Quercetin derivatives as novel antihypertensive agents: Synthesis and physiological characterization.

The antihypertensive flavonol quercetin (Q1) is endowed with a cardioprotective effect against myocardial ischemic damage. Q1 inhibits angiotensin converting enzyme activity, improves vascular relaxation, and decreases oxidative stress and gene expression. However, the clinical application of this flavonol is limited by its poor bioavailability and low stability in aqueous medium. In the aim to overcome these drawbacks and preserve the cardioprotective effects of quercetin, the present study reports on the preparation of five different Q1 analogs, in which all OH groups were replaced by hydrophobic functional moieties. Q1 derivatives have been synthesized by optimizing previously reported procedures and analyzed by spectroscopic analysis. The cardiovascular properties of the obtained compounds were also investigated in order to evaluate whether chemical modification affects their biological efficacy. The interaction with ß-adrenergic receptors was evaluated by molecular docking and the cardiovascular efficacy was investigated on the ex vivo Langendorff perfused rat heart. Furthermore, the bioavailability and the antihypertensive properties of the most active derivative were evaluated by in vitro studies and in vivo administration (1month) on spontaneously hypertensive rats (SHRs), respectively. Among all studied Q1 derivatives, only the ethyl derivative reduced left ventricular pressure (at 10(-8)M÷10(-6)M doses) and improved relaxation and coronary dilation. NOSs inhibition by L-NAME abolished inotropism, lusitropism and coronary effects. Chronic administration of high doses of this compound on SHR reduced systolic and diastolic pressure. Differently, the acetyl derivative induced negative inotropism and lusitropism (at 10(-10)M and 10(-8)÷10(-6)M doses), without affecting coronary pressure. Accordingly, docking studies suggested that these compounds bind both ß1/ß2-adrenergic receptors. Taking into consideration all the obtained results, the replacement of OH with ethyl groups seems to improve Q1 bioavailability and stability; therefore, the ethyl derivative could represent a good candidate for clinical use in hypertension.

Sericin/Poly(ethylcyanoacrylate) Nanospheres by Interfacial Polymerization for Enhanced Bioefficacy of Fenofibrate: In Vitro and In Vivo Studies.

Fenofibrate is a lipophilic drug used in hypercholesterolemia and hypertriglyceridemia as a lipid-regulating agent; however, it is characterized by poor water solubility and low dissolution rate, which result in a low oral bioavailability. In the present study, sericin/poly(ethylcyanoacrylate) nanospheres are synthesized by interfacial polymerization in aqueous media and investigated as a novel sericin-based delivery system for improved and enhanced oral bioefficacy of fenofibrate. The incorporation of sericin into the prepared cyanoacrylate nanoparticles and their spherical shape are confirmed by Lowry assay and scanning electron microscopy, respectively. Hydrophilic and mucoadhesive properties of the synthesized nanospheres are also evaluated. Finally, both in vitro release and in vivo studies are performed and the oral absorbable amount of fenofibrate is calculated to be higher than 70% when incorporated into the polymeric material, reducing the levels of total cholesterol (TC), triacylglycerols (TG), very low-density lipoproteins (VLDL), and low-density lipoproteins (LDL) compared to fenofibrate alone.

Caffeic Acid-PLGA Conjugate to Design Protein Drug Delivery Systems Stable to Irradiation.

This work reports the feasibility of caffeic acid grafted PLGA (g-CA-PLGA) to design biodegradable sterile microspheres for the delivery of proteins. Ovalbumin (OVA) was selected as model compound because of its sensitiveness of γ-radiation. The adopted grafting procedure allowed us to obtain a material with good free radical scavenging properties, without a significant modification of Mw and Tg of the starting PLGA (Mw PLGA = 26.3 ± 1.3 kDa vs. Mw g-CA-PLGA = 22.8 ± 0.7 kDa; Tg PLGA = 47.7 ± 0.8 °C vs. Tg g-CA-PLGA = 47.4 ± 0.2 °C). By using a W1/O/W2 technique, g-CA-PLGA improved the encapsulation efficiency (EE), suggesting that the presence of caffeic residues improved the compatibility between components (EEPLGA = 35.0% ± 0.7% vs. EEg-CA-PLGA = 95.6% ± 2.7%). Microspheres particle size distribution ranged from 15 to 50 µm. The zeta-potential values of placebo and loaded microspheres were -25 mV and -15 mV, respectively. The irradiation of g-CA-PLGA at the dose of 25 kGy caused a less than 1% variation of Mw and the degradation patterns of the non-irradiated and irradiated microspheres were superimposable. The OVA content in g-CA-PLGA microspheres decreased to a lower extent with respect to PLGA microspheres. These results suggest that g-CA-PLGA is a promising biodegradable material to microencapsulate biological drugs.

Brewing effect on levels of biogenic amines in different coffee samples as determined by LC-UV.

Eight biogenic amines (spermine, spermidine, putrescine, histamine, tyramine, phenylethylamine, cadaverine and serotonin) were determined by LC-UV after derivatization with dansyl-chloride in both ground coffee and coffee beverages obtained by different methods. In ground coffee, the most relevant amine was PUT, followed by SPD, HIS, TYR, CAD, SPM, PHE, and SER, with the total BAs content decreasing as the roasting degree increased. In coffee brews, the order was PUT, SPM, TYR, CAD, SPD, PHE, HIS, and SER, but at a very low level in comparison with the amount of BAs determined in roasted ground coffee. Beverages prepared by espresso, capsule, and pod machines had the lowest BAs contents, as a result of the thermal and physical stress imposed on ground coffee by these methods, while mocha contained the highest BAs amounts owing to lower pressure and longer brewing time.

Olive oil/policosanol organogels for nutraceutical and drug delivery purposes.

Low molecular weight organogels are semisolid systems structured by the assembly of molecules that crystallise under suitable process conditions. The inner microstructure of organogels is made up of a 3-D network, in which both an organic liquid solvent and other dispersed particles can be entrapped. In this work, olive oil organogels, structured from policosanol (a nutraceutical mixture of fatty alcohols), were studied in order to obtain the best formulation for producing a support for delivery of lipophilic agents (namely ferulic Acid) via oral administration. A rheological optimisation of the olive oil-policosanol organogel was first of all performed with Step Shear Rate Temperature Ramp tests. This provided important information on the policosanol fraction to be added to the system and on the onset of crystallisation temperature, an indirect measurement of the system melting point. It was found that a policosanol weight percentage of 0.03 was suitable to obtain contemporary semisolid organogels, consistent enough and thermally stable for human ingestion. In vitro tests on organogels loaded with ferulic acid were also carried out in order to simulate the oral intake of the nutraceutical compound. This evidenced a release mechanism determined by both erosion and diffusion; a good performance of gels and their ability to control the release rate through the degree of structuration were also observed.

A new method for the determination of biogenic amines in cheese by LC with evaporative light scattering detector.

This paper presents a new LC method with evaporative light scattering detection (ELSD), for the separation and determination of the biogenic amines (histamine, spermidine, spermine, tyramine, putrescine and ß-phenylethylamine) which are commonly present in cheese, as their presence and relative amounts give useful information about freshness, level of maturing, quality of storage and cheese authentication. The LC-ELSD method is validated by comparison of the results with those obtained through LC-UV determination, based on a pre-column dansyl chloride derivatisation step. The obtained data demonstrate that both methods can be interchangeably used for biogenic amines determination in cheese. The new LC-ELSD method shows good precision and permits to achieve, for standard solutions, limit of detection (LOD) values ranging from 1.4 to 3.6 mg L(-1) and limit of quantitation (LOQ) values ranging from 3.6 to 9.3 mg L(-1). The whole methodology, comprehensive of the homogenization-extraction process and LC-ELSD analysis, has been applied in the analysis of a typical Calabria (Southern Italy) POD cheese, known as Caciocavallo Silano. The most aboundant amine found was histamine, followed, in decreasing order, by tyramine, spermine, putrescine, ß-phenylethylamine and spermidine, for a total amount of 127 mg kg(-1). This value does not represent a possible risk for consumer health, according to the toxicity levels reported in literature and regarded as acceptable.

Antioxidant activity of a Mediterranean food product: "fig syrup".

In this work, the efficacy of fig syrup, a Mediterranean fig derivative, as a nutraceutical supplement, was demonstrated. Fig syrup is a fruit concentrate used as a common ingredient in the preparation of typical foods, and particularly in cakes. In vitro assays were performed to determine the amount of nutraceutical ingredients, such as phenolic compounds (3.92 mg equivalent of gallic acid per g) and flavonoids (0.35 mg equivalent of catechin per g), while HPLC analyses provided specific information about the composition of antioxidants in the syrup. Furthermore, total antioxidant activity, scavenging properties against DPPH and peroxyl radicals, and the anticholinesterase activity, clearly showed the efficacy of the syrup in preventing damage induced by free radicals and, thus, the applicability of this food derivative as a nutraceutical supplement.

Grafted thermo-responsive gelatin microspheres as delivery systems in triggered drug release.

In this paper, a novel class of microspheric hydrogels was synthesized by grafting of N-isopropyacrylamide (NIPAAm) with gelatin. The possibility of inserting commercial gelatin in a crosslinked structure bearing thermo-sensitive moieties, by radical process, represents an interesting innovation that significantly improves the device performance, opening new applications in biomedical and pharmaceutical fields. This synthetic approach allows a modification of the polymeric network composition, producing hydrogels with suitable physico-chemical properties and a transition temperature higher than NIPAAm homopolymers. The incorporation of monomers into the network was confirmed by infrared spectroscopy, and the composition of the polymerization feed was found to strictly influence the network density and the shape of hydrogels. Thermal analyses showed negative thermo-responsive behaviour with shrinking/swelling transition values in the temperature range 34.6-34.8 degrees C, according to the amount of the hydrophilic portions in the network. In order to test the preformed materials as drug carriers, diclofenac sodium salt was loaded into the spherical microparticles. After the determination of the drug entrapment percent, drug release profiles in media at different temperature were analysed. By using semi-empirical equations, the release mechanism was extensively studied and the diffusional contribution was evaluated.

Negative thermo-responsive microspheres based on hydrolyzed gelatin as drug delivery device.

This paper deals with the synthesis of thermo-responsive microspheres with proteic structure exhibiting a transition temperature close to the body temperature. Temperature-sensitive hydrogels have attracted extensive interest due to their potential and promising applications in drug delivery field since they can undergo a rapid and reversible phase transition from a swollen to a shrunken state depending on environmental temperature. The hydrogels were synthesized by free-radical polymerization of hydrolyzed methacrylated gelatin (HGel-MA) and N,N'-methylenebisacrylamide as pro-hydrophilic multifunctional macromer and crosslinker, respectively, and N-isopropylacrylamide as thermo-responsive monomer. Thermal analyses showed negative thermo-responsive behavior for all compositions and, by increasing the content of the hydrophilic moieties in the network, the transition temperature raised to 36.9 degrees C, close to the physiological values. In order to test the materials as drug carriers, diclofenac sodium salt was chosen as model drug. Drug release profiles, in phosphate buffer solution (pH 7.0, 10(-3) M) at 25 and 40 degrees C, depend on the hydrogel's crosslinking degree and hydrophilic/hydrophobic balance in the polymeric network. For all formulations, in the shrunken state, the drug release percent values ranged from 80% to 100% after 24 h, and after 3 h, more than 60% of therapeutics was delivered. On the contrary, the swelling of the loaded microparticles produces, even after 30 h, a drug release percent of about 75%. By using semi-empirical equations, the release mechanism was extensively studied and the diffusional contribute was evaluated. The physico-chemical characteristics of thermo-responsive materials confirm the applicability of the microspheres as drug delivery device.

Synthesis of methacrylic-ferulic acid copolymer with antioxidant properties by single-step free radical polymerization.

A novel, simple, and cheap method to synthesize antioxidant methacrylic-ferulic acid copolymer (PMAA-FA) by free radical polymerization was developed by employing a hydrogen peroxide-ascorbic acid pair to produce hydroxyl radicals acting as radical initiators. FT-IR spectra were performed to verify the insertion of ferulic acid into the polymeric chain, and the antioxidant activity of PMAA-FA was compared to that of a control polymer synthesized in the absence of antioxidant molecule. Good antioxidant activity was demonstrated by obtained materials, showing the efficiency of the polymerization method. This material could be useful in the pharmaceutical field and in the food industry (food packaging).