Trans-Epithelial Transport, Metabolism, and Biological Activity Assessment of the Multi-Target Lupin Peptide LILPKHSDAD (P5) and Its Metabolite LPKHSDAD (P5-Met)
Carmen Lammi,
Gilda Aiello,
Carlotta Bollati,
Jianqiang Li,
Martina Bartolomei,
Giulia Ranaldi,
Simonetta Ferruzza,
Enrico Mario Alessandro Fassi,
Giovanni Grazioso,
Yula Sambuy,
Anna Arnoldi
Affiliations
Carmen Lammi
Department of Pharmaceutical Sciences, University of Milan, 20122 Milan, Italy
Gilda Aiello
Department of Pharmaceutical Sciences, University of Milan, 20122 Milan, Italy
Carlotta Bollati
Department of Pharmaceutical Sciences, University of Milan, 20122 Milan, Italy
Jianqiang Li
Department of Pharmaceutical Sciences, University of Milan, 20122 Milan, Italy
Martina Bartolomei
Department of Pharmaceutical Sciences, University of Milan, 20122 Milan, Italy
Giulia Ranaldi
Food and Nutrition Research Centre, CREA, 00178 Rome, Italy
Simonetta Ferruzza
Food and Nutrition Research Centre, CREA, 00178 Rome, Italy
Enrico Mario Alessandro Fassi
Department of Pharmaceutical Sciences, University of Milan, 20122 Milan, Italy
Giovanni Grazioso
Department of Pharmaceutical Sciences, University of Milan, 20122 Milan, Italy
Yula Sambuy
Food and Nutrition Research Centre, CREA, 00178 Rome, Italy
Anna Arnoldi
Department of Pharmaceutical Sciences, University of Milan, 20122 Milan, Italy
P5 (LILPKHSDAD) is a hypocholesterolemic peptide from lupin protein with a multi-target activity, since it inhibits both 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCoAR) and proprotein convertase subtilisin/kexin type-9 (PCSK9). This work shows that, during epithelial transport experiments, the metabolic transformation mediated by intestinal peptidases produces two main detected peptides, ILPKHSDAD (P5-frag) and LPKHSDAD (P5-met), and that both P5 and P5-met are linearly absorbed by differentiated human intestinal Caco-2 cells. Extensive comparative structural, biochemical, and cellular characterizations of P5-met and the parent peptide P5 demonstrate that both peptides have unique characteristics and share the same mechanisms of action. In fact, they exert an intrinsically multi-target behavior being able to regulate cholesterol metabolism by modulating different pathways. The results of this study also highlight the dynamic nature of bioactive peptides that may be modulated by the biological systems they get in contact with.