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Worm body membrane bound short peptide ZL-4 with specific target and anti-schistosome function and application thereof

An anti-schistosomiasis, membrane-bound technology, applied in the application, resistance to vector-borne diseases, medical preparations containing active ingredients, etc., can solve the problems of unsafe use, toxic effects, low concentration, etc.

Inactive Publication Date: 2009-12-02
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] For a long time, there has been a common defect in the use of drugs for the treatment of tumors and pathogens: drugs are used in the body, widely distributed, metabolized quickly, and reach the target site at a low concentration, which not only makes the bioavailability of the drug very low, but more importantly Unfortunately, low-dose drugs will not work, and high-dose drugs will cause systemic toxicity. Therefore, people have long proposed the idea of ​​realizing drug-targeted therapy, that is, to develop molecules that specifically bind to tumors or pathogens as targets. It is expected to not only significantly improve the bioavailability of the drug, but more importantly, make the drug concentrate at the target site at a high concentration to exert its medicinal effect, reducing the drug's effect on the body. side effects of
The former two (antibodies and ligands) are antigenic, and they are antigenic when used in heterogeneous organisms, resulting in side effects of immune reactions, which are obviously unsafe for use in vivo, while the latter is a polypeptide molecule that is safe to use

Method used

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  • Worm body membrane bound short peptide ZL-4 with specific target and anti-schistosome function and application thereof
  • Worm body membrane bound short peptide ZL-4 with specific target and anti-schistosome function and application thereof
  • Worm body membrane bound short peptide ZL-4 with specific target and anti-schistosome function and application thereof

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Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1: the screening of short peptide ZL-4 molecule

[0030] Differential panning of live worms of Schistosoma japonicum with random 12 peptide library of commercial phage: release and collect cercariae from Schistosoma-positive oncomelania snails according to conventional methods, wash aseptically, and divide into two parts: one part is used for reverse adsorption peptide library ; The other was prepared by repeated aspiration with a 6-gauge needle syringe to prepare mechanically decapitated larvae for panning peptide libraries. Panning method: first take 5.7×10 10 pfu phage and 500±20 cercariae were added to a 15ml test tube and shaken at 37°C for 150min, centrifuged to collect the phage supernatant that did not bind to the cercariae, and then washed by TBS-T centrifugation for 10 times to discard the unbound phage, and then used The bound phages were collected by PEG / NaCl precipitation, and 2 μl of infected Escherichia coli was taken for abundance determinati...

Embodiment 2

[0033] Example 2: Specific binding and insect resistance of short peptide ZL-4

[0034] 1. Proving that ZL-4 specifically binds to schistosomiasis, the following methods are used.

[0035] 1. Detection of the specific binding ability of ZL-4 molecule and schistosomiasis off-tailed juvenile worm: Experimental comparison proves that ZL-6, ZL-4 and ZL-1 molecular phage clones and M13KE (phage standard control) and schistosome off-tailed live Specific binding ability of juvenile worms: method: take Schistosoma japonicum cercariae, mechanically remove tails, count worms, divide them into 1, 2, 3, 4 tubes (about 500 tailed larvae in each tube), add ZL- 6. For the phages of ZL-4, ZL-1 and M13KE, shake gently at 37°C for 2 hours, then centrifuge to discard the supernatant, collect the worms and wash them with TBS-T for 10 times to discard unbound phages, and then use The bound phages were collected by PEG / NaCl precipitation, and 2 μl of infected Escherichia coli was taken for abundan...

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Abstract

The invention provides a living worm body membrane bound short peptide with specific target and anti-schistosome function and application thereof. The research adopts a bacteriophage 12 peptide library to screen peptide ZL-4 from pellicle of tailing living schistosomulum; repeatedly proved by multi-technology identification, the ZL-4 polypeptide molecule has high membrane-binding rate for living schistosomulum or living adult; an enzyme immunohistochemistry positioning result shows that the ZL-4 polypeptide molecule can participate in specific binding reaction with heads and bodies of the tailing schistosomulum and the adult of schistosoma japonicum and with worm body membranes of slices thereof; and the three-batch anti-schistosome experiments in vitro and in vivo by using ZL-4 prove that the ZL-4 has obvious restraint influence on the survival and the growth of the schistosome. Therefore, the polypeptide molecule has targeting and anti-schistosome functions for the schistosome, and has potential practical value in the aspects of control research and application of schistosomiasis.

Description

technical field [0001] The invention belongs to the field of biological medicines or biological reagents, and specifically relates to a worm body membrane-bound short peptide ZL-4 with specific targeting and anti-schistosomiasis effects and its application. Background technique [0002] For a long time, there has been a common defect in the use of drugs for the treatment of tumors and pathogens: drugs are used in the body, widely distributed, metabolized quickly, and reach the target site at a low concentration, which not only makes the bioavailability of the drug very low, but more importantly Unfortunately, low-dose drugs will not work, and high-dose drugs will cause systemic toxicity. Therefore, people have long proposed the idea of ​​realizing drug-targeted therapy, that is, to develop molecules that specifically bind to tumors or pathogens as targets. It is expected to not only significantly improve the bioavailability of the drug, but more importantly, make the drug co...

Claims

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Application Information

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IPC IPC(8): C07K7/08C12N15/12A61K38/10A61P33/12G01N33/68
CPCY02A50/30
Inventor 曾庆仁刘彦魏琦
Owner CENT SOUTH UNIV
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