Tumor targeted polypeptide, and preparation method and application thereof

A technology targeting peptides and tumor markers, applied in the field of medicinal chemistry, can solve the problems of insufficient interaction and poor targeting effect, and achieve the effect of non-immunogenicity, small molecular weight and high purity

Inactive Publication Date: 2015-12-30
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
View PDF3 Cites 25 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it discloses that the polypeptide has a targeting effect for the treatment of cancer, howev

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
  • Tumor targeted polypeptide, and preparation method and application thereof
  • Tumor targeted polypeptide, and preparation method and application thereof
  • Tumor targeted polypeptide, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1 Construction of the polypeptide screening system of the present invention

[0063] 1) Experimental instruments and materials

[0064] N-methylmorpholine (NMM), piperidine, trifluoroacetic acid (TFA), dichloromethane (DCM), ninhydrin, vitamin C, phenol, tetramethyluronium hexafluorophosphate (HBTU), hexahydro Pyridine, triisopropylsilane (TIS), ethanedithiol (EDT), N,N dimethylformamide (DMF), anhydrous ether, resin, methanol, various Fmoc protected amino acids, PE-anti-HER1 Antibody, PE-anti-HER2 antibody, Cy5-Streptavidin (streptavidin), FITC-Streptavidin (streptavidin), Bio-AdembeadsStreptavidin (streptavidin-labeled magnetic nanospheres), peptide synthesis tube, Shaker, vacuum water pump, rotary evaporator, laser confocal microscope (ZEISSLSM710), the above reagents and materials were obtained from commercial sources.

[0065] 2) Synthesis of "one bead, one object" polypeptide library

[0066] The Fmoc solid-phase peptide synthesis method is used to s...

experiment example 1

[0075] Experimental example 1 detects the affinity of SEQ ID NO.1-3 polypeptides and HER1 and HER2 proteins by surface plasmon resonance (SPRi) method

[0076] Spot 1 mg / mL of SEQ ID NO.1-3 polypeptide and 1×PBS on the chip, incubate overnight at 4°C under humid conditions, then wash with 10×PBS for 10 minutes, then wash with 1×PBS for 10 minutes, and finally wash with deionized water Wash twice, 10min each time, immerse in 1×PBS containing 5% milk, incubate overnight at 4°C, then wash with 10×PBS for 10min, 1×PBS for 10min, and finally wash twice with deionized water, each Each time for 10 minutes, blow dry with nitrogen, and install the chip on the machine (Plexera HT surface plasmon resonance imaging system).

[0077] The mobile phase was sequentially passed through 1×PBS, 2×PBS, 0.625 μg / mL, 1.25 μg / mL, 2.5 μg / mL, 5 μg / mL and 10 μg / mL of human HER1 and HER2 purified proteins, and the SPRi signal was recorded and analyzed.

[0078] As can be seen from Figure 2(a)-(b), th...

experiment example 2

[0079] Experimental example 2 Interaction of SEQ ID NO.1-3 with HER1 and HER2 high expression cells MDA-MB-468 and SKBR3 and normal cell 293A

[0080] Breast cancer cell lines MDA-MB-468 and SKBR3 (HER2 highly expressed) cells were cultured in DMEM containing 20% ​​fetal bovine serum and RPMI1640 medium containing 10% fetal bovine serum, respectively, and normal human kidney fibroblasts 293A were cultured in In H-DMEM medium containing 10% fetal bovine serum, 1×10 5 / mL cell concentration into a round glass-bottom Petri dish (35mm), 37 ° C, 5% CO 2 After culturing in the cell incubator for 24 hours, discard the culture medium, add 50 μmol / L FITC-labeled H1P, H2P, and HMP polypeptides containing 1 μmol / L Hoechst33342 to the three types of cells, and incubate at 4°C for 0.5 hours in the dark, then discard the polypeptides respectively solution, and washed twice with pre-cooled 1×PBS, and added 5 μL PE-anti-HER1 antibody and PE-anti-HER2 antibody respectively. After incubating ...

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 invention relates to a tumor targeted polypeptide, and a preparation method and application thereof. The polypeptide is disclosed as the following general formula: X1X2X3X4X5WX6X7. The invention also relates to a nucleotide sequence for coding the polypeptide, an expression vector for expressing the polypeptide and a host cell. The invention also relates to a bivalent, multivalent or polypeptide coupling substance formed by the polypeptide, a pharmaceutical composition formed by the polypeptide used as a targeted polypeptide and a preparation/development preparation capable of killing cancer cells, and application thereof. The polypeptide is the most ideal tumor targeted polypeptide at present, has important application value in tumor molecular diagnosis and targeted therapy, provides important theoretical and practical basis for early diagnosis, targeted therapy and the like of mammary cancers, lung cancers and many other tumors, and thus, has wide application prospects.

Description

technical field [0001] The present invention relates to the field of medicinal chemistry, in particular to a polypeptide, a tumor-specific targeting polypeptide and applications thereof, in particular to a tumor-targeting polypeptide, its preparation method and application. Background technique [0002] Human epidermal growth factor receptor (EGFR) is the expression product of proto-oncogene c-erbB-1 (HER1), which belongs to receptor tyrosine protein kinase. Studies have shown that HER1 (EGFR) has relatively stable low expression in epithelial, mesenchymal, and neural tissues, and plays an important role in regulating the proliferation, growth, and differentiation of normal cells, but it is overexpressed in many malignant tumors. Such as non-small cell lung cancer, breast cancer, small cell carcinoma of the head and neck, gastric cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer, etc., HER1 overexpressed tumor cells have stronger invasion and metastasi...

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): C07K7/06A61K47/42A61K51/08A61K41/00A61K45/00A61P35/00
Inventor 胡志远王子华王蔚芝钱怡霞
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap