55-772-1929
Experimental General All the reagents were reagent grade and used without purification. THF was freshly distilled from sodium benzophenone ketyl. Column chromatography was performed using silica gel (Wakogel® C200, Wako Chem. Co., Osaka, Japan). The 1H and 13C NMR spectra were recorded in CDCl3 on 500 MHz and 126 MHz at Center for Research Facilities of Gyeongsang National University.
3,5-dimethoxytoluene. Hydroxy groups were converted to methoxy group following the previously reported method.1 A stirring mixture of orcinol monohydrate (14.2 g, 100 mmol), anhydrous K2CO3 41.4 g (300 mmol) and dimethyl sulfate 20.8 mL (220 mmol) in acetone (200 mL) was refluxed for 4 h. After adding 25 mL of NH4OH, the mixture was heated further 10 min. After cooling, the solvent was removed by evaporator. The residue was diluted with diethyl ether, washed with H2O, 2N NaOH, and brine, and dried over MgSO4. Distillation at 114 °C/ 20-25 mmHg gave desired compound (15.0 g, 99 mmol). Yield; 98.7%; Purity: 99%. 1H NMR (500 MHz, CDCl3, δ): 2.295 (3H, s), 3.759 (6H, s), 6.280 (1H, t, J=2.5 Hz), 6.331 (2H, dd, J1=2.5 Hz, J2=0.5 Hz);
13
C NMR (126 MHz, CDCl3, δ): 21.83,
55.23(×2), 97.54, 107.22 (×2), 140.23, 160.75 (×2).
2-Bromo-1,5-dimeth-3-methylbenzne. It was prepared by following the method of Mondal et al (2007).2 To a solution of 3,5-dimethoxytoluene (15.0 g, 99 mmol) in CHCl3 (150 mL) was added NBS (19.3 g, 109 mmol), and the reaction mixture was refluxed for 10 h. After cooling, water was added in the mixture and the organic layer was extracted with ether. After ethereal solution was washed with brine, it was dried over MgSO4. Silica gel chromatographic S1
purification using a mixture of 2% ether in hexane afforded the target compound (14.5 g, 63 mmol) as a solid. Yield: 63.9%; Purity: 98%; 1H NMR (500 MHz, CDCl3, δ): 6.429 (1H, d, J=2.5), 6.346 (1H, d, J=2.5), 3.861 (3H, s), 3.790 (3H, s), 2.392 (3H, s); 13C NMR (126 MHz, CDCl3, δ): 159.32, 156.65, 139.84, 107.23, 105.13, 97.25, 56.30, 55.49, 23.59.
Methyl 2,4-dimethoxy-6-methylbenzoate (DMB; 3). DMB 3 was synthesized by following the method of Mondal et al. (2007).2 To a stirring solution of 2-bromo-1,5-dimeth-3methylbenzne (14.5 g, 63 mmol) in THF 150 mL at -78 °C was added n-BuLi (1.6 M, 47.3 mL, 76 mmol). After stirring was continued for 1 h at that temperature, methyl chloroformate (15 mL, 190 mmol) was added slowly. After the mixture was stirred at -78 °C for further 1 h, the mixture was allowed to reach room temperature and kept stirring for 1 h. The reaction was quenched by addition of NH4Cl. Ether was added to the reaction mixture, and the separated ethereal solution washed with water and brined and dried over MgSO4. Silica gel chromatographic purification using a mixture of 2% ether in hexane gave 3 (10 g, 48 mmol) as a colorless oil. Yield: 76.5%; Purity: 95.0%; 1H NMR (500 MHz, CDCl3, δ): 6.305 (2H, br. S), 3.867 (3H, s), 3.773 (3H, s), 2.273 (3H, s);
13
C NMR (126 MHz, CDCl3, δ): 168.68,
161.37, 158.20, 138.21, 116.36, 106.66, 96.11, 55.83, 55.26, 51.95, 19.88.
Methyl orsellinate (methyl 2,4-dihydroxy-6-methylbenzoate; 2). To a stirring solution of AlCl3
(38.3 g, 288 mmol) in CH2Cl2 200 mL at 0 °C was added 3 (14.5 g, 63 mmol) in CH2Cl2 150 mL. After stirring for 12 h at room temperature, the reaction mixture was cooled to 0 °C and water was added carefully. After ether extraction, the organic layer was washed with water and brine and dried over MgSO4. Silica gel chromatographic purification using a mixture of 10% ether in hexane gave 1 (3.5 g, 18 mmol; Yield 37.3%, purity: 97.8%), and a mixture of S2
30% ether in hexane gave 2 (4.2 g, 23 mmol; Yield: 47.5%; Purity: 94.3%). Methyl orsellinate (2): 1H NMR (500 MHz, CDCl3, δ): 2.488 (3H, s), 3.925 (3H, s), 5.381 (1H, s), 6.228 (1H, m), 6.279 (1H, d, J=2.5), 11.753 (1H, s); 13C NMR (126 MHz, CDCl3, δ): 24.27, 51.91, 101.28, 105.70, 111.34, 144.03, 160.26, 165.32, 172.14.
Sparassol (methyl 2-hydroxy-4-methoxy-6-methylbenzoate; 1). A mixture of 2 (3.1 g, 17 mmol), K2CO3 (2.6 g, 19 mmol), and methyl iodide (4.8 g, 34 mmol) in acetone 150 mL was refluxed for 4 h. After cooling, the reaction mixture was filtered and the acetone was evaporated. After ether was added to the residue, the organic solution was washed with 2N NaOH, 2N HCl and brine, and dried over MgSO4. Silica gel chromatographic purification using a mixture of 10% ether in hexane gave 1 (2.7 g, 14 mmol). Yield; 82.0%; purity: 98.8%; 1
H NMR (500 MHz, CDCl3, δ): 2.494 (3H, s), 3.796 (3H, s), 3.922 (3H, s), 6.282 (1H, m),
6.330 (1H, d, J=2.5), 11.765 (1H, s);
13
C NMR (126 MHz, CDCl3, δ): 24.34, 51.83, 55.29,
98.71, 105.26, 111.17, 143.12, 163.95, 165.56, 172.22.
S3
Scheme 1. Synthetic scheme for sparassol (1), methyl orsellinate (2), and methyl 2,4dimethoxy-6-methylbenzoate (DMB; 3).
S4
Fig S1. NMR spectrum of sparassol (1).
S5
Fig S2. NMR spectrum of methyl orsellinate (2).
S6
Fig S3. NMR spectrum of methyl 2,4-dimethoxy-6-methylbenzoate (DMB, 3).
S7
References (1)
Mirrington, R. N.; Feutrill, G. I., Orcinol monomethyl ether. Org. Synth. 1988, Coll.
Vol. 6, 859-861. (2)
Mondal, M.; Puranik, V. G.; Argade, N. P., A facile phenol-driven intramolecular
diastereoselective thermal/base-catalyzed dipolar [2 + 2] annulation reactions: An easy access to complex bioactive natural and unnatural benzopyran congeners. J. Org. Chem. 2007, 72, 2068-2076.
S8
