Anticoagulant Activity Research Articles

An orthosteric/allosteric bivalent peptide agonist comprising covalently linked protease-activated receptor-derived peptides mimics in vitro and in vivo activities of activated protein C

BackgroundActivated protein C (APC) has anticoagulant and cytoprotective cell-signaling activities, which often require protease-activated receptor (PAR) 1 and PAR3 and PAR cleavages at noncanonical sites (R46-N47 and R41-G42, respectively). Some PAR1-derived (P1) peptides and PAR3-derived (P3) peptides, eg, P1-47-66 and P3-42-65, mimic APC’s cell signaling. In anti-inflammatory assays, these 2 peptides at low concentrations synergistically attenuate cellular inflammation. ObjectivesTo determine whether a P1 peptide covalently linked to a P3 peptide mimics APC’s anti-inflammatory and endothelial barrier stabilization activities. MethodsAnti-inflammatory assays employed stimulated THP-1 cells and caspase-1 measurements. Cultured human EA.hy926 or murine aortic endothelial cells (ECs) exposed to thrombin were monitored for transendothelial electrical resistance. Bivalent covalently linked P1:P3 peptides were studied for APC-like activities. ResultsIn anti-inflammatory assays, P1-47-55 was as active as P1-47-66 and some P3 peptides (eg, P3-44-54 and P3-51-65) were as active as P3-42-65. The bivalent P1:P3 peptide comprising P1-47-55-(Gly[10 residues])-P3-51-65 (designated “G10 peptide”) was more potently anti-inflammatory than the P1 or P3 peptide alone. In transendothelial electrical resistance studies of thrombin-challenged ECs, P1-47-55 and the G10 peptide mimicked APC’s protective actions. In dose-response studies, the G10 peptide was more potent than the P1-47-55 peptide. In murine EC studies, the murine PAR-sequence–derived G10 peptide mimicked murine APC’s activity. Anti-PAR1 and anti-PAR3 antibodies, but not anti–endothelial protein C receptor antibodies, abated G10’s cytoprotection, showing that G10’s actions involve PAR1:PAR3. G10 significantly increased survival in murine endotoxemia. ConclusionThe PAR-sequence–derived G10 peptide is a bivalent agonist that mimics APC’s cytoprotective, anti-inflammatory, and endothelial barrier–stabilizing actions and APC’s protection against endotoxemic mortality.

Systemic inflammation and delirium during critical illness.

The purpose of this study was to determine associations between markers of inflammation and endogenous anticoagulant activity with delirium and coma during critical illness. In this prospective cohort study, we enrolled adults with respiratory failure and/or shock treated in medical or surgical intensive care units (ICUs) at 5 centers. Twice per day in the ICU, and daily thereafter, we assessed mental status using the Richmond Agitation Sedation Scale (RASS) and the Confusion Assessment Method-Intensive Care Unit (CAM-ICU). We collected blood samples on study days 1, 3, and 5, measuring levels of C-reactive protein (CRP), interferon gamma (IFN-γ), interleukin (IL)-1 beta (IL-1β), IL-6, IL-8, IL-10, IL-12, matrix metalloproteinase-9 (MMP-9), tumor necrosis factor-alpha (TNF-α), tumor necrosis factor receptor 1 (TNFR1), and protein C using validated protocols. We used multinomial logistic regression to analyze associations between biomarkers and the odds of delirium or coma versus normal mental status the following day, adjusting for age, sepsis, Sequential Organ Failure Assessment (SOFA), study day, corticosteroids, and sedatives. Among 991 participants with a median age (interquartile range, IQR) of 62 [53-72] years and enrollment SOFA of 9 [7-11], higher concentrations of IL-6 (odds ratio [OR] [95% CI]: 1.8 [1.4-2.3]), IL-8 (1.3 [1.1-1.5]), IL-10 (1.5 [1.2-1.8]), TNF-α (1.2 [1.0-1.4]), and TNFR1 (1.3 [1.1-1.6]) and lower concentrations of protein C (0.7 [0.6-0.8])) were associated with delirium the following day. Higher concentrations of CRP (1.4 [1.1-1.7]), IFN-γ (1.3 [1.1-1.5]), IL-6 (2.3 [1.8-3.0]), IL-8 (1.8 [1.4-2.3]), and IL-10 (1.5 [1.2-2.0]) and lower concentrations of protein C (0.6 [0.5-0.8]) were associated with coma the following day. IL-1β, IL-12, and MMP-9 were not associated with mental status. Markers of inflammation and possibly endogenous anticoagulant activity are associated with delirium and coma during critical illness.

Chemical Synthesis of Fucosylated Chondroitin Sulfate Tetrasaccharide with Fucosyl Branch at the 6-OH of GalNAc Residue.

Fucosylated chondroitin sulfate is a unique glycosaminoglycan isolated from sea cucumbers, with excellent anticoagulant activity. The fucosyl branch in FCS is generally located at the 3-OH of D-glucuronic acid but, recently, a novel structure with α-L-fucose linked to the 6-OH of N-acetyl-galactosamine has been found. Here, using functionalized monosaccharide building blocks, we prepared novel FCS tetrasaccharides with fucosyl branches both at the 6-OH of GalNAc and 3-OH of GlcA. In the synthesis, the protective group strategy of selective O-sulfation, as well as stereoselective glycosylation, was established, which enabled the efficient synthesis of the specific tetrasaccharide compounds. This research enriches knowledge on the structural types of FCS oligosaccharides and facilitates the exploration of the structure-activity relationship in the future.

Protein disulfide isomerase cleaves allosteric disulfides in histidine-rich glycoprotein to regulate thrombosis

The essence of difference between hemostasis and thrombosis is that the clotting reaction is a highly fine-tuned process. Vascular protein disulfide isomerase (PDI) represents a critical mechanism regulating the functions of hemostatic proteins. Herein we show that histidine-rich glycoprotein (HRG) is a substrate of PDI. Reduction of HRG by PDI enhances the procoagulant and anticoagulant activities of HRG by neutralization of endothelial heparan sulfate (HS) and inhibition of factor XII (FXIIa) activity, respectively. Murine HRG deficiency (Hrg−/−) leads to delayed onset but enhanced formation of thrombus compared to WT. However, in the combined FXII deficiency (F12−/−) and HRG deficiency (by siRNA or Hrg−/−), there is further thrombosis reduction compared to F12−/− alone, confirming HRG’s procoagulant activity independent of FXIIa. Mutation of target disulfides of PDI leads to a gain-of-function mutant of HRG that promotes its activities during coagulation. Thus, PDI-HRG pathway fine-tunes thrombosis by promoting its rapid initiation via neutralization of HS and preventing excessive propagation via inhibition of FXIIa.

Semisynthesis of new sulfated heterorhamnan derivatives obtained from green seaweed Gayralia brasiliensis and evaluation of their anticoagulant activity

Marine green algae produce sulfated polysaccharides with diverse structures and a wide range of biological activities. This study aimed to enhance the biotechnological potential of sulfated heterorhamnan (Gb1) from Gayralia brasiliensis by chemically modifying it for improved or new biological functions. Using controlled Smith Degradation (GBS) and O-alkylation with 3-chloropropylamine, we synthesized partially water-soluble amine derivatives. GBS modification increase sulfate groups (29.3 to 37.5 %) and α-l-rhamnose units (69.9 to 81.2 mol%), reducing xylose and glucose, compared to Gb1. The backbone featured predominantly 3- and 2-linked α-l-rhamnosyl and 2,3- linked α-l-rhamnosyl units as branching points. Infrared and NMR analyses confirmed the substitution of hydroxyl groups with aminoalkyl groups. The modified compounds, GBS-AHCs and GBS-AHK, exhibited altered anticoagulant properties. GBS-AHCs showed reduced effectiveness in the APTT assay, while GBS-AHK maintained a similar anticoagulant activity level to Gb1 and GBS. Increased nitrogen content and N-alkylation in GBS-AHCs compared to GBS-AHK may explain their structural differences. The chemical modification proposed did not enhance its anticoagulant activity, possibly due to the introduction of amino groups and a positive charge to the polymer. This characteristic presents new opportunities for investigating the potential of these polysaccharides in various biological applications, such as antimicrobial and antitumoral activities.

Biosynthesis, characterization and antivenom activities of <i>Moringa oleifera</i> silver nanoparticles: an experimental approach

Moringa oleifera has been previously established to possess neutralizing potentials against Echis ocellatus venom. This study however, investigated the bioefficacy of silver nanoparticles biosynthesized from M. oleifera leaf extract aimed at improving its bioactivity against E. ocellatus venom-induced toxicities using in vivo and in vitro methods. The intrinsic characteristics of the produced M. oleifera-Silver nanoparticles (MO-AgNPs) were carried out using energy dispersive X-ray, X-ray diffraction, scanning electron microscopy, transmission electron microscopy and Fourier-transform infrared spectroscopy. Twenty-five male Wistar rats divided randomly into five groups (n=5) were used for the antivenom study. Group 1 received saline while groups 2 to 5 were envenomed intraperitoneally with 0.22mg/kg (LD50) of E. ocellatus venom. Group 2 was left untreated while groups 3 to 5 were treated with 0.2ml of antivenom, 5 and 10mg/kg MO-AgNPs post-envenomation, respectively. Blood and tissue of treated rats were analyzed for heamatological parameters and histopathology, respectively. The MO-AgNPs formation was confirmed with a colour change from light brown to yellowish-brown with maximum SPR band at 420nm from UV-Vis analysis, indicating a reflection of the bio-reduction of Ag+ to Ag0. The Transmission electron micrographs showed well dispersed spherical AgNPs with average particle size of 15.7nm. Treatment with MO-AgNPs caused a significant improvement of acute anemia, leucopenia and thrombocytopenia induced by the venom in the envenomed treated rats. Also, MO-AgNPs inhibited the haemorrhagic, haemolytic and anticoagulant activities of the venom. Tissue lesions observed in heart of envenomed untreated rats were attenuated after treatment with MO-AgNPs. The biosynthesized MO-AgNPs exhibited potent neutralizing potentials than M. oleifera crude extract against E. ocellatus venom toxicities.

Biochemical and biological characterization of the venoms of Naja kaouthia and Naja mandalayensis from Myanmar and neutralization effects of BPI cobra antivenom

Snakebite is a neglected public health issue, with many scientific and medical issues to be solved. Cobras are among the most common venomous snakes in Myanmar and are responsible for a considerable number of severe snakebite envenoming. There are three species of cobra (Naja kaouthia, Naja mandalayensis and Ophiophagus hannah) in Myanmar. The study aims to characterize the N. kaouthia and N. mandalayensis venoms and to investigate the efficacy of anti-cobra antivenom (BPI) against the two venoms. Protein components and fibrinogenolytic activity were determined by SDS-PAGE. Enzymatic activities for PLA2, protease and acetylcholinesterase were determined by spectrophotometric method. Anticoagulant activity was determined by recalcification time of citrated human plasma. Myotoxicity, necrotizing activity, median lethal dose (LD50) and median effective dose (ED50) were determined by WHO recommended methods. The SDS-PAGE displayed the proteins and enzymes containing in two venoms were different. N. kaouthia venom exhibited more in PLA2, acetylcholinesterase, anticoagulant, fibrinogenolytic and necrotizing activities than N. mandalayensis venom. N. mandalayensis venom had more protease activity and myotoxicity than N. kaouthia venom. The median lethal dose (LD50) of N. kaouthia and N. mandalayensis venom was 4.33 μg/mouse and 5.04 μg/mouse respectively. Both venoms induced fibrinogen Aα chain degradation in 30 min (N. kaouthia) and in 6 h (N. mandalayensis). The same median effective dose (ED50) (19.56 μg/mouse) showed that anti-NK antivenom can neutralize against lethal effect of N. mandalayensis venom. It can also neutralize the protease activity, anticoagulant activity and fibrinogenolytic activity of both venoms. Immunodiffusion and immunoblotting studies showed that the antivenom recognized its homologous venom (N. kaouthia) and cross-reacted against the heterologous venom (N. mandalayensis). The anti-NK antivenom is suitable to use for N. mandalayensis bite if monospecific antivenom is not available.

Anticoagulant properties and therapeutic potentials of wasp venom

AbstractWasp venom is rich in bioactive substances, such as proteins, peptides, and small molecules. The venom significantly affects the mammalian cardiovascular, nervous, and immune systems, causing mild to severe symptoms following stings. It exhibits both procoagulant and anticoagulant activities, and significant research has identified its ability to modulate the mammalian coagulation system. Active substances that inhibit clotting have been identified and purified through patient case reports and experimental studies. This study reviewed the findings on how wasp venom interacts with platelets and coagulation factors, such as fibrinogen and prothrombin, and demonstrated its dual influence on the coagulation cascade. This highlights the potential of the venom in therapeutic applications, especially as an anticoagulant, as evidenced by the inhibition of coagulation factors and prolonged clotting times after envenomation, suggesting its utility in developing novel anticoagulant therapies. This review focuses on the anticoagulant effects of social wasp venom, which is prevalent in sting incidents, summarizing the research and observations on its therapeutic potential. This emphasizes the significance of further studies to identify and utilize venom components as innovative anticoagulant treatments.

Polymers as candidates for broad‐spectrum antivirals—in vitro inhibition of Zika virus with sodium polyanethole sulfonate

AbstractZika virus (ZIKV), belonging to the family of flaviviruses, is an emerging mosquito‐borne pathogen causing microcephaly and other congenital abnormalities in newborns as well as various neurologic pathologies in adults. The climate change and the lifestyle of current societies, involving global mobility, result in a rapid broadening of ZIKV geographic distribution. There are no vaccines or drugs available to protect against or cure the diseases caused by that virus. Our in vitro studies have demonstrated that sodium polyanethole sulfonate (PASNa), a degradable polymer used in various biomedical applications, acts as an efficient ZIKV inhibitor, both in animal and human cells. The mechanistic studies indicated that PASNa bound to the ZIKV particles thus hindering their attachment to the host cells and also interfered in the late steps of viral replication. PASNa is characterized by very low cytotoxicity and its antiviral activity can be achieved at a concentration much lower than that at which its anticoagulant activity has to be considered. Isothermal titration calorimetry (ITC) studies have shown that the interaction of PASNa with human serum albumin (HSA), the most abundant blood protein, is the endothermic, entropy‐driven process resulting in the formation of stoichiometric (1:1) PASNa‐HSA complexes with a hydrodynamic diameter of 8.8 ± 0.8 nm and zeta potential (ζ) equal to −21.3 mV. The analysis of CD spectra indicated that PASNa did not significantly affect the HSA secondary structure on binding to that protein.

A Deep Overview of Anticoagulant Drugs: Recent Synthesis and Their Activity Assay

During the unprecedented COVID-19 pandemic, anticoagulant drugs have emerged as a crucial component of treatment alongside antivirus medications. Patients with severe COVID-19 frequently have critical conditions marked by blood clot development, necessitating the administration of anticoagulants. This review aims to provide a comprehensive overview of various anticoagulant drugs, their synthesis methods, and assays employed to predict their anticoagulant activity. Notable anticoagulant categories frequently utilized include oral anticoagulants heparin, non-vitamin K antagonists, and vitamin K antagonists. In recent years, the development of new anticoagulants has seen a shift towards a multifaceted approach that combines in silico prediction with in vitro and in vivo assays. In silico prediction techniques play a pivotal role in the initial screening process. This integrated approach has yielded promising results, paving the way for the synthesis of novel anticoagulant candidates, as substantiated by a battery of in vitro, in vivo, and ex-vivo tests.

Interference of oral anti-Xa anticoagulants during monitoring of unfractionated heparin treatments: practical and future attitudes

Contrary to direct oral anticoagulants (DOAC), unfractionated heparin (UFH) requires daily monitoring when administered at therapeutic dose. At present, UFH monitoring is preferably carried out by measuring plasma anti-Xa activity, however, in patients previously treated with an anti-Xa DOAC and switched to UFH, there is a high risk of DOAC interfering with the measurement of UFH anti-Xa activity. Residual anti-Xa DOAC in the sample can lead to an overestimation of the anticoagulant activity attributed to heparin and thus to incorrect anticoagulation. This risk of interference should not be overlooked because interference may occur even at concentration of DOAC below the hemostatic safety threshold and can last several days. To overcome this issue, several alternatives are being studied. This note provides an update on anti-Xa DOAC interference and different strategies available in current practice. It also underlines the importance of communication between biologists and clinicians on anticoagulant treatments received by patients.

Design and Development of Fast-dissolving Tablets of Apixaban using Single Coprocessed Excipient

Background: Apixaban is administered orally as an anticoagulant action and minimizes the chances of strokes and systemic embolism. The conventional film-coated tablet showed less bioavailability due to its minimal solubility in the gastrointestinal tract. Rationale: The goal of designing the current analysis is to prepare a prompt release tablet using a single Coprocessed excipient which minimizes multistep processing, time, and cost effectiveness. Methods: The prompt release tablets of Apixaban were developed by direct compression technique using multifunctional material like Prosolve ODT G2 and Prosolve Easytab. Drug excipients inertness was confirmed with FTIR and DSC. Flowing characteristics of Apixaban and co-processed excipients were evaluated in terms of Carr’s index, Angle of repose, and Hausner’s ratio. These co-processed excipients indicated exceptional flowing and compression characteristics. The ready tablets were assessed for weight variation, hardness, friability, dispersion time, disintegration, and dissolution studies. Results: Among 12 batches, the least disintegration time was shown with C6 with a cumulative drug release of 99.06 % within 25 min. The optimized batch C6 was highly stable when tested under stability guidelines at 40 0 C and at 75 % relative humidity. Conclusion: Among these co-processed excipients, Prosolve Easytab SP was found the best one for direct compression intended for the immediate or fast dissolving tablets.

Ex Vivo Antiplatelet Effects of Oral Anticoagulants.

The impact of non-vitamin K antagonist oral anticoagulants (NOACs) on platelet function is still unclear. We conducted a comprehensive ex vivo study aimed at assessing the effect of the four currently marketed NOACs on platelet function. We incubated blood samples from healthy donors with concentrations of NOACs (50, 150 and 250 ng/mL), in the range of those achieved in the plasma of patients during therapy. We evaluated generation of thrombin; light transmittance platelet aggregation (LTA) in response to adenosine diphosphate (ADP), thrombin receptor-activating peptide (TRAP), human γ-thrombin (THR) and tissue factor (TF); generation of thromboxane (TX)B2; and expression of protease-activated receptor (PAR)-1 and P-selectin on the platelet surface. All NOACs concentration-dependently reduced thrombin generation compared with control. THR-induced LTA was suppressed by the addition of dabigatran at any concentration, while TF-induced LTA was reduced by factor-Xa inhibitors. ADP- and TRAP-induced LTA was not modified by NOACs. TXB2 generation was reduced by all NOACs, particularly at the highest concentrations. We found a concentration-dependent increase in PAR-1 expression after incubation with dabigatran, mainly at the highest concentrations, but not with FXa inhibitors; P-selectin expression was not changed by any drugs. Treatment with the NOACs is associated with measurable ex vivo changes in platelet function, arguing for antiplatelet effects beyond the well-known anticoagulant activities of these drugs. There are differences, however, among the NOACs, especially between dabigatran and the FXa inhibitors.

Antiplatelet and Anticoagulant Effects of Two New Phenylpropanoid Sucrose Esters and Other Secondary Metabolites from the Aerial Part of Canna edulis.

This study isolated pure compounds from Canna edulis aerial parts and assessed their antiplatelet and anticoagulant potential. Structural elucidation resulted in the identification of two new compounds: caneduloside A (1) and caneduloside B (2), and eleven known compounds: 6'-acetyl-3,6,2'-tri-p-coumaroyl sucrose (3), 6'-acetyl-3,6,2'-triferuloyl sucrose (4), tiliroside (5), afzelin (6), quercitrin (7), 2-hydroxycinnamaldehyde (8), cinnamic acid (9), 3,4-dimethoxycinnamic acid (10), dehydrovomifoliol (11), 4-hydroxy-3,5-dimethoxybenzaldehyde (12), and (S)-(-)-rosmarinic acid (13). Compounds 3, 4, 6-9, 13 were previously reported for antithrombotic properties. Hence, antithrombotic tests were conducted for 1, 2, 5, 10-12. All tested compounds demonstrated a dose-dependent antiaggregatory effect, and 10 and 12 were the most potent for both ADP and collagen activators. Additionally, 10 and 12 showed anticoagulant effects, with prolonged prothrombin time and activated partial thromboplastin time. The new compound 1 displayed antiplatelet and anticoagulant activity, while 2 mildly inhibited platelet aggregation. C. edulis is a potential source for developing antithrombotic agents.

Green fabrication of ZnO nanoparticles via spirulina platensis and its efficiency against biofilm forming pathogens

Excessive consumption of antibiotics is considered one of the top public health threats, this necessitates the development of new compounds that can hamper the spread of infections. A facile green technology for the biosynthesis of Zinc oxide nanoparticles (ZnO NPs) using the methanol extract of Spirulina platensis as a reducing and stabilizing agent has been developed. A bunch of spectroscopic and microscopic investigations confirmed the biogenic generation of nano-scaled ZnO with a mean size of 19.103 ± 5.66 nm. The prepared ZnO NPs were scrutinized for their antibacterial and antibiofilm potentiality, the inhibition zone diameters ranged from 12.57 ± 0.006 mm to 17.33 ± 0.006 mm (at 20 µg/mL) for a variety of Gram-positive and Gram-negative pathogens, also significant eradication of the biofilms formed by Staphylococcus aureus and Klebsiella pneumoniae by 96.7% and 94.8% respectively was detected. The free radical scavenging test showed a promising antioxidant capacity of the biogenic ZnO NPs (IC50=78.35 µg/mL). Furthermore, the anti-inflammatory role detected using the HRBCs-MSM technique revealed an efficient stabilization of red blood cells in a concentration-dependent manner. In addition, the biogenic ZnO NPs have significant anticoagulant and antitumor activities as well as minimal cytotoxicity against Vero cells. Thus, this study offered green ZnO NPs that can act as a secure substitute for synthetic antimicrobials and could be applied in numerous biomedical applications.

CHEMICAL COMPOSITION, AMINO ACIDS PROFILE AND ANTIOXIDANT ACTIVITY OF ENZYMATIC PROTEIN HYDROLYSATE FROM Bohadschia marmorata SEA CUCUMBER

The sea cucumber is one of the potential sub-exports of fisheries. Sea cucumbers contain highly nutritious and bioactive compounds with antioxidant, antibacterial, anticoagulant, anti-inflammatory, and antiviral activities. Protein hydrolysates from sea cucumbers have the potential as antioxidants with the ability to reduce free radicals. The objective of this research is to examine the chemical characteristics and antioxidant activity of protein hydrolysates from Bohadschia marmorata sea cucumber. The research method used in this study was experimental and laboratory analysis. The results showed that the chemical composition of fresh Bohadschia marmorata sea cucumber had high moisture content (80.45 %), ash content (4.12 %), low fat content (6.17 %), and low protein content (8.96 %). This could be due to the environmental conditions and nutrient absorption by Bohadschia marmorata sea cucumbers. Fifteen types of amino acids with a total of 53.81 % were obtained from this study. The yield obtained for crude papain enzyme concentration was 16.26 %, and for pure papain enzyme concentration, it was 11.23 %. The antioxidant activity with IC50 > 200 indicates very weak antioxidant activity, and the resulting molecular weight reached 73.93 kDa

Evaluation of Antiplatelet Aggregation and Anticoagulant Effects of Oxalis Debilis Kunth Rhizome Extracts

Cardiovascular disease is one of the leading causes of death globally. The discovery and use of natural products in the prevention and treatment of cardiovascular diseases have achieved great interest. Oxalis debilis is traditionally used for the treatment of various ailments. This study aimed to investigate the potential antiplatelet and anticoagulant properties of extracts derived from O. debilis rhizome. The rhizome of O. debilis was extracted with methanol and then fractionated to obtain four extracts: the total methanol extract, the n-hexane, ethyl acetate, and water fraction. These four extracts were then tested for their ability to inhibit platelet aggregation and blood coagulation in human blood samples. This report demonstrated for the first time that O. debilis had inhibitory effects on platelet aggregation induced by both adenosine diphosphate and collagen as well as blood coagulation. Moreover, the total methanol extract, the ethyl acetate, and the water fraction had strong antiplatelet aggregation effects. All tested extracts had mild anticoagulant activities. This plant could be a valuable natural source for searching for antithrombotic substances, or for the development of supplementary products for the treatment and prevention of heart diseases and thrombosis.

Purification of an Acidic Polysaccharide with Anticoagulant Activity from the Marine Sponge Sarcotragus spinosulus.

Thromboembolic conditions are the most common cause of death in developed countries. Anticoagulant therapy is the treatment of choice, and heparinoids and warfarin are the most adopted drugs. Sulphated polysaccharides extracted from marine organisms have been demonstrated to be effective alternatives, blocking thrombus formation by inhibiting some factors involved in the coagulation cascade. In this study, four acidic glycan fractions from the marine sponge Sarcotragus spinosulus were purified by anion-exchange chromatography, and their anticoagulant properties were investigated through APTT and PT assays and compared with both standard glycosaminoglycans and holothurian sulphated polysaccharides. Moreover, their topographic localization was assessed through histological analysis, and their cytocompatibility was tested on a human fibroblast cell line. A positive correlation between the amount of acid glycans and the inhibitory effect towards both the intrinsic and extrinsic coagulation pathways was observed. The most effective anticoagulant activity was shown by a highly charged fraction, which accounted for almost half (about 40%) of the total hexuronate-containing polysaccharides. Its preliminary structural characterization, performed through infrared spectroscopy and nuclear magnetic resonance, suggested that it may consist of a fucosylated chondroitin sulphate, whose unique structure may be responsible for the anticoagulant activity reported herein for the first time.

Anticoagulant Activity Test of Ethanol Extract Coleus Scutellarioides ( L. ) Benth

Coleus scutellarioides (L.) Benth has been used as a traditional medicine as an antibacterial, anthelmintic, antidiabetic, and appetite enhancer and can treat various types of diseases such as coughs, hemorrhoids, boils, fever, diarrhea, and improves the menstrual cycle. C. scutellarioides plants grow in areas that have moist soil, such as rice fields, wild plants, or medicinal plants. This plant contains chemical compounds, including essential oils, flavonoids, phytosterols, alkaloids, and tannins. This study aims to determine the anticoagulant activity of the ethanol extract of C. scutellarioides. The method used in this research starts with sampling C. scutellarioides, making simplicia, extraction by maceration with ethanol solvent, and testing anticoagulant activity with two methods, namely the modified Lee-White method and the blood smear method. The modified Lee-White method was visually observed and tested on each type of human blood group, namely blood groups A, B, AB, and O, while the blood smear method was observed microscopically. The results of this research concluded that using the modified Lee-White method, C. scutellarioides extract with an extract concentration of 40 ppm had activity as an anticoagulant against various types of blood groups. In the directional smear method, preparations from blood samples were given. C. scutellarioides microscopically has anticoagulant activity because the blood cells were not related to each other, were still intact, and were separated from one another.

An up-to-date review of biomedical applications of serratiopeptidase and its biobetter derivatives as a multi-potential metalloprotease.

Serratiopeptidase is a bacterial metalloprotease used in a variety of medical applications. The multidimensional properties of serratiopeptidase make it noticeable as a miraculous enzyme. Anti-coagulant, anti-inflammatory and anti-biofilm activity of serratiopeptidase making it useful in reducing pain and swelling associated with various conditions including arthritis, diabetes, cancer, swelling, pain and also thrombolytic disorders. It breaks down fibrin, thins the fluids formed during inflammation and due to its anti-biofilm activity, can be used in the combination of antibiotics to reduce development of antibiotic resistance. However, some drawbacks like sensitivity to environmental conditions and low penetration into cells due to its large size have limited its usage as a potent pharmaceutical agent. To overcome such limitations, improved versions of the enzyme were introduced using protein engineering in our previous studies. Novel functional serratiopeptidases with shorter length and higher stability have seemingly created a hope for using this enzyme as a more effective therapeutic enzyme. This review explains the structural properties and functional aspects of serratiopeptidase, its main characteristics and properties, pre-clinical and clinical applications of the enzyme, improved qualities of the modified forms, different formulations of the enzyme, and the potential future developments.