A New Glucuronidated Metabolite of Andrographolide in Human

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【刊名】Chinese Chemical Letters


Andrographolide, one of the main bioactive constituents of Andrographis paniculata(Burm) Nees, a famous traditional Chinese medicine, was chemically designated as3-[2-[decahydro-6-hydroxy-5-(hydroxymethyl)-5,8a-dimethyl-2-methylene-1-napthalen-yl] ethylidene] dihydro-4-hydroxy-2(3H)-furanone. Andrographolide showed a widebiological activities, including antiinflammatory1,2,3, anti-allergic4'5, anti-plateletaggregation6, hepatoprotective7,8, anti-human immunodeficiency virus(HIV) activities9,10and was widely used in clinic for the treatment of fever, cold, inflammation, diarrhea andother infectious diseases. The pharmacokinetic studies showed that it absorbed andintensely metabolized in rats and human quickly11. Recently, our reasearch group foundthat one of the metabolites of andrographolide in rats after oral administration,14-deoxy-12(R)-sulfoandrographolide, was a antiinflammatory drug (Lianbizhi) used inclinic as an injection12,13. So the metabolites of andrographolide in human urine werefurther investigated. The present paper describes the structural elucidation of a newandrographolide metabolite (see Figure 1). Metabolite 114, white amorphous power, was positive for the Legal and Keddereactions, suggesting the presence of an α,β-unsaturated lactone. The negative ESI-MSshowed the quasi-molecular ion peak [M-H]- at m/z 523. Combined with the H-NMR 1and C-NMR spectral data, the molecular formula of 1 was determined to be C26H36O11. 13The MS2 spectrum of the [M-H]- ion m/z 523 provided a fragment ion at m/z 505[M-H-H2O]-. In the MS3 spectrum, the [M-H-H2O]- ion m/z 505 eliminated 176 amu(glucuronic acid -H2O) to give m/z 329, suggesting that 1 was a glucuronide conjugateof andrographolide. The 13C-NMR data also showed the existence of a glucuronic acid[δ 73.6 (C-4'),74.8 (C-2'), 76.1 (C-5'), 77.7 (C-3'), 104.8 (C-1'), 176.8 (C-6')]. Thelinkage site of the glucuronic acid moiety was determined to be at C-19 by analysis ofthe HMBC spectrum (see Figure 2), in which the signals of H-19 (5 4.56, d, 1H, 7=9.7Hz; 8 3.30, m, 1H) correlated with C-1' (δ 104.8) and the anomeric proton of glucuronide(δ 4.13, d, 1H, J=7.8 Hz) correlated with C-19 (δ 73.5). The β-form anomericconfiguration of the glucuronic acid was judged from its coupling constant of theanomeric proton (J=7.8 Hz). The carbon signals of the aglycone moiety of 1 and its parent drug andrographolidewere very similar except for the signals at C1~C5, C18 and C19, indicating that thevarieties of 1 occurred at ring A only. It was obvious that the hydroxyl-linked carbonsignal at 8 80.9 (C-3) of andrographolide had disappeared and a new carbon signal ofcarbonyl at 8 217.4 could be observed in 1, suggesting that the hydroxyl at C-3 ofandrographolide might be oxygenated to carbonyl. Due to the occurrence of theA New Glucuronidated Metabolite ofAndrographolide in Human 371oxygenation at C-3 and the glucuronidation at C-19, the signals of C-2, C-4, C-5 andC-19 of 1 shifted downfield from δ 29.0 to δ 39.8, δ 43.6 to δ 54.5, δ 56.3 to δ 58.4 and δ64.9 to δ 73.5, while the signals of C-1 and C-18 shifted upfield from δ 38.1 to 8 36.9and from δ 23.3 to δ 20.8, respectively. In the HMBC spectrum (also see Figure 2), thesignal of H-18 (δ 1.19, 3H, s) had peaks correlated with δ 217.4 (C-3), δ 73.5 (C-19), δ58.4 (C-5) and δ 54.5 (C-4), the signal of H-2β (δ 2.14, m, 1H) correlated with δ 217.4(C-3) and the signals of H-19 (δ 4.56, d, 1H, J=9.7 Hz; δ 3.30, m, 1H) had correlationswith δ 217.4 (C-3), δ 54.5 (C-4) and δ 20.8 (C-18). These correlated peaks furtherconfirmed that the carbonyl was located at C-3. Thus, the structure of the ring A couldbe established. Based on the above chemical and spectroscopic evidences, the structureof 1 was elucidated to be 3-carbonyl-andrographolide-19-O-β-D-glucuronide.A New Glucuronidated Metabolite of Andrographolide in Human1. Y. C. Shen, C. F. Chen, W. F. Chiou, Planta Med., 2000, 66, 314. 2. Y. C. Shen, C. F. Chen, W. F. Chiou, Br. J. Pharmacol., 2002, 135(2), 399. 3. S. Madav, S. K. Tandan, J. Lal, Fitoterapia, 1996, 67, 452. 4. S. Madav, C. F. Tripathi, S. K. Tandan, et al., Indian J. Pharm. Sci., 1998, 60, 176. 5. P. P. Gupta, J. S. Yandon, G. K. Patnaik, Pharm.Biol., 1998, 36, 72. 6. E. Amroyan, E. Gabrielian, A. Panossian, et al., Phytomedicine, 1999, 6, 27. 7. B. Shukla, P. K. S. Visen, G. K. Patnaik, B. N. Dhawan, Planta Med., 1992, 58, 146. 8. P. Pramyothin, W. Udomuksorn, S. Poungshompoo, C. Chaichantipyuth, Asia Pac. J. Pharmacol., 1994, 9, 73. 9. S. S. Handa, A. Sharma, Indian J. Med. Res., 1990, 92, 284. 10. A. Basak, S. Cooper, A G. Roberge, et al., Biochem. J., 1999, 338, 107. 11. A. Panossian, A. Hovhannisyan, G. Mamikonyan, et al., Phytomedicine, 2000, 7, 351 12. X. J. He, J. K. Li, H. Gao, et al., Tetrahedron, 2003, 59, 6603. 13. X. J. He, J. K. Li, H. Gao, et al., Chem. Pharm. Bull., 2003, 51, 586. 14. selected

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