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Application of circumsporozoite protein polypeptide CSP I-plus of plasmodium in preparing anti-malarial medicine

A circumsporozoite protein and Plasmodium technology, which is applied in the field of medical biology, can solve the problems that there are no reports on the anti-malarial effects of Plasmodium circumsporozoite protein polypeptides

Inactive Publication Date: 2014-03-05
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there is no relevant report on the anti-malarial effect of Plasmodium circumsporozoite protein polypeptide CSP I-plus

Method used

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  • Application of circumsporozoite protein polypeptide CSP I-plus of plasmodium in preparing anti-malarial medicine
  • Application of circumsporozoite protein polypeptide CSP I-plus of plasmodium in preparing anti-malarial medicine
  • Application of circumsporozoite protein polypeptide CSP I-plus of plasmodium in preparing anti-malarial medicine

Examples

Experimental program
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Effect test

Embodiment 1

[0016] S1. Solid-phase chemical synthesis of Plasmodium circumsporozoite protein polypeptide CSP I-plus: according to the sequence of amino acid residues of SEQ ID NO: 1, Beijing Zhongke Yaguang Co., Ltd. was entrusted to synthesize the polypeptide according to the conventional peptide synthesis method, and after purification Reach a purity of more than 95%.

[0017] S2. Research on the specific binding of CSP I-plus to hepatic cell surface molecule HSPG: HepG2.2.15 cells were used as the human hepatocyte model, and FITC-labeled CSP I-plus was added to a wet box at 37°C for 30 minutes, washed with PBS, mounted, and fluorescent Microscope observation. The results showed that CSP I-plus was mainly bound to the liver cell membrane. After treating hepatocytes with heparanase, which can specifically degrade HSPG, for 2 hours, it was found that it inhibited the binding of CSP I-plus to hepatocytes in a concentration-dependent manner ( figure 1 ), indicating that CSP I–plus ca...

Embodiment 2

[0024] Example 2 After the recombinant plasmid containing the gene encoding CSP I-plus was transfected into hepatocytes, the infection of hepatocytes with parasites was observed. Specific steps are as follows:

[0025] S1. Construction of the recombinant vector containing the coding gene of CSP I-plus:

[0026]Use the Primer Premier 5.0 biological software to design the oligonucleotide fragment containing the CSP I-plus coding gene, the sequence is shown in SEQ ID NO: 2 (including the HindⅢ restriction site, the BamHI restriction site and their respective restriction internal Dicer protected bases), and clone it into the pMD 20-T vector to obtain the recombinant plasmid pMD 20-T / CSP I-plus, transform the recombinant plasmid into E. After identification and DNA sequencing analysis, it was verified that the sequence of the target gene was completely correct. pMD 20-T / CSP I-plus was digested by HindⅢ and BamHI, and the target fragment was recovered and then connected to the lin...

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Abstract

The invention discloses novel application of the peptide fragment CSP I-plus of circumsporozoite protein CSP I-plus of plasmodium in treating plasmodium infection. Researches discover that the circumsporozoite protein CSP plays an important role in adhesion and hepatic cell invasion of plasmodium sporozoite, and the key point of malaria infection is that sporozoite of plasmodium invades liver cells, so that malaria infection can be prevented by stopping invasion of sporozoite. CSP I-plus is an important functional domain of CSP, can be specifically combined with a liver cell surface molecule, namely heparan sulfate proteoglycan (HSPG), and can stop combination of CSP and the liver cells. Therefore, the CSP I-plus is expected to be developed into a novel micro-molecule polypeptide medicine for treating plasmodium falciparum infection by stopping adhesion and liver cell invasion of CSP mediated plasmodium sporozoite, and the recombinant expression plasmid containing CSP I-plus coding genes is expected to be further developed into a novel anti-malarial genetic medicine, so that the circumsporozoite protein polypeptide CSP I-plus has an excellent clinical application prospect.

Description

Technical field [0001] The present invention is a pharmaceutical biotechnology field, which is specific to the application of Plasmid Circus spore protein polypeptide CSP I-Plus in preparing antimalarial drugs. Background technique [0002] It is a kind of infectious disease caused by Plasmodium parasitic in the human body. It is one of the five major parasitic diseases in my country and one of the six major tropical diseases in the world.109 countries around the world have malaria, about 3.3 billion people are threatened by malaria. About 500 million people infected with malaria each year, and 1 million of them died.WHO pointed out that the financial expenditure of malaria has risen from 2004 to $ 1.5 billion in 2009, and malaria has brought a heavy economic burden to some developing countries.Although there are currently treating drugs such as chlorine, phosphoric acid, and artemisinin, due to the complexity of life history, drug resistance, and high recurrence rates, the treat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/10A61K48/00A61P33/06
CPCY02A50/30
Inventor 朱家勇卢雪梅金小宝汪洁
Owner GUANGDONG PHARMA UNIV
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