Exploration of in vitro thrombolytic, anthelminthic, cytotoxic and in vivo anxiolytic potentials with phytochemical screening of flowers of Brassica nigra

Abstract Background Brassica nigra is a plant of Brassicaceae family, which possesses numerous medicinal values. Our present study is intended to assess the potential in vitro thrombolytic, anthelminthic, cytotoxic and in vivo anxiolytic properties of MCE of B. nigra flowers. MCE was fractioned for separating the compound on the basis of polarity by using chloroform, n-hexane and ethyl acetate solvent. Thrombolytic and anthelminthic activities were explained by collecting human erythrocytes and earthworms as test models, respectively. Anxiolytic activity was evaluated by elevated plus maze and hole board models while cytotoxic test was conducted through brine shrimp lethality bioassay. Results MCE revealed the presence of alkaloids, flavonoids, tannin, diterpenes, glycosides, carbohydrates, phenols, fixed oils and fat. In case of thrombolytic test, the MCE, CSF, ASF and n-HSF had produced maximum clot lysis activity at 5 and 10 mg/ml dose conditions. Two different concentrations (10 and 20 mg/ml) of MCE and its fractions showed significant (p < 0.05) anthelminthic activities in a dose-dependent manner. Significant anxiolytic activity was observed for all fractions which was comparable to the standard drug diazepam (p < 0.05). Again, the cytotoxic screening also presented good potentials for all fractions. Conclusion From the findings of present study, we can conclude that MCE of B. nigra flowers and its fraction possess significant anxiolytic, anthelmintic, anticancer and thrombolytic properties which may be a good candidate for treating these diseases through the determination of bio-active lead compounds.

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Therapeutic Potentials of Syzygium fruticosum Fruit (Seed) Reflected into an Array of Pharmacological Assays and Prospective Receptors-Mediated Pathways

Life ◽

10.3390/life11020155 ◽

2021 ◽

Vol 11 (2) ◽

pp. 155

Author(s):

Jannatul Nasma Rupa Moni ◽

Md. Adnan ◽

Abu Montakim Tareq ◽

Md. Imtiazul Kabir ◽

A.S.M. Ali Reza ◽

...

Keyword(s):

Bioactive Compounds ◽

Integrated Approach ◽

Gas Chromatography Mass Spectrometry ◽

Clot Lysis ◽

Lipinski’S Rule ◽

Lysis Activity ◽

In Silico Studies ◽

Immobility Time

Syzygium fruticosum (SF), a valuable Bangladeshi fruit, is considered an alternative therapeutic agent. Mainly, seeds are used as nutritional phytotherapy to ease physical and mental status by preventing chronic diseases. Here, we scrutinized the S. fruticosum seed’s fundamental importance in traditional medicine by following an integrated approach combining in vivo, in vitro, and in silico studies. The SF was fractionated with different solvents, and the ethyl acetate fraction of SF (EaF-SF) was further studied. Mice treated with EaF-SF (200 and 400 mg/kg) manifested anxiolysis evidenced by higher exploration in elevated plus maze and hole board tests. Similarly, a dose-dependent drop of immobility time in a forced swimming test ensured significant anti-depressant activity. Moreover, higher dose treatment exposed reduced exploratory behaviour resembling decreased movement and prolonged sleeping latency with a quick onset of sleep during the open field and thiopental-induced sleeping tests, respectively. In parallel, EaF-SF significantly (p < 0.001) and dose-dependently suppressed acetic acid and formalin-induced pain in mice. Also, a noteworthy anti-inflammatory activity and a substantial (p < 0.01) clot lysis activity (thrombolytic) was observed. Gas chromatography-mass spectrometry (GC–MS) analysis resulted in 49 bioactive compounds. Among them, 12 bioactive compounds with Lipinski’s rule and safety confirmation showed strong binding affinity (molecular docking) against the receptors of each model used. To conclude, the S. fruticosum seed is a prospective source of health-promoting effects that can be an excellent candidate for preventing degenerative diseases.

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Inhibition of Tumor Cell Proliferation In Vitro by Benzamide Derivatives

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.997.225 ◽

2014 ◽

Vol 997 ◽

pp. 225-228 ◽

Cited By ~ 1

Author(s):

Yan Ling Wu ◽

Li Wen Shen ◽

Yan Ping Ding ◽

Yoshimasa Tanaka ◽

Wen Zhang

Keyword(s):

Tumor Cell ◽

Cell Lines ◽

Malignant Tumors ◽

Dependent Manner ◽

Tumor Cell Lines ◽

Lead Compounds ◽

Proliferative Effect ◽

Benzamide Derivatives

Benzamide derivatives have been shown to have antitumor activity in various tumor cell lines in vitro as well as in vivo. In this study, we examined the anti-proliferative effect of four benzamide derivativeson Hela, H7402, and SK-RC-42 tumor cell lines in vitro by means of Real-Time cell assay (RTCA), and found that four benzamide derivatives suppressed proliferation of tumor cells in a time-and dose-dependent manner. The anti-proliferative activity of benzamide derivatives demonstrated that theycould be promising lead compounds for developing therapeutic agents for malignant tumors.

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Antiangiogenic kringles derived from human plasminogen and apolipoprotein(a) inhibit fibrinolysis through a mechanism that requires a functional lysine-binding site

Biological Chemistry ◽

10.1515/bc.2011.023 ◽

2011 ◽

Vol 392 (4) ◽

Cited By ~ 1

Author(s):

Jin-Hyung Ahn ◽

Ho-Jeong Lee ◽

Eun-Kyoung Lee ◽

Hyun-Kyung Yu ◽

Tae-Ho Lee ◽

...

Keyword(s):

Binding Site ◽

Binding Sites ◽

Safety Concern ◽

Tissue Type ◽

Clot Lysis ◽

Dependent Manner ◽

Kringle Domains ◽

Apolipoprotein A

AbstractMany proteins in the fibrinolysis pathway contain antiangiogenic kringle domains. Owing to the high degree of homology between kringle domains, there has been a safety concern that antiangiogenic kringles could interact with common kringle proteins during fibrinolysis leading to adverse effectsin vivo. To address this issue, we investigated the effects of several antiangiogenic kringle proteins including angiostatin, apolipoprotein(a) kringles IV9-IV10-V (LK68), apolipoprotein(a) kringle V (rhLK8) and a derivative of rhLK8 mutated to produce a functional lysine-binding site (Lys-rhLK8) on the entire fibrinolytic processin vitroand analyzed the role of lysine binding. Angiostatin, LK68 and Lys-rhLK8 increased clot lysis time in a dose-dependent manner, inhibited tissue-type plasminogen activator-mediated plasminogen activation on a thrombin-modified fibrinogen (TMF) surface, showed binding to TMF and significantly decreased the amount of plasminogen bound to TMF. The inhibition of fibrinolysis by these proteins appears to be dependent on their functional lysine-binding sites. However, rhLK8 had no effect on these processes owing to an inability to bind lysine. Collectively, these results indicate that antiangiogenic kringles without lysine binding sites might be safer with respect to physiological fibrinolysis than lysine-binding antiangiogenic kringles. However, the clinical signi-ficance of these findings will require further validationin vivo.

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A long-lasting, plasmin-activatable thrombin inhibitor aids clot lysis in vitro and does not promote bleeding in vivo

Thrombosis and Haemostasis ◽

10.1160/th08-08-0535 ◽

2009 ◽

Vol 101 (05) ◽

pp. 867-877 ◽

Cited By ~ 15

Author(s):

Louise Eltringham-Smith ◽

Sharon Gataiance ◽

Varsha Bhakta ◽

William Sheffield

Keyword(s):

Chimeric Protein ◽

Fold Increase ◽

Thrombin Inhibitor ◽

Clot Lysis ◽

Dependent Manner ◽

Recombinant Hirudin ◽

Concentration Dependent Manner ◽

Shed Blood

SummaryThe leech protein hirudin is a potent inhibitor of thrombin, but clinical use of recombinant hirudin is restricted by haemorrhagic risks, and complicated by hirudin’s rapid clearance from the circulation. We previously employed albumin fusion to slow hirudin variant 3 (HV3) clearance. In this study, we hypothesized that reconfiguration of the chimera, appending human serum albumin (HSA) to the N-terminus of HV3, with an intervening plasmin cleavage site, would create a slowly cleared, plasmin-activatable HV3. Potential plasmin cleavage sites were screened by expression in Escherichia coli, interposed between glutathione sulfotransferase and HV3 domains. The most reactive sequence (GSGIYR-ITY) was recreated in C-terminally His-tagged albumin fusion protein HSACHV3, expressed in Pichia pastoris yeast and purified by nickel-chelate affinity chromatography. HSACHV3 showed no thrombin inhibitory activity in the absence of plasmin, but liberated active HV3 in a time- and concentration-dependent manner in its presence. In a discontinuous clot assay involving clot-bound thrombin, HSACHV3 assisted clot lysis by limiting clot extension in a tPA- and concentration-dependent manner. Similar results were obtained in plasma at higher concentrations of HSACHV3. The chimeric protein exhibited much slower clearance in mice than unfused HV3, and indistinguishable pharmacokinetics from unfused recombinant HSA. In a mouse tail transection bleeding model, doses of HSACHV3 identical to those of HV3 that elicited a four-fold increase in the volume of shed blood were without effect. Our results suggest that HSACHV3 is a fully latent, plasmin activatable, long-lasting hirudin, of potential benefit in thrombotic disorders resistant to natural or pharmacological clot lysis.

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Ligand and Structure-Based Hybrid Screening for Anti-Parkinson Agents and their Pharmacological Evaluation

INTERNATIONAL JOURNAL OF PHARMACEUTICAL EDUCATION AND RESEARCH (IJPER) ◽

10.37021/ijper.v2i2.1 ◽

2020 ◽

Vol 2 (02) ◽

pp. 30-38

Author(s):

Mudita Mishra ◽

Pankaj K. Sonar ◽

Avinash C. Tripathi ◽

Shailendra K. Saraf ◽

Santosh Kumar Verma

Keyword(s):

Toxicity Assessment ◽

Dependent Manner ◽

Standard Drug ◽

Lipinski’S Rule ◽

Storage Vesicles ◽

Dose Dependent ◽

Hybrid Screening

The behavioral and biochemical antiparkinson effect of 7-hydroxyflavone (7-HF) was evaluated by using virtual screening with an e-pharmacophore and shape-based screening approach, and the compound was screened by using the Sigma Aldrich compound library. Screened hits were filtered based on Lipinski’s rule, absorption, distribution,metabolism,elimination, (software for evaluation) (ADME), and toxicity parameters. The best scoring hit, 7-hydroxy 2 phenyl-4H-chromen-4-one, i.e., 7-HF was selected based on shape similarity (> 0.7), g-score, and conserved interactions. Toxicity assessment of retrieved hits was carried out by Osiris and Lazar programs. This study aims to obtain some potential hits, against various antiparkinson category from reported literature and available online resources, and validate their potency by in vivo, in vitro methods. Reserpine 5 mg/kg produces Parkinson’s like condition by depleting presynaptic catecholamines, particularly dopamine through the process of degranulation of storage vesicles. 7-HF 25, 50, and 100 mg/kg was used as a test compound. Syndopa 275 mg/kg was used as a standard drug. The results demonstrate that treatment with 7-HF improved the total locomotor activity and muscular coordination in the rotarod test. In the open field test, enhanced rearing, grooming duration of mobility, and gripping strength in the chimney test, while a decrease in cataleptic scores in the bar test. 7-HF significantly increases catalase, superoxide dismutase, and reduces glutathione level, while reduced the Malondialdehyde (MDA) level. The total protein concentration was also increased in 7-HF treated groups. The behavioral and biochemical results obtained from this study disclosed a definite neuroprotective role of 7-HF in a dose-dependent manner. It is also clear that 7-HF showed potent and effective antiparkinson activity in a similar way as standard. Interestingly, in behavioral and biochemical studies, 7-HF showed approximately equivalent effects as compared to syndopa.

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Evaluation of ex vivo Thrombolytic Activity and in vitro Anti-inflammatory Activity of Thespesia populnea Leaf Extract

International Journal of Pharmaceutical Sciences and Drug Research ◽

10.25004/ijpsdr.2017.090511 ◽

2017 ◽

Vol 9 (05) ◽

Author(s):

Vineela Satuluri ◽

Vidyadhara Suryadevara ◽

Vijetha Pendyala ◽

Narasimhareddy M.

Keyword(s):

Leaf Extract ◽

Inflammatory Activity ◽

Clot Lysis ◽

Dependent Manner ◽

Thrombolytic Activity ◽

Standard Drug ◽

Alcohol Extract ◽

Thespesia Populnea ◽

Anti Inflammatory

Thrombotic disorders like myocardial and cerebral infarction are fatal blood clotting related diseases. Synthetic therapeutics used in such disorders has serious adverse effects, so there is a need to investigate some more safe natural thrombolytic agents. Present study is a preliminary work towards such endeavors. During this study analysis of thrombolytic activity and anti-inflammatory activity of Thespesia populnea leaf extract using a simple and quick in vitro clot lysis assay was performed. Various concentrations of leaf extract i.e. 200μg/ml; 400μg/ml and 600μg/ml were tested at various time intervals including; 24, 48 and 72 hours duration of incubation at 37°C for observing maximum clot lysis. The result findings indicated that concentrations of leaf extract enhanced the percentage of clot lysis in dose dependent manner along with the incubation time factor. However; streptokinase SK a reference standard and water were used as a positive and negative control showed clot lysis maximum 96.35% and 35.22% in 72 hours of incubation respectively. Alcohol extract of whole plant of Thespesia populnea (Family: Malvaceae) was assessed for its anti-inflammatory activity by in vitro methods. In vitro anti-inflammatory activity was evaluated using albumin denaturation assay at different concentrations. Diclofenac sodium was used as standard drug. The results showed that Thespesia populnea alcohol extract at a concentration range of 400-1600μl significantly protects from protein denaturation.

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Antioxidant and antiproliferative properties of Moringa oleifera Lam. leaf aqueous extract

Plant Science Today ◽

10.14719/pst.2020.7.4.936 ◽

2020 ◽

Vol 7 (4) ◽

Author(s):

D Athira Nair ◽

T J James ◽

S L Sreelatha ◽

Bibu John Kariyil

Keyword(s):

Lipid Peroxidation ◽

Antioxidant Activity ◽

Aqueous Extract ◽

Moringa Oleifera ◽

Dependent Manner ◽

Standard Drug ◽

In Vivo Analysis ◽

Moringa Oleifera Lam

Moringa oleifera Lam. is a highly valued medicinal plant in India, especially Kerala. In the present study, antioxidant activity of aqueous extract of leaves of M. oleifera was determined both in-vitro and in-vivo. Male Wistar rats of 3 age groups- 6, 12, and 18 months old were used for in-vivo analysis. In vitro anti-proliferative effect of the extract was carried out in Dalton’s Lymphoma Ascites (DLA) Cells. LCMS-QTOF analysis of the extract was also done to determine the bioactive components present in the extract. Antioxidant activity of M. oleifera leaf showed an IC 50 value of 10.47 ?g/ml and whereas for standard drug, ascorbic acid, it was 19.52 ?g/ml. In-vivo analysis of lipid peroxidation showed a significant reduction of lipid peroxidation in the brains of 12 and 18-months old treated groups. Up to 75% mortality of DLA cancerous cells was observed in-vitro in different concentrations of M. oleifera leaf water extract in a dose-dependent manner, demonstrating its anti-proliferative property. LCMS-QTOF analysis revealed the presence of emodin-8-glucoside in the extract. Molecular docking analysis (Auto Dock Vina) of emodin-8-glucoside with six cancer related proteins showed highest binding affinity with AKT-1 with a binding score of -10.4 kcal/mol, also showed good affinity with NF-kB (p65), Stat-3, Bcl-2, Bcl-xl and c-FLIP. This study helps to choose healthy diet practices to overcome free radical onslaught and cancerous cell proliferation especially in the later stages of life. This can also pave way for the emergence of diet based therapeutic cure for cancer.

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Molecular Docking, Computational and Antithrombotic Studies of Novel 1,3,4-oxadiazole Derivatives

10.20944/preprints201808.0463.v1 ◽

2018 ◽

Author(s):

Majda Batool ◽

Affifa Tajammal ◽

Firdous Farhat ◽

Francis Verpoort ◽

Zafar A. K. Khattak ◽

...

Keyword(s):

Factor Xa ◽

Negative Control ◽

Clot Lysis ◽

Standard Drug ◽

Reference Drug ◽

Docking Score ◽

Oxadiazole Derivatives ◽

Inhibitory Potential

A new series of 1,3,4-oxadiazoles derivatives was synthesized, characterized and evaluated for their in vitro and in vivo anti-thrombotic activity. Compounds (3a-3i) exhibited significant clot lysis with respect to negative control and reference drug streptokinase (30,000 IU) while enhanced clotting time (CT) values were observed (130-342 sec) for these tested compounds than the standard drug heparin (110 sec.). High affinity towards 1NFY with greater docking score was observed for the compounds (3a, 3i, 3e, 3d and 3h) than the control ligand RPR200095. In addition, very good inhibitory potential against factor Xa (F-Xa) was observed with higher docking scores (5612-6270) with ACE values (–189.68 to –352.28 kcal/mol) than the control ligand RPR200095 (Docking score 5192; ACE –197.81 kcal/mol. In vitro, in vivo and in silico results proposed that these newly synthesized compounds can be used as anti-coagulant agents.

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Scrutinizing pharmacological efficiency for Acacia auriculiformis by experimental and computational approach

Future Journal of Pharmaceutical Sciences ◽

10.1186/s43094-021-00221-7 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Shafinaz Nur ◽

Md. Mohotasin Hossain ◽

Nadia Islam ◽

Abu Montakim Tareq ◽

Nujhat Binte Hanif ◽

...

Keyword(s):

Protein Denaturation ◽

Acacia Auriculiformis ◽

Dependent Manner ◽

Binding Interaction ◽

Standard Drug ◽

Brine Shrimp Lethality Assay ◽

Anti Inflammatory ◽

Dose Dependent

Abstract Background The study sought to investigate the biological efficacy of methanol leave extract of Acacia auriculiformis (MEAA) via in vitro, in vivo, in silico approaches. The in vitro cytotoxicity was evaluated through brine shrimp lethality assay, and anti-inflammatory activity was determined by membrane stabilisation and protein denaturation methods (BSA and egg albumin). The in vivo antipyretic activity was examined via Brewer’s yeast induced pyrexia model. Results A. auriculiformis extract unveiled moderate cytotoxicity with significant anti-inflammatory efficacy (p < 0.001) compared to standard drug. This extract also exhibited dose-dependent time of paralysis and death for the worm (p < 0.001) in the anthelmintic test which was directly proportional to employed concentrations. A notable percentage of clot lysis effect (36.42 ± 1.95%, p < 0.001) was also observed for MEAA in human blood compared to control. However, this extract significantly (p < 0.05) reduced fever in a dose-dependent manner during the antipyretic experiment. Besides, in computer-aided investigations, two compounds (2,4-ditert-butylphenol and 3-hydroxy-β-damascone) revealed the best binding interaction with six proteins for cytotoxicity, inflammation, helminthic, thrombolytic and pyretic effect. Moreover, these two compounds satisfy Lipinski’s ‘Rule of Five’ and revealed drug-likeness profiles in the toxicological study. Conclusions These findings disclosed that methanol leaves extract of A. auriculiformis might be a potent source for anti-inflammatory, anti-helminthic, thrombolytic and antipyretic agents.

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Anti-thrombotic Effect of a PAI-1 Inhibitor in Rats Given Endotoxin

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650231 ◽

1996 ◽

Vol 75 (01) ◽

pp. 118-126 ◽

Cited By ~ 27

Author(s):

T Abrahamsson ◽

V Nerme ◽

M Strömqvist ◽

B Åkerblom ◽

A Legnehed ◽

...

Keyword(s):

Plasminogen Activator Inhibitor ◽

Rat Plasma ◽

Clot Lysis ◽

Fibrin Deposition ◽

Plasminogen Activator Inhibitor 1 ◽

Fab Fragment ◽

Dose Dependent Decrease ◽

Pai 1

SummaryThe aim of this study was to investigate the anti-thrombotic effects of an inhibitor of the plasminogen activator inhibitor-1 (PAI-1) in rats given endotoxin. In studies in vitro, PRAP-1, a Fab-fragment of a polyclonal antibody against human PAI-1, was shown to inhibit PAI-1 activity in rat plasma as well as to stimulate clot-lysis of the euglobulin fraction derived from rat plasma. Endotoxin administered to anaesthetised rats produced a marked increase in plasma PAI-1 activity. To study fibrin formation and lysis in vivo after intravenous (i. v.) injection of the coagulant enzyme batroxobin, 125I-fibrinogen was administered to the animals. The thrombi formed by batroxobin were rapidly lysed in control animals, while the rate of lysis was markedly attenuated in rats given endotoxin. PRAP-1 was administered i.v. (bolus + infusion) to rats given endotoxin and batroxobin and the PAI-1 inhibitor caused a dose-dependent decrease in the 125I-fibrin deposition in the lungs. An immunohistochemical technique was used to confirm this decrease in density of fibrin clots in the tissue. Furthermore, PRAP-1 decreased plasma PAI-1 activity in the rats and this reduction was correlated to the decrease in lung 125I-fibrin deposition at the corresponding time point. It is concluded that in this experimental model the PAI-1 antibody PRAP-1 may indeed inhibit thrombosis in animals exposed to endotoxin.

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