CN103263662A - Application of antiplatelet thrombolysin in preparation of medicament for treating thrombosis disease under condition of low shear force - Google Patents
Application of antiplatelet thrombolysin in preparation of medicament for treating thrombosis disease under condition of low shear force Download PDFInfo
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- CN103263662A CN103263662A CN2013102282192A CN201310228219A CN103263662A CN 103263662 A CN103263662 A CN 103263662A CN 2013102282192 A CN2013102282192 A CN 2013102282192A CN 201310228219 A CN201310228219 A CN 201310228219A CN 103263662 A CN103263662 A CN 103263662A
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Abstract
The invention relates to the field of medicaments, and in particular relates to application of antiplatelet thrombolysin in preparation of a medicament for treating a thrombosis disease under the condition of low shear force. Because the previous view considers that the inhibiting effect of the antiplatelet thrombolysin on platelet aggregation is realized by inhibiting the interaction of GPIb and VWF under the condition of high shear force, the previous research result does not consider that the antiplatelet thrombolysin under the condition of low shear force can play a role in inhibiting the platelet aggregation likewise, i.e., the application of the antiplatelet thrombolysin is different from the application and the acting mechanism published in the prior art. Experiments show that the antiplatelet thrombolysin inhibits platelet adhesion, platelet aggregation and thrombosis in whole blood of in-vitro infused indoor people or mice under the action of low shear force (300s<-1>).
Description
Technical field
The present invention relates to field of medicaments, relate in particular to the application of a kind of anti-platelet thrombolysin in preparing the medicine for the treatment of vessel embolism under the low-shearing force condition.
Background technology
Anti-platelet thrombolysin of the present invention (Antiplatelet thrombolysin, APT) be a kind of proteolytic enzyme of from the Agkistrodon acutus snake venom, separating, formed by α chain, two peptide chains of β chain, the α chain amino acid sequence is shown in SEQ ID NO.1, and the β chain amino acid sequence is shown in SEQ ID NO.2.
Present result of study shows, anti-platelet thrombolysin has the effect of the former and anticoagulant of tangible solution fibrin.Wherein, anti-platelet thrombolysin mainly is to interact by suppressing GPIb and VWF to the inhibitory action of platelet aggregation, and GPIb is one of topmost glycoprotein of platelet surface, and it only mediates the adhesion of platelet under high shear.VWF is the abbreviation of vWF ELISA, the common hemorrhage that the VWF quantity of autosomal inheritance and/or textural anomaly cause, descend with bleeding tendency, bleeding time prolongation, platelet adhering function, accompanying or not accompanying VIII factor quantity and/or dysfunction is main clinical characters.When shearing force refers to blood flow and the frictional force of blood vessel wall lining endothelium.The VWF of GPIb and blood plasma or vascular endothelial cell under high shear condition interacts, and reduces hematoblastic flow velocity.GPIb is combined with the VWF specificity, makes platelet adhesion on blood vessel wall, has activated the reaction of other receptor simultaneously, comprises activating GP II b/III α receptor, makes it and fibrinogenic combination, causes platelet aggregation, thereby causes the formation of thrombosis.Therefore, GPIb is combined with blood plasma VWF by blocking-up, can stop platelet adhesion on the vascular endothelial cell wall, anticoagulant.
As seen, existing result of study shows, anti-platelet thrombolysin to the inhibitory action of platelet aggregation by suppressing GPIb and the VWF realization that under high shear condition interacts.Therefore, do not think that anti-platelet thrombolysin under the low-shearing force condition can play the effect of anticoagulant equally.
Summary of the invention
In view of this, the technical problem to be solved in the present invention is to provide the application of a kind of anti-platelet thrombolysin in preparing the medicine for the treatment of vessel embolism under the low-shearing force condition, the present invention finds, anti-platelet thrombolysin is under the low-shearing force condition, still can play the effect of anticoagulant, thereby reach the purpose for the treatment of vessel embolism.
The invention provides the application of a kind of anti-platelet thrombolysin in preparing the medicine for the treatment of vessel embolism under the low-shearing force condition, anti-platelet thrombolysin is made up of α chain, two peptide chains of β chain, wherein, the α chain amino acid sequence is shown in SEQ ID NO.1, and the β chain amino acid sequence is shown in SEQ ID NO.2.
Because in the existing result of study, anti-platelet thrombolysin to the inhibitory action of platelet aggregation by suppressing GPIb and the VWF realization that under high shear condition interacts.Therefore, anti-platelet thrombolysin is not to realize by suppressing GPIb albumen and VWF interaction to the therapeutical effect of vessel embolism under the low-shearing force condition.
As preferably, vessel embolism is the vascular thromboembolism disease that venous return slows down and causes under the low-shearing force condition.
Preferably, vessel embolism is venous thrombosis under the low-shearing force condition.
Venous thrombosis refers to that blood condenses abnormally in dark venous lumen, it is the one of the main reasons of venous thrombosis that venous return slows down.The application's embodiment proves that this anti-platelet thrombolysin has anti thrombotic action under low-shearing force, thereby proves that it has the therapeutical effect of anti-venous thrombosis.
As preferably, the medicine of vessel embolism is chemicals or biological preparation under the treatment low-shearing force condition provided by the invention.
As preferably, treat the medicine of vessel embolism under the low-shearing force condition, comprise anti-platelet thrombolysin and acceptable accessories.
Preferably, the medicine of vessel embolism is oral formulations or injection under the treatment low-shearing force condition provided by the invention.
Preferred, the oral formulations of vessel embolism is tablet, capsule, pill, granule, drop pill, microcapsule or pellet under the treatment low-shearing force condition.
Preferred, under the treatment low-shearing force condition in the injection of vessel embolism the quality-volumetric concentration of anti-platelet thrombolysin be 6 μ g/mL.
The invention provides the application of a kind of anti-platelet thrombolysin in preparing the medicine for the treatment of vessel embolism under the low-shearing force condition.Since previous viewpoint think anti-platelet thrombolysin to the inhibitory action of platelet aggregation by suppressing GPIb and the VWF realization that under high shear condition interacts.Therefore, do not think in the result of study before this that anti-platelet thrombolysin under the low-shearing force condition can play the effect of anticoagulant equally.As seen, the purposes of anti-platelet thrombolysin provided by the invention, inequality with the purposes mechanism of action of announcing in the prior art.Experiment shows, low-shearing force (300s
-1) effect down, anti-platelet thrombolysin suppresses in vitro in the perfusion compartment platelet adhesion, gathering and thrombosis in people or the Mus whole blood.
Description of drawings
Fig. 1 shows Mus platelet adhesion in the negative control group, gathering and thrombosis effect, wherein, after Fig. 1 (a) shows perfusion 1min, Mus platelet adhesion, gathering and thrombosis effect in the negative control group, after Fig. 1 (b) shows perfusion 2min, Mus platelet adhesion, gathering and thrombosis effect in the negative control group, after Fig. 1 (c) shows perfusion 3min, Mus platelet adhesion, gathering and thrombosis effect in the negative control group;
Fig. 2 shows Mus platelet adhesion in the active medicine group, gathering and thrombosis effect, wherein, after Fig. 2 (a) shows perfusion 1min, Mus platelet adhesion, gathering and thrombosis effect in the active medicine group, after Fig. 2 (b) shows perfusion 2min, Mus platelet adhesion, gathering and thrombosis effect in the active medicine group, after Fig. 2 (c) shows perfusion 3min, Mus platelet adhesion, gathering and thrombosis effect in the active medicine group;
Fig. 3 shows in the perfusing course Mus platelet average fluorescent strength situation of change in time, standard error (SEM) is represented in the shadow region, wherein, curve 1 shows negative control group Mus platelet average fluorescent strength situation of change in time, and curve 2 shows active medicine group Mus platelet average fluorescent strength situation of change in time;
Fig. 4 shows human blood platelets adhesion in the negative control group, gathering and thrombosis effect, wherein, after Fig. 4 (a) shows perfusion 1min, human blood platelets adhesion in the negative control group, gathering and thrombosis effect, after Fig. 4 (b) shows perfusion 2min, human blood platelets adhesion in the negative control group, gathering and thrombosis effect, after Fig. 4 (c) shows perfusion 3min, human blood platelets adhesion in the negative control group, gathering and thrombosis effect;
Fig. 5 shows human blood platelets adhesion in the active medicine group, gathering and thrombosis effect, wherein, after Fig. 5 (a) shows perfusion 1min, human blood platelets adhesion in the active medicine group, gathering and thrombosis effect, after Fig. 5 (b) shows perfusion 2min, human blood platelets adhesion in the active medicine group, gathering and thrombosis effect, after Fig. 5 (c) shows perfusion 3min, human blood platelets adhesion in the active medicine group, gathering and thrombosis effect;
Fig. 6 shows in the perfusing course human blood platelets average fluorescent strength situation of change in time, SEM is represented in the shadow region, wherein, curve 1 shows negative control group human blood platelets average fluorescent strength situation of change in time, and curve 2 shows active medicine group human blood platelets average fluorescent strength situation of change in time.
The specific embodiment
The invention provides the application of a kind of anti-platelet thrombolysin in preparing the medicine for the treatment of vessel embolism under the low-shearing force condition, those skilled in the art can use for reference this paper content, suitably improve technological parameter and realize.Special needs to be pointed out is that all similarly replace and change apparent to those skilled in the art, they all are regarded as being included in the present invention.Method of the present invention and application are described by preferred embodiment, the related personnel obviously can change or suitably change and combination methods and applications as herein described in not breaking away from content of the present invention, spirit and scope, realizes and use the technology of the present invention.
The reagent that the present invention adopts is all common commercially available product, all can buy in market.
Below in conjunction with embodiment, further set forth the present invention:
Embodiment 1: anti-platelet thrombolysin is to platelet adhesion, gathering and thrombotic effect in the Mus whole blood in the perfusion compartment in vitro
Get the Mus whole blood and fluorescent dye DiOC6(1 μ M that contain anticoagulant heparin (25U/mL)) mix, at 37 ℃ of insulation 10min.After the insulation, control shearing force 300s
-1, (Harvard Apparatus, Holliston MA) along collagenic coating perifusion 3min, detect anti-platelet thrombolysin to platelet adhesion, gathering and thrombotic effect in the Mus whole blood in the perfusion compartment in vitro to use syringe pump.Wherein, the experiment of the Mus whole blood perfusion of handling without anti-platelet thrombolysin is negative to the group photograph, and the experiment of the Mus whole blood perfusion of handling through anti-platelet thrombolysin (6 μ g/mL) is the active medicine group.Adopt Zeiss Axiovert135 inverted fluorescence microscope (60X-W object lens), platelet adhesion, gathering and thrombosis in the real time record Mus whole blood perfusion process on the collagen.Experimental result is shown in Fig. 1～3.Light-colored part is shown the platelet aggregation part among Fig. 1 and Fig. 2.
Result of the test shows that under the low-shearing force flox condition, platelet adhesion and gathering are less than negative control group in the Mus whole blood that anti-platelet thrombolysin (6 μ g/mL) is handled, and show low-shearing force (300s
-1) time, anti-platelet thrombolysin suppresses in vitro in the perfusion compartment platelet adhesion, gathering and thrombosis in the Mus whole blood.
Embodiment 2: anti-platelet thrombolysin is to platelet adhesion, gathering and thrombotic effect in people's whole blood in the perfusion compartment in vitro
Get the people's whole blood and fluorescent dye DiOC6(1 μ M that contain anticoagulant heparin (25U/mL)) mix, at 37 ℃ of insulation 10min.After the insulation, control shearing force 300s
-1, (Harvard Apparatus, Holliston MA) along collagenic coating perifusion 3min, detect anti-platelet thrombolysin to platelet adhesion, gathering and thrombotic effect in people's whole blood in the perfusion compartment in vitro to use syringe pump.Wherein, the experiment of people's whole blood perfusion of handling without anti-platelet thrombolysin is negative to the group photograph, and the experiment of people's whole blood perfusion of handling through anti-platelet thrombolysin (6 μ g/mL) is the active medicine group.Adopt Zeiss Axiovert135 inverted fluorescence microscope (60X-W object lens), platelet adhesion, gathering and thrombosis in real time record people's whole blood perfusion process on the collagen.Experimental result is shown in Fig. 4～6.Light-colored part is shown the platelet aggregation part among Fig. 4 and Fig. 5.
Result of the test shows that under the low-shearing force flox condition, platelet adhesion and gathering are less than negative control group in people's whole blood that anti-platelet thrombolysin (6 μ g/mL) is handled, and show low-shearing force (300s
-1) time, anti-platelet thrombolysin suppresses in vitro in the perfusion compartment platelet adhesion, gathering and thrombosis in people's whole blood.
The above only is preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (8)
1. the application in the medicine of anti-platelet thrombolysin vessel embolism under preparation treatment low-shearing force condition, described anti-platelet thrombolysin is made up of α chain, two peptide chains of β chain, wherein, the α chain amino acid sequence is shown in SEQ ID NO.1, and the β chain amino acid sequence is shown in SEQ ID NO.2.
2. application according to claim 1 is characterized in that, vessel embolism is the vascular thromboembolism disease that venous return slows down and causes under the described low-shearing force condition.
3. application according to claim 1 and 2 is characterized in that, vessel embolism is venous thrombosis under the described low-shearing force condition.
4. application according to claim 1 is characterized in that, described medicine is chemicals or biological preparation.
5. application according to claim 1 is characterized in that, described medicine comprises anti-platelet thrombolysin and acceptable accessories.
6. application according to claim 1 is characterized in that, described medicine is oral formulations or injection.
7. application according to claim 6 is characterized in that, described oral formulations is tablet, capsule, pill, granule, drop pill, microcapsule or pellet.
8. application according to claim 6 is characterized in that, the quality-volumetric concentration of anti-platelet thrombolysin is 6 μ g/mL in the described injection.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105833255A (en) * | 2016-03-23 | 2016-08-10 | 兆科药业（合肥）有限公司 | Application of antiplatelet thrombolysin in preparation of medicine for treating thrombotic thrombocytopenic purpura (TTP) |
| WO2018068347A1 (en) * | 2016-10-14 | 2018-04-19 | 兆科药业（合肥）有限公司 | Method for preparing heterodimeric snake venom protein |
| CN109260464A (en) * | 2018-10-29 | 2019-01-25 | 兆科药业（合肥）有限公司 | The new application of anti-platelet thrombolysin |
| CN112618701A (en) * | 2020-12-25 | 2021-04-09 | 兆科药业（合肥）有限公司 | Application of antiplatelet thrombolysin in preparation of medicine for treating retinal vein obstructive diseases |
| CN113185595A (en) * | 2020-12-11 | 2021-07-30 | 兆科（广州）眼科药物有限公司 | Protein with activity of inhibiting growth of new blood vessels and inflammatory reaction and preparation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101838323A (en) * | 2010-02-03 | 2010-09-22 | 兆科药业（合肥）有限公司 | Anti-platelet thrombolysin and preparation method thereof |
-
2013
- 2013-06-08 CN CN201310228219.2A patent/CN103263662B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101838323A (en) * | 2010-02-03 | 2010-09-22 | 兆科药业（合肥）有限公司 | Anti-platelet thrombolysin and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| DM PETERSON 等: "Shear-induced platelet aggregation requires von Willebrand factor and platelet membrane glycoproteins Ib and IIb-IIIa", 《BLOOD》 * |
| KERSTIN JURK 等: "Platelets: Physiology and Biochemistry", 《SEMINARS IN THROMBOSIS AND HEMOSTASIS》 * |
| ROSEMARIE A REITER 等: "Platelets and new antiplatelet drugs", 《THERAPY》 * |
| YASUO IKEDA 等: "The role of von Willebrand factor and fibrinogen in platelet aggregation under varying shear stress", 《J. CLIN. INVEST.》 * |
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| CN105833255A (en) * | 2016-03-23 | 2016-08-10 | 兆科药业（合肥）有限公司 | Application of antiplatelet thrombolysin in preparation of medicine for treating thrombotic thrombocytopenic purpura (TTP) |
| WO2017161621A1 (en) * | 2016-03-23 | 2017-09-28 | 兆科药业（合肥）有限公司 | Use of antiplatelet thrombolysin in the preparation of medicine for treating thrombotic thrombocytopenic purpura |
| CN111035754A (en) * | 2016-03-23 | 2020-04-21 | 兆科药业（合肥）有限公司 | Application of antiplatelet thrombolysin in preparation of medicine for treating thrombotic thrombocytopenic purpura |
| WO2018068347A1 (en) * | 2016-10-14 | 2018-04-19 | 兆科药业（合肥）有限公司 | Method for preparing heterodimeric snake venom protein |
| US11485978B2 (en) | 2016-10-14 | 2022-11-01 | Zhaoke Pharmaceutical (Hefei) Company Limited | DNA and method for preparing heterodimer snake venom protein |
| JP2019531734A (en) * | 2016-10-14 | 2019-11-07 | 兆科薬業（合肥）有限公司Ｚｈａｏｋｅ Ｐｈａｒｍａｃｅｕｔｉｃａｌ（Ｈｅｆｅｉ） Ｃｏｍｐａｎｙ Ｌｉｍｉｔｅｄ | Method for producing heterodimeric snake venom protein |
| JP2022505919A (en) * | 2018-10-29 | 2022-01-14 | 兆科薬業（合肥）有限公司 | New use of antiplatelet thromboxane |
| CN109260464B (en) * | 2018-10-29 | 2021-10-15 | 兆科药业（合肥）有限公司 | New use of antiplatelet thrombolysin |
| CN109260464A (en) * | 2018-10-29 | 2019-01-25 | 兆科药业（合肥）有限公司 | The new application of anti-platelet thrombolysin |
| JP7260639B2 (en) | 2018-10-29 | 2023-04-18 | 兆科薬業（合肥）有限公司 | New use of antiplatelet thrombolysin |
| CN113185595A (en) * | 2020-12-11 | 2021-07-30 | 兆科（广州）眼科药物有限公司 | Protein with activity of inhibiting growth of new blood vessels and inflammatory reaction and preparation method thereof |
| CN112618701A (en) * | 2020-12-25 | 2021-04-09 | 兆科药业（合肥）有限公司 | Application of antiplatelet thrombolysin in preparation of medicine for treating retinal vein obstructive diseases |
| WO2022134133A1 (en) * | 2020-12-25 | 2022-06-30 | 兆科药业（合肥）有限公司 | Use of anti-platelet thrombolysin in preparation of drug for treating retinal vein occlusion disease |
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