Evaluation of an Undocumented Polyherbal (Faradin®) Used for the Treatment of Sickle Cell Disease in West Africa. Part I: Phytochemistry and Ex-vivo Anti-sickling Study

Background: An undocumented complementary and alternative medicine (CAM)- Faradin® - was screened for its phytochemicals, anti-sickling effect using a clinical protocol, and toxicity. Faradin is a polyherbal made up of Zanthoxylum zanthoxyloides, Alnus glutinosa and Alchornea cordofolia. Methods: The polyherbal and the individual components (coded V, M, and F) were separately screened for phytochemicals such as alkaloids, phenols, flavonoids, sterols and coumarins, using standard tests. Institutional Review Board approved clinicalprotocolinvolving4 female de-identified sickle cell patients was used for the ex-vivo anti-sickling evaluation. The negative control was phosphate-buffered saline (PBS)-washed erythrocytes treated with 2% sodium metabisulfite (SMB) to cause a hypoxic state and result in HBSS polymerization or sickling. As positive control, 2% w/v p-hydroxy benzoic acid (PHBA) solution was used. The anti-sickling effect was studied by incubating the washed erythrocytes in various concentrations of Faradin - Faradin: water v/v ratios (2.5:1 – 10:1) - for 4 hours followed by further incubation with SMB for 1½ hrs. The respective samples were examined under an optical microscope and the number of sickled cells counted. Cell viability to Faradin was also done using human primary hepatocytes (HPP) and Cell Titer-Glo Luminescent assay.

Results: Faradin tested positive for flavonoids, phenols, coumarins, alkaloids and antioxidants. The V component tested positive for coumarins, alkaloids and antioxidants. The M component was positive for phenols, alkaloids and antioxidants while the F component tested positive for flavonoids, phenols and antioxidants. For the anti-sickling study, the negative control (SMB treated erythrocytes) showed 20-28% increase in sickling relative to the PBS-washed. The positive control caused a reversal of sickling by decreasing the percent sickled cells from 21% to 8%. There was a dose-dependent decrease in percent sickled cells; the highest dose reduced the percent sickled cells from 21% to 2%. The activity was likely due to the phenols, polyphenols, flavonoids and antioxidants in the CAM. Cell viability of Faradin® was greater than 85%.

Conclusions: Thecombined phytochemicals in the polyherbal contributed to the reversal of sickling that is similar to the positive control. Faradin has a high potential for clinical effectiveness in the management of SCD.