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2.
Mar Drugs ; 18(1)2019 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-31861572

RESUMO

Heparin from mollusks with unique sulfated glycosaminoglycan exhibits strong anti-thrombotic activities. This study reports on a purified heparinoid from Coelomactra antiquata, which shows potent anticoagulant and fibrinolytic abilities. Its structure was characterized by infrared spectroscopy, high-performance liquid chromatography, and one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy. Its fibrinolytic activity was determined in vitro and in vivo. Its anticoagulant activity was determined by activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). The results indicated that clam heparinoid was a homogeneous glycosaminoglycan with a molecular weight of 30.99 kDa, mainly composed of →4)-α-IdoA2S-(1→4)-α-GlcNS3S6S (or GlcNS6S)-(1→4)-ß-GlcA-(1→4)-α-GlcNS6S (or GlcNAC)-(1→. Furthermore, this heparinoid showed a highly anticoagulant titer and fibrinolytic value of 149.63 IU/mg and 1.96 IU/mg, respectively. In summary, clam heparinoid shows great potential for application in the clinic and antithrombotic drugs industry.


Assuntos
Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Fibrinolíticos/química , Fibrinolíticos/isolamento & purificação , Heparina/química , Heparina/isolamento & purificação , Animais , Anticoagulantes/farmacologia , Bivalves , Feminino , Fibrinolíticos/farmacologia , Heparina/análogos & derivados , Heparina/farmacologia , Humanos , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Entorses e Distensões , Tempo de Trombina
3.
Fitoterapia ; 137: 104282, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31381956

RESUMO

Four new polyketides, alternatains A-D (1-4), along with 17 known compounds (5-21) were obtained from the solid substrate fermentation cultures of Alternaria alternata MT-47, an endophytic fungus isolated from the medicinal plant of Huperzia serrata. Their structures were elucidated by extensive spectroscopic and spectrometric techniques (1D and 2D NMR, IR, and HRESIMS) and calculated electronic circular dichroism (ECD) method. Compounds 4, 6, 15, and 21 exhibited inhibitory activities on ATP release of thrombin-activated platelets with IC50 values in the range of 18.2-68.8 µM.


Assuntos
Alternaria/química , Anticoagulantes/farmacologia , Plaquetas/efeitos dos fármacos , Huperzia/microbiologia , Policetídeos/farmacologia , Acetilcolinesterase , Trifosfato de Adenosina , Anticoagulantes/isolamento & purificação , Butirilcolinesterase , China , Inibidores da Colinesterase , Endófitos/química , Humanos , Estrutura Molecular , Plantas Medicinais/microbiologia , Policetídeos/isolamento & purificação
4.
Molecules ; 24(16)2019 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-31426507

RESUMO

Nowadays, pharmaceutical heparin is purified from porcine and bovine intestinal mucosa. In the past decade there has been an ongoing concern about the safety of heparin, since in 2008, adverse effects associated with the presence of an oversulfated chondroitin sulfate (OSCS) were observed in preparations of pharmaceutical porcine heparin, which led to the death of patients, causing a global public health crisis. However, it has not been clarified whether OSCS has been added to the purified heparin preparation, or whether it has already been introduced during the production of the raw heparin. Using a combination of different analytical methods, we investigate both crude and final heparin products and we are able to demonstrate that the sulfated contaminants are intentionally introduced in the initial steps of heparin preparation. Furthermore, the results show that the oversulfated compounds are not structurally homogeneous. In addition, we show that these contaminants are able to bind to cells in using well known heparin binding sites. Together, the data highlights the importance of heparin quality control even at the initial stages of its production.


Assuntos
Anticoagulantes/isolamento & purificação , Sulfatos de Condroitina/isolamento & purificação , Contaminação de Medicamentos , Heparina/isolamento & purificação , Animais , Anticoagulantes/química , Bovinos , Sulfatos de Condroitina/química , Heparina/química , Heparina Liase/química , Humanos , Hidrólise , Mucosa Intestinal/química , Espectroscopia de Ressonância Magnética , Polissacarídeo-Liase/química , Controle de Qualidade , Suínos
5.
Carbohydr Polym ; 222: 115010, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31320102

RESUMO

Two sulfated polysaccharides (SPs), F2 and F3, isolated from Codium isthmocladum were found to contain galactose, sulfate, and pyruvate. The apparent molecular weights of F2 and F3 were determined to be 62 and 61 kDa, respectively. NMR spectroscopy combined with chemical analysis showed that F2 and F3 have the same structural features. However, F3 showed higher sulfate/sugar ratio (1/2.6) than F2 (1/4). F2 and F3 are essentially (1 → 3)-ß-D-galactans with some branching at C6. Pyruvylation occurs at O3 and O4, forming 3,4-O-(1-carboxyethylidene)-ß-D-Galp residues; some of these pyruvylated residues contain sulfate groups at C6. Some non-branching residues contain sulfate at C4. None of the SPs exhibited antioxidant activity. MTT results indicated that 1 mg/mL of both SPs about 40% of PANC-1 cell viability. At 10 µg/mL, F2 and F3 had 1.7-fold longer clotting times compared to that of Clexane® at the same concentration. The higher sulfate content of F3 is not a determining factor for pharmacological activities of galactans, considering that both F2 and F3 exerted the effects.


Assuntos
Anticoagulantes/farmacologia , Antioxidantes/farmacologia , Clorófitas/química , Galactanos/farmacologia , Alga Marinha/química , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Sequência de Carboidratos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Galactanos/química , Galactanos/isolamento & purificação , Humanos , Piruvatos/química , Piruvatos/isolamento & purificação , Piruvatos/farmacologia , Ésteres do Ácido Sulfúrico/química , Ésteres do Ácido Sulfúrico/isolamento & purificação , Ésteres do Ácido Sulfúrico/farmacologia
6.
BMC Complement Altern Med ; 19(1): 148, 2019 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-31238930

RESUMO

BACKGROUND: Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson, SBT) is a valuable plant because of its medical and therapeutic potential. Different bioactive compounds in SBT berries are of special interest to various researchers. However, not only sea buckthorn berries, but also leaves of this plant (both fresh and dried) contain a lot of nutrients and bioactive compounds, including phenolic compounds. The present study was carried out in order to investigate antioxidant and anticoagulant properties of sea buckthorn twig and leaf extracts (0.5-50 µg/mL) by using various in vitro models. Moreover, the aim of present experiments was to compare the biological activity of SBT leaf extract and SBT twig extract with selected berry extracts (a rich source of phenolic compounds): SBT berry extract (flavonoids being the dominant components), a commercial extract from the berries of Aronia melanocarpa (Aronox®), and a grape seed extract. METHODS: We determined the effect of plant extracts on the oxidative stress using selected markers of this process, i.e. the level of carbonyl groups in proteins. Additionally, we analysed the potential mechanism of modulation of hemostatic properties of human plasma (using selected coagulation times). RESULTS: SBT twig and leaf extracts were observed to exhibit an antioxidant activity against two strong biological oxidants: hydrogen peroxide (H2O2) and H2O2/Fe (the donor of hydroxyl radicals), which induced human plasma lipid peroxidation and protein carbonylation. Both extracts also showed anticoagulant properties. CONCLUSIONS: Our present results have demonstrated that extracts from different parts of SBT, especially berries and twigs, in comparison to well-known berries (aronia and grape), may also be viewed as a good source of active substances - antioxidants for pharmacological or cosmetic applications. Moreover, it is very important from an economic point of view to know that there is a possibility of obtaining phenolic compounds not only from the berries or leaves, but also from twigs, which constitute a production waste.


Assuntos
Elaeagnaceae/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Caules de Planta/química , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Anticoagulantes/farmacologia , Antioxidantes/química , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Flavonoides/química , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Frutas/química , Humanos , Fenóis/química , Fenóis/isolamento & purificação , Fenóis/farmacologia , Photinia/química , Extratos Vegetais/isolamento & purificação , Tempo de Protrombina
7.
Mar Drugs ; 17(5)2019 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-31091758

RESUMO

(1) Background: Brown and red algal sulfated polysaccharides have been widely described as anticoagulant agents. However, data on green algae, especially on the Ulva genus, are limited. This study aimed at isolating ulvan from the green macroalga Ulva rigida using an acid- and solvent-free procedure, and investigating the effect of sulfate content on the anticoagulant activity of this polysaccharide. (2) Methods: The obtained ulvan fraction was chemically sulfated, leading to a doubling of the polysaccharide sulfate content in a second ulvan fraction. The potential anticoagulant activity of both ulvan fractions was then assessed using different assays, targeting the intrinsic and/or common (activated partial thromboplastin time), extrinsic (prothrombin time), and common (thrombin time) pathways, and the specific antithrombin-dependent pathway (anti-Xa and anti-IIa), of the coagulation cascade. Furthermore, their anticoagulant properties were compared to those of commercial anticoagulants: heparin and Lovenox®. (3) Results: The anticoagulant activity of the chemically-sulfated ulvan fraction was stronger than that of Lovenox® against both the intrinsic and extrinsic coagulation pathways. (4) Conclusion: The chemically-sulfated ulvan fraction could be a very interesting alternative to heparins, with different targets and a high anticoagulant activity.


Assuntos
Anticoagulantes/farmacologia , Fibroblastos/efeitos dos fármacos , Ulva/química , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Testes de Coagulação Sanguínea , Enoxaparina/farmacologia , Heparina/farmacologia , Humanos , Plasma/efeitos dos fármacos
8.
Biochimie ; 163: 12-20, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31059753

RESUMO

In parasites, cathepsins are implicated in mechanisms related to organism surveillance and host evasion. Some parasite cathepsins have fibrinogenolytic and fibrinolytic activity, suggesting that they may contribute to maintain blood meal fluidity for extended feeding periods. Here, it is shown that BmGTI (Rhipicephalus [Boophilus] microplus Gut Thrombin Inhibitor), a protein previously described as an inhibitor of fibrinogen hydrolysis and platelet aggregation by thrombin, and BmCL1 (Rhipicephalus [Boophilus] microplus Cathepsin-L like 1) are the same protein, hereinafter referred to using the earliest name (BmCL1). To further characterize BmCL1, Rhipicephalus microplus native and recombinant (rBmCL1) proteins were obtained. Native BmCL1 was isolated using thrombin-affinity chromatography, and it displays thrombin inhibition activity. We subsequently investigated rBmCL1 interaction with thrombin. We show that rBmCL1 and thrombin have a dissociation constant (ΚD) of 130.2 ±â€¯11.2 nM, and this interaction likely occurs due to a more electronegative surface of BmCL1 at pH 7.5 than at pH 5.0, which may favor an electrostatic binding to positively charged thrombin exosites. During BmCL1-thrombin interaction, thrombin is not degraded or inhibited. rBmCL1 impairs thrombin-induced fibrinogen clotting via a fibrinogenolytic activity. Fibrinogen degradation by BmCL1 occurs by the hydrolysis of Aα- and Bß-chains, generating products similar to those produced by fibrinogenolytic cathepsins from other organisms. In conclusion, BmCL1 likely has an additional role in R. microplus blood digestion, besides its role in hemoglobin degradation at acid pH. BmCL1 fibrinogenolytic activity indicates a proteolytic activity in the neutral lumen of tick midgut, contributing to maintain the fluidity of the ingested blood, which remains to be confirmed in vivo.


Assuntos
Catepsina L/metabolismo , Rhipicephalus/enzimologia , Trombina/metabolismo , Sequência de Aminoácidos , Animais , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Anticoagulantes/metabolismo , Catepsina L/química , Catepsina L/isolamento & purificação , Bovinos , Cinética , Modelos Moleculares , Proteólise
9.
Mar Drugs ; 17(5)2019 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-31100859

RESUMO

Therapeutic options for Alzheimer's disease, the most common form of dementia, are currently restricted to palliative treatments. The glycosaminoglycan heparin, widely used as a clinical anticoagulant, has previously been shown to inhibit the Alzheimer's disease-relevant ß-secretase 1 (BACE1). Despite this, the deployment of pharmaceutical heparin for the treatment of Alzheimer's disease is largely precluded by its potent anticoagulant activity. Furthermore, ongoing concerns regarding the use of mammalian-sourced heparins, primarily due to prion diseases and religious beliefs hinder the deployment of alternative heparin-based therapeutics. A marine-derived, heparan sulphate-containing glycosaminoglycan extract, isolated from the crab Portunus pelagicus, was identified to inhibit human BACE1 with comparable bioactivity to that of mammalian heparin (IC50 = 1.85 µg mL-1 (R2 = 0.94) and 2.43 µg mL-1 (R2 = 0.93), respectively), while possessing highly attenuated anticoagulant activities. The results from several structural techniques suggest that the interactions between BACE1 and the extract from P. pelagicus are complex and distinct from those of heparin.


Assuntos
Doença de Alzheimer/enzimologia , Secretases da Proteína Precursora do Amiloide/metabolismo , Ácido Aspártico Endopeptidases/metabolismo , Braquiúros/química , Ativação Enzimática/efeitos dos fármacos , Glicosaminoglicanos/farmacologia , Animais , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Anticoagulantes/farmacologia , Inibidores Enzimáticos/química , Inibidores Enzimáticos/isolamento & purificação , Inibidores Enzimáticos/farmacologia , Glicosaminoglicanos/química , Glicosaminoglicanos/isolamento & purificação
10.
Int J Biol Macromol ; 134: 405-412, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31071403

RESUMO

Chondroitin sulfate/dermatan sulfate (CS/DS) were isolated and purified for the first time from the bone of corb (Sciaena umbra) (CBG) and their chemical composition and anticoagulant activity were assessed. Infrared spectrum and agarose-gel electrophoresis for extracted CS/DS were also investigated. The results showed that the purified CS/DS obtained at a yield of 10% contains about 31.28% sulfate and an average molecular mass of 23.35 kDa. Disaccharide analysis indicated that CBG was composed of monosulfated disaccharides in positions 6 and 4 of the N-acetylgalactosamine (8.6% and 40.0%, respectively) and disulfated disaccharides in different percentages. The charge density was 1.4 and the ratio of 4:6 sulfated residues was equal to 4.64. Chondroitinase AC showed that the purified CS/DS contained mainly 74% CS and 26% DS. Moreover, the new CS/DS extracted from bone of corb showed a strong anticoagulant effect through activated partial thrombosis time (aPTT), thrombin time (TT) and prothrombin time (PT). In fact, CBG prolonged significantly (p < 0.05), aPTT and PT about 2.62 and 1.26 fold, respectively, greater than that of the negative control at a concentration of 1000 µg/mL. However, TT assay of CBG was prolonged 3.53 fold compared with the control at 100 µg/mL. The purified CS/DS displayed a promising anticoagulant potential, which may be used as a novel and soothing drug.


Assuntos
Anticoagulantes/química , Anticoagulantes/farmacologia , Osso e Ossos/química , Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacologia , Dermatan Sulfato/química , Dermatan Sulfato/farmacologia , Animais , Anticoagulantes/isolamento & purificação , Fracionamento Químico , Fenômenos Químicos , Sulfatos de Condroitina/isolamento & purificação , Dermatan Sulfato/isolamento & purificação , Peso Molecular , Umbridae
11.
Mar Drugs ; 17(4)2019 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-30934819

RESUMO

Sulfated polysaccharides from sea cucumbers possess distinct chemical structure and various biological activities. Herein, three types of polysaccharides were isolated and purified from Pattalus mollis, and their structures and bioactivities were analyzed. The fucosylated glycosaminoglycan (PmFG) had a CS-like backbone composed of the repeating units of {-4-d-GlcA-ß-1,3-d-GalNAc4S6S-ß-1-}, and branches of a sulfated α-l-Fuc (including Fuc2S4S, Fuc3S4S and Fuc4S with a molar ratio of 2:2.5:1) linked to O-3 of each d-GlcA. The fucan sulfate (PmFS) had a backbone consisting of a repetitively linked unit {-4-l-Fuc2S-α-1-}, and interestingly, every trisaccharide unit in its backbone was branched with a sulfated α-l-Fuc (Fuc4S or Fuc3S with a molar ratio of 4:1). Apart from the sulfated polysaccharides, two neutral glycans (PmNG-1 & -2) differing in molecular weight were also obtained and their structures were similar to animal glycogen. Anticoagulant assays indicated that PmFG and PmFS possessed strong APTT prolonging and intrinsic factor Xase inhibition activities, and the sulfated α-l-Fuc branches might contribute to the anticoagulant and anti-FXase activities of both PmFG and PmFS.


Assuntos
Anticoagulantes/química , Anticoagulantes/farmacologia , Polissacarídeos/química , Polissacarídeos/farmacologia , Pepinos-do-Mar/química , Animais , Anticoagulantes/isolamento & purificação , Coagulação Sanguínea/efeitos dos fármacos , Sequência de Carboidratos , Físico-Química , Cromatografia Líquida de Alta Pressão , Cisteína Endopeptidases , Humanos , Estrutura Molecular , Proteínas de Neoplasias/antagonistas & inibidores , Ressonância Magnética Nuclear Biomolecular , Polissacarídeos/isolamento & purificação , Relação Estrutura-Atividade , Sulfatos/química , Sulfatos/isolamento & purificação , Sulfatos/farmacologia
12.
Curr Pharm Biotechnol ; 20(6): 476-482, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30931857

RESUMO

BACKGROUND: Heparin is an essential drug used as an anticoagulant. Access to raw material suitable for heparin extraction is critical for creating a viable business opportunity. In Saudi Arabia, large amounts of raw material with potential for heparin extraction are wasted. OBJECTIVE: To extract heparin and low-molecular-weight heparin (LMWH) from the camel lung, and measure its potency and activity. METHODS: Heparin preparation included three steps: extraction, electrophoretic identification, and activity measurement. Fresh lung tissue (100 g) was minced and homogenized in a blender. Crude heparin extracts were prepared using Charles's or Volpi's method with slight modifications. Heparin was purified by electrophoresis using high-purity agarose gels in barium acetate buffer. The heparin activity of purified samples was assayed spectrophotometrically using commercial heparin kits. RESULTS: Charles's and Volpi's extraction methods were simple and easy to establish. The yield was 90 mg crude heparin per 100 g of camel lung tissue following Volpi's extraction protocol, whereas Charles's method did not yield any heparin. The separation of heparin and LMWH by gel electrophoresis resulted in sharp and clear product bands using material prepared according to Volpi's method. The heparin preparation had an anti-factor Xa activity of 37 IU/mg, indicating weak potency. CONCLUSION: Preparation of active heparin from camel lung tissue is a technology applicable in manufacturing. Further method development is needed to increase heparin purity and potency.


Assuntos
Anticoagulantes/isolamento & purificação , Camelus , Desenvolvimento de Medicamentos/métodos , Heparina de Baixo Peso Molecular/isolamento & purificação , Pulmão/química , Animais , Eletroforese em Gel de Ágar , Humanos
13.
Int J Biol Macromol ; 130: 664-674, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-30844450

RESUMO

The purpose of this study was to compare the application of several extraction methods, including cold extraction (CAE), hot extraction (HAE), ultrasonic-assisted extraction (UAE), and microwave-assisted extraction (MAE), all carried out in 0.1 M NaOH, under their respective best parameters, for obtaining products from leaf of Fragaria vesca L. The extracts with the highest anticoagulant activity were purified by multi-step extraction procedure and separated by gel permeation chromatography, giving rise to the macromolecular complexes. They were subsequently structurally characterized using colorimetric methods, FT-IR, GC-MS, and NMR (1H and 1H/13C HSQC), as polyphenolic-polysaccharide conjugates with the anticoagulant activity. The polysaccharide parts of the conjugates obtained by different extraction procedures were found to vary significantly. The most selective in their activity were the glycoconjugates extracted in UAE and MAE processes, i.e. arabinogalactan and pectin-like conjugates, respectively. In terms of their anticoagulant activity all of them were non-direct factor Xa inhibitors mediated by antithrombin.


Assuntos
Fragaria/química , Extratos Vegetais/química , Folhas de Planta/química , Polifenóis/química , Polifenóis/isolamento & purificação , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Anticoagulantes/farmacologia , Glicoconjugados/química , Glicoconjugados/isolamento & purificação , Glicoconjugados/farmacologia , Extração Líquido-Líquido/métodos , Micro-Ondas , Polifenóis/farmacologia , Polissacarídeos/farmacologia , Análise Espectral , Ondas Ultrassônicas
14.
Carbohydr Polym ; 214: 286-293, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30925999

RESUMO

In this paper, carrageenans having distinct sulfation patterns (κ-, ι-, ι/ν-, θ- and λ-carrageenans), were fully or partially oxidized at C-6 of the ß-d-Galp units using 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) and trichloroisocyanuric acid (TCCA) in bicarbonate buffer. The modified carrageenans were characterized by mono- and bidimensional 1H and 13C NMR spectroscopy. The influence of the sulfate and carboxyl groups onto anticoagulant activity was evaluated using Activated Partial Thromboplastin Time (aPTT) in vitro assay. The results showed a synergic effect of the carboxyl groups on the anticoagulant activity, which was dependent on the regiochemistry of the sulfate groups in the polysaccharide backbone. Sulfate groups at C2 of the ß-d-GalAp units appeared to positively influence the anticoagulant effect in comparison to C4-sulfate samples. Also, the partially oxidized κ-carrageenan derivative (κLO) showed better anticoagulant effect than the fully oxidized carrageenan (κHO).


Assuntos
Anticoagulantes/química , Carragenina/química , Ácidos Urônicos/química , Anticoagulantes/isolamento & purificação , Carragenina/isolamento & purificação , Estrutura Molecular , Oxirredução , Tempo de Tromboplastina Parcial , Rodófitas/química , Sulfatos/química , Ácidos Urônicos/síntese química
15.
Food Chem Toxicol ; 125: 614-620, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30738133

RESUMO

The objective of this study is to investigate the biological effects of phenolic compounds extracted from the sea buckthorn berries on oxidative stress and hemostasis. The sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) berries are rich in flavonoids and non-polar compounds. In this study, the activity of the phenolic fraction from the sea buckthorn berries was evaluated in vitro in comparison with three phenolic compounds: isorhamnetin (compound 1) and its two new derivatives: compound 2 (isorhamnetin 3-O-beta-glucoside-7-O-alfa-rhamnoside) and compound 3 (isorhamnetin 3-O-beta-glucoside-7-O-alfa-(3"'-isovaleryl)-rhamnoside). The impact of these phenolic compounds and the phenolic fraction against the effect of the donor of hydroxyl radicals - H2O2/Fe on proteins and lipids in human plasma was measured. Additionally, the aim of the study was to determine the effect of these phenolic compounds and the phenolic fraction on various typical hemostasis parameters. Our results show that the used derivatives of isorhamnetin possess different biological properties (e.g. antioxidant, anti-platelet and anticoagulant). The tested compounds can be seen as new natural beneficial compounds to be used in prevention and treatment of cardiovascular diseases.


Assuntos
Frutas/química , Hemostasia/efeitos dos fármacos , Hippophae/química , Estresse Oxidativo/efeitos dos fármacos , Quercetina/análogos & derivados , Anticoagulantes/isolamento & purificação , Anticoagulantes/farmacologia , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Humanos , Peróxido de Hidrogênio/efeitos adversos , Ferro/efeitos adversos , Peroxidação de Lipídeos/efeitos dos fármacos , Inibidores da Agregação de Plaquetas/isolamento & purificação , Inibidores da Agregação de Plaquetas/farmacologia , Carbonilação Proteica/efeitos dos fármacos , Quercetina/isolamento & purificação , Quercetina/farmacologia , Tempo de Trombina
16.
Int J Biol Macromol ; 121: 239-248, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30267823

RESUMO

World fisheries and aquaculture production totaled 167 million tons in 2014. This high fish production generates a lot of waste that could be used as raw material for extraction of substances of pharmacological interest. In this work, we extract and characterize glycosaminoglycans (GAGs) present in the viscera of Nile tilapia (Oreochromis niloticus) and Pacu (Piaractus mesopotamicus), which are among the most vastly produced fishes in inland aquaculture in Brazil. Moreover, the anticoagulant activity of the GAGs fractions was evaluated. GAGs were obtained from total defatted viscera, after proteolysis, precipitation with ethanol, anion exchange chromatography and treatment with chondroitinase. Chondroitin sulfate (CS), dermatan sulfate (DS) and heparan sulfate (HS) were identified by agarose gel electrophoresis and NMR analyses. CS, DS and HS were identified in equivalent fractions obtained from both fishes, and all GAGs fractions showed anticoagulant activity.


Assuntos
Anticoagulantes/isolamento & purificação , Anticoagulantes/farmacologia , Caraciformes/anatomia & histologia , Ciclídeos/anatomia & histologia , Glicosaminoglicanos/isolamento & purificação , Glicosaminoglicanos/farmacologia , Vísceras/química , Animais
17.
Int J Biol Macromol ; 123: 335-342, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30419328

RESUMO

A sulfated polysaccharide from Globularia alypum L. (GASP) was extracted with a yield of 14.2%. GASP is composed mostly of sulfate and total sugars (13.29% and 71.56%, respectively) with small amount of proteins and lipids. The chemical and structural characterization was studied by Infra-Red spectroscopic and gas chromatography-mass spectrometry (GC-MS). GASP composed of eight carbohydrates where galactose, glucose, and mannose are the major compounds (33.47%, 26.71% and 18.21%, respectively). The in vitro and in vivo anticoagulant activities in rats were tested using the standard coagulation assays activated partial thromboplastin time (aPTT), prothrombine time (TT) and thrombin time (PT) tests. Both doses of GASP (200 and 500 mg/kg b.w) displayed a significant in vitro (1.22 and 1.33-fold, 1.17 and 1.27-fold, and 1.21 and 1.26-fold, respectively) and in vivo (1.47 and 2.52-fold; 1.20 and 1.43-fold; 1.21 and 1.40-fold, respectively) compared with the control. Toxicity studies on liver performed by the catalytic activity of transaminases in plasma, oxidative stress markers and hepatic morphological changes indicated that GASP at both doses are not toxics. The important pharmacological and toxicological profile of GASP revealed that this compound may be used as a novel and effective drug.


Assuntos
Anticoagulantes/administração & dosagem , Coagulação Sanguínea/efeitos dos fármacos , Plantaginaceae/química , Polissacarídeos/administração & dosagem , Animais , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Testes de Coagulação Sanguínea , Cromatografia Gasosa , Humanos , Espectrometria de Massas , Tempo de Tromboplastina Parcial/métodos , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Ratos , Sulfatos/química , Tempo de Trombina/métodos
18.
Clin Chem Lab Med ; 57(5): 690-696, 2019 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-30427777

RESUMO

Background The aim of the study was to investigate the specificity of an activated charcoal-based product (DOAC Stop™) initially intended for the specific extraction of direct oral anticoagulants (DOACs) from test plasmas on a range of other anticoagulants. Methods Test plasmas were prepared by adding various anticoagulants to pooled normal plasma at concentrations prolonging an activated partial thromboplastin time (APTT) test by a factor of 1.5-3. These plasmas were treated with DOAC Stop™ for 5 and 20 min. Then APTTs were repeated and residual anticoagulant concentrations estimated from dose-response curves. Results The activated charcoal (AC)-based product was found to extract DOACs efficiently. It also bound the intravenous anticoagulants argatroban and lepirudin, but it had no effect on heparin, enoxaparin or danaparoid in plasma. Among other APTT-inhibiting agents that might be present in test plasmas from patients, it extracted protamine, aprotinin and polymyxin. It had no effect on annexin V, thrombomodulin, a typical lupus anticoagulant, a factor VIII antibody, activated protein C or its activator, but it did bind some cationic inhibitors of the APTT with molecular weight below approximately 30 kDa. Conclusions The AC-based product extracted DOACs efficiently with no effect on heparin-type anticoagulants. It did bind argatroban and hirudin-type anticoagulants, which might occur in plasmas from some inpatients, and APTT results obtained after its use should be interpreted after due consideration of patient medications.


Assuntos
Anticoagulantes/isolamento & purificação , Carvão Vegetal/química , Adsorção , Anticoagulantes/sangue , Anticoagulantes/química , Calibragem , Humanos , Tempo de Tromboplastina Parcial , Extração em Fase Sólida/métodos , Fatores de Tempo
19.
Int J Biol Macromol ; 124: 1230-1237, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30521914

RESUMO

Two novel polysaccharides were obtained from flowers of Apocynum venetum L., and named as Vp2a-II and Vp3. Their average molecular weights were 7 kDa and 9 kDa, respectively. Methods of monosaccharide analysis, acid hydrolysis and methylation analysis, Fourier transform-infrared spectroscopy (FT-IR), gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) spectroscopy were used to identify the structure of Vp2a-II and Vp3. Vp2a-II was composed of →6)-ß-d-Glcp-(1 → 6)-α-d-Galp-(1→ residues. Vp3 was composed of α-d-GlcpA-(3 → α-d-GalpA residues. The anticoagulant activity was evaluated by activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin time (PT), and fibrinogen (FIB) assays in vitro. Results indicated that Vp3 exhibited the anticoagulant activity.


Assuntos
Anticoagulantes/farmacologia , Apocynum/química , Coagulação Sanguínea/efeitos dos fármacos , Fibrinogênio/antagonistas & inibidores , Polissacarídeos/farmacologia , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Sequência de Carboidratos , Flores/química , Humanos , Hidrólise , Metilação , Estrutura Molecular , Peso Molecular , Tempo de Tromboplastina Parcial , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Tempo de Protrombina , Tempo de Trombina
20.
Int J Biol Macromol ; 125: 124-132, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30521917

RESUMO

The polyphenolic-polysaccharide complex was isolated from the dried aerial parts of the medicinal plant Agrimonia eupatoria L. using a multi-step process involving the degreasing of the plant material by extraction with organic solvents, followed by extraction with hot alkali, neutralization, further separation with organic solvents and dialysis. The complex was homogeneous with a molecular weight of about 55 × 103 g/mol and consisted mainly of carbohydrates and polyphenols matrix, composed of lignin-related units, with the dominance of dimethoxyphenyl structures. The carbohydrate moiety consists mostly of arabinogalactan associated with highly esterified rhamnogalacturonan. In vitro anticoagulant studies revealed the ability of the A. eupatoria complex to inhibit plasma clot formation, mainly in the intrinsic pathway of the blood coagulation cascade. Further studies on the mechanisms of this anticoagulant activity revealed that the isolate was primarily an indirect inhibitor of thrombin, mediated by antithrombin or by heparin cofactor II. Such mechanism of action is characteristic for highly sulfated glycosaminoglycans.


Assuntos
Agrimonia/química , Anticoagulantes/química , Anticoagulantes/farmacologia , Polifenóis/química , Polissacarídeos/química , Animais , Anticoagulantes/isolamento & purificação , Coagulação Sanguínea/efeitos dos fármacos , Humanos , Metilação , Peso Molecular , Compostos Fitoquímicos/química , Análise Espectral
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