CN103263662B - 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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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, particularly relate to the application in the medicine of a kind of anti-platelet thrombolysin vessel embolism under preparation treatment low shear conditions.
Background technology
Anti-platelet thrombolysin of the present invention (Antiplatelet thrombolysin, APT) be a kind of proteolytic enzyme separated from Agkistrodon acutus snake venom, be made up of α chain, β chain two peptide chains, α chain amino acid sequence is as shown in SEQ ID NO.1, and β chain amino acid sequence is as shown in SEQ ID NO.2.
Current result of study shows, anti-platelet thrombolysin has the effect of the former and anticoagulant of obvious solution fibrin.Wherein, anti-platelet thrombolysin is to the inhibitory action of platelet aggregation mainly by suppressing GPIb and VWF to interact, and GPIb is one of topmost glycoprotein of platelet surface, its mediating platelet adhesion 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, with bleeding tendency, prolonged bleeding time, platelet adhering function decline, companion or do not accompany VIII because of quantum count and/or dysfunction be Major Clinical feature.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 and VWF specific binding, makes platelet adhesion in blood vessel wall, have activated the reaction of other receptor simultaneously, comprises and activates GP II b/III α receptor, make it and fibrinogenic combination, cause platelet aggregation, thus cause the formation of thrombosis.Therefore, being combined with blood plasma VWF by blocking GPIb, can stoping platelet adhesion on vascular endothelial cell wall, anticoagulant.
Visible, existing result of study shows, anti-platelet thrombolysin under high shear condition to interact realization by suppressing GPIb and VWF to the inhibitory action of platelet aggregation.Therefore, do not think that anti-platelet thrombolysin can play the effect of anticoagulant equally under low shear conditions.
Summary of the invention
In view of this, the technical problem to be solved in the present invention is the application provided in the medicine of a kind of anti-platelet thrombolysin vessel embolism under preparation treatment low shear conditions, the present invention finds, anti-platelet thrombolysin is under low shear conditions, still can play the effect of anticoagulant, thus reach the object for the treatment of vessel embolism.
The invention provides the application in the medicine of a kind of anti-platelet thrombolysin vessel embolism under preparation treatment low shear conditions, anti-platelet thrombolysin is made up of α chain, β chain two peptide chains, wherein, α chain amino acid sequence is as shown in SEQ ID NO.1, and β chain amino acid sequence is as shown in SEQ ID NO.2.
Due in existing result of study, anti-platelet thrombolysin under high shear condition to interact realization by suppressing GPIb and VWF to the inhibitory action of platelet aggregation.Therefore, anti-platelet thrombolysin to the therapeutical effect of vessel embolism under low shear conditions not by suppressing GPIb albumen and VWF to interact to realize.
As preferably, under low shear conditions, vessel embolism is that venous return slows down the vascular embolic disease caused.
Preferably, under low shear conditions, vessel embolism is venous thrombosis.
Venous thrombosis refers to that blood condenses abnormally at Deep venou intracavity, and it is the one of the main reasons of venous thrombosis that venous return slows down.The embodiment of the application proves that this anti-platelet thrombolysin has anti thrombotic action at low shear, thus proves that it has the therapeutical effect of anti-venous thrombosis.
As preferably, under treatment low shear conditions provided by the invention, the medicine of vessel embolism is chemicals or biological preparation.
As preferably, under treatment low shear conditions, the medicine of vessel embolism, comprises anti-platelet thrombolysin and pharmaceutically acceptable adjuvant.
Preferably, under treatment low shear conditions provided by the invention, the medicine of vessel embolism is oral formulations or injection.
Preferred, under treatment low shear conditions, the oral formulations of vessel embolism is tablet, capsule, pill, granule, drop pill, microcapsule or pellet.
Preferred, under treatment low shear conditions vessel embolism injection in the quality-volumetric concentration of anti-platelet thrombolysin be 6 μ g/mL.
The invention provides the application in the medicine of a kind of anti-platelet thrombolysin vessel embolism under preparation treatment low shear conditions.Because previous viewpoint thinks that anti-platelet thrombolysin under high shear condition to interact realization by suppressing GPIb and VWF to the inhibitory action of platelet aggregation.Therefore, do not think in result of study before this that anti-platelet thrombolysin can play the effect of anticoagulant equally under low shear conditions.Visible, the purposes of anti-platelet thrombolysin provided by the invention is not identical with the purposes mechanism of action announced in prior art.Experiment shows, low-shearing force (300s
-1) under effect, anti-platelet thrombolysin suppresses in vitro platelet adhesion, gathering and thrombosis in people or Mus whole blood in perfusion compartment.
Accompanying drawing explanation
Fig. 1 shows Mus platelet adhesion in negative control group, gathering and thrombosis effect, wherein, after Fig. 1 (a) shows perfusion 1min, Mus platelet adhesion, gathering and thrombosis effect in negative control group, after Fig. 1 (b) shows perfusion 2min, Mus platelet adhesion, gathering and thrombosis effect in negative control group, after Fig. 1 (c) shows perfusion 3min, Mus platelet adhesion, gathering and thrombosis effect in negative control group;
Fig. 2 shows Mus platelet adhesion in active medicine group, gathering and thrombosis effect, wherein, after Fig. 2 (a) shows perfusion 1min, Mus platelet adhesion, gathering and thrombosis effect in active medicine group, after Fig. 2 (b) shows perfusion 2min, Mus platelet adhesion, gathering and thrombosis effect in active medicine group, after Fig. 2 (c) shows perfusion 3min, Mus platelet adhesion, gathering and thrombosis effect in active medicine group;
Fig. 3 shows Mus platelet average fluorescent strength situation of change in time in perfusing course; shadow region represents standard error (SEM); 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 that in negative control group, human blood platelets adheres to, assembles and thrombosis effect, wherein, after Fig. 4 (a) shows perfusion 1min, in negative control group, human blood platelets adheres to, assembles and thrombosis effect, after Fig. 4 (b) shows perfusion 2min, in negative control group, human blood platelets adheres to, assembles and thrombosis effect, and after Fig. 4 (c) shows perfusion 3min, in negative control group, human blood platelets adheres to, assembles and thrombosis effect;
Fig. 5 shows that in active medicine group, human blood platelets adheres to, assembles and thrombosis effect, wherein, after Fig. 5 (a) shows perfusion 1min, in active medicine group, human blood platelets adheres to, assembles and thrombosis effect, after Fig. 5 (b) shows perfusion 2min, in active medicine group, human blood platelets adheres to, assembles and thrombosis effect, and after Fig. 5 (c) shows perfusion 3min, in active medicine group, human blood platelets adheres to, assembles and thrombosis effect;
Fig. 6 shows human blood platelets average fluorescent strength situation of change in time in perfusing course, shadow region represents SEM, 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.
Detailed description of the invention
The invention provides the application in the medicine of a kind of anti-platelet thrombolysin vessel embolism under preparation treatment low shear conditions, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope methods and applications as herein described are changed or suitably change with combination, realize and apply 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, set forth the present invention further:
Embodiment 1: anti-platelet thrombolysin is to platelet adhesion, gathering and thrombotic effect in Mus whole blood in vitro perfusion compartment
Get the Mus whole blood containing anticoagulant heparin (25U/mL) and fluorescent dye DiOC6(1 μM) mix, at 37 DEG C of insulation 10min.After insulation, control shearing force 300s
-1, use syringe pump (Harvard Apparatus, Holliston, MA) along collagen coated surfaces perfusion 3min, detect anti-platelet thrombolysin to platelet adhesion, gathering and thrombotic effect in Mus whole blood in vitro perfusion compartment.Wherein, the experiment without the Mus whole blood perfusion of anti-platelet thrombolysin process is negative to group photograph, and the experiment of the Mus whole blood perfusion processed through anti-platelet thrombolysin (6 μ g/mL) is active medicine group.Adopt Zeiss Axiovert135 inverted fluorescence microscope (60X-W object lens), the platelet adhesion in real time record Mus whole blood perfusion process on collagen, gathering and thrombosis.Experimental result as shown in Figures 1 to 3.In Fig. 1 and Fig. 2, light-colored part shows platelet aggregation part.
Result of the test shows, and under low-shearing force flox condition, in the Mus whole blood that anti-platelet thrombolysin (6 μ g/mL) processes, platelet adhesion and gathering are less than negative control group, show low-shearing force (300s
-1) time, anti-platelet thrombolysin suppresses in vitro platelet adhesion, gathering and thrombosis in Mus whole blood in perfusion compartment.
Embodiment 2: anti-platelet thrombolysin is to platelet adhesion, gathering and thrombotic effect in people's whole blood in vitro perfusion compartment
Get the people's whole blood containing anticoagulant heparin (25U/mL) and fluorescent dye DiOC6(1 μM) mix, at 37 DEG C of insulation 10min.After insulation, control shearing force 300s
-1, use syringe pump (Harvard Apparatus, Holliston, MA) along collagen coated surfaces perfusion 3min, detect anti-platelet thrombolysin to platelet adhesion, gathering and thrombotic effect in people's whole blood in vitro perfusion compartment.Wherein, the experiment without people's whole blood perfusion of anti-platelet thrombolysin process is negative to group photograph, and the experiment of the people's whole blood perfusion processed through anti-platelet thrombolysin (6 μ g/mL) is active medicine group.Adopt Zeiss Axiovert135 inverted fluorescence microscope (60X-W object lens), the platelet adhesion in real time record people whole blood perfusion process on collagen, gathering and thrombosis.Experimental result as shown in figures 4-6.In Fig. 4 and Fig. 5, light-colored part shows platelet aggregation part.
Result of the test shows, and under low-shearing force flox condition, in people's whole blood that anti-platelet thrombolysin (6 μ g/mL) processes, platelet adhesion and gathering are less than negative control group, show low-shearing force (300s
-1) time, anti-platelet thrombolysin suppresses in vitro platelet adhesion, gathering and thrombosis in people's whole blood in perfusion compartment.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles 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 (7)
1. the application of anti-platelet thrombolysin in the medicine preparing vessel embolism under treatment low shear conditions, described anti-platelet thrombolysin is made up of α chain, β chain two peptide chains, wherein, α chain amino acid sequence is as shown in SEQ ID NO.1, and β chain amino acid sequence is as shown in SEQ ID NO.2.
2. application according to claim 1, is characterized in that, under described low shear conditions, vessel embolism is that venous return slows down the vascular embolic disease caused.
3. application according to claim 1 and 2, is characterized in that, under described low shear conditions, vessel embolism is venous thrombosis.
4. application according to claim 1, is characterized in that, described medicine is biological preparation.
5. application according to claim 1, is characterized in that, described medicine comprises anti-platelet thrombolysin and pharmaceutically acceptable adjuvant.
6. application according to claim 1, is characterized in that, described medicine is injection.
7. application according to claim 6, is characterized in that, in described injection, the quality-volumetric concentration of anti-platelet thrombolysin is 6 μ g/mL.
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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) |
| CN106854656A (en) * | 2016-10-14 | 2017-06-16 | 兆科药业（合肥）有限公司 | A kind of preparation method of heterodimer echidnotoxin |
| CN109260464B (en) * | 2018-10-29 | 2021-10-15 | 兆科药业（合肥）有限公司 | New use of antiplatelet thrombolysin |
| CN113185595B (en) * | 2020-12-11 | 2023-05-09 | 兆科（广州）眼科药物有限公司 | Protein with effects of inhibiting angiogenesis and inflammatory reaction activity 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 |
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| 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 |
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