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Identification of Coumarins from the Fruit of Citrus hystrix DC as Inhibitors of Nitric Oxide Generation in Mouse Macrophage RAW 264.7 Cells

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Department of Biotechnological Science, Faculty of Biology-Oriented Science and Technology, Kinki University, Iwade-Uchita, Wakayama 649-6493, Japan; Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan; Department of Citriculture, Fruit Tree Research Station, Okitsu, Shimizu, Shizuoka 424-0292, Japan; Faculty of Pharmacy, Meijo University, Tempaku, Nagoya 468-8503, Japan; and Environmental Science Unit, Central Laboratory and Greenhouse Complex, Kasetsart University, Nakorn-Pathom, Thailand
Cite this: J. Agric. Food Chem. 1999, 47, 1, 333–339
Publication Date (Web):December 10, 1998
Copyright © 1999 American Chemical Society

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    Three known coumarins have been isolated from Citrus hystrix DC as inhibitors of both lipopolysaccharide (LPS) and interferon-γ (IFN-γ)-induced nitric oxide (NO) generation in RAW 264.7 cells. The inhibitory activity of bergamottin (IC50 = 14 μM) was comparable to that of N-(iminoethyl)-l-ornithine (l-NIO) (IC50 = 7.9 μM), whereas oxypeucedanin and 5-[(6‘,7‘-dihydroxy-3‘,7‘-dimethyl-2-octenyl)oxy]psoralen, structurally different from bergamottin only in their side-chain moieties, were notably less active. Using 21 coumarins, we structurally classified various types of coumarins into groups A−C:  (A) bearing an isoprenyl (IP) or a geranyl (GR) group, highly active; (B) bearing an IP group cyclized to a coumarin ring, moderately active; (C) bearing an IP group modified with hydroxyl group(s) and/or having other functional groups except for the IP, completely inactive. Cellular uptake studies suggested that coumarins in group C are inactive because of poor permeability to the cell membrane.

    Keywords: Citrus hystrix DC; psoralen; coumarin; nitric oxide; RAW 264.7 cells; structure−activity relationships

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     Kinki University.

     Kyoto University.


     Fruit Tree Research Station.

     Meijo University.

     Kasetsart University.


     Author to whom correspondence should be addressed (telephone +81-75-753-6281; fax +81-75-753-6284; e-mail [email protected]).

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    30. T. K. Lim. Citrus hystrix. 2012, 634-643.
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    42. Marellapudi S. Ramachandra, Gottumukkala V. Subbaraju. Short Synthesis of 3‐Prenylcoumarins by an Unusual Prenylation. Synthetic Communications 2006, 36 (24) , 3723-3727.
    43. T. Wegener, H. Wagner. The active components and the pharmacological multi-target principle of STW 5 (Iberogast®). Phytomedicine 2006, 13 , 20-35.
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