The aim of this study was to extract, protect and increase the bioaccessibility of phenolic compounds extracted from the Chilean Tórtola common bean. For obtaining phenolic compounds, two methods ...were analyzed, focusing on their extraction and their drying. In the method where ultrasound-assisted extraction and spray drying were used, we obtained a yield five times greater than the conventional method. Thereafter, we evaluated an encapsulation system with two material shells, alginate and Eudragit L-100. The protective capacity of the capsules was verified by determining antioxidant capacity, total phenolic compounds content, as well as biomedical capacities of powder-encapsulated extract. Antithrombotic activity was evaluated as biomedical capacity, observing a 30% reduction in platelet aggregation. Additionally, quality characteristics were assessed using FITR, microscopic confocal, TGA, and hygrometry techniques. TGA analyses demonstrated the thermal protection of the capsule, reducing the degradation of encapsulated phenolic compounds by threefold compared to the non-encapsulated extract. Finally, two in vitro digestion models were used to simulate the physiological digestion. In both models, we observed an increase in TPC bioaccessibility even over 100%. These results would benefit biomedical applications and industrial processing of phenolic compounds extracted from common beans.
•Use of Tortóla beans as a source of phenolic compounds with biomedical capacity.•Development of an efficient extraction and drying method that significantly increases yield compared to conventional methods.•Development of an encapsulation system using alginate and Eudragit-L100•Demonstration that encapsulated phenolic compounds show a 60% higher antithrombotic capacity than non-encapsulated ones.•Increase in intestinal bioaccessibility in vitro of phenolic compounds.
The increasing incidence of cardiovascular diseases has created an urgent need for safe and effective antithrombotic agents. In this study, we aimed to elucidate the structural characteristics and ...antithrombotic activity of a novel polysaccharide isolated from Auriculariaauricula fruiting bodies. The purified polysaccharide AAP-b2 (12.02 kDa) was composed of mannose, glucuronic acid, glucose and xylose, with a molar ratio of 89.25:30.50:4.25:1.00. Methylation and NMR analyses showed that AAP-b2 primarily consisted of →2,3)-Manp-(1→, →3)-Manp-(1→, →4)-GlcAp-(1→ and Manp-(1→. A thrombus mouse model induced by carrageenan was used in this research to evaluate its antithrombotic effect. AAP-b2 significantly inhibited platelet aggregation, reduced the black tail length and prolonged the coagulation time, including activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT), exerting a good inhibitory effect on thrombosis in mice. The antithrombotic activity of AAP-b2 was found to be related to the inhibition of platelet activation by regulation of endothelial nitric oxide synthases (eNOs), endothelin-1 (ET-1), prostacyclin (PGI2) and thromboxane B2 (TXB2), along with the enhancement of anticoagulant activity by affecting antithrombin III (AT-III) and protein C (PC) pathways.
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•An intravascular catheter was coated with CMC layer exhibiting antimicrobial, antithrombotic, and low friction properties.•Whole manufacturing process was water-based, hence ...eco-friendly and biocompatible.•Functionalities of CMC layer were enhanced by engineering the porous structure.•Sufficient lubricity of porous CMC coated catheter was verified via trackability test.
Antimicrobial, antithrombotic activity and low-friction functions are essential for the surface of intravascular catheters (ICs). However, the multifunctional surface, encompassing all of the above properties has not yet been realized. Here, we report a novel strategy for biocompatible and eco-friendly surface modification of the ICs with multifunctional polysaccharide, O-carboxymethyl chitosan (CMC). Micro- and nanoscale porous CMC (p-CMC) layer was simply fabricated via a selective elimination of the water-soluble polyethylene glycol (PEG) from heterogeneous CMC/PEG composite. The p-CMC structure exhibited a significantly enhanced hydration rate and superhydrophilic property. In particular, the antifouling property of superhydrophilic surface showed excellent anti-adhesion of Escherichia coli and platelets along with the intrinsic multifunctionality of CMC, indicating a dual effect of our p-CMC layer: (1) intrinsic antibacterial and antithrombotic properties of CMC and (2) anti-adhesion of substances on superhydrophilic surface. Meanwhile, despite the rough surface of the p-CMC layer, it showed high lubricity and durability under continuous wet friction conditions. Furthermore, we demonstrated that the actual p-CMC coated intravascular catheter (IC) provides superior trackability in a curved artificial blood vessel. The potential of the proposed coating strategy can be offered not only ICs, but also wide range of polymer-based applications including vascular filters, grafts, pacemakers and soft robots.
The aim of this study was to investigate the multifunctional properties of specific synthetic peptides previously identified in milk fermented with Lactococcus lactis NRRL B-50571 and NRRL B-50572 on ...the inhibition of enzymes involved in hypertension (ACE), type 2 diabetes (DPP-IV) and thrombosis (thrombin). Moreover, in vitro gastrointestinal digestion was performed in order to know the stability of peptides and its effect on the biological activities. Also, in silico analysis was employed for the prediction of sequences after digestion. Results showed that peptide inhibitory activities on ACE, DPP-IV and thrombin were enhanced after digestion. After in vitro digestion, peptide NAVPITPTLN showed the best efficiency to inhibit DPP-IV. Likewise, NAVPITPTLN and QEPVLGPVRGPFIIV showed high thrombin inhibitory activities, and HPHPHLSFMAIPP and SLPQNIPPL presented high ACE inhibitory activities. In vitro and in silico studies indicated that these peptides are precursors of multifunctional peptide sequences. Therefore, this study suggests that the specific synthetic peptides previously identified in milk fermented with Lactococcus lactis NRRL B-50571 and NRRL B-50572 offer potential beneficial health effects in the management of hypertension, thrombosis and diabetes.
•Peptides studied presented inhibitory activity on ACE, thrombin and DPP-IV.•YPSYGL is a new DPP-IV inhibitor peptide released by L. lactis NRRL B-50572.•HPHPHLSFMAIPP and SLPQNIPPL showed ACE and DPP-IV inhibition.•Novel potential thrombin inhibiting peptides were identified by in silico analysis.
Amaranth protein isolate and fractions were obtained and subjected to proteolysis in order to evaluate its potential antithrombotic activity. The proteins were first hydrolyzed with alcalase (pH 10, ...37 °C) and then with trypsin (pH 8, 37 °C). The samples were characterized physicochemically and antithrombotic activity was evaluated using clotting tests (PT, TT and APTT) and the microplates assay. The fractions compared to the hydrolysates exhibited different electrophoretic profiles (tricine-SDSPAGE) and gel filtration chromatograms, evidencing the presence of different molecular species. The hydrolysis improved in every sample the bioactivity detected, excepting for the glutelin fraction, which exhibited the highest antithrombotic activity, significantly superior (p < 0.05) compared to the other fractions and the isolate. This behavior was observed in the two assays that analyzed the common path of the coagulation cascade at similar concentrations: TT (81.0 ± 8.5 s with a control of 19.5 ± 0.7 s) and microplate test (IC50 80 μg/mL), indicating a possible mechanism of action that involves the thrombin activity or the polymerization of fibrin monomers. The glutelin fraction showed a potential capacity to inhibit coagulation, appearing as a promising ingredient to formulate functional foods.
•Amaranth seed proteins and hydrolysates were obtained and characterized.•Antithrombotic proteins from amaranth were detected by two different assays.•The glutelin fraction exhibited the highest antithrombotic activity.•Hydrolysis improved the bioactivity detected, excepting for the glutelin fraction.
Thrombin is one of the key enzymes of the blood coagulation system and a promising target for the development of anticoagulants. One of the most specific natural thrombin inhibitors is hirudin, ...contained in the salivary glands of medicinal leeches. The medicinal use of recombinant hirudin is limited because of the lack of sulfation on Tyr63, resulting in a 10-fold decrease in activity compared to native (sulfated) hirudin. In the present work, a set of hirudin derivatives was tested for affinity to thrombin: phospho-Tyr63, Tyr63(carboxymethyl)Phe, and Tyr63Glu mutants, which mimic Tyr63 sulfation and Gln65Glu mutant and lysine-succinylated hirudin, which enhance the overall negative charge of hirudin, as well as sulfo-hirudin and desulfo-hirudin as references. Using steered molecular dynamics simulations with subsequent umbrella sampling, phospho-hirudin was shown to exhibit the highest affinity to thrombin among all hirudin analogs, including native sulfo-hirudin; succinylated hirudin was also prospective. Phospho-hirudin exhibited the highest antithrombotic activity in in vitro assay in human plasma. Taking into account the modern methods for obtaining phospho-hirudin and succinylated hirudin, they are prospective as anticoagulants in clinical practice.
A previously undescribed open-loop decarbonizing cembranolide, sarcocinerenolide A, and eight undescribed cembranolides, sarcocinerenolides B–I, characterized by poly-membered oxygen ring fragments ...were isolated from the soft coral Sarcophyton cinereum collected from the South China Sea. The structures and absolute configurations of these previously undescribed compounds were precisely determined by analysis of NMR data, DP4+ and ECD spectra. The bioactivities of the compounds were evaluated using zebrafish models and sarcocinerenolides C and H exhibited anti-thrombotic activity.
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•One decarbonizing and eight polyoxygenated cembranolides were reported.•The structures were determined by analysis of NMR data, DP4+ and ECD spectra.•Compounds exhibited anti-thrombotic activity.
The interest in bioactive compounds from natural sources, such as marine organisms, has increased considerably in recent years. Among these compounds, sulfated polysaccharides from seaweed exhibit a ...broad spectrum of biological activities. Sulfated polysaccharides from green algae are still poorly investigated. For this reason, in this study, using unusual methodologies, such as extraction conditions, FACE, and RAMAN, we investigated the structural features of F50UL and F70UL sulfated polysaccharides from Ulva lactuca L. and their distinct in vitro anticoagulant and in vivo antithrombotic activities. Sulfated polysaccharides of U. lactuca were obtained by enzymatic proteolysis with ALCALASE® and fractionated by acetone precipitation. F50Ul and F70Ul sulfated polysaccharides with higher yield were partially chemically characterized by Fluorophore-assisted carbohydrate electrophoresis (FACE) and RAMAN spectroscopy analysis and submitted to an in vitro screening by APTT, PT, TT, and anti-factor Xa and IIa tests. The venae cavae ligature experimental model for the analysis of in vivo antithrombotic activity of F50Ul and F70Ul sulfated polysaccharides were also performed. The U. lactuca L. sulfated polysaccharides characterization by FACE and RAMAN showed a typical ulvans structure that contains as principal component rhamnose, but other monosaccharides (uronic acid, glucose, and galactose) are present. F50Ul (0.1–1.0 μg/μl) showed anticoagulant activity in vitro. However, F70Ul that has a similar composition did not present these effects. Also, only F50Ul sulfated polysaccharides (≥5 μg/g) showed a great in vivo antithrombotic concentration-dependent and time-dependent activity. In summary, we demonstrate the use of unusual extraction and characterization analysis procedures for U. lactuca L. sulfated polysaccharides and the ability of F50Ul to reduce the weight of thrombus in rats probably by the association with factors Xa and IIa inhibition. These results provide strong evidence of the anticoagulant potential of these sulfated polysaccharides isolated from Ulva lactuca L.
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•Sulfated polysaccharides from Ulvalactuca L. obtained by proteolytic procedures•Raman structural analysis of seaweed sulfated polysaccharides•U. lactuca sulfated polysaccharides were able to inhibit the coagulation Factor Xa.•In vivo inhibition of clot formation by sulfated polysaccharides from U.lactuca L.
A series of pyrrolo3,2-dpyrimidineone derivatives have been designed and synthesized as novel antithrombotic agents. Thereinto, compound 17a is a strong FXa inhibitor with excellent in vivo ...antithrombotic activity.
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•Pyrrolo3,2-dpyrimidineone compounds were designed and synthesized.•All of tested compounds possess effective anticoagulant potency in vitro.•Compound 17a is an in vivo highly antithrombotic and strong FXa inhibitor.•Large conjugate system is an ideal skeleton of FXa inhibitors.
A series of pyrrolo3,2-dpyrimidineone compounds have been designed and synthesized as novel FXa inhibitors. Bioassay of the tested compounds showed moderate to excellent anticoagulant potency in vitro. Further FXa inhibitory and bioactivity evaluation in rats, the FeCl3-induced venous thrombosis model, showed that the compound 17a has good FXa inhibitory activity (IC50 = 1.57 nM) and in vivo antithrombotic potency. The anticoagulant effects of compound 17a were dose dependent whether in vitro or in vivo. The results further confirmed our hypothesis that the large conjugated structure is an ideal skeleton binding FXa.
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