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

Medical use of inhibitors of glutaminyl and glutamate cyclases

A technology of glutaminyl cyclase and amino acid, applied in medical preparations containing active ingredients, amine active ingredients, drug combinations, etc.

Inactive Publication Date: 2012-01-04
VIVORYON THERAPEUTICS NV
View PDF47 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Not yet determined in vivo [pGlu 3 ] Cyclization mechanism of the 3-position N-terminal glutamic acid of the natural precursor of Aβ (Shirotani, K., Tsubuki, S., Lee, H.J., Maruyama, K., and Saido, T.C. (2002) Neurosci Lett 327, 25 -28)

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
  • Medical use of inhibitors of glutaminyl and glutamate cyclases
  • Medical use of inhibitors of glutaminyl and glutamate cyclases
  • Medical use of inhibitors of glutaminyl and glutamate cyclases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0281] Example 1: Preparation of human and papaya QC

[0282] Host strain and medium

[0283] The Pichia pastoris strain X33 (AOX1, AOX2) for human QC expression was grown, transformed and analyzed according to the manufacturer's instructions (Invitrogen). Prepare the medium required for P. pastoris according to the manufacturer's recommendations, namely buffered glycerol (BMGY) complex or methanol (BMMY) complex medium and fermentation basal salt medium.

[0284] Molecular cloning of plasmid vector encoding human QC

[0285] All cloning steps are performed using standard molecular biology techniques. The vector pPICZαB (Invitrogen) was used for expression in yeast. The pQE-31 vector (Qiagen) is used to express human QC in E. coli. The mature QC cDNA starting from codon 38 was fused in frame with the 6×histidine tag encoded by the plasmid. After amplification and subcloning with primers pQCyc-1 and pQCyc-2, the fragments were inserted into the expression vector using the restric...

Embodiment 2

[0298] Example 2: Determination of glutaminyl cyclase activity

[0299] Fluorescence analysis

[0300] All determinations were performed at 30°C with BioAssay Reader HTS-7000Plus Microplate Reader (Perkin Elmer). QC activity was evaluated by fluorescence measurement using H-Gln-βNA. The sample is composed of 0.2mM fluorescent substrate, 0.2M Tris / HCl containing 20mM EDTA, 0.25U pyroglutamylaminopeptidase (Unizyme, Denmark) and appropriately diluted QC aliquots with a final volume of 250 μl. The excitation / emission wavelength is 320 / 410nm. The analytical reaction is started by adding glutaminyl cyclase. The QC activity is determined by the standard curve of β-naphthylamine under the analytical conditions. One unit is defined as the amount of QC that catalyzes the production of 1 μmol pGlu-βNA from H-Gln-βNA per minute under the conditions.

[0301] In the second fluorescence analysis, H-Gln-AMC was used as a substrate to determine the activity of QC. The reaction was carried o...

Embodiment 3

[0307] Example 3: MALDI-TOF mass spectrometry analysis

[0308] A Hewlett-Packard G2025LD-TOF System with a linear time-of-flight analyzer was used for matrix-assisted laser desorption / ionization mass spectrometry analysis. The device is equipped with a 337nm nitrogen laser, a voltage acceleration source (5kV) and a 1.0m flying pipeline. The detector is in positive ion mode, and a LeCroy 9350M digital storage oscilloscope connected to a personal computer is used to record and filter the signal. The sample (5 μl) is mixed with an equal volume of matrix solution. For the matrix solution, we use an aqueous solution (1 / 1, v / ) by dissolving 30mg 2',6'-dihydroxyacetophenone (Aldrich) and 44mg diammonium hydrogen citrate (Fluka) in 1ml acetonitrile / 0.1% TFA. v) DHAP / DAHC prepared in. A small volume (approximately 1 μl) of the matrix-analyte-mixture was transferred to the probe tip and immediately evaporated in the vacuum chamber (Hewlett-Packard G2024A sample preparation accessory) t...

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 novel physiological substrates of mammalian glutaminyl cyclase (QC, EC 2.3.2.5), new effectors of QC and the use of such effectors and pharmaceutical compositions comprising such effectors for the treatment of diseases that can be treated by modulation of QC-activity, e.g. diseases selected from the group consisting of duodenal cancer with or w / o Heliobacter pylori infections, colorectal cancer, Zolliger-Ellison syndrome, Familial British Dementia and Familial Danish Dementia.

Description

[0001] This application is a division of a Chinese patent application whose application date is October 15, 2004, the application number is 200480030202.5, and the invention title is "Application of Glutaminyl Cyclase Effector and Glutamate Cyclase Effector" Application. Technical field [0002] The present invention relates to glutaminyl cyclase (QC, EC2.3.2.5), which catalyzes the conversion of N-terminal glutamine residues into pyroglutamic acid (5-oxo-proline, pGlu*) Intramolecular cyclization that simultaneously releases ammonia and intramolecular cyclization that converts N-terminal glutamic acid residues into pyroglutamic acid while releasing water molecules. [0003] The present invention confirms that QC of mammals is a metalloenzyme, and provides a new physiological substrate of mammalian QC, a new type of QC effector, and QC effector and pharmaceutical composition containing QC effector can be used for treatment by adjusting QC. Active use in the treatment of diseases. ...

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 Applications(China)
IPC IPC(8): A61K45/00A61K45/06A61K31/131A61K31/417A61K31/4174A61K31/4178A61P25/28A61K31/00A61K31/13A61K31/4184A61K38/08
CPCA61K31/416A61K38/07A61K31/00A61K31/13A61K38/06A61K31/4184A61K31/4178A61K38/08A61P1/04A61P25/00A61P25/28A61P31/04A61P35/00A61P43/00
Inventor 斯特凡·席林托尔斯滕·霍夫曼安德烈·约翰内斯·尼斯特罗杰汉斯-乌尔里希·德穆特乌尔里希·海泽
Owner VIVORYON THERAPEUTICS NV
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