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@#Anticoagulant drugs are widely used in the prevention and treatment of thromboembolic diseases. In recent years, coagulation factor Xa inhibitors have become a hot spot for the development of new anticoagulant drugs. However, the anticoagulant drugs currently used still have side-effects such as increased bleeding risk. This paper analyzes the patent applications of coagulation factor Xa inhibitors, and summarizes the development route of patent application of Bristol-Myers Squibb Company and Guangdong Dongyang Pharmaceutical Co. , Ltd. as representatives of key enterprises, in order to provide some useful references and recommendations for the development, patent protection countermeasure and intellectual property strategy of coagulation factor Xa for domestic pharmaceutical enterprises.
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Antithrombus is one of the effective methods to prevent and treat cardiovascular diseases. Based on the theory of traditional Chinese medicine,the author's previous research and relevant literature,it was found that the alkaloids in Houttuynia cordata has potential antithrombotic effect. However,the pharmacological substance basis and antithrombotic mechanism of H. cordata have not been clarified. In this study,molecular docking was used for virtual screening of antithrombotic alkaloids from H. cordata. Seventy alkaloids selected from H. cordata were screened in the docking ligand data-base with teen thrombosis targets with known crystal structures as the receptors. In addition,the small-molecule approved or to be approved drugs of targets from Drug Bank database were set as a positive reference with minimum score(S value) of each target's approved or to be approved drugs as threshold. The Dock module in Molecular Operating Environment(Version 2016) software was applied to screen the potential active compounds which their scores(S value) were lower than the minimum score of reference. At last the mechanism of antithrombotic effect was preliminarily revealed by compared the main active sites of the test alkaloids with original ligands and references. This study provided some useful information to development of antithrombus drugs.
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Alkaloids , Pharmacology , Fibrinolytic Agents , Pharmacology , Houttuynia , Chemistry , Molecular Docking Simulation , Phytochemicals , PharmacologyABSTRACT
AIM To explore the protective effects of Huoxue Kangnao Granules (Astragali Radix,Morindae officinalis Radix,Puerariae lobatae Radix,etc.) on cerebral ischemia-reperfusion injury in rats.METHODS The rat model for middle cerebral artery occlusion (MCAO) was established,SD rats were randomly divided into sham operation group,model group,positive control group,Huoxue Kangnao Granules low-,middle-and highdose groups.Nerve function score was tested.Volume of cerebral infarction was calculated by TTC staining.SOD,MDA,6-keto-PGF1α,TXB2,IL-1 β and TNF-α were detected by ELISA.The expression levels of MMP-9 and ICAM-1 were detected by RT-PCR.To observe the influence of Huoxue Kangnao Granules on platelet aggregative function,clotting time was analyzed by automatic blood coagulation analyzer,plasma T-PA and PAI activities were determined by ELISA,and hemorheology indices were detected by automatic biochemical analyzer.RESULTS Huoxue Kangnao Granules could significantly decrease cerebral infarction volume,improve nerve function,reduce MDA content,increase SOD activity;increase 6-keto-PGF1 α level,reduce TXB2 content;decrease TNF-α and IL-1β expressions;inhibit MMP-9 and ICAM-1 expressions;extend KPTT and PT;elevate t-PA activity,inhibit PAI activity;decrease whole blood viscosity,enhance erythrocyte deformation ability.Each dosage group of Huoxue Kangnao Granules had a positive correlation between dose and response.CONCLUSION Huoxue Kangnao Granules can protect cerebral ischemia-reperfusion injury in rats,whose mechanisms may be related to anti-free radical injury,anti-thrombus,anti-cerebral vasospasm,improving cerebrovascular permeability,anti-inflammatory,anti-coagulation and improving abnormal changes of hemorheology.
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The antithrombotic effect of Caragana jubata (Pall.)Poir.ethanolic extract (TE)was evaluated by inferior vena cava thrombosis in rats and acute pulmonary thrombosis in mice. To search for the bioactive fractions of TE, comparison on acute pulmonary thrombosis was made between the two main fractions of TE (TE-1 and TE-2). Besides, pharmacological effects of TE, TE-1 and TE-2 on bleeding time and clotting time were also studied. Reference substances combined with UPLC/DAD-q-TOF-MS were applied to identify the main six compounds and other chemical constituents of the TE. The results showed that TE could significantly reduce the rat thrombosis weight in all doses (P<0.01) and improve the protective rate to mice in medium and high doses (P<0.05). TE-2 showed a stronger effect on protecting the mice from paralysis or death and prolonging the bleeding time and clotting time than TE-1. Chemical constituents in TE mainly include isoflavones, pterocarpans and stilbenoids. Constituents in TE-2 were mainly isoflavones and pterocarpans, while those in TE-1 were mainly stilbenoids, which could be inferred that all of these three kinds of constituents may be responsible for the antithrombotic effects of Caragana jubata.
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Objective To analyze the profile of dermcidin (DCD) changes in different stages of acute coronary syndrome (ACS) by quantifying the serum 4 183Da DCD peptide fragment deriving from different ACS patients treated with early antithrombotic therapy.Methods A total of 118 patients with confirmed diagnosis of ACS were enrolled. Immediately after visiting a doctor, the venous blood was collected and afterwards instantly the patient was given orally 300 mg of aspirin and 300 mg clopidogrel, and according to the patient's condition and the consent of his/her or acknowledgement of family members achieved, emergency percutaneous coronary interference (PCI) or thrombolysis or conservative treatment was adopted separately. After anti-thrombotic treatment, at 2, 4, 6, 8, 10, 12, 16, 20, 24, 32, 40, 48, 60 and 72 hours, venous blood was collected and serum isolated respectively. The concentration of 4 183Da DCD fragment in serum was determined by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Simultaneously, the myoglobin (Myo), cardiac troponin I (cTnI) and MB isoenzyme of creatine kinase (CK-MB) were also detected.Results The mean relative strength of nature logarithmic transformations of 4 183Da DCD fragment of 118 patients with ACS was 2.75±1.02 before treatment on admission, and after intervention therapy (mainly antithrombotic therapy) it was decreased to 1.84±1.19 (P = 0.005) and 1.74±1.12 (P = 0.000) at 2 hours and 4 hours, respectively, and then after 4 hours it was slightly elevated. 4 183Da polypeptide increased earlier than myocardial injury markers.Conclusion Aspirin and clopidogrel can significantly decrease the concentration of 4 183Da DCD peptide fragment in serum in patients with ACS, which indicates that the DCD fragment could be used as one of the indexes for observation on early efficacy of antithrombotic therapy.
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Objective To evaluate the level of international normalized ratio (INR) stability and explore the relationship between INR and complication.Methods One hundred and twenty-nine patients with nonvalvular atrial fibrillation (NVAF) and receiving warfarin anticoagulation treatment were collected from May 2005 to May 2010.INR stability in all patients was calculated.INR stability was defined as the ratio of INR check times within the target range over the total INR check times in the same time period.Results The median time of follow-up was 1.8 years.During the follow-up period,the total effective monitoring value of INR was 2574 times (INR was 0.9-6.0),in which 1085 times (42.2％) was outside the anticoagulation target range (INR was 2.0-3.0).INR stability was (60.9 ± 14.9)％.INR stability had significant difference between patients with or without stroke [(61.2 ± 15.0)％ vs.(53.3 ± 4.9)％] (P ＜ 0.05).The factors effecting warfarin anticoagulation intensity were analyzed:unexplained reason (573 times,52.8％),poor medication compliance (214 times,19.7％),changing food structure (143 times,13.2％),the western medicine(109 times,10.0％),the traditional Chinese medicine (12 times,1.1％) and intaking of spirits (34 times,3.1％).Conclusions During warfarin anticoagulation treatment,INR stability ＞ 60％ is safe and effective.Among the known factors which effect anticoagulation stability of warfarin,medication compliance is still the main factor.
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Objective To construct the eukaryotic expression vector of human fibrinolysin for further study on anti-thrombus and thrombolysis.Methods Human fibrinolysin gene was amplified from human fetal liver tissue with PCR,and was cloned into eukaryotic expression plasmid pCI-neo and then sequenced.After the correct identification by sequencing,the eukaryotic expression recombinant plasmid of human fibrinolysin was constructed and identified with PCR and enzyme digestion.Results The PCR amplification product of fibrinolysin gene was 1 740 bp.The result of sequencing was the same as that registered in GenBank.The results of PCR and enzyme digestion showed that the recombinant eukaryotic expression plasmid had correct codogenic gene fragment.Conclusion The eukaryotic expression recombinant plasmid of human fibrinolysin gene is successfully constructed and identified.
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Objective To investigate the effects of Ziyin Huoxue Jiedu Chinese herbs(ZYHXJD) on vascular endothelial cells injured by glucose,insulin and low-density lipoprotein.Methods Human umbilical vein endothelial cell line ECV-304 was exposed to different concentration of glucose,insulin and oxidized low-density lipoprotein(Ox-LDL),and the effects of ZYHXJD on the injured vascular endothelial cells were investigated.The cells in each group were cultured for additional 48 h.And then,cell viability was examined,and the supernatants were used to determine the contents of tissue plaminogen activator(tPA),plasminogen activator inhibitor(PAI),and intercellular adhesion molecule-1(ICAM-1) respectively.Results The viability of the cells in the model group decreased markedly(P
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Objective To study the part of Anti-thrombus and its property from Ground Beetle. Methods To dispart the Ground beetle into four segments which were shell, head, chest and abdomen, and adopt the method of frozen and melt to do the further test. Result Extracts of shell and chest were not active, extracts of head and abdomen were active. The activity of abdomen extracts was much better than that of the insect extracts. Its active component was glycoproteins. Conclusion The activity of abdomen extracts (Tf) is relatively high, and demonstrates wider value.
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Free flap transplantation with microvascular anastomosis has been successfully performed by development of surgical technique, materials and postoperative monitoring equipments of flap. But success rate of microvascular anastomosis is influenced by various factors, and failure rate is about 5-10%. The most influential factor for success rate is surgical technique and other factors that influence failure of microvascular anastomosis are ischemic time of free flap, thrombus formation of anastomosis region and vascular spasm. Many studies has been published in microvascular anastomosis with histologic effect for irrigating solution. But local irrigation solution has been used clinically in microvascular anastomosis, the comparison with each solution, microhistological study for endothelial cell repair and vascular patency has not been reported. The heparin which is anti-thrombotic agent, and urokinase which is fibrinolytic agent are used for this study. Vascular patency and thrombus formation in experimental micro-arterial anastomosis, and endothelial repair were observed with histologic analysis, scanning electron microscopy, transmission electron microscopic examination. The results were obtained as follows: 1. In vascular patency test in 30 minute and 7 days after micro-arterial anastomosis, equal effects of good vascular patency were obtained in group of local irrigation with heparin and urokinase. 2. In thrombus formation in 7 days after micro-arterial anastomosis, equal effects of minimal thrombus formation were obtained in group of local irrigation with heparin and urokinase. 3. In toluidin blue staining in 7 days after micro-arterial anastomosis, local destruction of endothelial cell and inner elastic lamina were seen and endothelial repair was not seen. 4. In scanning electron microscope examination in 7 days after micro-arterial anastomosis, endothelial cell was not seen in peripheral to suture materials, thrombus associated fibrin network was observed. 5. In transmission electron microscope examination in 7 days after microarterial anastomosis, inflammatory cell was seen within smooth muscle cells in site of endothelial cell destruction, smooth muscle cell around suture material were arranged irregularly, some collagenous change were seen. From the results obtained in this study, same results of good vascular patency and anti-thrombotic effect of heparin and urokinase were obtained as a local irrigation solution, and repair of endothelial cell was not seen in 7 days after micro-arterial anastomosis.
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Collagen , Endothelial Cells , Fibrin , Free Tissue Flaps , Heparin , Microscopy, Electron, Scanning , Myocytes, Smooth Muscle , Spasm , Sutures , Thrombosis , Urokinase-Type Plasminogen Activator , Vascular PatencyABSTRACT
Free flap transplantation with microvascular anastomosis has been successfully performed by development of surgical technique, materials and postoperative monitoring equipments of flap. But success rate of microvascular anastomosis is influenced by various factors, and failure rate is about 5-10%. The most influential factor for success rate is surgical technique and other factors that influence failure of microvascular anastomosis are ischemic time of free flap, thrombus formation of anastomosis region and vascular spasm. Many studies has been published in microvascular anastomosis with histologic effect for irrigating solution. But local irrigation solution has been used clinically in microvascular anastomosis, the comparison with each solution, microhistological study for endothelial cell repair and vascular patency has not been reported. The heparin which is anti-thrombotic agent, and urokinase which is fibrinolytic agent are used for this study. Vascular patency and thrombus formation in experimental micro-arterial anastomosis, and endothelial repair were observed with histologic analysis, scanning electron microscopy, transmission electron microscopic examination. The results were obtained as follows: 1. In vascular patency test in 30 minute and 7 days after micro-arterial anastomosis, equal effects of good vascular patency were obtained in group of local irrigation with heparin and urokinase. 2. In thrombus formation in 7 days after micro-arterial anastomosis, equal effects of minimal thrombus formation were obtained in group of local irrigation with heparin and urokinase. 3. In toluidin blue staining in 7 days after micro-arterial anastomosis, local destruction of endothelial cell and inner elastic lamina were seen and endothelial repair was not seen. 4. In scanning electron microscope examination in 7 days after micro-arterial anastomosis, endothelial cell was not seen in peripheral to suture materials, thrombus associated fibrin network was observed. 5. In transmission electron microscope examination in 7 days after microarterial anastomosis, inflammatory cell was seen within smooth muscle cells in site of endothelial cell destruction, smooth muscle cell around suture material were arranged irregularly, some collagenous change were seen. From the results obtained in this study, same results of good vascular patency and anti-thrombotic effect of heparin and urokinase were obtained as a local irrigation solution, and repair of endothelial cell was not seen in 7 days after micro-arterial anastomosis.
Subject(s)
Collagen , Endothelial Cells , Fibrin , Free Tissue Flaps , Heparin , Microscopy, Electron, Scanning , Myocytes, Smooth Muscle , Spasm , Sutures , Thrombosis , Urokinase-Type Plasminogen Activator , Vascular PatencyABSTRACT
A protein designated as Flammulina Velutipes (FV) protein was isolated from the cultured mycelium of FV and purified to homogeneity by CM cellulose CM 52 and gel filtration chromatography. The molecular weight of FV protein were found to be 1.75?10~4u with pI 4.3. Amino acid analysis of FV protein indicated the presence of 14 kinds of amino acids, especially rich in Glu, Asp and Thr.Potent inhibitory effect of FV protein on thrombus formation, in the blood was observed in vitro when rats were given intravenously. The length of thrombus was shortened and the dry weight of the thrombus decreased significantly at the dose of 7.5?10~(-7) mol/kg. Thrombus formation was completely inhibited at the dose of 10~()-8 mol/kg. The IC_(50) was found to be 4.4?10~(-7) mol/kg. FV protein was shown to inhibit collegen induced aggregation of rat platelets both in vitro and in vivo. Its concentration for complete inhibition in vitro was 7.5?10~(-5) mol/L. The IC_(50) was found to be 4?10~(-5)mol/ L. However, FV protein dose of 1.25?10~(-5) mol/kg was required for 60% inhibition in vivo