NO-, TNF-, and IL-12-inducing activity of fractions of EPs® 7630

The efficacy of EPs® 7630, a herbal medicinal product for the treatment of upper respiratory tract infections, has been demonstrated in a number of clinical studies [1,2]. Despite considerable efforts, the remedial effects cannot be related to a chemically defined principle. Using several functional bioassays, EPs® 7630 has shown appreciable immunomodulatory activities in vitro [3].

In continuation of our previous work regarding the search for immunologically active constituents, EPs® 7630 was subjected to a variety of extraction procedures (EtOAc, CH₂Cl₂, H₂O) giving a range of subfractions. In a modified protocol, EPs® 7630 was initially separated into a MeOH-soluble (MSP) and -insoluble (MIP) fraction which were subsequently similarly treated as above. Each sample was tested for its NO-, TNF- and IL-12-inducing capacity using non-infected and GFP-transfected-Leishmania major-infected bone marrow-derived macrophages (BMMΦ). Following a 48h incubation period, significant TNF levels (ca 79 pg/ml vs. 200 pg/ml with IFN+LPS as positive control) were induced by EPs® 7630 and the H₂O phase of MIP, respectively, while only negligible NO and IL-12 productions were determined in all the samples. In flow cytometric (FACS) analysis the GFP-signal indicating viable parasites decreased to 29 and 21%, when cells were treated with EPs® 7630 and the H₂O phase of MIP, respectively. Untreated cells were used as a control. The conspicuous intracellular killing of parasites, commonly related to NO, suggested that extracellular NO radicals were non-detectable in the Griess assay, while these cytotoxic effector molecules were effective intracellularly. This may be attributed to radical scavengers present in this fraction or to relatively low but efficient NO levels. Attention is currently paid to the chemical constituents of this active fraction.

Acknowledgements: EPs® 7630 was kindly provided by Dr Willmar Schwabe GmbH & Co, Karlsruhe, Germany.

References: [1] Matthys, A. et al. (2007) Curr. Med. Res. Opin. 23: 323–31. [2] Chuchalin, A.G. et al. (2005) Explore (NY) 1: 437–45 [3] Kolodziej, H., Kiderlen, A.F. (2007) Phytomedicine; 14 Suppl 1: 18–26