Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Polypeptide with brain targeted medicine delivery characteristic and preparation method thereof

A brain-targeting and characteristic technology, applied in the field of protein peptides, can solve the problems such as the research and report on targeting functional groups, and achieve the effects of good brain-targeting effect, small molecular weight, and convenient synthesis.

Inactive Publication Date: 2014-07-09
FUDAN UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] But so far, there have been no related studies and reports on the screening of targeted functional groups injected intravenously into the brain by phage display technology.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Polypeptide with brain targeted medicine delivery characteristic and preparation method thereof
  • Polypeptide with brain targeted medicine delivery characteristic and preparation method thereof
  • Polypeptide with brain targeted medicine delivery characteristic and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 In vivo screening of phage display random cyclic heptapeptide library

[0032] The present invention uses a phage display random peptide library to screen polypeptides with blood-brain barrier affinity, which includes the following steps:

[0033] Ph.D-C7C of NEW ENGLAND Biolabs TM Peptide library diluted to 1 x 10 10 pfu titer, the diluent is TBS buffer (tris, Tris) (50 mM Tris, 150 mM sodium chloride), injected into the rat body through the tail vein, and the rat was sacrificed after 60 min. Cardiac perfusion with saline to remove nonspecific phages from the systemic circulation. Remove the rat brain tissue in the ultra-clean bench, homogenize it under sterile conditions, and add 1 mM phenylsulfonyl fluoride (PMSF), 20 μg / mL protease inhibitor (Aprotin) and 1 μg / mL leupeptin at the same time (Leupeptin) in TBS; wash three times with washing solution TBST (0.1% Tween-20); add glycine elution buffer (0.2M Glycine-HCl, pH 2.2) to elute and recover the speci...

Embodiment 2

[0038] Example 2 In vivo screening of phage display random dodecapeptide library

[0039] Choose Ph.D-12 from NEW ENGLAND Biolabs TM peptide library. The first round of screening method is as follows: the Ph.D-C7C of NEW ENGLAND Biolabs TM Peptide library diluted to 1 x 10 10 pfu titer, the diluent is TBS buffer (tris, Tris; 50 mM Tris, 150 mM sodium chloride), and the tail vein of rats is injected. After 60 min, the rats were anesthetized, and 100 μL of cerebrospinal fluid was extracted under sterile conditions. Take an appropriate amount of cerebrospinal fluid and use ER2738 Escherichia coli in logarithmic growth phase to determine the titer of phage by titration method, and the rest will be used for the next round of screening after amplification. In this way, four rounds of screening were carried out, and the titer of phage obtained in each round of screening was determined, and a single phage clone was randomly selected from the results of the last round for use.

...

Embodiment 3

[0041] Example 3 Determination of phage titer

[0042] The phages obtained in each round of screening in Example 1 and Example 2 were diluted 10-fold with LB medium, and 10 μL of the diluted phage was mixed with 200 μL of E. coli ER2738 in logarithmic growth phase, and added to LB top layer agar (top layer agar: each liter contains 10 g tryptone, 5 g yeast extract, 5 g sodium chloride, 1 g magnesium chloride and 7 g agar powder, which is preheated at 50 °C, and is ready for use after autoclaving) quickly Pour into LB plates containing isopropyl-β-D-thiogalactopyranoside and 5-bromo-4-chloro-3-indolegalactopyranoside (IPTG / Xgal), overnight, and count phage coeruleus .

[0043] Titer calculation formula: phage titer (pfu) = 100 × number of plaques × dilution factor, that is, phage forming units per 1 mL.

[0044] Enrichment of phages displaying brain-targeting peptides: As shown in Table 1 and Table 2, after four rounds of screening, the brain recovery rates of phages obtaine...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
particle sizeaaaaaaaaaa
particle sizeaaaaaaaaaa
Login to View More

Abstract

The invention belongs to the technical field of protein polypeptides, and relates to a polypeptide with brain-targeted drug delivery properties, including the molecular structure of the peptide and its application in preparing brain-targeted drug delivery. The present invention uses phage display technology to obtain a phage monoclonal that has the function of penetrating the blood-brain barrier and can be enriched in the brain through multiple rounds of screening in Sprague Dawley outbred rats (SD rats); DNA) sequencing confirmed that these phage display polypeptides contained a common sequence. At the same time, the obtained polypeptide has brain-targeted drug delivery properties confirmed by immunohistochemical method and comparison of recovery rate in vivo. The polypeptide and its derived peptides can be used to modify drug-loaded nanoparticles, liposomes, vesicles or micelles to construct a brain-targeted drug delivery system. It can improve the intracerebral delivery of drugs, enhance the therapeutic effect on brain diseases, and reduce systemic toxic and side effects.

Description

technical field [0001] The invention belongs to the technical field of protein polypeptides, and relates to a polypeptide with brain-targeted drug delivery properties, including the molecular structure of the peptide and its application in preparing brain-targeted drug delivery. Background technique [0002] With the aging of human society, the incidence of brain diseases is increasing year by year, which is seriously endangering human life and health. Brain diseases usually include central nervous system diseases (Parkinson's disease, Alzheimer's disease), brain tumors, cerebrovascular diseases, brain virus and bacterial infections and other diseases. [0003] The prior art discloses that the blood-brain barrier (BBB) ​​is a barrier system between blood and brain tissue, which can isolate unnecessary or harmful substances in the brain tissue and protect the brain tissue. Brain capillary endothelial cells and glial cells on the blood-brain barrier together constitute a tigh...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06A61K47/42
Inventor 李婧炜蒋新国冯亮
Owner FUDAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com