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Signal conotoxin mutant polypeptides compound lt14a-7, preparation and use thereof

A conotoxin and peptide compound technology, applied in the field of polypeptide compounds, can solve the problem of relatively few researches on the venom of the insect-eating cono snail, and achieve the effects of just the right binding strength, specific action site, and exact drug target.

Inactive Publication Date: 2009-02-25
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the currently known conotoxins are isolated from fish-eating cone snails, and there are relatively few studies on the venom of insectivorous cone snails.

Method used

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  • Signal conotoxin mutant polypeptides compound lt14a-7, preparation and use thereof
  • Signal conotoxin mutant polypeptides compound lt14a-7, preparation and use thereof
  • Signal conotoxin mutant polypeptides compound lt14a-7, preparation and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1: Solid-phase synthesis and separation and purification of signal conotoxin lt14a mutant lt14a-7

[0052] 1. After accurately weighing 0.1mmol of Rink resin (total 0.2g), as for the manual synthesis reactor, wash it with DMF six times, then add DMF about 2 to 3 times the volume of the resin and soak for 30 minutes, and then blow nitrogen to make the resin Can flip up and down in the solution.

[0053] 2. Add 20% piperidine in an amount twice the volume of the resin to the manual synthesis reactor, and react with nitrogen gas for 10 minutes. Then rinse twice with DMF.

[0054] 3. Add 20% piperidine in an amount twice the volume of the resin to the manual synthesis reactor again, and react with nitrogen gas for 20 minutes. Then rinse six times with DMF.

[0055] 4. Weigh 0.45mmol of Fmoc-AA, HOBt and HBTU, first dissolve HOBT and HBTU with 1ml DMF, then add the solution into Fmoc-AA and activate for 15 minutes.

[0056] 5. Add the activated Fmoc-AA solution a...

Embodiment 2

[0071] Example 2: The analgesic activity of lt14a-7 after intracerebroventricular injection is better than that of lt14a and MVIIA.

[0072] The analgesic activity of Lt14a-7 was determined by the acetic acid writhing method. KM mice weighing about 18 g, half male and half male, were divided into random groups with 8 mice in each group. The intracerebroventricular injection concentration was 5 μg / μl, and each mouse was injected with 10 μl. At intervals of 1.5h, 3h, and 6h, 0.7% acetic acid was injected intraperitoneally, 20 μl per mouse. The number of writhing times of mice within 15 min was recorded. Take the average value for each group. And compare the significance of writhing frequency difference with normal saline control group. The result is as figure 2 , as shown in 3. Experiments showed that the analgesic activity of lt14a and its mutant lt14a-7 was better than that of MVIIA at 3 hours, while the drug activity of lt14a mutant was weaker than that of MVIIA at 1.5 ...

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Abstract

The invention relates to a mutant polypeptide compound lt14a-7 of South China Sea conus littertus linnaeus toxin lt14a, the preparation technology and application of the polypeptide. The peptide is obtained by replacing seventh lysine with lactamine through mature conotoxin lt14a, so as to be called as lt14a-7 (A is used for replacing the seventh K); the amino acid sequence direction is MCPPLCAPSCTNC from N end to C end. The lt14a-7 is prepared by the chemosynthesis method, and the polypeptide has the activity of antagonism neuron type acetylcholine receptor, important value in the development of analgesics, and good effect in the treatment of chronic pain caused by some diseases and receptor study.

Description

technical field [0001] The invention relates to a polypeptide compound, its preparation method and its application as medicine, belonging to the field of biology. Background technique [0002] Cono snails belong to the phylum Molluscs, Gastropoda, and Conidae. Most of them inhabit the shallow waters of tropical oceans. They are named for their conical or taro-shaped shapes. Conus snails are relatively young organisms. Fossil records prove that the genus Conus first appeared in the Eocene (Eocene). The disappearance of ammonites, marine predatory molluscs during the Mesozoic (mesozoic) objectively promoted the first large-scale conus snails. of speciation. The second large-scale radiation of cone snails began in the Miocene and basically lasted until now (Terlau and Olivera, 2004). [0003] There are about 700 species of cone snails in the world, and the cone snail is one of the most successful evolutionary organisms among marine invertebrates. Although there are many kind...

Claims

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Application Information

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IPC IPC(8): C07K7/08C12N15/12A61K38/10A61P25/04G01N33/48
Inventor 徐安龙任政华尤驭文王磊潘武广孙丹丹官景慈周茂军吴赟曾夏芸周亮
Owner SUN YAT SEN UNIV
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