A. M. Iannuzzi, M. De Leo, F. Camangi, M. P. Germanò, V. D’Angelo, N. De Tommasi, A. Braca

In Italy, there is a long-held tradition concerning the cultivation of Prunus avium L. (Rosaceae) fruits used both for eating and for making juices, jams, and drinks (1). P. avium, commonly called “sweet cherry”, is a vigorous tree extensively distributed in temperate areas of the world (2). Its fruits, which ripen between May and July, are very popular not only for their taste, colour, and sweetness but also for their nutritional properties consisting in a considerable amount of phenolic compounds (3). Despite the wide-spread interest in cherries, little is known about the chemical composition and bioactive properties of P. avium petioles which are considered plant waste without any commercial interest (2,4). The aim of this work was to compare six ancient Italian cultivars both for the phenolic and anthocyanin content of fruits (pulp and peel) and chemical constituents of petioles by HPLC-PDA-ESI-MS/MS analyses. Furthermore, the antitumor activity of phenolic compounds with a particular focus on apoptosis-inducing and potential cytotoxic effects was investigated. As far as we know, no research on the potential anti-angiogenic activity of P.avium has been yet reported. Therefore, the biological effect of fruit and petiole extracts of each cultivar on the formation of new blood vessels using as an in vivo model the chick embryo chorioallantoic membrane (CAM) was explored. Among Italian cherries, the following cultivars were selected: Ferrovia (from the Puglia Region), Capellina, Morellina, Ciambellana, Napoletana, and Bianca (from the Casentino area in Tuscany, see http:/germoplasma.regione.toscana.it/) (5). The quali-quantitative analysis of fruit extracts evidenced a similar phenolic profile with coumaroyl quinic acids, chlorogenic acid, rutin, kaempferol 3-O-rutinoside, and kaempferol 3-O--D-glucopyranoside, as the most representative compounds for all cultivars. Moreover, cyanidin-3-O-rutinoside was the only anthocyanin detected in all the cultivars (except for Bianca where anthocyanins were completely absent), being the most abundant in Capellina and Morellina cultivars. All cultivars petiole extracts resulted in a similar phenolic profile, among which the major constituents were catechin, taxifolin 7-O--D-glucopyranoside, apigenin 4-O--D-glucopyranoside, apigenin 5-O--D-glucopyranoside, naringenin 7-O--D-glucopyranoside, pinocembrin 7-O--D-glucopyranoside, chrysin 7-O--D-glucopyranoside, dihydrowogonin 7-O--D-glucopyranoside, and rutin, being Capellina, Napoletana and Bianca characterized also by the presence of procyanidin B type dimers and trimers. Biological results showed that among fruit extracts, Ferrovia, Morellina, and Capellina cultivars exhibited the best antiangiogenic response inducing a marked reduction of the microvasculature of the CAMs (30.7-43.5% of inhibition at 100 and 160 µg/egg). In addition, the biological screening showed a good inhibition on the capillary growth even after treatment of the CAMs with Morellina, Ferrovia, and Ciambellana petiole extracts (30.6- 42.4% of inhibition at 100 and 160 µg/egg). In conclusion, the extracts from P. avium cultivars here presented can be considered a valuable source of angiomodulators.