Mimic peptide of human plasminogen activator and preparation thereof

A technology of human plasminogen and peptidomimetic, applied in the fields of biochemical equipment and methods, peptides, material testing products, etc., can solve the problems of inappropriate thromboprophylactic drugs, incomplete recanalization, delayed initial reperfusion, etc.

Inactive Publication Date: 2005-10-12
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still some defects: UK and SK are non-fibrin-specific thrombolytic agents; t-PA and scu-PA require large doses in clinical application; there is initial reperfusion delay or failure, incomplete recanalization, bleeding, and rethrombosis and other issues; although SAK is a fibrin-specific thrombolytic agent, it has strong immunogenicity, which affects the safety of its clinical use; in addition, the current thrombolytic drugs are not suitable for use as preventive drugs for thrombotic diseases
Collen D’s group in Belgium used a random peptide library to study the interaction between Sak mutants and Plg (Jespers et al., 1999), but there has been no international report on the interaction between the two at the level of mimic peptides

Method used

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  • Mimic peptide of human plasminogen activator and preparation thereof
  • Mimic peptide of human plasminogen activator and preparation thereof
  • Mimic peptide of human plasminogen activator and preparation thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Expression of human microplasminogen (μPm)

[0031] Transform Pichia pastoris (purchased from Invitregin, USA) with the inducible expression plasmid pPIC9K / μPm into GS115 strain (GS115 strain, pPIC9K plasmid purchased from Invitregin, USA), and transform by electroporation, with a 0.2 cm size electroporation cuvette, with a voltage of 1500V, The field strength is 7500V / cm, the pulse time is 10ms, and His+ transformants are screened. Transformed clones with multi-copy exchange recombination at the AOX1 gene locus of GS115 were screened with G418. Mark the YPD plate for later use and prepare glycerol bacteria for preservation.

[0032] Pick the expression strain from the above YPD plate and inoculate it in the BMGY medium to amplify and culture for about 24 hours, measure the OD 600 =0.2-0.6, centrifuge, collect the bacteria, change the BMMY induction medium, use methanol as a carbon source to induce expression for 72-96h. The supernatant was collected after centrifuga...

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Abstract

Human plasminogen activator analog peptide and its production are disclosed. It is based on protein molecule interaction critical theory and uses solvent of streptokinase and glycosylzyme as mechanism. It is carried out by taking human plasminogen as target protein by surface displaying technology of phagicin random polypeptide base, biological elutriating, screening, obtaining polypeptide sequence with mu Pg high affinity, and solid-phase synthesizing the analog peptide. Its advantages include small molecular weight, low immunity, simple process and easy control of product output, purity and activity. It can be used to prepare oral preparation and prevent thrombogenesis.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a human plasminogen activator mimic peptide and its preparation method and application. Background technique [0002] Thromboembolic disease seriously threatens human health and is one of the main causes of death and disability. The formation of thrombus reduces or blocks blood supply to tissues, leading to local hypoxia, cell necrosis and loss of tissue function. Thrombolytic drugs dissolve thrombus, recanalize blood vessels, restore tissue blood supply, reduce cell death, reduce infarct size, maintain tissue function, and reduce early and late mortality. Thrombolytic therapy is an important means of treatment of thromboembolic diseases. [0003] Thrombolytic drugs are mostly plasminogen activators, which can activate the conversion of Pg in plasma into Pm, and Pm catalyzes the hydrolysis of fibrin, the main matrix of thrombus, so as to dissolve thrombus and re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C12N15/81C12Q1/25C12Q1/68G01N33/68
Inventor 宋后燕张宇杲宋钢
Owner FUDAN UNIV
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