In-silico, in-vitro and ex-vivo evidence of combining silymarin phytopharmaceutical with piperine, and fulvic acid for enhancing its solubility and permeability

Pharm Dev Technol. 2023 Sep;28(7):595-610. doi: 10.1080/10837450.2023.2227966. Epub 2023 Jun 24.


Non-alcoholic fatty liver disease is one of the leading causes of death worldwide. Even if with such a high mortality there is no definite treatment approved. Thus, there is a need to develop a formulation which can have multiple pharmacological activities. Herbal drugs are among the most promising compounds that act by different pharmacological actions. For increasing the bio-activity of Silymarin we had isolated five active biomarker molecules from silymarin extract (as a Phytopharmaceutical) in our previous work. It possesses lower bioavailability due to poor solubility, lesser permeability and first pass metabolism effect. Therefore, from the literature we had screened two bioavailability enhancers i.e. piperine and fulvic acid for overcoming the drawbacks associated with silymarin. Hence, in this study we had first explored the ADME-T parameters and then evaluated their in-silico activity for different enzymes involved in inflammation and fibrosis. Interestingly, it was found that besides the bioavailability enhancing property, piperine and fulvic acid also shown anti-inflammatory and anti-fibrotic action, particularly more activity was demonstrated by fulvic acid than piperine. Furthermore, the concentration of the bioavailability enhancers i.e. 20% FA and 10% PIP were optimized by QbD assisted solubility studies. Moreover, the percentage release and apparent permeability coefficient of the optimized formulation was found to be 95% and 90%, respectively as compared to 6.54*106 and 1.63*106 respectively by SM suspension alone. Furthermore, it was found that plain rhodamine solution penetrated only up to 10 um whereas, formulation penetrated up to 30 um. Thus, combining these three, can not only increase the bioavailability of silymarin, but might also, increase the physiological action synergistically.

Keywords: Nonalcoholic fatty liver disease; anti-fibrotic; anti-inflammatory; fulvic acid; piperine; silymarin.

MeSH terms

  • Biological Availability
  • Permeability
  • Silymarin* / pharmacology
  • Solubility


  • Silymarin
  • piperine
  • fulvic acid