Competitive ELISA-Based Screening of Plant Derivatives for the Inhibition of VEGF Family Members Interaction with Vascular Endothelial Growth Factor Receptor 1

Abstract

The activation of vascular endothelial growth factor receptor-1 (VEGFR-1, also known as Flt-1) is crucial in many physiological and pathological conditions, like angiogenesis, cancer, inflammation, hematopoiesis, bone marrow precursors/stem cells recruitment in tumor angiogenesis, and metastasis formation. Many recent reports indicate that molecules able to antagonize Flt-1 activity have gained a strong interest in the view of therapeutic approaches. In order to identify new compounds able to interfere in the Flt-1 recognition by VEGFs family members, we have developed a highly sensitive competitive ELISA-based screening to study plant extracts and derivatives. Several fractions of the n-butanol extract of Pteleopsis suberosa leaves and of the chloroform extract of Parinari campestris leaves demonstrated by a bioassay-guided fractionation an evident inhibition of VEGF-A or placental growth factor (PlGF) interaction with Flt-1, with an inhibition over 50 % in particular for the VEGF-A/Flt-1 interaction at a concentration of 100 μg/mL. This activity seems be due to the presence of a combination of compounds acting synergistically.

Key words

Angiogenesis - VEGF-PlGF and Flt-1 - flavonoids - terpenes - Pteleopsis suberosa - Combretaceae - Parinari campestris - Chrysobalanaceae

References

• 1 Yancopoulos G D, Davis S, Gale N W, Rudge J S, Wiegand S J, Holash J. Vascular-specific growth factors and blood vessel formation.  Nature. 2000;  407 242-8
• 2 Shibuya M. Differential roles of vascular endothelial growth factor receptor-1 and receptor-2 in angiogenesis.  J Biochem Mol Biol. 2006;  39 469-8
• 3 Carmeliet P, Moons L, Luttun A, Vincenti V, Compernolle V, De Mol M. et al . Synergism between vascular endothelial growth factor and placental growth factor contributes to angiogenesis and plasma extravasation in pathological conditions.  Nat Med. 2001;  7 575-3
• 4 Bellomo D, Headrick J P, Silins G U, Paterson C A, Thomas P S, Gartside M. et al . Mice lacking the vascular endothelial growth factor-B gene (Vegfb) have smaller hearts, dysfunctional coronary vasculature, and impaired recovery from cardiac ischemia.  Circ Res. 2000;  86 E29-35
• 5 Carmeliet P. Angiogenesis in health and disease.  Nat Med. 2003;  9 653-60
• 6 Luttun A, Tjwa M, Carmeliet P. Placental growth factor (PlGF) and its receptor Flt-1 (VEGFR-1): novel therapeutic targets for angiogenic disorders.  Ann N Y Acad Sci. 2002;  979 80-93
• 7 Kaplan R N, Riba R D, Zacharoulis S, Bramley A H, Vincent L, Costa C. et al . VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre-metastatic niche.  Nature. 2005;  438 820-7
• 8 Fischer C, Jonckx B, Mazzone M, Zacchigna S, Loges S, Pattarini L. et al . Anti-PlGF inhibits growth of VEGF(R)-inhibitor-resistant tumors without affecting healthy vessels.  Cell. 2007;  131 63-75
• 9 Lyden D, Hattori K, Dias S, Costa C, Blaikie P, Butros L. et al . Impaired recruitment of bone-marrow-derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth.  Nat Med. 2001;  7 1194-201
• 10 Fiore G, Nencini C, Cavallo F, Capasso A, Bader A, Giorgi G. et al . In vitro antiproliferative effect of six Salvia species on human tumor cell lines.  Phytotherapy Res. 2006;  20 01-3
• 11 De Leo M, Braca A, Sanogo R, Cardile V, De Tommasi N, Russo A. Antiproliferative activity of Pteleopsis suberosa leaf extract and its flavonoid components in human prostate carcinoma cells.  Planta Med. 2006;  72 604-10
• 12 Siciliano T, Bader A, De Tommasi N, Morelli I, Braca A. Sulfated pregnane glycosides from Periploca graeca. .  J Nat Prod. 2005;  68 1164-8
• 13 Braca A, Abdel-Razik A F, Mendez J, Morelli I. A new kaurane diterpene dimer from Parinari campestris. .  Fitoterapia. 2005;  76 614-9
• 14 Otsuka H, Hirata E, Shinzato T, Takeda Y. Stereochemistry of megastigmane glucosides from Glochidion zeylanicum and Alangium premnifolium. .  Phytochemistry. 2003;  62 763-8
• 15 Xu H X, Kadota S, Wang H, Kurokawa M, Shiraki K, Matsumoto T. et al . A new hydrolyzable tannin from Geum japonicum and its antiviral activity.  Heterocycles. 1994;  38 167-75
• 16 Agrawal P K. Carbon-13 NMR of flavonoids. Amsterdam; Elsevier 1989
• 17 Braca A, Armenise A, Morelli I, Mendez J, Mi Q, Chai H -B. et al . Structure of kaurane-type diterpenes from Parinari sprucei and their potential anticancer activitiy.  Planta Med. 2004;  70 540-50
• 18 Ballesta-Acosta M C, Pascual-Villalobos M J, Rodriguez B. A new 24-nor-oleanane triterpenoid from Salvia carduacea. .  J Nat Prod. 2002;  65 1513-5
• 19 De Felice A, Bader A, Leone A, Sosa S, Della Loggia R, Tubaro A. et al . New polyhydroxylated triterpenes and anti-inflammatory activity of Salvia hierosolymitana. .  Planta Med. 2006;  72 643-9
• 20 Morikawa T, Zhang Y, Nakamura S, Matsuda H, Muraoka O, Yoshikawa M. Bioactive contituents from Chinese natural medicines. XXII. Absolute structures of new megastigmane glycosides, sedumosides E₁, E₂, E₃, F₁, F₂, and G, from Sedum sarmentosum (Crassulaceae).  Chem Pharm Bull. 2007;  55 435-41
• 21 Mayer R. Three lupane derivates from Leptospermum scoparium. .  Arch Pharm. 1996;  329 447-50
• 22 McInnes A G, Walter J A, Wright J LC. Carbon-13 NMR spectra of Δ 24(28)-phytosterols.  Org Magn Reson. 1980;  13 302-3
• 23 Bae D G, Kim T D, Li G, Yoon W H, Chae C B. Anti-flt1 peptide, a vascular endothelial growth factor receptor 1-specific hexapeptide, inhibits tumor growth and metastasis.  Clin Cancer Res. 2005;  11 651-61.
• 24 Errico M, Riccioni T, Iyer S, Pisano C, Acharya K R, Persico M G. et al . Identification of placenta growth factor determinants for binding and activation of Flt-1 receptor.  J Biol Chem. 2004;  279 43 929-39