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

Application of a fusion polypeptide of ziconotide and tat peptide in the preparation of medicines

A fusion technology of peptides and ziconotide, which is applied in the direction of peptides containing positioning/targeting motifs, drug combinations, medical preparations with non-active ingredients, etc., can solve the problem that the analgesic effect and shelf life cannot reach clinical application, Unable to carry out good quality control, unable to carry out large-scale promotion and other problems, to achieve the effect of long-term drug effect, excellent analgesic effect, and low clinical risk

Active Publication Date: 2019-12-24
SHENZHEN RUIJIAN BIOSCI TECH LTD
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are technical methods in the prior art to use cell-penetrating peptides to assist conotoxins to cross the blood-brain barrier. For example, ziconotide is packaged in virus particles, and TAT polypeptide is linked to the surface of virus particles. The virus particles Ziconotide can be delivered through the blood-brain barrier. However, the preparation process of this method is complicated, and the virus packaging process cannot perform good quality control, and it is difficult to apply it on a large scale in the industry.
As another example, the N-terminus of the TAT peptide and conotoxin is used as a linking unit to prepare a fusion polypeptide. The fusion polypeptide can pass through the blood-brain barrier and is suitable for intravenous injection. However, the fusion polypeptide connected to the N-terminal Through intravenous injection, the analgesic effect and existence time in the body cannot meet the requirements of clinical application, and large-scale promotion cannot be carried out.

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
  • Application of a fusion polypeptide of ziconotide and tat peptide in the preparation of medicines
  • Application of a fusion polypeptide of ziconotide and tat peptide in the preparation of medicines
  • Application of a fusion polypeptide of ziconotide and tat peptide in the preparation of medicines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The preparation of embodiment 1 different types of ziconotide fusion peptides

[0042] Four different types of fusion peptides were prepared and named as protected polypeptides MVIIA-a, MVIIA-b, MVIIA-c, and MVIIA-d, respectively. At the same time, ziconotide was prepared and named MVIIA as a control. In this experiment, the F-moc automatic solid-phase synthesis method was adopted, and the specific steps are as follows:

[0043] Synthesis of polypeptides: Protected polypeptides and their derivatives were assembled on resin using a model of the 433A automatic synthesizer (ABI, Foster City, CA). The peptide resin was incubated in suspension for 2.5 hours at room temperature to deprotect the group. The suspension system is composed of 10 ml of TFA, 0.75 g of phenol, 0.25 ml of 1,2-ethanedithiol, 0.5 ml of sulfide anisole and 0.5 ml of water. (Wat methoxycarbonyl (Fmoc), a common alkoxycarbonyl amino protecting group). The resin was separated from the peptide deprotecte...

Embodiment 2

[0046] Example 2: Chemical properties and structural characterization of different types of ziconotide fusion peptides

[0047] 1. Chemical properties of MVIIA and its variants

[0048] The buffer-treated linear peptides were treated for 24-48 hours at 4°C, and then analyzed by high-performance liquid chromatography. It was found that the folding of the linear peptides resulted in the appearance of a major peak and several minor peaks. The buffer system includes 1mM glutathione, 0.1mM oxidized glutathione, 1mM EDTA, and 0.2mg / mL linear peptide, and the pH of the solution is 7.9. The major product was purified and evaluated by analytical reversed-phase high-performance liquid chromatography, and the purity of the peptide was determined to be greater than 98%. Determination was performed with an Ultraflex III TOF / TOF mass spectrometer (Bruker). The prepared polypeptide sequence is shown in Table 1, and its one-step oxidative folding HPLC analysis profile is shown in figure 1 ...

Embodiment 3

[0057] Embodiment 3: Electrophysiological experiments of different types of ziconotide fusion peptides

[0058] In order to further study the electrophysiological effects of different types of improved ziconotide and the inhibitory effect on calcium ion (CaV2.2) channels, the following experiments were carried out:

[0059] HEK293T cells (capable of expressing SV40 large T antigen) were cultured in DMEM high-glucose medium (Gibco) containing 10% fetal bovine serum, 1% penicillin and streptomycin. The incubator environment is 37°C, 5% CO 2 . Dr. Diane Lipscombe provided the alpha of the rat CaV2.2 channel 1B Splice variant e37a, auxiliary subunit α 2 δ 1 and beta 3 Plasmids (Addgene plasmid #26569, #26575, #26574). Then three kinds of plasmids (3μg), 0.4μg enhanced green fluorescent protein gene and liposome were transiently transfected into HEK293T cells. 24 hours after transfection, the cells were seeded on glass slides and incubated in an incubator (37°C, 5% CO 2 ) f...

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

No PUM Login to View More

Abstract

The present invention provides an application of a fusion polypeptide of ziconotide and TAT peptide in preparation of a medication. In the present invention, a fusion peptide is obtained by connecting the C terminal of ziconotide with the N terminal of a cell membrane penetrating peptide, and thus, the defects that intramuscular injection cannot be realized and the like are overcome. The fusion polypeptide can pass through a blood-brain barrier and is applicable for intravenous administration, intraperitoneal administration, or intranasal administration; the operation is convenient and clinical risk is low; after being intravenously, intraperitoneally or intranasally applied, the fusion polypeptide has long pharmacodynamic action time in vivo, has a good analgesic effect and fewer peptide side effect, and is applicable for large-scale clinical application. The fusion polypeptide of the present invention is simple to prepare, a preparation process and quality during preparation are controllable, and the fusion polypeptide is applicable for large-scale industrial production.

Description

technical field [0001] The invention belongs to the technical field of polypeptide medicines, and in particular relates to the application of a ziconotide fusion polypeptide in the preparation of medicines. Background technique [0002] Ziconotide (trade name PrialtTM, Elan Pharmaceuticals) is the first conotoxin drug approved by the US Food and Drug Administration (FDA) in 2004. Its target action site is N-type voltage-gated calcium ion channel, and it is the first-line drug for subarachnoid space (intrathecal) compound analgesia. Ziconotide is the artificial synthesis of the hydrophilic polypeptide ω-MVIIA in the venom peptide of the Pacific fish-eating snail—the conch snail—and it is the first new type of non-morphine analgesic used in clinical practice. Its molecular formula is C 102 h 172 N 36 o 32 S 7 , Structural formula: H-Cys-Lys-Gly-Lys-Gly-Ala-Lys-Cys-Ser-Arg-Leu-Met-Tyr-Asp-Cys-Cys-Thr-Gly-S er-Cys-Arg-Ser- Gly-Lys-Cys-NH 2 . [0003] Ziconotide can be us...

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): C07K19/00A61K38/17A61K47/64A61P25/00A61P35/00
CPCA61K38/1767A61K47/64A61P25/00A61P35/00C07K14/43504C07K2319/10
Inventor 李书鹏周强
Owner SHENZHEN RUIJIAN BIOSCI TECH LTD
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