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Platelet envelope glycoprotein oligopeptide or trim/derivate thereof and application thereof

A technology of membrane glycoproteins and modifiers, applied in the fields of synthetic membrane-penetrating oligopeptides, molecular biology and hematology, can solve problems such as thrombocytopenia, and achieve small molecular weight, low immunogenicity and stable molecular structure. Effect

Inactive Publication Date: 2010-06-16
BEIHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, antiplatelet drugs such as aspirin and ticlopidine are commonly used clinically at home and abroad, all of which inhibit the binding and adhesion of platelets and vWF. The biologically active substances released by activated platelets will also negatively affect the physiological balance of bleeding and coagulation, and there are major disadvantages such as causing thrombocytopenia, bleeding and embolism after use.

Method used

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  • Platelet envelope glycoprotein oligopeptide or trim/derivate thereof and application thereof
  • Platelet envelope glycoprotein oligopeptide or trim/derivate thereof and application thereof
  • Platelet envelope glycoprotein oligopeptide or trim/derivate thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Preparation and identification of anti-platelet GPIbα 559Ser phosphorylated antibody and detection of platelet GPIbα.

[0025] 1. Materials and reagents: Synthetic oligopeptides, New Zealand white rabbits, Freund's adjuvant, MBS, and KLH were purchased from Sigma. Goat anti-rabbit IgG-HRP was purchased from Zhongshan Company. ECL reaction reagents were purchased from Prelix Company, and the rest were domestic analytical reagents.

[0026] 2. Method

[0027] (1) Platelet GPIbα559Ser phosphorylated antibody preparation test: Synthetic oligopeptides were cross-linked with MBS and KLH, used as immune antigens, fully mixed with Freund’s adjuvant and emulsified, and then subcutaneously immunized at multiple points on the back. After three immunizations, collected and separated for immunization serum. Synthetic oligopeptide sequences such as figure 1 shown.

[0028] (2) Platelet GPIbα559Ser phosphorylated antibody titer and specificity detection ELISA test: ELI...

Embodiment 2

[0030] Example 2: Research on the effects of membrane-penetrating oligopeptides on platelet adhesion and aggregation

[0031] 1. Materials and reagents:

[0032] Membrane-penetrating oligopeptide (sequence such as Figure 4 shown), ristocetin, ADP, and RGDS are Sigma products; the rest are domestic analytical reagents.

[0033] 2. Method

[0034] (1) Membrane-penetrating oligopeptides inhibit ristocetin-induced platelet aggregation test: citrate anticoagulant whole blood from healthy people, separate platelet-rich plasma (PRP), adjust the platelet concentration to 3×10 8 / ml, take 3 μl of membrane-penetrable oligopeptide and add it to 300 μl of PRP, and take 3 μl of oligopeptide solvent DMSO as a control (the other two synthetic oligopeptides are the same as the control), incubate at 22°C for 5 minutes, and then incubate at 37°C for 5 minutes. Platelet aggregation was induced by adding ristocetin (final concentration 1.25 mg / ml). Aggregation results were detected by a plat...

Embodiment 3

[0038] Example 3: Membrane-penetrating oligopeptides reduce platelet adhesion to vWF-coated surfaces in flow chamber devices.

[0039] 1. Materials and reagents:

[0040] Membrane-penetrating oligopeptide, vWF (prepared in this laboratory), whole blood of fresh healthy people.

[0041] 2. Method

[0042] vWF (concentration: 30 μg / ml) coated glass thin tube (GC120-7.5, Harvard Apparatus Inc.), incubated overnight at 4°C in a humid chamber, washed with PBS for 3 times, blocked with 5% BSA (PBS) at room temperature for 2 hours, rinsed with PBS 3 spares. Whole blood of healthy people was anticoagulated with ACD (2.5% trisodium citrate, 2.0% D-glucose, 1.5% citric acid), and platelet-rich plasma (PRP) was separated. After washing with platelet washing solution CGS (0.12M NaCl, 0.0129M trisodium citrate, 0.03M D-glucose, 0.1% bovine serum albumin, pH 6.5), Modified Tyrode's Buffer (2.5mM Hepese, 150mM NaCl, 2.5mM KCl , 12mMNaHCO3, 5.5mM D-glucose, 1mM CaCl2, 1mM MgCl2, 1mg / ml BS...

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PUM

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Abstract

The invention provides an oligopeptide R-G-S(p)-L-P capable of influencing platelet function or trim / derivate thereof and application of the oligopeptide R-G-S(p)-L-P or trim / derivate thereof in preventing and / or treating arterial thrombus disease and / or related disease interacted by platelet GPIbalpha and ABP. The platelet treated by the oligopeptide R-G-S(p)-L-P or trim / derivate thereof can effectively inhibit adhesion function of the platelet including ristomycin induced platelet aggregation function without influencing platelet aggregation function including ADP induced platelet aggregation function, thus inhibiting thrombopoiesis while basically maintaining the integrity of platelet functional status. The invention can be directly used for research and development of clinical anti-thrombus medicine.

Description

technical field [0001] The present invention relates to a novel synthetic membrane-penetrating oligopeptide, its amino acid sequence and its ability to inhibit platelet function. [0002] The present invention belongs to molecular biology and hematology. Background technique [0003] Thromboembolic diseases, such as myocardial infarction, ischemic cerebral infarction, and venous thromboembolism, are currently one of the causes that seriously endanger human health and lead to high mortality. Prevention and treatment of thrombosis has become an important clinical practice. Prevention methods. Platelets play a very important role in the process of thrombus formation. During normal blood circulation, platelets are in a quiescent state, and at the site of vessel wall damage, especially in the high shear stress environment of blood vessels, platelets will receive Ib in the surface membrane glycoprotein (GP) Ib-IX complex. The body binds to the plasma von Willebrand factor (vWF)...

Claims

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Application Information

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IPC IPC(8): C07K7/06A61K38/08A61P7/02A61P9/10
Inventor 戴克胜
Owner BEIHANG UNIV
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