A kind of trans-blood-brain barrier protein transporter and preparation method and application

A blood-brain barrier and transporter technology, applied in the field of biotechnology drugs, can solve the problems of difficult to reach the therapeutic concentration of drugs and cannot pass through, and achieve the effect of promoting regeneration and neurological function recovery, and good brain targeting

Active Publication Date: 2019-08-20
XIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a protein transporter across the blood-brain barrier, to solve the problem that polypeptides with more than 6 amino acids generally cannot pass through the blood-brain and blood-spine barriers, and it is difficult to achieve effective drug treatment concentrations

Method used

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  • A kind of trans-blood-brain barrier protein transporter and preparation method and application
  • A kind of trans-blood-brain barrier protein transporter and preparation method and application
  • A kind of trans-blood-brain barrier protein transporter and preparation method and application

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preparation example Construction

[0049] 3.2. Preparation of MCAO model (referring to the occlusion method of the rat middle cerebral artery established by Zea Longa et al.): 10% chloral hydrate solution was injected intraperitoneally to anesthetize the animal. The rats were fixed in the supine position, the skin was incised in the middle of the neck, and all layers of tissue were bluntly separated to expose the right common carotid artery. Separate to the section after the bifurcation of internal carotid artery and external carotid artery, and place the line for later use. The internal carotid and common carotid arteries were clipped with arterial clips, and the proximal and distal ends of the external carotid arteries were ligated. Pull the free end of the external carotid artery to be in a straight line with the internal carotid artery, insert and ligate the nylon thread through the external carotid artery, open the arterial clip at the internal carotid artery, insert the nylon thread into the internal caro...

Embodiment 1

[0066] Using DMF as solvent, dissolve 10 mg DHA in a glove box filled with argon; add 5.8 mg EDCI and 4.1 mg HOBT dropwise with stirring at 100 rpm in an ice bath; activate the carboxyl group for 8 hours. In a glove box filled with argon, BDNF was added dropwise with stirring at 100 rpm under ice bath conditions. GFP+ (DHA and BDNF GFP+ The molar ratio is 100:1) acid amine condensation reaction occurs, react for 8 hours; centrifuge at 3000rpm to concentrate the protein transporter, add PBS buffer, wash the modified protein, remove EDCI, HOBT and unreacted DHA.

Embodiment 2

[0068] Using DMF as a solvent, dissolve 30 mg of DHA in an argon-filled glove box; add 11.6 mg of EDCI and 9.8 mg of HOBT dropwise with stirring at 100 rpm in an ice bath; activate the carboxyl group for 6 hours. In a glove box filled with argon, BDNF was added dropwise with stirring at 100 rpm under ice bath conditions. GFP+ (DHA and BDNF GFP+ The molar ratio is 100:1) acid amine condensation reaction occurs, react for 10 hours; centrifuge at 3000rpm to concentrate the protein transporter, add PBS buffer, wash the modified protein, remove EDCI, HOBT and unreacted DHA.

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Abstract

The invention discloses an across-blood-brain-barrier protein transporter, and a preparation method and application thereof. Carboxyl of activated DHA (docosahexaenoic acid) and amino on the side chain of BDNFGFP+amino acid residue are subjected to acidamide condensation reaction to prepare the brain-targeted protein transporter. The protein transporter can act on the injured neurons via the blood-brain barrier, and has multiple actions of transportation, tracing and treatment; and the DHA-modified BDNF provides a non-injury high-efficiency administration mode for central nervous system injury.

Description

technical field [0001] The invention belongs to the field of biotechnology drugs, and in particular relates to a protein transporter across the blood-brain barrier; the invention also relates to a preparation method and application of the protein transporter. Background technique [0002] The number of new patients with brain-related diseases such as cerebral infarction and cerebral hemorrhage in my country is between 2 million and 2.5 million, and the annual death toll is as high as 1.5 million, and it is still increasing at an annual rate of 8.7%. More than 5 million brain injury recovery survivors (accounting for about 75% of patients in the country) are accompanied by severe disabilities such as intellectual impairment, sensory impairment, and motor impairment in varying degrees. According to the survey of the Economic Research Institute of the Ministry of Health of my country, the annual cost of brain injury treatment in my country is more than 100 billion, which brings...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C07K1/107A61K38/18A61P25/00
Inventor 沙保勇冯浩景晓红苏兴利
Owner XIAN MEDICAL UNIV
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