scholarly journals IN VITRO STUDY ON PROTEASE AND THROMBOLYTIC ACTIVITY OF AQUEOUS EXTRACT FROM LEUCAS ASPERA (L.) LEAVES

2021 ◽  
pp. 73-76
Author(s):  
NAGAMANI J. E. ◽  
USHA SAH ◽  
RAVINDRANATH H. A.
Keyword(s):  
Molecular Weight ◽  
Aqueous Extract ◽  
Lytic Activity ◽  
Crude Enzyme ◽  
Clot Lysis ◽  
Thrombolytic Activity ◽  
Reference Drug ◽  
Leucas Aspera ◽  
Caseinolytic Activity

Objective: The current study is an attempt to screen for the in vitro clot lysis and proteolytic activity of aqueous extract of Leucas aspera leaves. Methods: Thrombolytic activity and protease activity of the crude enzyme obtained by ammonium sulfate precipitation and dialysis were assayed using blood clot and casein as substrates respectively. Native PAGE and gel documentation studies were performed to calculate the molecular weight of the enzyme. Results: In the study, 40% salt fractioned crude enzyme sample exhibited significant thrombolytic and caseinolytic activity. Further dose-dependent increased activity was observed with the maximum lytic activity of 52.11±1.04 % at 1 mg/ml of the sample when compared to the reference drug streptokinase (71.39±0.32%). Also, 68.72±0.62 U/hr of caseinolytic activity was observed for 1 mg/ml of the sample fraction. Conclusion: The study highlights and validates the efficacy of Leucas aspera leaves extract for thrombolytic and proteolytic actions. Enzyme with an approximate molecular weight, 19.89 KDa could be responsible for the significant lytic activity.

Effect of Standard Heparin and a Low Molecular Weight Heparin on Thrombolytic and Fibrinolytic Activity of Single-Chain Urokinase Plasminogen Activator In Vitro

1989 ◽  
Vol 62 (03) ◽  
pp. 923-926 ◽  
Author(s):  
D M Lourenço ◽  
A M Dosne ◽  
A Kher ◽  
M Samama
Keyword(s):  
Molecular Weight ◽  
Low Molecular Weight Heparin ◽  
Clot Lysis ◽  
Low Molecular Weight ◽  
Single Chain ◽  
Plasma Clot ◽  
Thrombolytic Activity ◽  
Significant Difference ◽  
Plasma Factor

SummaryThe effect of unfractioned heparin (UH) and low molecular weight heparin (LMWH) (Kabi 2165 - Fragmin®) on in vitro scu-PA thrombolytic and fibrinogenolytic activity was investigated. Thrombolytic activity was evaluated by following lysis of radiolabeled plasma clot immersed in plasma in presence of scu-PA alone or with either form of heparin. A 200 IU/ml scu-PA concentration produced clot lysis within 7 hr. UH or LMWH led to a slightly faster clot lysis which was statistically significant only at the 2nd and 3rd hour. No significant difference could be evidenced between UH and LMWH effect. During clot lysis, plasmin, generated within the clot led to a gradual transformation of scu-PA to tcu-PA, specially after a 4-hr incubation. Appearance of tcu-PA activity in the plasma surrounding the clot was significantly inhibited by either form of heparin. This finding contrasts with results observed in purified systems and suggests the presence of heparin-dependent plasma factor(s) inhibiting tcu-PA formation or its activity. Possible candidates might be anti-thrombin III and PAI-3.No fibrinogen breakdown was observed when plasma was incubated for 7 hr at 37° C in presence of scu-PA alone (200 IU/ ml) or with either form of heparin. However, in presence of a plasma clot, an important fibrinogen breakdown was observed during clot lysis reflecting the action of plasmin and/or tcu-PA generated within the clot, in the surrounding plasma. Fibrinogenolysis was less pronounced in the presence of both heparin preparations possibly as a consequence of the reduction in the tcu-PA level. These results underline the importance of plasma factors in the interaction of heparin with plasminogen activators such as scu-PA.

scholarly journals In silico and antithrombotic studies of 1,3,4-oxadiazoles derived from benzimidazole

2015 ◽  
Vol 11 (1) ◽  
pp. 67 ◽  
Author(s):  
B. Vishwanathan ◽  
B. M. Gurupadayya ◽  
K. Venkata Sairam
Keyword(s):  
In Silico ◽  
Factor Xa ◽  
Docking Studies ◽  
Negative Control ◽  
Clot Lysis ◽  
Drug Candidate ◽  
Thrombolytic Activity ◽  
Reference Drug ◽  
Oxadiazole Derivatives

<p class="Abstract">In the present study, a series of 1,3,4-oxadiazole derivatives (4a-4k) derived from benzimidazole were docked onto factor Xa (PDB: 1NFY) protein using SYBYLX 2.1. and also evaluated for <em>in vitro</em> clot lysis for thrombolytic activity. The synthesized molecules were also screened for in silico ADME studies. The molecular docking studies highlighted that the molecules showed high affinity towards 1NFY with higher docking score and the <em>in silico</em> ADME results were promising and indicated that the molecules holds great potential as a drug candidate. The thrombolytic evaluation was performed for decrease in solid clot weight by the clot lysis study at a concentration of 6.25, 12.5 and 25 µM strengths, respectively. The results of in vitro clot lysis for thrombolytic evaluation revealed that the tested compounds 4a-4k exhibited significant clot lysis with respect to negative control phosphate buffered saline and in comparison to the reference drug streptokinase (30,000 IU). Among all the tested compounds, compound 4j, 4d and 4g exhibited potent thrombolytic activity with EC<sub>50</sub> value of 16.2, 18.1 and 23.7 µM, respectively. The thrombolytic efficacy investigation highlights that the synthesized compound 4j could be considered for further clinical studies to ascertain its possible hit as thrombolytic agents.</p><p> </p>

Comparative Study of the Activity of High and Low Molecular Weight Urokinase in the Presence of Fibrin

1982 ◽  
Vol 47 (01) ◽  
pp. 036-040 ◽  
Author(s):  
M Samama ◽  
M Castel ◽  
O Matsuo ◽  
M Hoylaerts ◽  
H R Lijnen
Keyword(s):  
Molecular Weight ◽  
Human Plasma ◽  
Test System ◽  
Clot Lysis ◽  
Low Molecular Weight ◽  
Plate Method ◽  
Thrombolytic Activity ◽  
Fibrin Plate

SummaryThe fibrinolytic or thrombolytic activity of low molecular weight urokinase (LMW-UK) and high molecular weight urokinase (HMW-UK) is not significantly different when measured in a bovine fibrin plate method, in a circulating plasma system containing a 125I-labelled human fibrin clot, or on 125I-fibrin films in culture plates using normal or α2-antiplasmin depleted human plasma.In a human fibrin plate method however HMW-UK was found to be more active than LMW-UK. In a purified system on human 125I-fibrin films the activation of native or modified human plasminogen by HMW-UK was also found to be more effective than by LMW-UK.Using a clot lysis test system we did not observe a different inhibition of LMW-UK and HMW-UK upon incubation in human plasma. This is in contrast with previous reports that HMW-UK is inhibited more rapidly in human plasma than LMW-UK.In a purified system the inhibition rate of LMW-UK and HMW-UK by α2-antiplasmin is the same (rate constants at 25ΰC of 167 ± 9 M−1s−1 and 171 ± 5 M−1s−1 respectively).The clinical trials available at present used doses of urokinase which were in excess of those required to obtain a maximal fibrinolytic effect. This might explain why in these trials no difference was observed between the thrombolytic effect of LMW-UK and HMW-UK, while in vitro HMW-UK appeared to be more effective. However, one should always be careful to extrapolate in vitro observations as such to the in vivo situation encountered during thrombolytic therapy.

scholarly journals Thrombolytic Activity of Alkaline Protease Purified from a Mutant Strain Bacillus licheniformis MZK05M9

2018 ◽  
Vol 21 (1) ◽  
pp. 63-70
Author(s):  
Md Asad Uz Zaman ◽  
Taqiyah Akhtar ◽  
ATM Zafrul Azam ◽  
Md Arafat Al Mamun ◽  
Md Mozammel Hoq ◽  
...  
Keyword(s):  
Mutant Strain ◽  
Bacillus Licheniformis ◽  
Serine Proteases ◽  
Specific Activity ◽  
Pharmaceutical Journal ◽  
Crude Enzyme ◽  
Clot Lysis ◽  
Bacillus Species ◽  
Thrombolytic Activity

Investigations were performed to find out new microbial enzymes as thrombolytics having better efficacy and specificity. Mutant strain of Bacillus species, B. licheniformis MZK05M9 was cultured in modified urea-glucose media followed by purification using ammonium sulphate precipitation and ultrafiltration through centricon tube of specific MWCO value. The production method yielded 823.42 units/mg of the crude enzyme from mutant strain MZK05M9 and after purification 37695.64 units/mg. The molecular weight of the purified enzyme was estimated as 27.2 kDa and purification increased its specific activity to 16.5 fold with a recovery of 10%. The purified proteases were identified as serine proteases by irreversible inhibition of activity with phenylmethylsulfonyl fluoride (PMSF) and it exhibited 32.84% thrombolytic activity, by in vitro clot lysis assay. Stability studies showed that crude enzyme from mutant strain MZK05M9 remained stable up to a temperature of 45˚C and showed maximum stability at pH range 7.5 to 8.5. Our observation indicates that proteases produced by Bacillus licheniformis mutant have the potential to be developed as a viable thrombolytic agent.Bangladesh Pharmaceutical Journal 21(1): 63-70, 2018

scholarly journals Molecular Docking, Computational, and Antithrombotic Studies of Novel 1,3,4-Oxadiazole Derivatives

2018 ◽  
Vol 19 (11) ◽  
pp. 3606 ◽  
Author(s):  
Majda Batool ◽  
Affifa Tajammal ◽  
Firdous Farhat ◽  
Francis Verpoort ◽  
Zafar Khattak ◽  
...  
Keyword(s):  
Factor Xa ◽  
Clot Lysis ◽  
Clotting Time ◽  
Reference Drug ◽  
Docking Score ◽  
High Affinity ◽  
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 reference drug streptokinase (30,000 IU), and enhanced clotting time (CT) values (130–342 s) than heparin (110 s). 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, impressive inhibitory potential against factor Xa (F-Xa) was observed with higher docking scores (5612–6270) with Atomic Contact Energy (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 might be used as anticoagulant agents.

scholarly journals SYNTHESIS OF BENTONITE NANOCLAY AND INCORPORATION OF CASSIA FISTULA LEAF EXTRACT TO FORM ORGANOBENTONITE: CHARACTERIZATION AND ITS BIOMEDICAL APPLICATIONS

2018 ◽  
Vol 11 (9) ◽  
pp. 381
Author(s):  
Anand Raj Lfa ◽  
Jeslin J
Keyword(s):  
Biomedical Applications ◽  
Leaf Extract ◽  
Spherical Shape ◽  
Xrd Analysis ◽  
Antioxidant Property ◽  
Clot Lysis ◽  
Cassia Fistula ◽  
Thrombolytic Activity ◽  
Lysis Activity

Objective: In this work, methanolic leaf extract from Cassia fistula (known as aragvadha) was incorporated into bentonite nanoclay to form organobentonite. This organobentonite of nanosize was further used for its effective biomedical applications since medicinal clay finds its own advantage over decades.Methods: The bentonite nanoclay was produced by energetic stirring followed by centrifugation and was characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FTIR). The organobentonite was produced using freeze and thaw method. Antioxidant property was studied using Molyneux method, and thrombolytic activity was analyzed using in vitro clot lysis method.Results: The nanosize of bentonite nanoclay between 57 and 82 nm with irregular to spherical shape was confirmed using SEM analysis. The sharp diffraction peak in XRD analysis shows the crystalline nature of bentonite nanoclay, and FTIR results revealed the successful incorporation of the methanolic extract within the bentonite nanoclay. The organobentonite exhibits 84.5% antioxidant property as well as 31% clot lysis activity when compared to the extract and the bentonite nanoclay.Conclusion: Thus, the non-toxic and inexpensive bentonite nanoclay have a high aspect ratio with multifarious applications in medicine, food, cosmetics, and health products. Through this study, the bentonite nanoclay modified using plant alkaloid (organobentonite) is found to possess good biomedical property.

scholarly journals In vitro Antioxidant, Membrane Stabilizing and Thrombolytic Activities of Glycosmis arborea

2012 ◽  
Vol 15 (2) ◽  
pp. 141-143 ◽  
Author(s):  
Sayed Mamur Hossain ◽  
Farhana Islam ◽  
Tasnuva Sharmin ◽  
Hasib Sheikh ◽  
AM Rashedul Hasan ◽  
...  
Keyword(s):  
Carbon Tetrachloride ◽  
Methanol Extract ◽  
Soluble Fraction ◽  
Pharmaceutical Journal ◽  
Clot Lysis ◽  
Hypotonic Solution ◽  
Rat Erythrocytes ◽  
Thrombolytic Activity ◽  
Negative Controls

The methanol extract of leaves and stems of Glycosmis arborea and their pet-ether, carbon tetrachloride, chloroform and aqueous soluble partitionates were subjected to assays for antioxidant activity by Folin-Ciocalteau reagent, membrane stabilizing and thrombolytic activities. The carbon tetrachloride soluble fraction of stems of G. arborea demonstrated the presence of significant amount of phenolic compounds (36.95 ± 0.54 mg of GAE/g of extract). The extractives inhibited heat as well as hypotonic solution-induced haemolysis of rat erythrocytes in vitro. The pet-ether soluble fraction of leaves and stems of G. arborea showed 20.46 and 38.24% and 22.50 and 48.25%. inhibition of hemolysis of RBC caused by heat and hypotonic solution, as compared to 30.55 and 72.91% inhibition of hemolysis of RBC caused by the standard Acetyl salicylic acid at 0.01 mg/ml concentration, respectively. Among the four fractions, the chloroform soluble materials of the stems of G. arborea revealed highest thrombolytic activity with clot lysis value of 36.50% while standard streptokinase and water used as positive and negative controls, showed 64.25±0.26 and 2.35±0.35 % lysis of clot respectively DOI: http://dx.doi.org/10.3329/bpj.v15i2.12579 Bangladesh Pharmaceutical Journal 15(2): 141-143, 2012

scholarly journals In vitro and in vivo evaluation of pharmacological potentials of Campsis radicans L

2019 ◽  
Vol 5 (1) ◽  
Author(s):  
Mirazul Islam ◽  
Tabassum Jannat ◽  
Md. Ruhul Kuddus ◽  
Mohammad Abdur Rashid ◽  
Mohammad Rashedul Haque
Keyword(s):  
Castor Oil ◽  
Methanol Extract ◽  
Radical Scavenging ◽  
Free Radical Scavenging ◽  
Clot Lysis ◽  
Thrombolytic Activity ◽  
Tail Immersion ◽  
Soluble Fractions

Abstract Background Campsis radicans L. is a flowering plant in Bangladesh, traditionally used for the treatment of several human diseases. In this study, in vitro antioxidant, thrombolytic, membrane stabilizing and in vivo analgesic, hypoglycemic, anti-diarrheal and CNS antidepressant activities of organic soluble fractions of crude methanol extract of C. radicans leaf were investigated using appropriate experimental models. Methods The leaves of C. radicans were collected, authenticated, dried and extracted with methanol at room temperature for 30 days. The concentrated methanol extract was partitioned to petroleum-ether (PESF), dichloromethane (DMSF) and ethyl acetate (EASF) soluble fractions. The antioxidant activity of these fractions was determined by DPPH free radical scavenging method. Total phenolic content was determined by the Folin-Ciocalteau’s spectrophotometric method. The thrombolytic activity was assessed by measuring clot lysis ability whereas the membrane stabilizing activity was evaluated by heat- and hypotonic solution-induced hemolysis assay. Tail immersion procedure and acetic acid- induced writhing model were used to measure the analgesic activity of C. radicans. The hypoglycemic, anti-diarrheal and CNS antidepressant activities were determined by oral glucose tolerance test, castor oil-induced diarrheal model and thiopental-sodium induced sleeping time test in mice, respectively. Results All the organic soluble fractions of C. radicans contained phenolic compounds varying from 6.38 to 60.13 mg of GAE/gm of extractive, while in DPPH assay, EASF showed the highest free radical scavenging activity with IC50 is 4.69 μg/ml. The PESF exhibited highest thrombolytic activity (57.14% clot lysis) and the DMSF showed maximum 53.95% inhibition of heat-induced hemolysis of human RBCs. In both tail immersion and acetic acid induced writhing models, the PESF, DMSF, EASF at the doses of 200 and 400 mg/kg body weight, induced a significant (P < 0.001) decrease in the painful sensation in mice. Substantial (P < 0.05) anti-hyperglycemic activity of test samples was found in mice loaded with glucose at the same doses mentioned earlier. Castor oil induced diarrheal test of the plant extractives has shown significant effect in comparison to control group. In CNS antidepressant activity assay, the test samples were able to reduce the duration of sleep in mice caused by thiopental administration. Conclusion All these findings revealed that C. radicans possess significant antioxidant, thrombolytic, membrane stabilizing, analgesic, hypoglycemic, anti-diarrheal and CNS antidepressant activities.

scholarly journals Antifungal activity of Ocimum gratissimum L., Lantana camara L. & Pteleopsis suberosa Engl. & Dies used in the treatment of vulvovaginal candidiasis in Benin

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Jean Robert Klotoe ◽  
Brice Armand Fanou ◽  
Eric Agbodjento ◽  
Arnaud Houehou ◽  
Lauris Fah ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Antifungal Activity ◽  
Aqueous Extract ◽  
Lantana Camara ◽  
Vulvovaginal Candidiasis ◽  
Reference Drug ◽  
Clinical Strains ◽  
Ocimum Gratissimum

Abstract Background Vulvovaginal candidiasis is a widespread mycotic infection that affects a large proportion of women of childbearing age. Its management in traditional medicine is based on the use of medicinal plants. This study aimed to evaluate the antifungal activity of Ocimum gratissimum L., Lantana camara L. and Pteleopsis suberosa Engl. & Diels used in the treatment of vulvovaginal candidiasis in Benin. Results The data obtained from the in vitro antifungal test show that the strains tested (ATCC 90028 and two clinical strains: 1MA and 3MA) were more sensitive to aqueous extracts with a better effect for Pteleopsis suberosa. This potential of the tested extracts correlated with their richness in total polyphenols. The extract of the Pteleopsis suberosa was very active on the inhibition of the reference strain ATCC 90028. On the clinical strains (1MA and 3MA) the aqueous extract of Pteleopsis suberosa showed a better MIC on the 1MA strain. In vivo model, inoculation of 100 µL of the concentrated Candida albicans suspension 1.5 × 105 UFC/mL induced the candidiasis of the female Wistar rat. The treatment with the aqueous extract of Pteleopsis suberosa, like fluconazole (reference drug), significantly reduced Candida albicans infection at a dose of 100 mg/kg after 1, 7 and 13 days of treatment. Conclusion This study revealed the potential antifungal of the Ocimum gratissimum, Lantana camara and Pteleopsis suberosa. Pteleopsis suberosa has better antifungal activity in vitro and in vivo. These observations justify the use of their medicinal plant in the traditional treatment of vulvovaginal candidiasis in Benin.

scholarly journals Extraction and Preformulation Study of Deer Antler Velvet Extract:Physical Characterization of Aqueous and Ethanol Extract

2019 ◽  
Vol 9 (02) ◽  
Author(s):  
Hariyadi D M ◽  
Setyawan D ◽  
Suciati D1 ◽  
Widyowati R ◽  
Chang H-I ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Physicochemical Properties ◽  
Protein Content ◽  
Aqueous Extract ◽  
Ethanol Extract ◽  
Drug Formulation ◽  
Deer Antler ◽  
Ethanol Extracts ◽  
Preformulation Study

Objective: The aim of the research was to extract the deer antler velvet from Kalimantan Indonesia and to study physical characteristics between 70% ethanol and aqueous extract. Materials and methods: Ethanol extracts was extracted from deer antler velvet using maceration and modified maceration method. Ethanol extracts were compared to aqueous extract which produced using maceration technique. The extract profiles were determined by screening test and physicochemical properties as preformulation study were characterized using Thin Layer Chromatography (TLC), Nuclear Magnetic Resonance (NMR), X-ray diffractometer, differential thermal analysis (DTA), solubility test, BCA protein content, and molecular weight using SDS PAGE assay. Results: Extracts were successfully prepared and determined. Physicochemical properties of 70% ethanol extract and aqueous extract resulted different characteristics in melting point, solubility, crystallinity and protein content. Both ethanol extract and aqueous deer antler velvet extract contained group compounds of terpenoids and steroids and contains high amount of proteins at molecular weight of 17 to 43 kDa. In terms of crystallinity, ethanol and aqueous extracts had different crystal lattices. Conclusion: The aqueous extract of deer antler velvet was then recommended for further in vitro drug formulation and characterization. The molecular weight of majority protein inside aqueous deer antler velvet extract was 17 kDa and this datas will be useful for further drug formulation

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