Synthesis and cytotoxic activity evaluation of 2,3-thiazolidin-4-one derivatives on human breast cancer cell lines

It is well known that resveratrol (RSV) displayed cancer-preventing and anticancer properties but its clinical application is limited because of a low bioavailability and a rapid clearance from the circulation. Aim of this work was to synthesize pharmacologically active resveratrol analogs with an enhanced structural rigidity and bioavailability. In particular, we have synthesized a library of 2,3-thiazolidin-4-one derivatives in which a thiazolidinone nucleus connects two aromatic rings. Some of these compounds showed strong inhibitory effects on breast cancer cell growth. Our results indicate that some of thiazolidin-based resveratrol derivatives may become a new potent alternative tool for the treatment of human breast cancer

Sintesi di nuovi dimetilcarbazoli, (Dimetil) primidocarbazoli, Benzofurochinazolinoni e benzotienochinazolinoni ad attività antitumorale

Dottorato di Ricerca in: Ambiente salute e processi ecosostenibili, Ciclo XXI, a.a. 2007-2008Università della Calabri

Wild plants like nutraceuticals sources: INNOVATIVE EXTRACTION, ENCAPSULATION IN W/O AND O/W EMULSION AND ANTIOXIDANT ACTIVITY OF BIOACTIVE PIGMENTS FROM OPUNTIA FICUS INDICA FRUIT

The use and exploitation of plants widely disseminated, which contain bioactive compounds (nutraceuticals), could be the best way to have beneficial effects on health; this also could reduces the environmental impact of transport on long distance of fruits and vegetables, that often involving the loss of nutritional properties of the products. Opuntia ficus-indica is a plant endemic of South America, wild and invasive and it produces an edible fruit, that it grows on the flat pads of the cactus (Fig.1). The fruit is a berry and varying in colour(Fig.2), it develops green (less sweet), becomes orange-yellow and at the end the colour is reddish purple (very sweet). This is reach in betalains, has an application in the low acid foods as natural colorants [1]. It is often used to make candies and jelly or added on drinks such lemonade; its pulp and juice have been used to treat a lot of diseases, such as wounds and inflammations of the digestive and urinary tract in folk medicine. Betalains are water-soluble nitrogenous pigments. They can be divided into two structural groups, the red to red-violet betacyanins (Latin-Beta, beet and Greek-kyanos, blue color) and the yellow betaxanthins (Latin-Beta, beet and Greek-xanthos, yellow) (Fig. 3). The most important source of betanin (Fig. 4) as colouring agent is the red beet (Beta vulgaris subsp. vulgaris) root. 1. Plant Opuntia ficus indica (L.) Mill. 2. Different cultivar of Opuntia ficus-indica [1] Maran J.P. and Manikandan S. (2012). “Response surface modeling and optimization of process parameters for aqueous extraction of pigments from prickly pear (Opuntia ficus-indica) fruit”. Dyes and Pigments, 95: 465-472. [2] Romo-Hualde A.; Yetano-Cunchillos A.I.; González-Ferrero C.; M.J. Sáiz-Abajo; González-Navarro C.J. (2012). Supercritical fluid extraction and microencapsulation of bioactive compounds from red pepper (Capsicum annum L.) by-products. Food Chemistry, 133, 1045–1049. [3] Fernández-López J.A., Almela L. (2001). Application of high-performance liquid chromatography to the characterization of the betalain pigments in prickly pear fruits. Journal of Chromatography, A (913) 415–420. [4] Spizzirri G.U., Cirillo G., Curcio M., Altimari I., Picci N., Iemma F. (2013). “Stabilization of oxidable vitamins by flavonoid-based hydrogels”. Reactive & Functional Polymers, 73: 1030–1037. [5] Abdalla A.E., Roozen J.P. (1999). “Effect of plant extracts on the oxidative stability of sunfower oil and emulsion”. Food Chemistry, 64: 323-329. Wild plants like nutraceuticals sources: INNOVATIVE EXTRACTION, ENCAPSULATION IN W/O AND O/W EMULSION AND ANTIOXIDANT ACTIVITY OF BIOACTIVE PIGMENTS FROM OPUNTIA FICUS INDICA FRUIT In this work has been investigated the possibility of phytochemicals extraction by using the supercritical fluids extraction (SFE) with CO2 from fruits, optimizing the selectivity between them and other substances presents in the vegetal matrix. Supercritical fluid technology is nowadays the most innovative method for preparing bioactive products used as supplements and for the preparation of products for children since the extract is free of solvent traces (the CO2 diffuses at room temperature). The extracts obtained by SFE-CO2 are “Generally Recognized As Safe (GRAS)” by the American Food and Drugs Administration, they can be added to any food [2]. Materials and Methods 5. Collection and peel of Opuntia ficus-indica fruits Opuntia ficus-indica Fruits First Harvesting (November 2012) Second Harvesting (December 2012) Third Harvesting (October 2013) pH 5,68±0,08 5,73±0,04 5,71±0,02 aw 0,928±0,002 0,934±0,005 0,921±0,003 °Bx 15,0±0,1 14,8±0,2 15,2±0,1 UW.b 83,98±0,17 84,37±0,24 83,62±0,08 10. Pulp homogenized from fruit Different solvent extraction Stirred centrifugation filtration 3. The chromophore of betalains (1), the aglycone of most of betacyanins (2), and a proline-containing betaxanthin (3). Spectrophotometric analysis of extracts Betacyanins content was expressed as betanin content (molar mass M = 551,48 g/mol). Wavelength was setted to 527 nm (Fig. 8). HPLC analysis of extract The analysis of all extracts both from solvent extraction and belonged from SFE were carried out by means of HPLC using a Smartline HPLC system (Fig. 9). Chromatographic separation was carried out using a 2,0 mm ID ×150 mm L, with precolumn, C-18 TSKgel ODS-100 V, 21810. Extraction of betalains with supercritical fluids All samples after collection were previously cleaned and peeled, homogenized and frozen; at the time of the analysis; some of them were centrifuged, other partially dried and added with different sand Ottawa and Diatomee, in order to maximize the yields of extraction. The supercritical extractions by CO2 were performed on a Spe-ed SFE 4 extractor (Fig. 7). Flow CO2= 1.5 L/min. • extra virgin olive oil 20w/w% • Tween 60 • buffer solution • CaCl2; • pectin L.M. • ethanol EMU CPFE EMU STD + extract purified of Opuntia ficus-indica EMU STD EMU VIT E EMU STD + Vitamin E • extra virgin olive oil 70w/w% • mono- and diglycerides of • fatty acids • NaCl 0,1 M EMU CPFE EMU STD + extract purified of Opuntia ficus-indica EMU STD EMU VIT E EMU STD + Vitamin E Preparaions of emulsion oil in water (O/W) and water in oil (W/O) Purification of extract (CPFE): the pulp fruit was homogenized, centrifuged and precipitated with acetone; then filtrated with a PTFE siringe. The emulsification was performed using a rotor stator system (Ultra Turrax T 50). Antioxidant activity The antioxidant properties of all emulsions (O/W and W/O), at four different times after preparations (t0= 1 day; t1= 5 days; t2= 10 days; t3= 15 days), were evaluated by measurement of the scavenging activity towards DPPH and ABTS radicals. DPPH Assay (2,2′-diphenyl-1- picrylhydrazyl) λ=517 nm ABTS Assay [2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)] λ=734 nm DPPH and ABTS radicals inhibition in O/W emulsions DPPH and ABTS radicals inhibition in W/O emulsions The homogenised mix (Fig. 10) was prepared to measure the level of acidity of the primary source, brix degrees, water activity and humidity. The measurements of water activity and humidity are a key parameter in the quality control of moisture-sensitive products. Preliminary analysis of pulp 4. Molecular structure of betanin present in red beet roo

Wild plants like nutraceuticals sources: INNOVATIVE EXTRACTION, ENCAPSULATION IN W/O AND O/W EMULSION AND ANTIOXIDANT ACTIVITY OF BIOACTIVE PIGMENTS FROM OPUNTIA FICUS INDICA FRUIT

The use and exploitation of plants widely disseminated, which contain bioactive compounds (nutraceuticals), could be the best way to have beneficial effects on health; this also could reduces the environmental impact of transport on long distance of fruits and vegetables, that often involving the loss of nutritional properties of the products. Opuntia ficus-indica is a plant endemic of South America, wild and invasive and it produces an edible fruit, that it grows on the flat pads of the cactus (Fig.1). The fruit is a berry and varying in colour(Fig.2), it develops green (less sweet), becomes orange-yellow and at the end the colour is reddish purple (very sweet). This is reach in betalains, has an application in the low acid foods as natural colorants [1]. It is often used to make candies and jelly or added on drinks such lemonade; its pulp and juice have been used to treat a lot of diseases, such as wounds and inflammations of the digestive and urinary tract in folk medicine. Betalains are water-soluble nitrogenous pigments. They can be divided into two structural groups, the red to red-violet betacyanins (Latin-Beta, beet and Greek-kyanos, blue color) and the yellow betaxanthins (Latin-Beta, beet and Greek-xanthos, yellow) (Fig. 3). The most important source of betanin (Fig. 4) as colouring agent is the red beet (Beta vulgaris subsp. vulgaris) root. 1. Plant Opuntia ficus indica (L.) Mill. 2. Different cultivar of Opuntia ficus-indica [1] Maran J.P. and Manikandan S. (2012). “Response surface modeling and optimization of process parameters for aqueous extraction of pigments from prickly pear (Opuntia ficus-indica) fruit”. Dyes and Pigments, 95: 465-472. [2] Romo-Hualde A.; Yetano-Cunchillos A.I.; González-Ferrero C.; M.J. Sáiz-Abajo; González-Navarro C.J. (2012). Supercritical fluid extraction and microencapsulation of bioactive compounds from red pepper (Capsicum annum L.) by-products. Food Chemistry, 133, 1045–1049. [3] Fernández-López J.A., Almela L. (2001). Application of high-performance liquid chromatography to the characterization of the betalain pigments in prickly pear fruits. Journal of Chromatography, A (913) 415–420. [4] Spizzirri G.U., Cirillo G., Curcio M., Altimari I., Picci N., Iemma F. (2013). “Stabilization of oxidable vitamins by flavonoid-based hydrogels”. Reactive & Functional Polymers, 73: 1030–1037. [5] Abdalla A.E., Roozen J.P. (1999). “Effect of plant extracts on the oxidative stability of sunfower oil and emulsion”. Food Chemistry, 64: 323-329. Wild plants like nutraceuticals sources: INNOVATIVE EXTRACTION, ENCAPSULATION IN W/O AND O/W EMULSION AND ANTIOXIDANT ACTIVITY OF BIOACTIVE PIGMENTS FROM OPUNTIA FICUS INDICA FRUIT In this work has been investigated the possibility of phytochemicals extraction by using the supercritical fluids extraction (SFE) with CO2 from fruits, optimizing the selectivity between them and other substances presents in the vegetal matrix. Supercritical fluid technology is nowadays the most innovative method for preparing bioactive products used as supplements and for the preparation of products for children since the extract is free of solvent traces (the CO2 diffuses at room temperature). The extracts obtained by SFE-CO2 are “Generally Recognized As Safe (GRAS)” by the American Food and Drugs Administration, they can be added to any food [2]. Materials and Methods 5. Collection and peel of Opuntia ficus-indica fruits Opuntia ficus-indica Fruits First Harvesting (November 2012) Second Harvesting (December 2012) Third Harvesting (October 2013) pH 5,68±0,08 5,73±0,04 5,71±0,02 aw 0,928±0,002 0,934±0,005 0,921±0,003 °Bx 15,0±0,1 14,8±0,2 15,2±0,1 UW.b 83,98±0,17 84,37±0,24 83,62±0,08 10. Pulp homogenized from fruit Different solvent extraction Stirred centrifugation filtration 3. The chromophore of betalains (1), the aglycone of most of betacyanins (2), and a proline-containing betaxanthin (3). Spectrophotometric analysis of extracts Betacyanins content was expressed as betanin content (molar mass M = 551,48 g/mol). Wavelength was setted to 527 nm (Fig. 8). HPLC analysis of extract The analysis of all extracts both from solvent extraction and belonged from SFE were carried out by means of HPLC using a Smartline HPLC system (Fig. 9). Chromatographic separation was carried out using a 2,0 mm ID ×150 mm L, with precolumn, C-18 TSKgel ODS-100 V, 21810. Extraction of betalains with supercritical fluids All samples after collection were previously cleaned and peeled, homogenized and frozen; at the time of the analysis; some of them were centrifuged, other partially dried and added with different sand Ottawa and Diatomee, in order to maximize the yields of extraction. The supercritical extractions by CO2 were performed on a Spe-ed SFE 4 extractor (Fig. 7). Flow CO2= 1.5 L/min. • extra virgin olive oil 20w/w% • Tween 60 • buffer solution • CaCl2; • pectin L.M. • ethanol EMU CPFE EMU STD + extract purified of Opuntia ficus-indica EMU STD EMU VIT E EMU STD + Vitamin E • extra virgin olive oil 70w/w% • mono- and diglycerides of • fatty acids • NaCl 0,1 M EMU CPFE EMU STD + extract purified of Opuntia ficus-indica EMU STD EMU VIT E EMU STD + Vitamin E Preparaions of emulsion oil in water (O/W) and water in oil (W/O) Purification of extract (CPFE): the pulp fruit was homogenized, centrifuged and precipitated with acetone; then filtrated with a PTFE siringe. The emulsification was performed using a rotor stator system (Ultra Turrax T 50). Antioxidant activity The antioxidant properties of all emulsions (O/W and W/O), at four different times after preparations (t0= 1 day; t1= 5 days; t2= 10 days; t3= 15 days), were evaluated by measurement of the scavenging activity towards DPPH and ABTS radicals. DPPH Assay (2,2′-diphenyl-1- picrylhydrazyl) λ=517 nm ABTS Assay [2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)] λ=734 nm DPPH and ABTS radicals inhibition in O/W emulsions DPPH and ABTS radicals inhibition in W/O emulsions The homogenised mix (Fig. 10) was prepared to measure the level of acidity of the primary source, brix degrees, water activity and humidity. The measurements of water activity and humidity are a key parameter in the quality control of moisture-sensitive products. Preliminary analysis of pulp 4. Molecular structure of betanin present in red beet roo

Benzothienoquinazolinones as new inhibitor of Topoisomerases and Tubulin

Ellipticine is a naturally occurring alkaloid with a carbazole backbone, well known for their antitumor activities, exerted through DNA intercalation, high DNA binding affinity and inhibition of the topoisomerases [1]. However, because of onset of some side effects the therapeutic application of Ellipticine and its derivatives used in clinic therapy, still remain limited [2]. Accordingly, with the aim to reduce side effects and improving their biologic activities, the search for new Ellipticine analogues is always demanded. This work is focused on the creation of new library of benzothienoquinazolinones (4-9), analogues of Ellipticine, in which both the carbazole moiety and the pyridine ring were replaced by a dibenzothiopheneand a pyrimidine moiety, respectively. The synthesis of these 3-(alkyl(dialkyl)amino)benzothieno[2,3- f]quinazolin-1(2H)-ones) was realized in a simple one-pot reaction using 3-aminodibenzothiophene as a starting material (Figure 1). These have shown an interesting anti-proliferative activity on two breast cancer cell lines, MCF-7 and MDAMB- 231. Molecular docking of these compounds was performed on the crystallographic enzyme structures of Tubulin, Topoisomerase I and II. The simulation results showed that the compounds investigated bind the enzymes with a relatively high affinity, at least in the low micromolar range. Other in vitro assays are in progress to confirm the in silico results

Phosphonium Salt Displays Cytotoxic Effects Against Human Cancer Cell Lines

Aims/Objective: Phosphonium salts are compounds whose structural characteristics enable them to cross the plasma and mitochondrial membrane with ease. Cancer cells have higher plasma membrane potentials than normal cells; phosphonium salts selectively accumulate in the mitochondria of neoplastic cells and inhibit mitochondrial function. Method: In the present work, we investigated the cytotoxic activity of lipophilic phosphonium salt (11- methoxy11-oxo-undecyl) triphenylphosphonium bromide (MUTP) as well as of the two new phosphine oxide salts, 3,3'-(methylphosphoryl) dibenzenaminium chloride (SBAMPO) and 3,3' (phenylphosphoryl) dibenzenaminium chloride (SBAPPO) on the proliferation of breast cancer cell line (MCF-7) and human uterin cervix adenocarcinoma cells (HeLa). Result: We showed that only MUTP exhibits antiproliferative effects on both cell lines, without affecting the normal breast epithelial cell proliferation. More specifically, we demonstrated that MUTP treatment of breast cancer cells is associated with impaired cell-cycle progression and metabolically induces mitochondrial damage and triggers apoptotic cell death in MCF-7 and HeLa cells. Taken together, these findings suggest that MUTP may be capable of selectively targeting neoplastic cell growth and therefore has potential applications as anticancer agent

Modeling, preparation and characterization of a dipole moment switch driven by Z/E photoisomerization

We report the results of a multidisciplinary research effort where the methods of computational photochemistry and retrosynthetic analysis/synthesis have contributed to the preparation of a novel N-alkylated indanylidene-pyrroline Schiff base featuring an exocyclic double bond and a permanent zwitterionic head. We show that, due to its large dipole moment and efficient photoisomerization, such a system may constitute the prototype of a novel generation of electrostatic switches achieving a reversible light-induced dipole moment change on the order of 30 D. The modeling of a peptide fragment incorporating the zwitterionic head into a conformationally rigid side chain shows that the switch can effectively modulate the fluorescence of a tryptophan probe

α–ω Alkenyl‐bis‐ S

Cancer is going to be the first cause of mortality worldwide in the 21th century. It is considered a multifactorial disease that results from the combined influence of many genetic aberrations, leading to abnormal cell proliferation. As microtubules are strongly implicated with cellular growth, they represent an important target for cancer treatment. The well-known Microtubule Targeting Agents (MTAs) including paclitaxel, colchicine and vinca alkaloids are commonly used in the treatment of various cancers. However, adverse effects and drug resistance are the major limitation in their clinical use. With the aim to find new candidates able to induce microtubule alteration with reduced toxic effects or drug resistance, we studied a new small series of derivatives that present imidazolinic, guanidinic, thioureidic and hydrazinic groups (1-9). All the compounds were tested for their antitumor activity against a panel of six tumoral cell models. Particularly, compound 8 (1,9-nonanediyl-bis-Samidinothiourea-dihydrobromide) showed the lower IC50 value against HeLa cells, together with a low cytotoxicity on the normal cells. This compound was able to induce the apoptotic mitochondrial pathway and inhibited the tubulin polymerization with a similar efficacy of Vinblastine and Nocodazole. Taken together, these promising biological properties make compound 8 useful for the development of novel therapeutic approaches in cancer treatment