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Novel brain targeting preparation for preventing and treating neurodegenerative disease

A new type of preparation technology, applied in the field of new brain-targeted preparations for the prevention and treatment of neurodegenerative diseases, can solve the problems of lack of research on differences and immature mechanism of action

Active Publication Date: 2013-10-30
BIOPHARM RES & DEV CENT JINAN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the research on the systemic therapeutic effect of nasal administration entering the systemic circulation, the research on nasal administration targeting the central nervous system is still in the exploratory stage, such as the mechanism of action is still immature, and there is a lack of different animal (rat, dog, sheep). , monkeys) and between animals and humans; it is still in its infancy in terms of formulations, and only a few preparations are used in clinical practice, such as Pharmacia's nicotine for alternative smoking cessation, Sandoz's for Dihydroergotamine nasal preparation for treating migraine

Method used

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  • Novel brain targeting preparation for preventing and treating neurodegenerative disease
  • Novel brain targeting preparation for preventing and treating neurodegenerative disease
  • Novel brain targeting preparation for preventing and treating neurodegenerative disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Construction, expression and purification of TAT-aFGF fusion protein

[0039] The DNA fragment encoding TAT-aFGF (SEQ ID No: 8) was cloned into the prokaryotic expression vector pET-3c (the vector was purchased from invitrogen), and transformed into Escherichia coli BL21 (DE3) (the cells were purchased from invitrogen), and the ampicillin resistance was screened Recombinant. The expression was induced with a final concentration of 1 mM IPTG, and then combined with cation exchange (CM-Sepharose FF) chromatography, heparin affinity chromatography (Heparin-Sepharose CL-6B) and gel exclusion chromatography (Sephadex G-25). The fusion protein was isolated and purified using the method. MTT method was used to detect the proliferative activity of the fusion protein on Balb / c 3T3 cells.

Embodiment 2

[0041] Construction, expression and purification of TAT-bFGF / OX26 fusion protein

[0042] Conventional gene cloning and PCR overlapping techniques were used to construct the Pichia pastoris pAO815α-TAT-bFGF / OX26 transfer plasmid containing the αfactor signal peptide gene to express the fusion protein TAT-bFGF / OX26 (the sequence of TAT-bFGF / OX26 is shown in SEQ ID No: 10, plasmid pAO815α was purchased from Invitrogen Company), methanol was used as the only carbon source for yeast growth, and the fusion protein was directly secreted into the yeast in vitro culture medium. The fusion protein was isolated and purified by cation exchange (CM-Sepharose FF) chromatography, heparin affinity chromatography (Heparin-Sepharose CL-6B) and gel exclusion chromatography (Sephadex G-25).

Embodiment 3

[0044] Construction, expression and purification of bio-TAT-aFGF / OX26-SA fusion protein

[0045] Bio (biotin)-TAT-aFGF was expressed in fusion, and bio-TAT-aFGF / OX26-SA was formed by covalently combining it with OX26-SA (purchased from the British abcam company) linked with antibiotics. After purification, it is prepared into a freeze-dried preparation, which is dissolved with physiological saline before use, and administered intravenously or nasally.

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Abstract

The invention provides a novel brain targeting preparation for preventing and treating a neurodegenerative disease. The novel brain targeting preparation comprises effective dose of fusion protein of cell growth factor and TAT (transcriptional activator) cell-penetrating peptide, or fusion protein of the cell growth factor, TAT and a transferrin receptor monoclonal antibody, a protein activity protecting agent, and proper auxiliary material, wherein the fusion protein of cell growth factor and TAT cell-penetrating peptide or the fusion protein of the cell growth factor, TAT and transferrin receptor monoclonal antibody is an active ingredient. The novel brain targeting preparation disclosed by the invention has excellent performance of breaking through a blood brain barrier, and is specifically targeted to the brain; the defect that single TAT does not have cell selectivity can be overcome; a brain neurons microenvironment is regulated and controlled by the effects of neurotrophy, paracrine and the like, so as to play the roles of preventing and treating the neurodegenerative disease. The novel brain targeting preparation can be prepared into a freeze-drying preparation, a liquid preparation or an adhesive preparation and applied in an intravenous injection or nasal delivery manner.

Description

technical field [0001] The present invention relates to a novel brain-targeting preparation for preventing and treating neurodegenerative diseases, specifically, the brain-targeting preparation comprises an effective amount of fusion protein of cell growth factor and TAT penetrating peptide or cell growth factor and Fusion protein of TAT and transferrin receptor monoclonal antibody, protein activity protector and suitable auxiliary materials, wherein the fusion protein of cell growth factor and TAT penetrating peptide or cell growth factor and TAT and transferrin receptor monoclonal The fusion protein of the antibody is the active ingredient. The brain-targeting preparation can be prepared into a freeze-dried preparation, a liquid preparation or an adhesive preparation for intravenous injection or nasal administration. Background technique [0002] Neurodegenerative disease (Neurodegenerative disease, ND) is a group of chronic progressive neurological diseases based on prim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K47/42A61K47/48A61P25/28A61P25/16A61P25/14
Inventor 黄亚东项琪张齐好苏志坚郑青
Owner BIOPHARM RES & DEV CENT JINAN
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