Antifungal, Antiprotozoal, Cytotoxic and Piscicidal Properties of Justicidin B and a New Arylnaphthalide Lignan from Phyllanthus piscatorum

Abstract

Using activity against Candida albicans as a lead, the bioactivity-guided fractionation of the dichloromethane extract of Phyllanthus piscatorum led to the isolation of the arylnaphthalide lignan justicidin B (1), which was found to be present in high amounts, and a new C-11 hydroxylated derivative that we named piscatorin (2). We provide evidence that justicidin B (1) is the main active principle of P. piscatorum, showing the same biological effects that were found for the raw extracts. Justicidin B inhibited the growth of the pathogenic fungi Aspergillus fumigatus (MIC ≥ 1 μg/mL), Aspergillus flavus (MIC ≥ 12 μg/mL), and Candida albicans (MIC ≥ 4 μg/mL), but was not effective against Cryptococcus neoformans and Blastoschizomyces capitatus. Justicidin B also exhibited strong activity against the trypomastigote form of Trypanosoma brucei rhodesiense (IC₅₀ = 0.2 μg/mL) and moderate activity against Trypanosoma cruzi (IC₅₀ = 2.6 μg/mL). Testing against Plasmodium falciparum showed only weak activity. This is the first report on the in vitro fungicidal and antiprotozoal effects of justicidin B. In addition, both compounds exhibited a non-specific cytotoxicity in neoplastic and primary cell cultures. No antibacterial effects were detected. Both compounds were piscicidal against zebra fish and it is shown for the first time that piscatorin (2) and justicidin B (1) are the piscicidal principles of P. piscatorum, exhibiting a potency that is comparable to rotenone.

Key words

Phyllanthus piscatorum - Euphorbiaceae - justicidin B - lignan - antifungal - Candida albicans - fish poison

References

• 1 Gertsch J, Niomawë , Gertsch-Roost K, Sticher O. Phyllanthus piscatorum ethnopharmacological studies on a women's medicinal plant of the Yanomamï Amerindians. Journal of Ethnopharmacology 2002; accepted
• 2 Schultes R E, Raffauf R F. The Healing Forest, Medicinal and Toxic Plants of the Northwest Amazonia. Dioscorides Press Oregon; pp. 1990: 164-66
• 3 Çalis I, Zor M, Saracoglu I, Isimir A, Ruegger H. Four novel cycloartane glycosides from Astragalus oleifolius .  Journal of Natural Products. 1996;  59 1019-23
• 4 Rios J L, Recio M C, Villar A. Screening methods for natural products with antimicrobial activity: a review of the literature.  Journal of Ethnopharmacology. 1998;  23 127-49
• 5 Thöni-Tobler R. Phyllanthus piscatorum Kunth in H.B.K. Isolierung von Arylnaphthalid-Lignanen und Prüfung auf ihre biologische Aktivität. Diploma thesis, Swiss Federal Institute of Technology Zurich Switzerland; 2001: pp. 52
• 6 Heilmann J, Mayr S, Brun R, Rali T, Sticher O. Antiprotozoal activity and cytotoxicity of novel 1,7-dioxa-dispiro[5.1.5.2]pentadeca-9,12-dien-11-one derivatives from Amomum aculeatum .  Helvetica Chimica Acta. 2000;  83 2939-45
• 7 Ankli A, Heinrich M, Bork P, Lutz W, Bauerfeind P, Brun R, Schmid C, Weiss C, Bruggisser R, Gertsch J, Wasescha M, Sticher O. Yucatec Mayan medicinal plants: Evaluation based on indigenous uses.  Journal of Ethnopharmacology. 2002;  79 43-52
• 8 Liu H, Orjala J, Sticher O, Rali T. Acylated flavonol glycosides from leaves of Stenochlaena palustris .  Journal of Natural Products. 1999;  62 70-5
• 9 Heilmann J, Wasescha M R, Schmidt T J. The influence of glutathione and cysteine levels on the cytotoxicity of helenanolide type sesquiterpene lactones against KB cells.  Bioorganic and Medicinal Chemistry. 2001;  9 2189-94
• 10 Gertsch J, Güttinger M, Sticher O, Heilmann J. Relative quantification of mRNA levels in Jurkat T cells with RT - real time - PCR: New possibilities for the screening of anti-inflammatory and cytotoxic natural compounds.  Pharmaceutical Research. 2002;  19 1235-42
• 11 Teixeira J R, Lapa A J, Souccar C, Valle J R. Timbos: ichthyotoxic plants used by Brazilian Indians.  Journal of Ethnopharmacology. 1984;  10 311-8
• 12 Ohta K, Chen Y L, Marumo S, Munakata K. Studies on the piscicidal components of Justicia hayatai var. decumbens. I. Isolation and piscicidal activities of justicidin A and B.  Agricultural and Biological Chemistry. 1969;  33 610-14
• 13 Pettit G R, Schaufelberger D E. Isolation and structure of the cytostatic lignan glycoside phyllanthostatin A.  Journal of Natural Products. 1988;  51 1104-12
• 14 Bachmann T L, Ghia F, Thorssell K GB. Identification of lignans and γ-lactones from the ichthyotoxic Ecuadorian plant Phyllanthus anisolobus .  Phytochemistry. 1993;  32 189
• 15 Day S H, Chiu N Y, Won S J, Lin C N. Cytotoxic lignans of Justicia ciliata .  Journal of Natural Products. 1999;  62 056-58
• 16 Joseph H, Gleye J, Moulis C, Mensah L J, Roussakis C, Gratas C. Justicidin B, a cytotoxic principle from Justicia pectoralis .  Journal of Natural Products. 1988;  51 599-600
• 17 Asano J, Chiba K, Tada M, Yoshii T. Antiviral activity of lignans and their glycosides from Justicia procumbens .  Phytochemistry. 1996;  42 713-17
• 18 Baba A, Oda T, Taketomi S, Notoya K, Nishimura A, Makino H, Sohda T. Studies on disease-modifying antirheumatic drugs. III. Bone resorption inhibitory effects of ethyl 4-(3,4-dimethoxyphenyl)-6,7-dimethoxy-2-(1,2,4-triazol-1-ylmethyl) quinoline-3-carboxylate (TAK-603) and related compounds.  Chemical and Pharmaceutical Bulletin. 1999;  47 369-74
• 19 Chen C C, Hsin W C, Ko F N, Huang Y L, Ou J C, Teng C M. Antiplatelet arylnaphthalide lignans from Justicia procumbens .  Journal of Natural Products. 1996;  59 1149-50
• 20 Munoz V, Sauvain M, Bourdy G, Callapa J, Rojas I, Vargas L, Tae A, Deharo E. The search for natural bioactive compounds through a multidisciplinary approach in Bolivia. Part II. Antimalarial activity of some plants used by Mosetene indians.  Journal of Ethnopharmacology. 2000;  69 139-55
• 21 Tona L, Mesia K, Ngimbi N P, Chrimwami B, Okond'ahoka  , Cimanga K, de Bruyne T, Apers S, Hermans N, Totte J, Pieters L, Vlietinck A J. In-vivo antimalarial activity of Cassia occidentalis, Morinda morindoides and Phyllanthus niruri .  Annuals of Tropical Medicine and Parasitology. 2001;  95 47-57