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

SDF-1-based glycosaminoglycan antagonists and methods of using same

A technology of SDF-1, 1.SDF-1, applied in chemical instruments and methods, medical preparations containing active ingredients, drug combinations, etc.

Inactive Publication Date: 2010-09-15
PROTAFFIN BIOTECHNOLOGIE AG
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite an equally distant relationship to two large families of chemokines, mature SDF-1 shows a high degree of similarity between different species

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
  • SDF-1-based glycosaminoglycan antagonists and methods of using same
  • SDF-1-based glycosaminoglycan antagonists and methods of using same
  • SDF-1-based glycosaminoglycan antagonists and methods of using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1: Improve GAG ​​binding affinity

[0068] Measured by isothermal fluorescence titration, compared with wild-type SDF-1a, the GAG ​​binding affinity of the SDF-1a mutant was improved after modification (see figure 2 ). Perkin Elmer (Baconsfield, UK) LS50B fluorometer was used for steady-state fluorescence measurement. The 180nM pre-equilibrated solution of SDF-1a wild-type or mutant in PBS (pH 7.2, 125mM NaCl) has an emission peak in the range of 300-390nm (excitation wavelength 282nm). Put the cuvette holder into the incubator, add aliquot of heparan sulfate to equilibrate for 2 minutes, and obtain the binding isotherm. The slit width for excitation and emission was set to 12nm, and the spectral data was recorded at a speed of 200nm / min. Set a wavelength threshold of 290nm to avoid scattered light. The fluorescence spectrum is corrected with reference to the background value, and each region within the range of 300 to 390 nm is integrated. The normalized mea...

Embodiment 2

[0069] Example 2: Knock down GPCR activity

[0070] A 48-well Boyden chamber system (Neuroprobe) equipped with a 5 μm PVP-coated polycarbonate membrane was used to study SDF-1a wild-type and mutant-directed cell migration. Put wild-type SDF-a1 and mutants in RPMI 1640 containing 20mM HEPES pH 7.3 and 1mg / ml BSA with 5nM~20nM dilutions in triplicate into the bottom row of the chamber, including those containing only buffer hole. Put 50μl of 2×10 in the same medium 6 Cells / ml THP-1 cell suspension (European collection of cell cultures) is placed in the upper row of wells. At 37°C and 5% CO 2 After incubating for 2 hours, the upper surface of the filter membrane was washed with Hank's balanced salt solution. Migrating cells are fixed in methanol and used (Merck) for dyeing. Count 5 migrating cells in the field of view with 400 times magnification in each well. After correcting the mean values ​​of three independent experiments with reference to the background, plot the chemokin...

Embodiment 3

[0071] Example 3: Inhibition of breast cancer metastasis

[0072] The murine xenograft model was used to study the inhibitory activity of SDF-1a mutant on cancer metastasis. In the in vivo experiment, 10-week-old female immunodeficiency SCID mice were used. Feed them separately throughout the experiment. After the animals were transported and settled down, they were allowed to rest for a few days before the experiment started.

[0073] LMD-231 cells are used in this xenograft model. These cells were obtained from lung metastasis of mice vaccinated with MDA-MB-231. These cells are known to express high levels of CXCR4 and are commonly used in xenotransplantation experiments. These adherent cells were cultured in the following medium, which was a medium prepared with complete minimal essential medium and complete RPMI medium at a ratio of 50:50, and the cells were divided into bottles at a ratio of 1:3. At 37°C and 5% CO 2 Culture the above-mentioned cells in a humidified atmo...

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 relates to novel mutants of human stromal cell-derived factor-1 which exhibit increased glycosaminoglycan (GAG) binding affinity and inhibited or down-regulated GPCR activity compared to wild type SDF-1, methods for producing these mutants and to their use for preparing medicaments for the treatment of cancer.

Description

Technical field [0001] The present invention relates to human stromal cell-derived factor-1 (human stromal cell-derived factor-1), that is, new mutants of SDF-1α, SDF-1β, SDF-1γ or any of their variants, which are comparable to wild-type SDF-1 In comparison, the new mutant showed the following characteristics: (i) glycosaminoglycan (GAG) binding affinity was enhanced and (ii) GPCR activity was inhibited or down-regulated. The present invention also relates to the use of the new mutant to treat cancer. . Background technique [0002] Chemokines are small (8-11kD), soluble chemoattractant cytokine (chemoattractantcytokine). According to the relative positions of their cysteine ​​residues, chemokines are divided into four categories. Alpha chemokine has a non-conservative amino acid separating the first two cysteine ​​residues (CXC-motif), while the first two cysteine ​​residues of β-chemokine are adjacent to each other (CC-based sequence). Fractalkine(CX 3 C chemokine) is the o...

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
IPC IPC(8): C07K14/52A61P35/00A61K38/19
CPCA61K38/00C07K14/522A61P35/00A61P35/04
Inventor 安德烈亚斯·孔格尔艾萨·沃纳贾森·斯林斯比西米·阿利约翰·A·柯比
Owner PROTAFFIN BIOTECHNOLOGIE AG
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